minibatch.py

#!/usr/bin/env python
# coding: utf8
"""Example of training spaCy's named entity recognizer, starting off with an
existing model or a blank model.

For more details, see the documentation:
* Training: https://spacy.io/usage/training
* NER: https://spacy.io/usage/linguistic-features#named-entities

Compatible with: spaCy v2.0.0+
Last tested with: v2.1.0
"""
from __future__ import unicode_literals, print_function

import random
from pathlib import Path
import spacy
from spacy.util import minibatch, compounding
import sys


# training data
TRAIN_DATA = [('Hydrocarbons catalysed by TmCYP4G122 and TmCYP4G123 in Tenebrio molitor modulate the olfactory response of the parasitoid Scleroderma guani. Hydrocarbons (HCs) present on the epicuticle of terrestrial insects are not only used to reduce water loss but are also used as chemical signals. The cytochrome p450 CYP4G gene is essential for HC biosynthesis in some insects. However, its function in Tenebrio molitor is unknown. Moreover, it is not yet known whether CYP4G of a host can modulate the searching behaviours of its parasitoid. Here, we explore the function of the TmCYP4G122 and CYP4G123 genes in T. molitor. The TmCYP4G122 and CYP4G123 transcripts could be detected in all developmental stages. Their expression was higher in the fat body and abdominal cuticle than in the gut. Their transcript levels in mature larvae under desiccation stress [relative humidity (RH) < 5%] was significantly higher than that in the control (RH = 70%). Injection of dsCYP4G122 and dsCYP4G123 caused a reduction in HC biosynthesis and was associated with increased susceptibility to desiccation. Individuals of the parasitoid Scleroderma guani that emerged from mealworm pupae showed host preference for normal pupae whereas S. guani that emerged from pupae lacking CYP4G122 or/and CYP4G123 lost this searching preference. The current results confirm that CYP4G122 and CYP4G123 regulate the biosynthesis of HCs and modulate the olfactory response of its parasitoid S. guani.',{'entities': [(26,36,'Gene'),(41,51,'Gene'),(55,71,'Species'),(122,139,'Species'),(307,312,'Gene'),(393,409,'Species'),(460,465,'Gene'),(570,580,'Gene'),(585,593,'Gene'),(603,613,'Species'),(619,629,'Gene'),(634,642,'Gene'),(1115,1132,'Species'),(1214,1222,'Species'),(1255,1263,'Gene'),(1271,1279,'Gene'),(1345,1353,'Gene'),(1358,1366,'Gene'),(1454,1462,'Species')]}),('Chitin deacetylase 1 and 2 are indispensable for larval-pupal and pupal-adult molts in Heortia vitessoides (Lepidoptera: Crambidae). Heortia vitessoides Moore is a notorious defoliator of Aquilaria sinensis (Lour.) Gilg trees. Chitin deacetylases (CDAs) catalyze the N-deacetylation of chitin, which is a crucial process for chitin modification. Here, we identified and characterized HvCDA1 and HvCDA2 from H. vitessoides. HvCDA1 and HvCDA2 possess typical domain structures of CDAs and belong to the Group I CDAs. HvCDA1 and HvCDA2 were highly expressed before and after the larval-larval molt. In addition, both exhibited relatively high mRNA expression levels during the larval-pupal molt, the pupal stage, and the pupal-adult molt. HvCDA1 and HvCDA2 transcript expression levels were highest in the body wall and relatively high in the larval head. Significant increases in the HvCDA1 and HvCDA2 transcript expression levels were observed in the larvae upon exposure to 20-hydroxyecdysone. RNA interference-mediated HvCDA1 and HvCDA2 silencing significantly inhibited HvCDA1 and HvCDA2 expression, with abnormal or nonviable phenotypes being observed. Post injection survival rates of the larvae injected with dsHvCDA1 and dsHvCDA2 were 66.7% and 46.7% (larval-pupal) during development and 23.0% and 6.7% (pupal-adult), respectively. These rates were significantly lower than those of the control group insects. Our results suggest that HvCDA1 and HvCDA2 play important roles in the larval-pupal and pupal-adult transitions and represent potential targets for the management of H. vitessoides.',{'entities': [(0,20,'Gene'),(87,106,'Species'),(133,152,'Species'),(188,206,'Species'),(227,245,'Gene'),(384,390,'Gene'),(395,401,'Gene'),(407,421,'Species'),(423,429,'Gene'),(434,440,'Gene'),(515,521,'Gene'),(526,532,'Gene'),(736,742,'Gene'),(747,753,'Gene'),(882,888,'Gene'),(893,899,'Gene'),(1020,1026,'Gene'),(1031,1037,'Gene'),(1072,1078,'Gene'),(1083,1089,'Gene'),(1442,1448,'Gene'),(1453,1459,'Gene'),(1583,1597,'Species')]}),('Hunchback knockdown induces supernumerary segment formation in Bombyx. Insect segment number within species appears to be fixed irrespective of germ types: long vs. short/intermediate. The present study showed induction of supernumerary segment formation by the knockdown of Bombyx hunchback (Bm-hb), presumably by terminal segment addition, a short/intermediate-like-segmentation mode that is not observed in normal Bombyx embryogenesis. This suggests that Bm-hb suppresses segmentation. The results obtained also suggest that the gap gene Bm-Kr (Bombyx Kruppel) provides a permissive environment for the progression of segmentation by suppressing the expression Bm-hb, which terminates segmentation. This indicates a novel mechanism by which the gap gene is involved in segmentation. It appears that Bm-Kr and Bm-hb are involved in segment counting and their interplay contributes to the correct number of segments being formed in Bombyx. Similar mechanisms may be operating in insects that employ the non-Drosophilan mode of segmentation such as in short/intermediate-germ insects.',{'entities': [(0,9,'Gene'),(63,69,'Species'),(275,281,'Species'),(282,291,'Gene'),(293,298,'Gene'),(417,423,'Species'),(458,463,'Gene'),(541,546,'Gene'),(548,554,'Species'),(555,562,'Gene'),(664,669,'Gene'),(812,817,'Gene'),(933,939,'Species')]}),('Apoptosis-mediated vasa down-regulation controls developmental transformation in Japanese Copidosoma floridanum female soldiers. Copidosoma floridanum is a polyembryonic, caste-forming, wasp species. The ratio of investment in different castes changes with environmental stressors (e.g. multi-parasitism with competitors). The vasa gene was first identified in Drosophila melanogaster as a germ-cell-determining factor, and C. floridanum vasa (Cf-vas) gene positive cells have been known to develop into reproductive larvae. Cf-vas seems to control the ratio of investment in C. floridanum larval castes. In this study, we identified environmental factors that control Cf-vas mRNA expression in Japanese C. floridanum by examining Cf-vas mRNA expression under competitor (Meteorus pulchricornis) venom stress; we treated the male and female morulae with M. pulchricornis venom. We also assessed the effects of multi-parasitism of Japanese C. floridanum with M. pulchricornis and found an increasing number of female soldier larvae. The results showed that several amino acid sequences differ between the Japanese and US Cf-vas. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) showed that Japanese Cf-vas mRNA is expressed in both male and female larvae and pupae, but mRNA expression decreases in adults. Cf-vas mRNA expression significantly decreased, while C. floridanum dronc (Cf-dronc) mRNA expression increased, in female morulae after M. pulchricornis venom treatment at 20  h and 0  h of the culture period, respectively. Females and males showed different Cf-vas or Cf-dronc mRNA expression after M. pulchricornis venom treatment. Therefore, M. pulchricornis venom could affect the ratio of investment in different female castes of Japanese C. floridanum by decreasing Cf-vas mRNA expression via apoptosis.',{'entities': [(19,23,'Gene'),(90,111,'Species'),(129,150,'Species'),(327,331,'Gene'),(361,384,'Species'),(424,437,'Species'),(438,442,'Gene'),(444,450,'Gene'),(525,531,'Gene'),(576,589,'Species'),(669,675,'Gene'),(704,717,'Species'),(731,737,'Gene'),(772,794,'Species'),(854,870,'Species'),(939,952,'Species'),(958,974,'Species'),(1120,1126,'Gene'),(1220,1226,'Gene'),(1328,1334,'Gene'),(1382,1395,'Species'),(1396,1401,'Gene'),(1403,1411,'Gene'),(1464,1480,'Species'),(1587,1593,'Gene'),(1600,1605,'Gene'),(1628,1644,'Species'),(1673,1689,'Species'),(1772,1785,'Species'),(1800,1806,'Gene')]}),('MiR-219 represses expression of dFMR1 in Drosophila melanogaster. AIMS: Fragile X mental retardation protein (FMRP) plays a vital role in mRNA trafficking and translation inhibition to regulate the synthesis of local proteins in neuronal axons and dendritic terminals. However, there are no reports on microRNA (miRNA)-mediated regulation of FMRP levels in Drosophila. Here, we aimed to identify miRNAs regulating FMRP levels in Drosophila. MAIN METHODS: Using online software, we predicted and selected 11 miRNAs potentially acting on the Drosophila fragile X mental retardation 1 (dFMR1) transcript. These candidates were screened for modulation of dFMR1 transcript levels at the cellular level using a dual luciferase reporter system. In addition, we constructed a transgenic Drosophila model overexpressing miR-219 in the nervous system and quantified dFMRP by western blotting. The neuromuscular junction phenotype in the model was studied by immunofluorescence staining. KEY FINDINGS: Among the 11 miRNAs screened, miR-219 and miR-960 reduced luciferase gene activity by binding to the 3 -UTR of the dFMR1 transcript. Mutation of the miR-219 or miR-960 binding sites on the transcript resulted in complete or partial elimination of the miRNA-induced repression. Western blots revealed that dFMRP expression was decreased in the miR-219 overexpression model (Elav>miR-219). Drosophila larvae overexpressing miR-219 showed morphological abnormalities at the neuromuscular junction (increased synaptic boutons and synaptic branches). This finding is consistent with some phenotypes observed in dfmr1 mutants. SIGNIFICANCE: Our results suggest that miR-219 regulates dFMR1 expression in Drosophila and is involved in fragile X syndrome pathogenesis. Collectively, these findings expand the current understanding of miRNA-mediated regulation of target molecule-related functions.',{'entities': [(0,7,'Gene'),(32,37,'Gene'),(41,64,'Species'),(72,108,'Gene'),(110,114,'Gene'),(342,346,'Gene'),(357,367,'Species'),(414,418,'Gene'),(429,439,'Species'),(540,550,'Species'),(551,581,'Gene'),(583,588,'Gene'),(651,656,'Gene'),(779,789,'Species'),(811,818,'Gene'),(856,861,'Gene'),(1021,1028,'Gene'),(1033,1040,'Gene'),(1106,1111,'Gene'),(1140,1147,'Gene'),(1151,1158,'Gene'),(1296,1301,'Gene'),(1334,1341,'Gene'),(1369,1376,'Gene'),(1412,1419,'Gene'),(1597,1602,'Gene'),(1651,1658,'Gene'),(1669,1674,'Gene'),(1689,1699,'Species')]}),('WSV181 inhibits JAK/STAT signaling and promotes viral replication in Drosophila. The Janus kinase/signal transducers and activators of transcription (JAK/STAT) pathway plays a critical role in host defense against viral infections. Here, we report the use of the Drosophila model system to investigate the modulation of the JAK/STAT pathway by the white spot syndrome virus (WSSV) protein WSV181. WSV181 overexpression in transgenic flies resulted in the downregulation of STAT92E and STAT92E-targeted genes. This result indicates that WSV181 can suppress JAK/STAT signaling by controlling STAT92E expression. An infection experiment was carried out on transgenic Drosophila infected with Drosophila C virus and on Litopenaeus vannamei injected with recombinant WSV181 and WSSV. The increased viral load and suppressed transcript levels of JAK/STAT pathway components indicate that WSV181 can promote viral proliferation by inhibiting the JAK/STAT pathway. This study provided evidence for the role of WSV181 in viral replication and revealed a new mechanism through which WSSV evades host immunity to maintain persistent infection.',{'entities': [(0,6,'Gene'),(16,19,'Gene'),(20,24,'Gene'),(69,79,'Species'),(150,153,'Gene'),(154,158,'Gene'),(263,273,'Species'),(324,327,'Gene'),(328,332,'Gene'),(348,373,'Species'),(375,379,'Species'),(389,395,'Gene'),(397,403,'Gene'),(473,480,'Gene'),(485,492,'Gene'),(536,542,'Gene'),(556,559,'Gene'),(560,564,'Gene'),(590,597,'Gene'),(715,735,'Species'),(762,768,'Gene'),(773,777,'Species'),(840,843,'Gene'),(844,848,'Gene'),(882,888,'Gene'),(939,942,'Gene'),(943,947,'Gene'),(1002,1008,'Gene'),(1073,1077,'Species')]}),('Biochemical characterization of three midgut chitin deacetylases of the Lepidopteran insect Bombyx mori. Peritrophic membrane (PM) is a chitin and protein-containing extracellular matrix that lines the midgut in most insect species, functioning as a barrier to exogenous toxins and pathogens. Midgut chitin deacetylases (CDAs) are chitin-modifying enzymes known to alter the mechanical property and permeability of PM. However, biochemical properties and specific roles of these enzymes remain elusive. In this study, the midgut-expressed CDAs (BmCDA6, BmCDA7 and BmCDA8) from Bombyx mori were cloned, recombinantly expressed and purified and their enzymatic activities toward PM chitin were determined. Of the three enzymes, BmCDA7 exhibited the highest activity (0.284  mol/min/ mol), while BmCDA8 showed lower activity of 0.061  mol/min/ mol. BmCDA6 was inactive towards PM chitin. Gene expression patterns indicated that although all three CDA genes were specifically expressed in the anterior midgut, they differed in their temporal expression patterns. BmCDA6 was expressed almost exclusively at the mid-molt stage, the stage when the PM was thick and with multiple chitin layers. Unlike BmCDA6, high expression levels of BmCDA7 and BmCDA8 were observed only at the feeding stage, the stage when the PM is thin and with fewer chitin layers. The different gene expression patterns and biochemical characteristics provide new information about the functional specialization among BmCDA6, BmCDA7 and BmCDA8 proteins.',{'entities': [(45,63,'Gene'),(92,103,'Species'),(300,318,'Gene'),(321,324,'Gene'),(545,551,'Gene'),(553,559,'Gene'),(564,570,'Gene'),(577,588,'Species'),(726,732,'Gene'),(793,799,'Gene'),(846,852,'Gene'),(944,947,'Gene'),(1059,1065,'Gene'),(1194,1200,'Gene'),(1228,1234,'Gene'),(1239,1245,'Gene'),(1484,1490,'Gene'),(1492,1498,'Gene'),(1503,1509,'Gene')]}),('Suppression of Gene Juvenile Hormone Diol Kinase Delays Pupation in <i>Heortia vitessoides</i> Moore. Juvenile hormone diol kinase (JHDK) is a critical enzyme involved in juvenile hormone degradation in insects. In this study, <i>HvJHDK</i> in the <i>Heortia vitessoides</i> Moore (Lepidoptera: Crambidae) transcriptional library was cloned. Stage-specific expression patterns of <i>HvJHDK</i>, <i>HvJHEH</i>, and <i>HvJHE</i> as well as juvenile hormone titers were determined. The three tested enzymes participated in juvenile hormone degradation. Moreover, juvenile hormone titers peaked after larval-larval molts, consistent with a role for juvenile hormone in inhibition of metamorphosis. <i>HvJHDK</i> was subsequently suppressed using RNA interference (RNAi) to reveal its functions. Different concentrations of ds<i>JHDK</i> elicited the optimal interference efficiency at different life stages of <i>H. vitessoides</i>. Suppression of <i>HvJHDK</i> decreased HvJHDK content and increased the juvenile hormone titer, thereby resulting in reduced triglyceride content, sharply declined survival rate, clearly lethal phenotypes, and extended larval growth. Moreover, suppression of <i>HvJHDK</i> upregulated <i>HvJHEH</i> and <i>HvJHE</i> expression levels, suggesting that there is feedback regulation in the juvenile hormone metabolic pathway. Taken together, our findings provide molecular references for the selection of novel insecticidal targets.',{'entities': [(20,48,'Gene'),(71,90,'Species'),(102,130,'Gene'),(132,136,'Gene'),(230,236,'Gene'),(251,270,'Species'),(383,389,'Gene'),(398,404,'Gene'),(417,422,'Gene'),(697,703,'Gene'),(909,923,'Species'),(947,953,'Gene'),(968,974,'Gene'),(1191,1197,'Gene'),(1217,1223,'Gene'),(1235,1240,'Gene')]}),('Changes in the expression of four ABC transporter genes in response to imidacloprid in Bemisia tabaci Q (Hemiptera: Aleyrodidae). Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae), a globally invasive species complex that causes serious damage to field crops, has developed resistance to imidacloprid and many other pesticides. Insect detoxify to pesticides may partially depend on ABC transporters, which contribute to the detoxification of xenobiotics. To determine whether genes in the ABCG subfamily are involved in imidacloprid detoxification in B. tabaci Q, we cloned four ABCG subfamily genes based on the published MED/Q genome and on our previous study of the transcriptional response of ABC transporters in B. tabaci Q adults to imidacloprid. As indicated by the quantification of mRNA levels after a 6-h exposure, the expression level of ABCG3 was 3.3-fold higher in B. tabaci Q adults exposed to 100   g/mL imidacloprid rather than to the buffer control. The expression level of ABCG3 was higher in females than in males but did not significantly differ among eggs or nymphal stages and did not significantly differ among head, thorax, and abdomen tissues of adults. Knockdown of ABCG3 via RNA interference significantly increased the mortality of imidacloprid-treated laboratory and field-collected adults of B. tabaci Q. These results indicate that the ABCG3 gene may be involved in imidacloprid detoxification by B. tabaci Q.',{'entities': [(34,49,'Gene'),(87,101,'Species'),(130,144,'Species'),(384,399,'Gene'),(553,562,'Species'),(719,728,'Species'),(851,856,'Gene'),(880,889,'Species'),(993,998,'Gene'),(1194,1199,'Gene'),(1324,1333,'Species'),(1369,1374,'Gene'),(1430,1439,'Species')]}),('An Overview of Embryogenesis: External Morphology and Transcriptome Profiling in the Hemipteran Insect <i>Nilaparvata lugens</i>. During embryogenesis of insects, the morphological and transcriptional changes are important signatures to obtain a better understanding of insect patterning and evolution. The brown planthopper <i>Nilaparvata lugens</i> is a serious insect pest of rice plants, but its embryogenesis has not uncovered. Here, we described embryonic development process of the pest and found it belongs to an intermediate-germ mode. The RNA-seq data from different times (6, 30, 96, and 150 h, after egg laying) of embryogenesis were then analyzed, and a total of 10,895 genes were determined as differentially expressed genes (DEGs) based on pairwise comparisons. Afterward, 1,898 genes, differentially expressed in at least two comparisons of adjacent embryonic stages were divided into 10 clusters using K means cluster analysis (KMCA). Eight-gene modules were established using a weighted gene co-expression network analysis (WGCNA). Gene expression patterns in the different embryonic stages were identified by combining the functional enrichments of the stage-specific clusters and modules, which displayed the expression level and reprogramming of multiple developmental genes during embryogenesis. The "hub" genes at each embryonic stage with possible crucial roles were identified. Notably, we found a "center" set of genes that were related to overall membrane functions and might play important roles in the embryogenesis process. After parental RNAi of the <i>MSTRG.3372</i>, the hub gene, the embryo was observed as abnormal. Furthermore, some homologous genes in classic embryonic development processes and signaling pathways were also involved in embryogenesis of this insect. An improved comprehensive finding of embryogenesis within the <i>N. lugens</i> reveals better information on genetic and genomic studies of embryonic development and might be a potential target for RNAi-based control of this insect pest.',{'entities': [(106,124,'Species'),(307,324,'Species'),(328,346,'Species'),(379,383,'Species'),(1584,1594,'Gene'),(1869,1878,'Species')]}),('<i>NlATG1</i> Gene Participates in Regulating Autophagy and Fission of Mitochondria in the Brown Planthopper, <i>Nilaparvata lugens</i>. Autophagy plays multiple roles in regulating various physiological processes in cells. However, we currently lack a systematic analysis of autophagy and the autophagy-related gene 1 <i>ATG1</i> in the brown planthopper (BPH, <i>Nilaparvata lugens</i>), one of the most destructive of the insect pests of rice. In this study, the full-length cDNA of an autophagy-related gene, <i>NlATG1</i>, was cloned from BPH. Real-time qPCR (RT-qPCR) revealed that this <i>NlATG1</i> gene was expressed differently across developmental stages, at higher levels in nymphs but lower levels in adults. RNA interference with dsNlATG1 significantly decreased the mRNA level of the target gene to 14.6% at day 4 compared with that of the dsGFP control group. The survival of the dsNlATG1-treated group decreased significantly from day 4 onward, dropping to 48.3% on day 8. Examination using transmission electron microscopy (TEM) showed that epithelial cells of the BPH s midgut in the dsNlATG1-treated group had less autophagic vacuoles than did the dsGFP control, and knockdown of <i>NlATG1</i> clearly inhibited the starvation-induced autophagy response in this insect. RNA interference of <i>NlATG1</i> upregulated the <i>NlFis1</i> gene involved in mitochondrial fission, leading to reductions in mitochondrial width and area. Furthermore, knockdown of <i>NlATG1</i> also decreased the ATP content and accumulation of glycogen. Together, these results demonstrate that the <i>NlATG1</i> gene participates in regulating autophagy and fission of mitochondria in the brown planthopper, making it a potentially promising target for pest control given its key role in autophagy, including maintaining the normal structure and function of mitochondria.',{'entities': [(3,9,'Gene'),(91,108,'Species'),(113,131,'Species'),(338,355,'Species'),(357,360,'Species'),(365,383,'Species'),(441,445,'Species'),(516,522,'Gene'),(544,547,'Species'),(596,602,'Gene'),(1083,1086,'Species'),(1203,1209,'Gene'),(1313,1319,'Gene'),(1478,1484,'Gene'),(1598,1604,'Gene'),(1686,1703,'Species')]}),('RNA interference of tyrosine hydroxylase caused rapid mortality by impairing cuticle formation in Nilaparvata lugens (Hemiptera: Delphacidae). BACKGROUND: The application of RNA interference (RNAi) technique in controlling agricultural insect pests has been receiving much attention since the discovery of RNAi. The brown planthopper (BPH) Nilaparvata lugens, a notorious pest of rice, has evolved a high level of resistance to many kinds of insecticides. Tyrosine hydroxylase (Th) is an indispensable survival gene in holometabolous insects, playing key roles in cuticle tanning and immunity. In this study, we investigated whether Th could be used as a potential target in controlling N. lugens. RESULTS: Here, we demonstrated that NlTh had a periodical expression pattern during molting with the highest level observed in epidermis. Dysfunction of NlTH by dsNlTh microinjection or 3-IT feeding similarly caused rapid death of N. lugens. Compared with dsGFP control BPHs, dsNlTh injected BPHs (i) had cuticle pigmentation and sclerotizaton defects; (ii) had less endocuticle lamella in tergum integument; (iii) showed higher mortality during the molting process as a result of defective cuticle shedding; (iv) showed feeding disorders indicated by a low number of probe wound dots on rice; (v) had more vulnerable cuticle. CONCLUSION: This study demonstrated that TH orthologues play a conservative and crucial role for exocuticle tanning in both holometabolous and hemimetabolous insects, and NlTh could be targeted for RNAi-mediated BPH control. The rapid lethal phenotype of NlTH dysfunction BPHs partly induced by cuticle formation defects.  2020 Society of Chemical Industry.',{'entities': [(20,40,'Gene'),(98,116,'Species'),(316,333,'Species'),(335,338,'Species'),(340,358,'Species'),(380,384,'Species'),(456,476,'Gene'),(478,480,'Gene'),(633,635,'Gene'),(687,696,'Species'),(734,738,'Gene'),(851,855,'Gene'),(929,938,'Species'),(1286,1290,'Species'),(1366,1368,'Gene'),(1496,1500,'Gene'),(1537,1540,'Species'),(1580,1584,'Gene')]}),('The histone deacetylase NlHDAC1 regulates both female and male fertility in the brown planthopper, Nilaparvata lugens. Histone acetylation is a specific type of chromatin modification that serves as a key regulatory mechanism for many cellular processes in mammals. However, little is known about its biological function in invertebrates. Here, we identified 12 members of histone deacetylases (NlHDACs) in the brown planthopper (BPH), Nilaparvata lugens. RNAi-mediated silencing assay showed that NlHdac1, NlHdac3 and NlHdac4 played critical roles in female fertility via regulating ovary maturation or ovipositor development. Silencing of NlHdac1 substantially increased acetylation level of histones H3 and H4 in ovaries, indicating NlHDAC1 is the main histone deacetylase in ovaries of BPH. RNA sequencing (RNA-seq) analysis showed that knockdown of NlHdac1 impaired ovary development via multiple signalling pathways including the TOR pathway. Acoustic recording showed that males with NlHdac1 knockdown failed to make courtship songs, and thus were unacceptable to wild-type females, resulting in unfertilized eggs. Competition mating assay showed that wild-type females overwhelmingly preferred to mate with control males over NlHdac1-knockdown males. These findings improve our understanding of reproductive strategies controlled by HDACs in insects and provide a potential target for pest control.',{'entities': [(4,23,'Gene'),(24,31,'Gene'),(80,97,'Species'),(99,117,'Species'),(373,393,'Gene'),(395,402,'Gene'),(411,428,'Species'),(436,454,'Species'),(498,505,'Gene'),(507,514,'Gene'),(519,526,'Gene'),(641,648,'Gene'),(736,743,'Gene'),(756,775,'Gene'),(854,861,'Gene'),(991,998,'Gene'),(1234,1241,'Gene')]}),('Molecular features and expression profiles of octopamine receptors in the brown planthopper, Nilaparvata lugens. BACKGROUND: Octopamine, the invertebrate counterpart of adrenaline and noradrenaline, regulates and modulates many physiological and behavioral processes in insects. It mediates its effects by binding to specific octopamine receptors, which belong to the superfamily of G-protein coupled receptors (GPCRs). The expression profiles of octopamine receptor genes have been well documented in different developmental stages and multiple tissue types in several different insect orders. However, little work has addressed this issue in Hemiptera. RESULTS: In this study, we cloned four octopamine receptor genes from brown planthopper. The deduced amino acid sequences share high identity with other insect homologues and have the characteristic GPCRs domain architecture: seven transmembrane domains. These genes were expressed in all developmental stages and examined tissues. The expression of NlOA2B3 and NlOA3 was relatively higher in egg and first instar nymph stage than in other stages and other receptor genes. All of these receptor genes were more highly expressed in brain than other tissues. CONCLUSION: The identification of octopamine receptor genes in this study will provide a foundation for investigating the diverse roles played by NlOARs and for exploring specific target sites for chemicals that control agricultural pests.  2019 Society of Chemical Industry.',{'entities': [(46,66,'Gene'),(74,91,'Species'),(93,111,'Species'),(447,466,'Gene'),(694,713,'Gene'),(725,742,'Species'),(1005,1012,'Gene'),(1017,1022,'Gene'),(1246,1265,'Gene')]}),('Genome-Wide Screening and Functional Analysis Reveal That the Specific microRNA nlu-miR-173 Regulates Molting by Targeting Ftz-F1 in Nilaparvata lugens. Background: Molting is a crucial physiological behavior during arthropod growth. In the past few years, molting as well as chitin biosynthesis triggered by molting, is subject to regulation by miRNAs. However, how many miRNAs are involved in insect molting at the genome-wide level remains unknown. Results: We deeply sequenced four samples obtained from nymphs at the 2nd-3rd and 4th-5th instars, and then identified 61 miRNAs conserved in the Arthropoda and 326 putative novel miRNAs in the brown planthopper Nilaparvata lugens, a fearful pest of rice. A total of 36 mature miRNAs with significant different expression levels at the genome scale during molting, including 19 conserved and 17 putative novel miRNAs were identified. After comparing the expression profiles, we found that most of the targets of 36 miRNAs showing significantly differential expression were involved in energy and hormone pathways. One of the 17 putative novel miRNAs, nlu-miR-173 was chosen for functional study. nlu-miR-173 acts in 20-hydroxyecdysone signaling through its direct target, N. lugens Ftz-F1(NlFtz-F1), a transcription factor. Furthermore, we found that the transcription of nlu-miR-173 was promoted by Broad-Complex (BR-C), suggesting that its involvement in the 20-hydroxyecdysone pathway contributes to proper molting function. Conclusion: We provided a comprehensive resource of miRNAs associated with insect molting and identified a novel miRNA as a potential target for pest control.',{'entities': [(80,91,'Gene'),(123,129,'Gene'),(133,151,'Species'),(646,663,'Species'),(664,682,'Species'),(702,706,'Species'),(1103,1114,'Gene'),(1148,1159,'Gene'),(1224,1233,'Species'),(1234,1240,'Gene'),(1241,1249,'Gene'),(1324,1335,'Gene')]}),('Identification and functional analysis of a novel chorion protein essential for egg maturation in the brown planthopper. In insect eggs, the chorion has the essential function of protecting the embryo from external agents during development while allowing gas exchange for respiration. In this study, we found a novel gene, Nilaparvata lugens chorion protein (NlChP), that is involved in chorion formation in the brown planthopper, Nilaparvata lugens. NlChP was highly expressed in the follicular cells of female adult brown planthoppers. Knockdown of NlChP resulted in oocyte malformation and the inability to perform oviposition, and electron microscopy showed that the malformed oocytes had thin and rough endochorion layers compared to the control group. Liquid chromatography with tandem mass spectrometry analysis of the eggshell components revealed four unique peptides that were matched to NlChP. Our results demonstrate that NlChP is a novel chorion protein essential for egg maturation in N. lugens, a hemipteran insect with telotrophic meroistic ovaries. NlChP may be a potential target in RNA interference-based insect pest management.',{'entities': [(50,65,'Gene'),(102,119,'Species'),(324,342,'Species'),(343,358,'Gene'),(360,365,'Gene'),(413,430,'Species'),(432,450,'Species'),(452,457,'Gene'),(552,557,'Gene'),(898,903,'Gene'),(934,939,'Gene'),(999,1008,'Species'),(1066,1071,'Gene')]}),('Analysis of Homologs of Cry-toxin Receptor-Related Proteins in the Midgut of a Non-Bt Target, Nilaparvata lugens (St l) (Hemiptera: Delphacidae). The brown planthopper (BPH) Nilaparvata lugens is one of the most destructive insect pests in the rice fields of Asia. Like other hemipteran insects, BPH is not susceptible to Cry toxins of Bacillus thuringiensis (Bt) or transgenic rice carrying Bt cry genes. Lack of Cry receptors in the midgut is one of the main reasons that BPH is not susceptible to the Cry toxins. The main Cry-binding proteins (CBPs) of the susceptible insects are cadherin, aminopeptidase N (APN), and alkaline phosphatase (ALP). In this study, we analyzed and validated de novo assembled transcripts from transcriptome sequencing data of BPH to identify and characterize homologs of cadherin, APN, and ALP. We then compared the cadherin-, APN-, and ALP-like proteins of BPH to previously reported CBPs to identify their homologs in BPH. The sequence analysis revealed that at least one cadherin, one APN, and two ALPs of BPH contained homologous functional domains identified from the Cry-binding cadherin, APN, and ALP, respectively. Quantitative real-time polymerase chain reaction used to verify the expression level of each putative Cry receptor homolog in the BPH midgut indicated that the CBPs homologous APN and ALP were expressed at high or medium-high levels while the cadherin was expressed at a low level. These results suggest that homologs of CBPs exist in the midgut of BPH. However, differences in key motifs of CBPs, which are functional in interacting with Cry toxins, may be responsible for insusceptibility of BPH to Cry toxins.',{'entities': [(24,58,'Gene'),(83,85,'Species'),(94,112,'Species'),(150,167,'Species'),(169,172,'Species'),(174,192,'Species'),(244,248,'Species'),(296,299,'Species'),(336,358,'Species'),(360,362,'Species'),(378,382,'Species'),(392,394,'Species'),(414,427,'Gene'),(474,477,'Species'),(525,545,'Gene'),(547,551,'Gene'),(584,592,'Gene'),(594,610,'Gene'),(612,615,'Gene'),(622,642,'Gene'),(644,647,'Gene'),(759,762,'Species'),(804,812,'Gene'),(814,817,'Gene'),(823,826,'Gene'),(849,857,'Gene'),(860,863,'Gene'),(870,873,'Gene'),(891,894,'Species'),(918,922,'Gene'),(953,956,'Species'),(1007,1015,'Gene'),(1021,1024,'Gene'),(1034,1038,'Gene'),(1042,1045,'Species'),(1118,1126,'Gene'),(1128,1131,'Gene'),(1137,1140,'Gene'),(1286,1289,'Species'),(1316,1320,'Gene'),(1332,1335,'Gene'),(1340,1343,'Gene'),(1399,1407,'Gene'),(1477,1481,'Gene'),(1505,1508,'Species'),(1548,1552,'Gene'),(1650,1653,'Species')]}),('Double-stranded RNA targeting calmodulin reveals a potential target for pest management of Nilaparvata lugens. BACKGROUND: Calmodulin (CaM) is an essential protein in cellular activity and plays important roles in many processes in insect development. RNA interference (RNAi) has been hypothesized to be a promising method for pest control. CaM is a good candidate for RNAi target. However, the sequence and function of CaM in Nilaparvata lugens are unknown. Furthermore, the double-stranded RNA (dsRNA) target to CaM gene in pest control is still unavailable. RESULTS: In the present study, two alternatively spliced variants of CaM transcripts, designated NlCaM1 and NlCaM2, were cloned from N. lugens. The two cDNA sequences exhibited 100% identity to each other in the open reading frame (ORF), and only differed in the 3  untranslated region (UTR). NlCaM including NlCaM1 and NlCaM2 mRNA was detectable in all developmental stages and tissues of N. lugens, with significantly increased expression in the salivary glands. Knockdown of NlCaM expression by RNAi with different dsRNAs led to an inability to molt properly, increased mortality, which ranged from 49.7 to 92.5%, impacted development of the ovaries and led to female infertility. There were no significant reductions in the transcript levels of vitellogenin and its receptor or in the total vitellogenin protein level relative to the control group. However, a significant reduction in vitellogenin protein was detected in ovaries injected with dsNlCaM. In addition, a specific dsRNA of NlCaM for control of N. lugens was designed and tested. CONCLUSION: NlCaM plays important roles mainly in nymph development and uptake of vitellogenin by ovaries in vitellogenesis in N. lugens. dsRNA derived from the less conserved 3 -UTR of NlCaM shows great potential for RNAi-based N. lugens management.  2018 Society of Chemical Industry.',{'entities': [(30,40,'Gene'),(91,109,'Species'),(123,133,'Gene'),(135,138,'Gene'),(341,344,'Gene'),(420,423,'Gene'),(427,445,'Species'),(514,517,'Gene'),(630,633,'Gene'),(658,664,'Gene'),(669,675,'Gene'),(694,703,'Species'),(854,859,'Gene'),(870,876,'Gene'),(881,887,'Gene'),(951,960,'Species'),(1039,1044,'Gene'),(1551,1556,'Gene'),(1572,1581,'Species'),(1619,1624,'Gene'),(1734,1743,'Species'),(1793,1798,'Gene'),(1836,1845,'Species')]}),('Characterization of NlHox3, an essential gene for embryonic development in Nilaparvata lugens. Hox genes encode transcriptional regulatory proteins that control axial patterning in all bilaterians. The brown planthopper (BPH), Nilaparvata lugens (Hemiptera: Delphacidae), is a destructive insect pest of rice plants in Asian countries. During analysis of the N. lugens transcriptome, we identified a Hox3-like gene (NlHox3) that was highly and specifically expressed in the embryonic stage. We performed functional analysis on the gene to identify its roles in embryonic development and its potential use as a target in RNA interference (RNAi) based pest control. The sequence analysis showed that NlHox3 was homologous to the Hox3 gene and was most closely related with zen of Drosophila. There were no significant differences in oviposition between the treated and control females after injecting double-stranded RNA of NlHox3 (dsNlHox3) into newly emerged female adult BPHs; however, there was a significant difference in the hatchability of those eggs laid, which no egg from the treated group hatched normally. Injecting female adult BPHs with dsNlHox3 led to necrosis of these offspring embryos, with eye reversal and undeveloped organs, suggesting that NlHox3 was an essential gene for embryonic development and might be a potential target for RNAi-based control of this insect pest.',{'entities': [(20,26,'Gene'),(75,93,'Species'),(202,219,'Species'),(221,224,'Species'),(227,245,'Species'),(304,308,'Species'),(359,368,'Species'),(400,409,'Gene'),(416,422,'Gene'),(698,704,'Gene'),(922,928,'Gene'),(1260,1266,'Gene')]}),('An adenylyl cyclase like-9 gene (NlAC9) influences growth and fecundity in the brown planthopper, Nilaparvata lugens (St l) (Hemiptera: Delphacidae). The cAMP/PKA intracellular signaling pathway is launched by adenylyl cyclase (AC) conversion of adenosine triphosphate (ATP) to 3 , 5 -cyclic AMP (cAMP) and cAMP-dependent activation of PKA. Although this pathway is very well known in insect physiology, there is little to no information on it in some very small pest insects, such as the brown planthopper (BPH), Nilaparvata lugens St l. BPH is a destructive pest responsible for tremendous crop losses in rice cropping systems. We are investigating the potentials of novel pest management technologies from RNA interference perspective. Based on analysis of transcriptomic data, the BPH AC like-9 gene (NlAC9) was up-regulated in post-mating females, which led us to pose the hypothesis that NlAC9 is a target gene that would lead to reduced BPH fitness and populations. Targeting NlAC9 led to substantially decreased soluble ovarian protein content, yeast-like symbiont abundance, and vitellogenin gene expression, accompanied with stunted ovarian development and body size. Eggs laid were decreased and oviposition period shortened. Taken together, our findings indicated that NlAC9 exerted pronounced effects on female fecundity, growth and longevity, which strongly supports our hypothesis.',{'entities': [(3,26,'Gene'),(33,38,'Gene'),(79,96,'Species'),(98,116,'Species'),(210,226,'Gene'),(489,506,'Species'),(508,511,'Species'),(514,532,'Species'),(539,542,'Species'),(607,611,'Species'),(785,788,'Species'),(805,810,'Gene'),(894,899,'Gene'),(983,988,'Gene'),(1053,1072,'Species'),(1088,1100,'Gene'),(1281,1286,'Gene')]}),('Identification of a sugar gustatory receptor and its effect on fecundity of the brown planthopper Nilaparvata lugens. In insects, the gustatory system plays a crucial role in multiple physiological behaviors, including feeding, toxin avoidance, courtship, mating and oviposition. Gustatory stimuli from the environment are recognized by gustatory receptors. To date, little is known about the function of gustatory receptors in agricultural pest insects. In this study, we cloned a sugar gustatory receptor gene, NlGr11, from the brown planthopper (BPH), Nilaparvata lugens (St l), a serious pest of rice in Asia; we then identified its ligands, namely, fructose, galactose and arabinose, by calcium imaging assay. After injection of NlGr11 double-stranded RNA, we found that the number of eggs laid by BPH decreased. Moreover, we found that NlGr11 inhibited the phosphorylation of adenosine monophosphate-activated protein kinase (AMPK) and promoted the phosphorylation of protein kinase B (AKT). These findings demonstrated that NlGr11 could accelerate the fecundity of BPH through AMPK- and AKT-mediated signaling pathways. This is the first report to indicate that a gustatory receptor modulates the fecundity of insects and that the receptor could be a potential target for pest control.',{'entities': [(26,44,'Gene'),(80,97,'Species'),(98,116,'Species'),(337,356,'Gene'),(405,424,'Gene'),(488,506,'Gene'),(513,519,'Gene'),(530,547,'Species'),(549,552,'Species'),(555,573,'Species'),(600,604,'Species'),(734,740,'Gene'),(803,806,'Species'),(842,848,'Gene'),(1031,1037,'Gene'),(1072,1075,'Species'),(1171,1189,'Gene')]}),('Influence of the RDL A301S mutation in the brown planthopper Nilaparvata lugens on the activity of phenylpyrazole insecticides. We discovered the A301S mutation in the RDL GABA-gated chloride channel of fiprole resistant rice brown planthopper, Nilaparvata lugens populations by DNA sequencing and SNP calling via RNASeq. Ethiprole selection of two field N. lugens populations resulted in strong resistance to both ethiprole and fipronil and resulted in fixation of the A301S mutation, as well as the emergence of another mutation, Q359E in one of the selected strains. To analyse the roles of these mutations in resistance to phenylpyrazoles, three Rdl constructs: wild type, A301S and A301S+Q359E were expressed in Xenopus laevis oocytes and assessed for their sensitivity to ethiprole and fipronil using two-electrode voltage-clamp electrophysiology. Neither of the mutant Rdl subtypes significantly reduced the antagonistic action of fipronil, however there was a significant reduction in response to ethiprole in the two mutated subtypes compared with the wild type. Bioassays with a Drosophila melanogaster strain carrying the A301S mutation showed strong resistance to ethiprole but not fipronil compared to a strain without this mutation, thus further supporting a causal role for the A301S mutation in resistance to ethiprole. Homology modelling of the N. lugens RDL channel did not suggest implications of Q359E for fiprole binding in contrast to A301S located in transmembrane domain M2 forming the channel pore. Synergist bioassays provided no evidence of a role for cytochrome P450s in N. lugens resistance to fipronil and the molecular basis of resistance to this compound remains unknown. In summary this study provides strong evidence that target-site resistance underlies widespread ethiprole resistance in N. lugens populations.',{'entities': [(43,60,'Species'),(61,79,'Species'),(168,199,'Gene'),(221,225,'Species'),(226,243,'Species'),(245,263,'Species'),(355,364,'Species'),(717,731,'Species'),(1089,1112,'Species'),(1362,1371,'Species'),(1599,1608,'Species'),(1824,1833,'Species')]}),('MicroRNA and dsRNA targeting chitin synthase A reveal a great potential for pest management of the hemipteran insect Nilaparvata lugens. BACKGROUND: Two RNA silencing pathways in insects are known to exist that are mediated by short interfering RNAs (siRNAs) and microRNAs (miRNAs), which have been hypothesised to be promising methods for insect pest control. However, a comparison between miRNA and siRNA in pest control is still unavailable, particularly in targeting chitin synthase gene A (CHSA). RESULTS: The dsRNA for Nilaparvata lugens CHSA (dsNlCHSA) and the microR-2703 (miR-2703) mimic targeting NlCHSA delivered via feeding affected the development of nymphs, reduced their chitin content and led to lethal phenotypes. The protein level of NlCHSA was downregulated after female adults were injected with dsNlCHSA or the miR-2703 mimic, but there were no significant differences in vitellogenin (NlVg) expression or in total oviposition relative to the control group. However, 90.68 and 46.13% of the eggs laid by the females injected with dsNlCHSA and miR-2703 mimic were unable to hatch, respectively. In addition, a second-generation miRNA and RNAi effect on N. lugens was observed. CONCLUSION: Ingested miR-2703 seems to be a good option for killing N. lugens nymphs, while NlCHSA may be a promising target for RNAi-based pest management. These findings provide important evidence for applications of small non-coding RNAs (snRNAs) in insect pest management.  2016 Society of Chemical Industry.',{'entities': [(29,46,'Gene'),(117,135,'Species'),(495,499,'Gene'),(525,543,'Species'),(544,548,'Gene'),(568,579,'Gene'),(581,589,'Gene'),(607,613,'Gene'),(752,758,'Gene'),(832,840,'Gene'),(1064,1072,'Gene'),(1173,1182,'Species'),(1218,1226,'Gene'),(1265,1274,'Species'),(1289,1295,'Gene')]}),('Molecular characterization, expression analysis and RNAi knock-down of elongation factor 1a and 1y from Nilaparvata lugens and its yeast-like symbiont. In the present paper, four cDNAs encoding the alpha and gamma subunits of elongation factor 1 (EF-1) were cloned and sequenced from Nilaparvata lugens, named NlEF-1a, NlEF-1y, and its yeast-like symbiont (YLS), named YsEF-1a and YsEF-1y, respectively. Comparisons with sequences from other species indicated a greater conservation for EF-1a than for EF-1y. NlEF-1a has two identical copies. The deduced amino acid sequence homology of NlEF-1a and NlEF-1y is 96 and 64%, respectively, compared with Homalodisca vitripennis and Locusta migratoria. The deduced amino acid sequence homology of YsEF-1a and YsEF-1y is 96 and 74%, respectively, compared with Metarhizium anisopliae and Ophiocordyceps sinensis. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis revealed that the expression level of NlEF-1a and NlEF-1y mRNA in hemolymph, ovary, fat body and salivary glands were higher than the midgut and leg tissue. YsEF-1a and YsEF-1y was highly expressed in fat body. The expression level of NlEF-1a was higher than that of NlEF-1y. Through RNA interference (RNAi) of the two genes, the mortality of nymph reached 92.2% at the 11th day after treatment and the ovarian development was severely hindered. The RT-qPCR analysis verified the correlation between mortality, sterility and the down-regulation of the target genes. The expression and synthesis of vitellogenin (Vg) protein in insects injected with NlEF-1a and NlEF-1y double-stranded RNA (dsRNA) was significantly lower than control groups. Attempts to knockdown the YsEF-1 genes in the YLS was unsuccessful. However, the phenotype of N. lugens injected with YsEF-1a dsRNA was the same as that injected with NlEF-1a dsRNA, possibly due to the high similarity (up to 71.9%) in the nucleotide sequences between NlEF-1a and YsEF-1a. We demonstrated that partial silencing of NlEF-1a and NlEF-1y genes caused lethal and sterility effect on N. lugens. NlEF-1y shares low identity with that of other insects and therefore it could be a potential target for RNAi-based pest management.',{'entities': [(71,90,'Gene'),(104,122,'Species'),(131,150,'Species'),(226,245,'Gene'),(247,251,'Gene'),(284,302,'Species'),(310,316,'Gene'),(319,325,'Gene'),(336,355,'Species'),(357,360,'Species'),(509,515,'Gene'),(587,593,'Gene'),(599,605,'Gene'),(805,827,'Species'),(832,855,'Species'),(975,981,'Gene'),(987,993,'Gene'),(1172,1178,'Gene'),(1204,1210,'Gene'),(1586,1592,'Gene'),(1598,1604,'Gene'),(1725,1728,'Species'),(1773,1782,'Species'),(1846,1852,'Gene'),(1947,1953,'Gene'),(2010,2016,'Gene'),(2022,2028,'Gene'),(2074,2083,'Species'),(2085,2091,'Gene')]}),('Discovery and functional identification of fecundity-related genes in the brown planthopper by large-scale RNA interference. Recently, transcriptome and proteome data have increasingly been used to identify potential novel genes related to insect phenotypes. However, there are few studies reporting the large-scale functional identification of such genes in insects. To identify novel genes related to fecundity in the brown planthopper (BPH), Nilaparvata lugens, 115 genes were selected from the transcriptomic and proteomic data previously obtained from high- and low-fecundity populations in our laboratory. The results of RNA interference (RNAi) feeding experiments showed that 91.21% of the genes were involved in the regulation of vitellogenin (Vg) expression and may influence BPH fecundity. After RNAi injection experiments, 12 annotated genes were confirmed as fecundity-related genes and three novel genes were identified in the BPH. Finally, C-terminal binding protein (CtBP) was shown to play an important role in BPH fecundity. Knockdown of CtBP not only led to lower survival, underdeveloped ovaries and fewer eggs laid but also resulted in a reduction in Vg protein expression. The novel gene resources gained from this study will be useful for constructing a Vg regulation network and may provide potential target genes for RNAi-based pest control.',{'entities': [(74,91,'Species'),(420,437,'Species'),(439,442,'Species'),(445,463,'Species'),(738,750,'Gene'),(752,754,'Gene'),(785,788,'Species'),(940,943,'Species'),(954,980,'Gene'),(982,986,'Gene'),(1027,1030,'Species'),(1055,1059,'Gene'),(1171,1173,'Gene'),(1276,1278,'Gene')]}),('Ran Involved in the Development and Reproduction Is a Potential Target for RNA-Interference-Based Pest Management in Nilaparvata lugens. UNASSIGNED: Ran (RanGTPase) in insects participates in the 20-hydroxyecdysone signal transduction pathway in which downstream genes, FTZ-F1, Kruppel-homolog 1 (Kr-h1) and vitellogenin, are involved. A putative Ran gene (NlRan) was cloned from Nilaparvata lugens, a destructive phloem-feeding pest of rice. NlRan has the typical Ran primary structure features that are conserved in insects. NlRan showed higher mRNA abundance immediately after molting and peaked in newly emerged female adults. Among the examined tissues ovary had the highest transcript level, followed by fat body, midgut and integument, and legs. Three days after dsNlRan injection the NlRan mRNA abundance in the third-, fourth-, and fifth-instar nymphs was decreased by 94.3%, 98.4% and 97.0%, respectively. NlFTZ-F1 expression levels in treated third- and fourth-instar nymphs were reduced by 89.3% and 23.8%, respectively. In contrast, NlKr-h1 mRNA levels were up-regulated by 67.5 and 1.5 folds, respectively. NlRan knockdown significantly decreased the body weights, delayed development, and killed >85% of the nymphs at day seven. Two apparent phenotypic defects were observed: (1) Extended body form, and failed to molt; (2) The cuticle at the notum was split open but cannot completely shed off. The newly emerged female adults from dsNlRan injected fifth-instar nymphs showed lower levels of NlRan and vitellogenin, lower weight gain and honeydew excretion comparing with the blank control, and no offspring. Those results suggest that NlRan encodes a functional protein that was involved in development and reproduction. The study established proof of concept that NlRan could serve as a target for dsRNA-based pesticides for N. lugens control.',{'entities': [(0,3,'Gene'),(117,135,'Species'),(149,152,'Gene'),(154,163,'Gene'),(270,276,'Gene'),(278,295,'Gene'),(297,302,'Gene'),(308,320,'Gene'),(347,350,'Gene'),(357,362,'Gene'),(380,398,'Species'),(437,441,'Species'),(443,448,'Gene'),(527,532,'Gene'),(792,797,'Gene'),(1046,1053,'Gene'),(1121,1126,'Gene'),(1508,1513,'Gene'),(1518,1530,'Gene'),(1652,1657,'Gene'),(1782,1787,'Gene'),(1843,1852,'Species')]}),('Silencing a sugar transporter gene reduces growth and fecundity in the brown planthopper, Nilaparvata lugens (St l) (Hemiptera: Delphacidae). UNASSIGNED: The brown planthopper (BPH), Nilaparvata lugens, sugar transporter gene 6 (Nlst6) is a facilitative glucose/fructose transporter (often called a passive carrier) expressed in midgut that mediates sugar transport from the midgut lumen to hemolymph. The influence of down regulating expression of sugar transporter genes on insect growth, development, and fecundity is unknown. Nonetheless, it is reasonable to suspect that transporter-mediated uptake of dietary sugar is essential to the biology of phloem-feeding insects. Based on this reasoning, we posed the hypothesis that silencing, or reducing expression, of a BPH sugar transporter gene would be deleterious to the insects. To test our hypothesis, we examined the effects of Nlst6 knockdown on BPH biology. Reducing expression of Nlst6 led to profound effects on BPHs. It significantly prolonged the pre-oviposition period, shortened the oviposition period, decreased the number of eggs deposited and reduced body weight, compared to controls. Nlst6 knockdown also significantly decreased fat body and ovarian (particularly vitellogenin) protein content as well as vitellogenin gene expression. Experimental BPHs accumulated less fat body glucose compared to controls. We infer that Nlst6 acts in BPH growth and fecundity, and has potential as a novel target gene for control of phloem-feeding pest insects.',{'entities': [(12,29,'Gene'),(71,88,'Species'),(90,108,'Species'),(158,175,'Species'),(177,180,'Species'),(183,201,'Species'),(203,227,'Gene'),(229,234,'Gene'),(770,773,'Species'),(885,890,'Gene'),(904,907,'Species'),(940,945,'Gene'),(1154,1159,'Gene'),(1393,1398,'Gene'),(1407,1410,'Species')]}),('The insect ecdysone receptor is a good potential target for RNAi-based pest control. RNA interference (RNAi) has great potential for use in insect pest control. However, some significant challenges must be overcome before RNAi-based pest control can become a reality. One challenge is the proper selection of a good target gene for RNAi. Here, we report that the insect ecdysone receptor (EcR) is a good potential target for RNAi-based pest control in the brown planthopper Nilaparvata lugens, a serious insect pest of rice plants. We demonstrated that the use of a 360 bp fragment (NlEcR-c) that is common between NlEcR-A and NlEcR-B for feeding RNAi experiments significantly decreased the relative mRNA expression levels of NlEcR compared with those in the dsGFP control. Feeding RNAi also resulted in a significant reduction in the number of offspring per pair of N. lugens. Consequently, a transgenic rice line expressing NlEcR dsRNA was constructed by Agrobacterium- mediated transformation. The results of qRT-PCR showed that the total copy number of the target gene in all transgenic rice lines was 2. Northern blot analysis showed that the small RNA of the hairpin dsNlEcR-c was successfully expressed in the transgenic rice lines. After newly hatched nymphs of N. lugens fed on the transgenic rice lines, effective RNAi was observed. The NlEcR expression levels in all lines examined were decreased significantly compared with the control. In all lines, the survival rate of the nymphs was nearly 90%, and the average number of offspring per pair in the treated groups was significantly less than that observed in the control, with a decrease of 44.18-66.27%. These findings support an RNAi-based pest control strategy and are also important for the management of rice insect pests.',{'entities': [(11,28,'Gene'),(370,387,'Gene'),(389,392,'Gene'),(456,473,'Species'),(474,492,'Species'),(519,523,'Species'),(583,588,'Gene'),(615,620,'Gene'),(627,632,'Gene'),(727,732,'Gene'),(868,877,'Species'),(906,910,'Species'),(927,932,'Gene'),(1092,1096,'Species'),(1229,1233,'Species'),(1271,1280,'Species'),(1303,1307,'Species'),(1348,1353,'Gene'),(1774,1778,'Species')]}),('The expression of Spodoptera exigua P450 and UGT genes: tissue specificity and response to insecticides. Cytochrome P450 and UDP-glucosyltransferase (UGT) as phase I and phase II metabolism enzymes, respectively, play vital roles in the breakdown of endobiotics and xenobiotics. Insects can increase the expression of detoxification enzymes to cope with the stress from xenobiotics including insecticides. However, the molecular mechanisms for insecticide detoxification in Spodoptera exigua remain elusive, and the genes conferring insecticide metabolisms in this species are less well reported. In this study, 68 P450 and 32 UGT genes were identified. Phylogenetic analysis showed gene expansions in CYP3 and CYP4 clans of P450 genes and UGT33 family of this pest. P450 and UGT genes exhibited specific tissue expression patterns. Insecticide treatments in fat body cells of S. exigua revealed that the expression levels of P450 and UGT genes were significantly influenced by challenges of abamectin, lambda-cyhalothrin, chlorantraniliprole, metaflumizone and indoxacarb. Multiple genes for detoxification were affected in expression levels after insecticide exposures. The results demonstrated that lambda-cyhalothrin, chlorantraniliprole, metaflumizone and indoxacarb induced similar responses in the expression of P450 and UGT genes in fat body cells; eight P450 genes and four UGT genes were co-up-regulated significantly, and no or only a few CYP/UGT genes were down-regulated significantly by these four insecticides. However, abamectin triggered a distinct response for P450 and UGT gene expression; more P450 and UGT genes were down-regulated by abamectin than by the other four compounds. In conclusion, P450 and UGT genes from S. exigua were identified, and different responses to abamectin suggest a different mechanism for insecticide detoxification.',{'entities': [(18,35,'Species'),(36,40,'Gene'),(45,48,'Gene'),(105,120,'Gene'),(125,148,'Gene'),(150,153,'Gene'),(474,491,'Species'),(615,619,'Gene'),(627,630,'Gene'),(702,706,'Gene'),(711,715,'Gene'),(725,729,'Gene'),(740,745,'Gene'),(767,771,'Gene'),(776,779,'Gene'),(877,886,'Species'),(926,930,'Gene'),(935,938,'Gene'),(1319,1323,'Gene'),(1328,1331,'Gene'),(1363,1367,'Gene'),(1383,1386,'Gene'),(1454,1457,'Gene'),(1579,1583,'Gene'),(1588,1591,'Gene'),(1614,1618,'Gene'),(1623,1626,'Gene'),(1715,1719,'Gene'),(1724,1727,'Gene'),(1739,1748,'Species')]}),('Carboxylesterase genes in pyrethroid resistant house flies, Musca domestica. Carboxylesterases are one of the major enzyme families involved in the detoxification of pyrethroids. Up-regulation of carboxylesterase genes is thought to be a major component of insecticide resistant mechanisms in insects. Based on the house fly transcriptome and genome database, a total of 39 carboxylesterase genes of different functional clades have been identified in house flies. In this study, eleven of these genes were found to be significantly overexpressed in the resistant ALHF house fly strain compared with susceptible aabys and wild-type CS strains. Eight up-regulated carboxylesterase genes with their expression levels were further induced to a higher level in response to permethrin treatments, indicating that constitutive and inductive overexpression of carboxylesterases are co-responsible for the enhanced detoxification of insecticides. Spatial expression studies revealed these up-regulated genes to be abundantly distributed in fat bodies and genetically mapped on autosome 2 or 3 of house flies, and their expression could be regulated by factors on autosome 1, 2 and 5. Taken together, these results demonstrate that multiple carboxylesterase genes are co-upregulated in resistant house flies, providing further evidence for their involvement in the detoxification of insecticides and development of insecticide resistance.',{'entities': [(0,16,'Gene'),(60,75,'Species'),(196,212,'Gene'),(315,324,'Species'),(374,390,'Gene'),(569,578,'Species'),(663,679,'Gene'),(1232,1248,'Gene')]}),('Molecular characterization of glutamate-gated chloride channel and its possible roles in development and abamectin susceptibility in the rice stem borer, Chilo suppressalis. Glutamate-gated chloride channels (GluCls) mediate fast inhibitory neurotransmission in invertebrate nervous systems, and are of considerable interest in insecticide discovery. The full length cDNA encoding CsGluCl was cloned from the rice stem borer Chilo suppressalis (Walker). Multiple cDNA sequence alignment revealed three variants of CsGluCl generated by alternative splicing of exon 3 and exon 9. While all the transcripts were predominantly expressed in both nerve cord and brain, the expression patterns of these three variants differed among other tissues and developmental stages. Specifically, the expression level of CsGluCl C in cuticle was similar to that in nerve cord and brain, and was the predominant variant in late pupae and early adult stages. Both injection and oral delivery of dsGluCl significantly reduced the mRNA level of CsGluCl. Increased susceptibility to abamectin and reduced larvae growth and pupation rate were observed in dsGluCl-treated larvae. Thus, our results provide the evidence that in addition to act as the target of abamectin, GluCls also play important physiological roles in the development of insects.',{'entities': [(30,62,'Gene'),(137,141,'Species'),(154,172,'Species'),(209,214,'Gene'),(381,388,'Gene'),(409,413,'Species'),(425,443,'Species'),(514,521,'Gene'),(804,811,'Gene'),(1024,1031,'Gene')]}),('The regulation of crecropin-A and gloverin 2 by the silkworm Toll-like gene 18 wheeler in immune response. The innate immune system is conserved among different insect species in its response to microorganism infection. The transmembrane receptors of the Toll superfamily play an important role in activating immune response, however, the function of silkworm Toll family member 18 Wheeler (18 W) remained unclear. Here, the 18w gene in silkworm was characterized. A relatively high transcription level of Bm18w mRNA was found in Malpighian tubules, and in eggs, larvae pre-molt to fourth instar, pupae and adults. When silkworm larvae were infected with E. coli or S. aureus, Bm18w showed a significant response, especially to E. coli, but did not have antibacterial activity. To further identify the downstream antimicrobial peptide genes of Bm18w, expression of Bm18w was knocked down with siRNA in vitro, resulting in significant decreases of cecropin-A, gloverin 2, and moricin B3. The overexpression of Bm18w was carried out using pIZT/V5-His-mCherry insect vector in BmN cells and significant upregulation of cecropin-A and gloverin 2 was detected, as well as upregulation of attacin and defensin. Based on the results, we concluded that Bm18w is involved in response to bacterial infection by selectively inducing the expression of antimicrobial peptide genes, especially cecropin-A and gloverin 2. This study provides valuable data to supplement understanding of the immune pathway of the silkworm.',{'entities': [(34,44,'Gene'),(52,60,'Species'),(76,86,'Gene'),(351,359,'Species'),(379,389,'Gene'),(391,395,'Gene'),(425,428,'Gene'),(437,445,'Species'),(506,511,'Gene'),(620,628,'Species'),(655,662,'Species'),(666,675,'Species'),(677,682,'Gene'),(728,735,'Species'),(844,849,'Gene'),(865,870,'Gene'),(947,957,'Gene'),(959,969,'Gene'),(1009,1014,'Gene'),(1116,1126,'Gene'),(1131,1141,'Gene'),(1183,1190,'Gene'),(1195,1203,'Gene'),(1245,1250,'Gene'),(1380,1390,'Gene'),(1395,1405,'Gene'),(1498,1506,'Species')]}),('Bombyxin/Akt signaling in relation to the embryonic diapause process of the silkworm, Bombyx mori. Our previous study showed that phosphorylation of glycogen synthase kinase (GSK)-3b is related to the embryonic diapause process in Bombyx. However, the upstream signaling pathway was not clearly understood. In the present study, we examined bombyxin/Akt signaling in relation to the embryonic diapause process of B. mori. Results showed that GSK-3b phosphorylation stimulated by dechorionation was blocked by LY294002, a specific phosphatidylinositol 3-kinase (PI3K) inhibitor, indicating involvement of PI3K in GSK-3b phosphorylation in dechorionated eggs. Direct determination of Akt phosphorylation showed that dechorionation stimulated Akt phosphorylation. The Akt phosphorylation was blocked by LY294002. Temporal changes in Akt phosphorylation showed that different changing patterns exist between diapause and developing eggs. Relatively higher phosphorylation levels of Akt were detected between days 3 and 5 after oviposition in non-diapause eggs compared to those at the same stages in diapause eggs. Upon treatment with HCl, which prevents diapause initiation, Akt phosphorylation levels exhibited a later and much broader peak compared to diapause eggs. Examination of expression levels of the bombyxin-Z1 gene showed that in diapause eggs, a major peak occurred 1  day after oviposition, and its level then sharply decreased on day 2. However, in both non-diapause and HCl-treated eggs, a major broad peak was detected between days 1 and 4 after oviposition. These temporal changes in bombyxin-Z1 gene expression levels during embryonic stages coincided with changes in Akt phosphorylation, indicating that bombyxin-Z1 is likely an upstream signaling component for Akt phosphorylation. Taken together, our results indicated that PI3K/Akt is an upstream signaling pathway for GSK-3b phosphorylation and is associated with the diapause process of B. mori eggs. To our knowledge, this is the first study to demonstrate the potential correlation between bombyxin/Akt signaling and the embryonic diapause process.',{'entities': [(0,8,'Gene'),(9,12,'Gene'),(76,84,'Species'),(86,97,'Species'),(231,237,'Species'),(341,349,'Gene'),(350,353,'Gene'),(413,420,'Species'),(530,559,'Gene'),(682,685,'Gene'),(740,743,'Gene'),(765,768,'Gene'),(830,833,'Gene'),(978,981,'Gene'),(1172,1175,'Gene'),(1306,1317,'Gene'),(1598,1609,'Gene'),(1683,1686,'Gene'),(1720,1731,'Gene'),(1778,1781,'Gene'),(1847,1850,'Gene'),(1958,1965,'Species'),(2063,2071,'Gene'),(2072,2075,'Gene')]}),('Tudor knockdown disrupts ovary development in Bactrocera dorsalis. One of the main functions of the piwi-interacting RNA pathway is the post-transcriptional silencing of transposable elements in the germline of many species. In insects, proteins belonging to the Tudor superfamily proteins belonging to the Tudor superfamily play an important role in to play an important role in this mechanism. In this study, we identified the tudor gene in the oriental fruit fly, Bactrocera dorsalis, investigated the spatiotemporal expressional profile of the gene, and performed a functional analysis using RNA interference. We identified one transcript for a tudor homologue in the B. dorsalis transcriptome, which encodes a protein containing the typical 10 Tudor domains and an Adenosine triphosphate (ATP) synthase delta subunit signature. Phylogenetic analysis confirmed the identity of this transcript as a tudor homologue in this species. The expression profile indicated a much higher expression in the adult and pupal stages compared to the larval stages (up to a 60-fold increase), and that the gene was mostly expressed in the ovaries, Malpighian tubules and fat body. Finally, gene knockdown of tudor in B. dorsalis led to clearly underdeveloped ovaries in the female adult and reductions in copulation rate and amount of oviposition, indicating its important role in reproduction. The results of this study shed more light on the role of tudor in ovary development and reproduction.',{'entities': [(0,5,'Gene'),(46,65,'Species'),(263,268,'Gene'),(307,312,'Gene'),(429,434,'Gene'),(447,465,'Species'),(467,486,'Species'),(649,654,'Gene'),(672,683,'Species'),(749,754,'Gene'),(902,907,'Gene'),(1196,1201,'Gene'),(1205,1216,'Species'),(1440,1445,'Gene')]}),('Characterization of three heat shock protein 70 genes from Liriomyza trifolii and expression during thermal stress and insect development. Heat shock proteins (HSPs) participate in diverse physiological processes in insects, and HSP70 is one of the most highly conserved proteins in the HSP family. In this study, full-length cDNAs of three HSP70 genes (Lthsc70, Lthsp701, and Lthsp702) were cloned and characterized from Liriomyza trifolii, an important invasive pest of vegetable crops and horticultural crops worldwide. These three HSP70s exhibited signature sequences and motifs that are typical of the HSP70 family. The expression patterns of the three Lthsp70s during temperature stress and in different insect development stages were studied by real-time quantitative PCR. Lthsp701 was strongly induced by high- and low-temperature stress, but Lthsc70 and Lthsp702 were not very sensitive to temperature changes. All three Lthsp70s were expressed during insect development stages, but the expression patterns were quite different. The expression of Lthsc70 and Lthsp702 showed significant differences in expression during leafminer development; Lthsc70 was most highly expressed in female adults, whereas Lthsp702 was abundantly expressed in larvae and prepupae. Lthsp701 expression was not significantly different among leafminer stages. These results suggest that functional differentiation within the LtHSP70 subfamily has occurred in response to thermal stress and insect development.',{'entities': [(26,47,'Gene'),(59,77,'Species'),(139,158,'Gene'),(160,164,'Gene'),(229,234,'Gene'),(341,346,'Gene'),(354,361,'Gene'),(363,371,'Gene'),(377,385,'Gene'),(422,440,'Species'),(607,612,'Gene'),(658,666,'Gene'),(780,788,'Gene'),(851,858,'Gene'),(863,871,'Gene'),(930,938,'Gene'),(1056,1063,'Gene'),(1068,1076,'Gene'),(1152,1159,'Gene'),(1212,1220,'Gene'),(1270,1278,'Gene')]}),('Knockdown of NADPH-cytochrome P450 reductase and CYP6MS1 increases the susceptibility of Sitophilus zeamais to terpinen-4-ol. Terpinen-4-ol showed highly insecticidal activity to stored-grain pest Sitophilus zeamais, and cytochrome P450s were strongly induced in response to terpinen-4-ol fumigation. Understanding of the function of P450 enzyme system in the susceptibility to terpinen-4-ol in S. zeamais will benefit the potential application of terpinen-4-ol in controlling stored-grain pests. In the present study, the synergist piperonyl butoxide increased the toxicity of terpinen-4-ol to S. zeamais, with a synergism ratio of 3.5-fold. Two isoforms of NADPH-cytochrome P450 reductase (SzCPR) were identified, with the difference at the N-terminal. SzCPR contained an N-terminal membrane anchor, FMN, FAD, and NADP binding domains. Expression levels of SzCPR were upregulated by tea tree oil (TTO) and its main constituent terpinen-4-ol under different concentrations and time periods. RNAi was generated for S. zeamais by feeding adults dsRNA and the knockdown of SzCPR increased the susceptibility of S. zeamais to terpinen-4-ol, with higher mortality of adults than control under terpinen-4-ol fumigation. Further RNAi analysis showed that P450 gene CYP6MS1 mediated the susceptibility of S. zeamais to terpinen-4-ol. These results revealed that cytochrome P450 enzyme system, especially CYP6MS1 participated in the susceptibility of S. zeamais to terpinen-4-ol. The findings provided a foundation to clarify the metabolic mechanisms of terpinen-4-ol in stored-grain pests.',{'entities': [(13,44,'Gene'),(49,56,'Gene'),(89,107,'Species'),(197,215,'Species'),(221,236,'Gene'),(395,405,'Species'),(595,605,'Species'),(659,690,'Gene'),(692,697,'Gene'),(755,760,'Gene'),(859,864,'Gene'),(885,893,'Species'),(1015,1025,'Species'),(1071,1076,'Gene'),(1109,1119,'Species'),(1259,1266,'Gene'),(1298,1308,'Species'),(1355,1370,'Gene'),(1397,1404,'Gene'),(1443,1453,'Species')]}),('Decapentaplegic function in wing vein development and wing morph transformation in brown planthopper, Nilaparvata lugens. The decapentaplegic (dpp) gene plays a variety of roles in diverse cellular and molecular processes of the growth and development. In insects, dpp is mainly required for dorsal-ventral patterning and appendage formation. The brown planthopper (BPH) Nilaparvata lugens, a major pest of rice, possesses two distinct wing morphs described as long-winged (LW) and short-winged (SW) morphs. With our lab-maintained stable strains of LW and SW BPH, RNA interference (RNAi) was used to research the functions of N. lugens dpp (Nldpp) on wing development. Silencing of Nldpp in the SW strain led to the significant lengthening of the forewing, while Nldpp-knockdown in the LW strain resulted in distorted wings. Moreover, knockdown of Nldpp caused the complete absence of wing veins. During the development of wing-pads, the Nldpp abundance in the terga of the SW strain was significantly higher than that of the LW strain. Through controlling the direction of wing morph transformation, we found that the expression level of Nldpp increased in the NlInR1-knockdown BPH (tending to SW) and abundance of Nldpp declined after dsNlInR2 injection (tending to LW). Our results showed that Nldpp is mainly responsible for the formation and development of veins in BPH. Also, Nldpp can be regulated by NlInR1/2 and participate in the wing morph transformation.',{'entities': [(0,15,'Gene'),(83,100,'Species'),(102,120,'Species'),(126,141,'Gene'),(143,146,'Gene'),(265,268,'Gene'),(347,364,'Species'),(366,369,'Species'),(371,389,'Species'),(407,411,'Species'),(560,563,'Species'),(627,636,'Species'),(637,640,'Gene'),(642,647,'Gene'),(683,688,'Gene'),(764,769,'Gene'),(849,854,'Gene'),(939,944,'Gene'),(1140,1145,'Gene'),(1180,1183,'Species'),(1217,1222,'Gene'),(1298,1303,'Gene'),(1372,1375,'Species'),(1383,1388,'Gene')]}),('Rh6 gene modulates the visual mechanism of host utilization in fruit fly Bactrocera minax. BACKGROUND: Vision plays a critical role in host location and oviposition behavior for herbivorous insects. However, the molecular mechanisms underlying visual regulation in host recognition and oviposition site selection in insects remains unknown. The aim of this study was to explore the key visual genes that are linked to the host plant location of the fruit fly, Bactrocera minax. RESULTS: Using a host specialist fruit fly, B. minax, which lays eggs only into immature green citrus fruit, we undertook behavioral, transcriptomic, and RNAi research to identify the molecular basis for host fruit color recognition. In laboratory and field assays we found that adults prefer green over other colors, and this preference is significantly increased in sexually mature over immature flies. Furthermore, we identified that the Rh6 gene, responsible for green spectral sensitivity, has elevated expression in mature flies over immature flies. RNAi suppression of Rh6 eliminated the preference for green, resulting in a significant decrease in the number of eggs laid by B. minax in green unripe citrus. CONCLUSION: These results show that the Rh6 gene modulates the visual mechanism of host utilization in B. minax, providing a genetic basis for visual host location in a non-model insect herbivore.  2018 Society of Chemical Industry.',{'entities': [(0,3,'Gene'),(63,72,'Species'),(73,89,'Species'),(449,458,'Species'),(460,476,'Species'),(511,520,'Species'),(522,530,'Species'),(919,922,'Gene'),(1054,1057,'Gene'),(1161,1169,'Species'),(1234,1237,'Gene'),(1297,1305,'Species')]}),('Dysfunction of LSD-1 induces JNK signaling pathway-dependent abnormal development of thorax and apoptosis cell death in Drosophila melanogaster. Perilipins are evolutionarily conserved from insects to mammals. Lipid storage droplet-1 (LSD-1) is a member of the lipid droplet s surface-binding protein family and counterpart to mammalian perilipin 1. The role of LSD-1 has already been reported in lipid metabolism of Drosophila. However, the function of this gene during specific tissue development is still under investigation. Here, we found that LSD-1 is expressed in the notum of the wing imaginal disc, and notum-specific knockdown of Lsd-1 by pannir-GAL4 driver leads to split thorax phenotype in adults, suggesting an essential role of LSD-1 in development of Drosophila thorax. As overexpression of JNK homolog, bsk (basket) suppresses Lsd-1 knockdown phenotype, the role of LSD-1 in thorax development was proved to be dependent on the activity of the Drosophila c-Jun N-terminal kinase (JNK). The puckered (puc) expression led to significant decrease in the JNK activity in wing discs of Lsd-1 knockdown flies. In addition, we also detected that depletion of Lsd-1 enhances apoptotic cell death in the wing notum area. Taken together, these data demonstrated that LSD-1 functions in Drosophila thorax development by regulating JNK pathway.',{'entities': [(15,20,'Gene'),(29,32,'Gene'),(120,143,'Species'),(210,233,'Gene'),(235,240,'Gene'),(327,336,'Species'),(337,348,'Gene'),(362,367,'Gene'),(417,427,'Species'),(549,554,'Gene'),(640,645,'Gene'),(743,748,'Gene'),(767,777,'Species'),(807,810,'Gene'),(820,823,'Gene'),(825,831,'Gene'),(844,849,'Gene'),(883,888,'Gene'),(961,971,'Species'),(972,995,'Gene'),(997,1000,'Gene'),(1007,1015,'Gene'),(1017,1020,'Gene'),(1068,1071,'Gene'),(1098,1103,'Gene'),(1169,1174,'Gene'),(1274,1279,'Gene'),(1293,1303,'Species'),(1337,1340,'Gene')]}),('The midgut V-ATPase subunit A gene is associated with toxicity to crystal 2Aa and crystal 1Ca-expressing transgenic rice in Chilo suppressalis. Insecticidal crystal (Cry) proteins produced by the bacterium Bacillus thuringiensis (Bt) are toxic to a diverse range of insects. Transgenic rice expressing Cry1A, Cry2A and Cry1C toxins have been developed that are lethal to Chilo suppressalis, a devastating insect pest of rice in China. Identifying the mechanisms underlying the interactions of Cry toxins with susceptible hosts will improve both our understanding of Cry protein toxicology and long-term efficacy of Bt crops. In this study, we tested the hypothesis that V-ATPase subunit A contributes to the action of Cry1Ab/1Ac, Cry2Aa and Cry1Ca toxins in C. suppressalis. The full-length V-ATPase subunit A transcript was initially cloned from the C. suppressalis larval midgut and then used to generate double-stranded RNA (dsRNA)-producing bacteria. Toxicity assays using transgenic rice lines TT51 (Cry1Ab and Cry1Ac fusion genes), T2A-1 (Cry2Aa), and T1C-19 (Cry1Ca) in conjunction with V-ATPase subunit A dsRNA-treated C. suppressalis larvae revealed significantly reduced larval susceptibility to T2A-1 and T1C-19 transgenic rice, but not to TT51 rice. These results suggest that the V-ATPase subunit A plays a crucial role in mediating Cry2Aa and Cry1Ca toxicity in C. suppressalis. These findings will have significant implications on the development of future resistance management tools.',{'entities': [(11,29,'Gene'),(116,120,'Species'),(124,142,'Species'),(166,169,'Gene'),(206,228,'Species'),(230,232,'Species'),(286,290,'Species'),(302,307,'Gene'),(309,314,'Gene'),(319,324,'Gene'),(371,389,'Species'),(420,424,'Species'),(493,496,'Gene'),(566,569,'Gene'),(615,617,'Species'),(670,688,'Gene'),(718,724,'Gene'),(730,736,'Gene'),(741,747,'Gene'),(758,773,'Species'),(791,809,'Gene'),(851,866,'Species'),(988,992,'Species'),(1005,1011,'Gene'),(1045,1051,'Gene'),(1066,1072,'Gene'),(1094,1112,'Gene'),(1127,1142,'Species'),(1234,1238,'Species'),(1256,1260,'Species'),(1293,1311,'Gene'),(1346,1352,'Gene'),(1357,1363,'Gene'),(1376,1391,'Species')]}),('Functional characterization of odorant-binding proteins from the scarab beetle Holotrichia oblita based on semiochemical-induced expression alteration and gene silencing. With the advent of next-generation sequencing, it is now possible to rapidly identify the entire repertoire of olfactory genes likely to be involved in chemical communication of an insect species. It remains, however, a challenge to identify olfactory proteins, such as odorant receptors and odorant-binding proteins (OBPs), vis- -vis the odorants they detect. It has been reported that exposing the olfactory system to a physiologically relevant odorant alters the transcript levels of odorant receptor(s) involved in the detection of the tested odorant. We applied this paradigm in an attempt to identify putative OBPs from the scarab beetle Holotrichia oblita involved in the reception of plant-derived kairomones. Twenty-nine OBP genes were identified in the H. oblita transcriptome, 20 of which were enriched in antennae compared with nonolfactory tissues. Of these, 2 OBP genes, HoblOBP13 and HoblOBP9, were upregulated upon exposure to one of the female attractants (E)-2-hexenol and phenethyl alcohol; none of the OBP transcripts changed upon exposure to methyl anthranilate, which does not attract H. oblita females. Binding assays showed that HoblOBP13 and HoblOBP9 have high affinity for (E)-2-hexenol and phenethyl alcohol, respectively. RNAi treatment showed that transcripts of both HoblOBP13 and HoblOBP9 declined in a time-course manner 24-72  h postinjection. OBP-dsRNA-treated female beetles showed significantly lower attraction to (E)-2-hexenol and phenethyl alcohol than did water-injected beetles and those treated with GFP-dsRNA. We, therefore, concluded that HoblOBP13 and HoblOBP9 are essential for H. oblita reception of the plant-derived kairomones (E)-2-hexenol and phenethyl alcohol.',{'entities': [(31,55,'Gene'),(79,97,'Species'),(463,487,'Gene'),(489,493,'Gene'),(787,791,'Gene'),(815,833,'Species'),(901,904,'Gene'),(934,943,'Species'),(1045,1048,'Gene'),(1056,1065,'Gene'),(1070,1078,'Gene'),(1193,1196,'Gene'),(1278,1287,'Species'),(1324,1333,'Gene'),(1338,1346,'Gene'),(1468,1477,'Gene'),(1482,1490,'Gene'),(1548,1551,'Gene'),(1754,1763,'Gene'),(1768,1776,'Gene'),(1795,1804,'Species')]}),('Characterization and expression profiling of odorant-binding proteins in Anoplophora glabripennis Motsch. In insects, olfaction plays a critical role in locating hosts, recognizing mates, and selecting oviposition sites. The Asian long-horned beetle (Anoplophora glabripennis Motschulsky) feeds on 43 species of trees in 15 families, but its chemosensory mechanisms are poorly understood. Herein, genes encoding 61 odorant-binding proteins (OBPs) were identified from the published genome and our previous A. glabripennis transcriptomic data. To investigate their physiological functions, we performed expression profiling of all AglaOBPs in the antennae, legs, and maxillary palps of both sexes. Phylogenetic analysis clustered A. glabripennis OBPs into four subgroups, comprising 29 Minus-C OBPs, 15 Antennae-binding proteins (ABPIIs), 10 Classic OBPs, and one Plus-C OBP. 12 AglaOBP genes were expressed specifically in antennae, and AglaOBP3, AglaOBP18, AglaOBP21, AglaOBP33, AglaOBP41, AglaOBP45, and AglaOBP47 were particularly highly expressed in male antennae. These proteins may function in the detection of female sex pheromones. AglaOBP23 and AglaOBP44 were preferentially expressed in maxillary palps. Expression profiling suggests that many OBPs may be involved in olfaction and gustation, in addition to carrying hydrophobic molecules. The AglaOBPs family has acquired functional diversity concurrently with functional constraints, and further investigation could provide insight into the roles of OBPs in chemoreception.',{'entities': [(45,69,'Gene'),(73,97,'Species'),(251,275,'Species'),(415,439,'Gene'),(441,445,'Gene'),(506,521,'Species'),(630,638,'Gene'),(729,744,'Species'),(745,749,'Gene'),(793,797,'Gene'),(849,853,'Gene'),(878,885,'Gene'),(937,945,'Gene'),(947,956,'Gene'),(958,967,'Gene'),(969,978,'Gene'),(980,989,'Gene'),(991,1000,'Gene'),(1006,1015,'Gene'),(1140,1149,'Gene'),(1154,1163,'Gene'),(1254,1258,'Gene'),(1354,1362,'Gene'),(1512,1516,'Gene')]}),('Insecticide Exposure Triggers a Modulated Expression of ABC Transporter Genes in Larvae of Anopheles gambiae s.s. Insecticides remain a main tool for the control of arthropod vectors. The urgency to prevent the insurgence of insecticide resistance and the perspective to find new target sites, for the development of novel molecules, are fuelling the study of the molecular mechanisms involved in insect defence against xenobiotic compounds. In this study, we have investigated if ATP-binding cassette (ABC) transporters, a major component of the defensome machinery, are involved in defence against the insecticide permethrin, in susceptible larvae of the malaria vector Anopheles gambiae sensu stricto. Bioassays were performed with permethrin alone, or in combination with an ABC transporter inhibitor. Then we have investigated the expression profiles of five ABC transporter genes at different time points following permethrin exposure, to assess their expression patterns across time. The inhibition of ABC transporters increased the larval mortality by about 15-fold. Likewise, three genes were up-regulated after exposure to permethrin, showing different patterns of expression across the 48 h. Our results provide the first evidences of ABC transporters involvement in defence against a toxic in larvae of An. gambiae s.s. and show that the gene expression response is modulated across time, being continuous, but stronger at the earliest and latest times after exposure.',{'entities': [(56,71,'Gene'),(91,108,'Species'),(503,506,'Gene'),(672,703,'Species'),(779,794,'Gene'),(864,879,'Gene'),(1009,1012,'Gene'),(1246,1249,'Gene'),(1315,1326,'Species')]}),('Systemic RNAi of V-ATPase subunit B causes molting defect and developmental abnormalities in Periplaneta fuliginosa. The vacuolar (H+ )-ATPases (V-ATPases) are ATP-driven proton pumps with multiple functions in many organisms. In this study, we performed structural and functional analysis of vha55 gene that encodes V-ATPase subunit B in the smokybrown cockroach Periplaneta fuliginosa (Blattodea). We observed a high homology score of the deduced amino acid sequences between 10 species in seven orders. RNAi of the vha55 gene in P. fuliginosa caused nymphal/nymphal molting defects with incomplete shedding of old cuticles, growth inhibition, as well as bent and wrinkled cuticles of thoraxes and abdominal segments. Since growth inhibition caused by vha55 RNAi did not interfere in the commencement of cockroach molting, molting timing and body growth might be controlled by independent mechanism. Our study suggested V-ATPases might be a good candidate molecule for evolutionary and developmental studies of insect molting.',{'entities': [(17,35,'Gene'),(93,115,'Species'),(121,143,'Gene'),(145,154,'Gene'),(293,298,'Gene'),(317,335,'Gene'),(343,363,'Species'),(364,386,'Species'),(518,523,'Gene'),(532,545,'Species'),(754,759,'Gene'),(922,931,'Gene')]}),('Environmental dissemination of mcr-1 positive Enterobacteriaceae by Chrysomya spp. (common blowfly): An increasing public health risk. Until recently, the role of insects, and particularly flies, in disseminating antimicrobial resistance (AMR) has been poorly studied. In this study, we screened blowflies (Chrysomya spp.) from different areas near the city of Phitsanulok, Northern Thailand, for the presence of AMR genes and in particular, mcr-1, using whole genome sequencing (WGS). In total, 48 mcr-1-positive isolates were recovered, consisting of 17 mcr-1-positive Klebsiella pneumoniae (MCRPKP) and 31 mcr-1-positive Escherichia coli (MCRPEC) strains. The 17 MCRPKP were shown to be clonal (ST43) with few single poly nucleomorphs (SNPs) by WGS analysis. In in-vitro models, the MCRPKP were shown to be highly virulent. In contrast, 31 recovered MCRPEC isolates are varied, belonging to 12 different sequence types shared with those causing human infections. The majority of mcr-1 gene are located on IncX4 plasmids (29/48, 60.42%), sharing an identical plasmid backbone. These findings highlight the contribution of flies to the AMR contagion picture in low- and middle-income countries and the challenges of tackling global AMR.',{'entities': [(31,36,'Gene'),(68,77,'Species'),(84,98,'Species'),(307,316,'Species'),(442,447,'Gene'),(499,504,'Gene'),(556,561,'Gene'),(571,592,'Species'),(609,614,'Gene'),(624,640,'Species'),(948,953,'Species'),(982,987,'Gene')]}),('Comparative analysis of C-type lectin domain proteins in the ghost moth, Thitarodes xiaojinensis (Lepidoptera: Hepialidae). Insects have a large family of C-type lectins involved in cell adhesion, pathogen recognition and activation of immune responses. In this study, 32 transcripts encoding C-type lectin domain proteins (CTLDPs) were identified from the Thitarodes xiaojinensis transcriptome. According to their domain structures, six CTLDPs with one carbohydrate-recognition domain (CRD) were classified into the CTL-S subfamily. The other 23 CTLDPs with two CRDs were grouped into the immulectin (IML) subfamily. The remaining three with extra regulatory domains were sorted into the CTL-X subfamily. Phylogenetic analysis showed that CTL-S and CTL-X members from different insects could form orthologous groups. In contrast, no T. xiaojinensis IML orthologues were found in other insects. Remarkable lineage-specific expansion in this subfamily was observed reflecting that these CTLDPs, as important receptors, have evolved diversified members in response to a variety of microbes. Prediction of binding ligands revealed that T. xiaojinensis, a cold-adapted species, conserved the ability of CRDs to combine with Ca2+ to keep its receptors from freezing. Comparative analysis of induction of CTLDP genes after different immune challenges indicated that IMLs might play critical roles in immune defenses. This study examined T. xiaojinensis CTLDPs and provides a basis for further studies of their characteristics.',{'entities': [(24,53,'Gene'),(61,71,'Species'),(73,96,'Species'),(293,322,'Gene'),(324,330,'Gene'),(357,380,'Species'),(438,444,'Gene'),(547,553,'Gene'),(834,849,'Species'),(986,992,'Gene'),(1133,1148,'Species'),(1299,1304,'Gene'),(1431,1446,'Species'),(1447,1453,'Gene')]}),('Two delta class glutathione S-transferases involved in the detoxification of malathion in Bactrocera dorsalis (Hendel). BACKGROUND: The oriental fruit fly Bactrocera dorsalis (Hendel), a widespread agricultural pest, has evolved resistance to many insecticides, including organophosphorus compounds. Glutathione S-transferases (GSTs) are involved in xenobiotic detoxification and insecticide resistance in many insects. However, the role of delta class GSTs in detoxifying malathion in B. dorsalis is unknown. Here, we evaluated the roles of two delta class GSTs in malathion detoxification in this species. RESULTS: Two delta class GSTs genes, BdGSTd1 and BdGSTd10, were characterized in B. dorsalis. They were highly expressed in 5-day-old adults, as well as in midgut and Malpighian tubules. Upon malathion exposure, the two genes were upregulated by 2.63- and 2.85-fold, respectively. Injection of double-stranded RNA targeting BdGSTd1 or BdGSTd10 significantly reduced their mRNA levels in adults and also significantly increased adult susceptibility to malathion. The expression of these two GSTs in Escherichia coli helped the host to endure malathion stress at a concentration of 10  g  mL-1 according to a Cell Counting Kit-8 assay. High-performance liquid chromatography analyses indicated that malathion could be significantly depleted by the two delta GSTs. The role of BdGSTd10 in malathion sequestration was also discussed. CONCLUSION: BdGSTd1 and BdGSTd10 play important roles in the detoxification of malathion in B. dorsalis.  2019 Society of Chemical Industry.',{'entities': [(16,41,'Gene'),(90,109,'Species'),(136,154,'Species'),(155,174,'Species'),(300,326,'Gene'),(328,332,'Gene'),(453,457,'Gene'),(486,497,'Species'),(558,562,'Gene'),(633,637,'Gene'),(645,652,'Gene'),(657,665,'Gene'),(689,700,'Species'),(932,939,'Gene'),(943,951,'Gene'),(1098,1102,'Gene'),(1106,1122,'Species'),(1364,1368,'Gene'),(1382,1390,'Gene'),(1450,1457,'Gene'),(1462,1470,'Gene'),(1530,1541,'Species')]}),('Clustered miR-2, miR-13a, miR-13b and miR-71 coordinately target Notch gene to regulate oogenesis of the migratory locust Locusta migratoria. MicroRNAs (miRNAs),  22-nt small noncoding RNAs with a crucial role in various biological processes of organisms, are usually clustered in the genome. However, little is known about the miRNA clusters involved in insect reproduction. By small RNA sequencing and quantification followed by qRT-PCR, we found that the expression of invertebrate-specific miR-2/13/71 cluster including miR-2, miR-13a, miR-13b and miR-71 significantly decreased after adult ecdysis of the migratory locust, Locusta migratoria. Luciferase reporter assay and RNA immunoprecipitation demonstrated that miR-2/13/71 bound to the protein coding sequence of Notch and downregulated its expression. Injection of miR-2/13/71 agomiRs led to significant decrease of Notch expression as well as markedly reduced levels of Vitellogenin mRNA, suppressed oocyte maturation and impaired ovarian growth. Moreover, the expression of miR-2/13/71 was repressed by juvenile hormone (JH). Our results thus point to a previously unidentified mechanism by which JH-repressed miR-2/13/71 coordinately downregulates Notch to modulate insect reproduction. The increase of JH and decrease of miR-2/13/71 expression in both previtellogenic and vitellogenic stages of adult females ensure a high level of Notch expression, critically contributing to JH-dependent vitellogenesis and oogenesis.',{'entities': [(10,15,'Gene'),(17,24,'Gene'),(26,33,'Gene'),(38,44,'Gene'),(65,70,'Gene'),(105,121,'Species'),(122,140,'Species'),(494,499,'Gene'),(524,529,'Gene'),(531,538,'Gene'),(540,547,'Gene'),(552,558,'Gene'),(610,626,'Species'),(628,646,'Species'),(720,725,'Gene'),(772,777,'Gene'),(825,830,'Gene'),(876,881,'Gene'),(1036,1041,'Gene'),(1172,1177,'Gene'),(1211,1216,'Gene'),(1285,1290,'Gene'),(1396,1401,'Gene')]}),('Physiological functions of a methuselah-like G protein coupled receptor in Lymantria dispar Linnaeus. Insect G protein coupled receptors (GPCRs) have been identified as a highly attractive target for new generation insecticides discovery due to their critical physiological functions. However, few insect GPCRs have been functionally characterized. Here, we cloned the full length of a methuselah-like GPCR gene (Ldmthl1) from the Asian gypsy moth, Lymantria dispar. We then characterized the secondary and tertiary structures of Ldmthl1. We also predicted the global structure of this insect GPCR protein which is composed of three major domains. RNA interference of Ldmthl1 resulted in a reduction of gypsy moths  resistance to deltamethrin and suppressed expression of downstream stress-associated genes, such as P450s, glutathione S transferases, and heat shock proteins. The function of Ldmthl1 was further investigated using transgenic lines of Drosophila melanogaster. Drosophila with overexpression of Ldmthl1 showed significantly longer lifespan than control flies. Taken together, our studies revealed that the physiological functions of Ldmthl1 in L. dispar are associated with longevity and resistance to insecticide stresses. Potentially, Ldmthl1 can be used as a target for new insecticide discovery in order to manage this notorious forest pest.',{'entities': [(29,71,'Gene'),(75,91,'Species'),(109,136,'Gene'),(138,143,'Gene'),(305,310,'Gene'),(386,406,'Gene'),(413,420,'Gene'),(431,447,'Species'),(449,465,'Species'),(530,537,'Gene'),(593,597,'Gene'),(668,675,'Gene'),(703,714,'Species'),(816,821,'Gene'),(823,849,'Gene'),(855,874,'Gene'),(892,899,'Gene'),(951,974,'Species'),(976,986,'Species'),(1010,1017,'Gene'),(1148,1155,'Gene'),(1159,1168,'Species'),(1252,1259,'Gene')]}),('Two functionally distinct CYP4G genes of Anopheles gambiae contribute to cuticular hydrocarbon biosynthesis. Cuticular hydrocarbon (CHC) biosynthesis is a major pathway of insect physiology. In Drosophila melanogaster the cytochrome P450 CYP4G1 catalyses the insect-specific oxidative decarbonylation step, while in the malaria vector Anopheles gambiae, two CYP4G paralogues, CYP4G16 and CYP4G17 are present. Analysis of the subcellular localization of CYP4G17 and CYP4G16 in larval and pupal stages revealed that CYP4G16 preserves its PM localization across developmental stages analyzed; however CYPG17 is differentially localized in two distinct types of pupal oenocytes, presumably oenocytes of larval and adult developmental specificity. Western blot analysis showed the presence of two CYP4G17 forms, potentially associated with each oenocyte type. Both An. gambiae CYP4Gs were expressed in D. melanogaster flies in a Cyp4g1 silenced background in order to functionally characterize them in vivo. CYP4G16, CYP4G17 or their combination rescued the lethal phenotype of Cyp4g1-knock down flies, demonstrating that CYP4G17 is also a functional decarbonylase, albeit of somewhat lower efficiency than CYP4G16 in Drosophila. Flies expressing mosquito CYP4G16 and/or CYP4G17 produced similar CHC profiles to  wild-type  flies expressing the endogenous CYP4G1, but they also produce very long-chain dimethyl-branched CHCs not detectable in wild type flies, suggesting that the specificity of the CYP4G enzymes contributes to determine the complexity of the CHC blend. In conclusion, both An. gambiae CYP4G enzymes contribute to the unique Anopheles CHC profile, which has been associated to defense, adult desiccation tolerance, insecticide penetration rate and chemical communication.',{'entities': [(26,31,'Gene'),(41,58,'Species'),(194,217,'Species'),(222,237,'Gene'),(238,244,'Gene'),(335,352,'Species'),(358,363,'Gene'),(376,383,'Gene'),(388,395,'Gene'),(453,460,'Gene'),(465,472,'Gene'),(514,521,'Gene'),(598,604,'Gene'),(792,799,'Gene'),(860,871,'Species'),(897,912,'Species'),(924,930,'Gene'),(1003,1010,'Gene'),(1012,1019,'Gene'),(1073,1079,'Gene'),(1117,1124,'Gene'),(1202,1209,'Gene'),(1213,1223,'Species'),(1251,1258,'Gene'),(1266,1273,'Gene'),(1351,1357,'Gene'),(1494,1499,'Gene'),(1586,1597,'Species'),(1598,1603,'Gene'),(1637,1646,'Species')]}),('DDC plays vital roles in the wing spot formation, egg production, and chorion tanning in the brown planthopper. The gene dopa decarboxylase (Nlddc) of the brown planthopper (BPH, Nilaparvata lugens) was identified in the genome and transcriptome of the insect. The open reading frame of Nlddc is 1,434 bp in length and, it has the potential to encode a protein of 477 amino acid with a conserved pyridoxal-dependent decarboxylase domain and a typical motif, NFNPHKW. Real-time quantification polymerase chain reaction analyses revealed that this gene was highly expressed in the integument and brain, and transcript level peaked in the late stages of egg period and each nymph instar with periodicity. RNA interference results revealed that Nlddc played essential roles in pigment synthesis, formation of wing spot, egg production, and tanning of the chorion. A rapid accumulation of Nlddc transcripts was detected, and it coincided with the injection of the hormone 20-hydroxyecdysone (20E), suggesting that Nlddc transcription was regulated by 20E.',{'entities': [(0,3,'Gene'),(93,110,'Species'),(121,139,'Gene'),(141,146,'Gene'),(155,172,'Species'),(174,177,'Species'),(179,197,'Species'),(287,292,'Gene'),(741,746,'Gene'),(884,889,'Gene'),(1009,1014,'Gene')]}),('Structural glycoprotein LmAbd-9 is required for the formation of the endocuticle during locust molting. Insect cuticle is a composite made of chitin filaments embedded in a proteinaceous matrix, consisting mainly of structural cuticular proteins. In the present study, an endocuticle structural glycoprotein gene, LmAbd-9, was characterized based on the Locusta migratoria transcriptome. LmAbd-9 encodes a glycoprotein with a chitin binding domain 4, belonging to RR-1 subclass of the CPR family, which has two potential O-linked glycosylation sites (S115 and T137) at which glycosylation modification may occur. LmAbd-9 was highly expressed in the integument and showed periodic expression during molting. The expression levels of LmAbd-9 were significantly down-regulated after injection with 20-hydroxyecdysone (20E) for 6, 12, and 24  h, whereas it was upregulated after double-stranded RNA-mediated RNA interference of the 20E receptor gene LmEcR and LmFTZ-F1beta at day 2 of fifth instar nymphs for 48  h. After injection of dsLmAbd-9 on day 2 of fifth instar nymphs, the insects could normally molt to adults and showed no macroscopic phenotype; however, the cuticle of the adults was thinner, and there were significantly fewer endocuticular lamellae than in the control. Thus, LmAbd-9 that negatively regulated by the 20E signaling pathway was involved in the formation of the endocuticle in L. migratoria.',{'entities': [(24,31,'Gene'),(314,321,'Gene'),(354,372,'Species'),(388,395,'Gene'),(613,620,'Gene'),(732,739,'Gene'),(946,951,'Gene'),(956,968,'Gene'),(1286,1293,'Gene'),(1401,1414,'Species')]}),('Characterization of a broad-based mosquito yeast interfering RNA larvicide with a conserved target site in mosquito semaphorin-1a genes. BACKGROUND: RNA interference (RNAi), which has facilitated functional characterization of mosquito neural development genes such as the axon guidance regulator semaphorin-1a (sema1a), could one day be applied as a new means of vector control. Saccharomyces cerevisiae (baker s yeast) may represent an effective interfering RNA expression system that could be used directly for delivery of RNA pesticides to mosquito larvae. Here we describe characterization of a yeast larvicide developed through bioengineering of S. cerevisiae to express a short hairpin RNA (shRNA) targeting a conserved site in mosquito sema1a genes. RESULTS: Experiments conducted on Aedes aegypti larvae demonstrated that the yeast larvicide effectively silences sema1a expression, generates severe neural defects, and induces high levels of larval mortality in laboratory, simulated-field, and semi-field experiments. The larvicide was also found to induce high levels of Aedes albopictus, Anopheles gambiae and Culex quinquefasciatus mortality. CONCLUSIONS: The results of these studies indicate that use of yeast interfering RNA larvicides targeting mosquito sema1a genes may represent a new biorational tool for mosquito control.',{'entities': [(43,48,'Species'),(116,129,'Gene'),(297,310,'Gene'),(312,318,'Gene'),(380,404,'Species'),(406,419,'Species'),(600,605,'Species'),(652,665,'Species'),(744,750,'Gene'),(792,805,'Species'),(835,840,'Species'),(872,878,'Gene'),(1082,1098,'Species'),(1100,1117,'Species'),(1122,1144,'Species'),(1219,1224,'Species'),(1271,1277,'Gene')]}),('Copper-induced H2O2 accumulation confers larval tolerance to xanthotoxin by modulating CYP6B50 expression in Spodoptera litura. In the plant-insect arms race, plants synthesize toxic compounds to defend against herbivorous insects, whereas insects employ cytochrome P450 monooxygenases (P450s) to detoxify these phytotoxins. As ubiquitous environmental contaminants, heavy metals can be easily absorbed by plants and further accumulated in herbivorous insects through the food chains, resulting in tangible consequences for plant-insect interactions. However, whether heavy metals can influence P450 activities and thereby cause further effects on larval tolerance to phytotoxins remains unknown. In this study, we shown that prior exposure to copper (Cu) enhanced larval tolerance to xanthotoxin in Spodoptera litura, a major polyphagous pest of agriculture. P450 activities were induced in larvae exposed to Cu or xanthotoxin, and a midgut specific expressed P450 gene, CYP6B50 was cross-induced after exposure to these two toxic xenobiotics. Knocking down CYP6B50 by RNA interference (RNAi) rendered the larvae more sensitive to xanthotoxin. As defense against oxidative stress following metal exposure has been demonstrated to affect insecticide resistance, the reactive oxygen species (ROS) generation and antioxidant enzyme activities were assessed. Cu exposure caused the accumulation of hydrogen peroxide (H2O2) and enhanced the activities of superoxide dismutase (SOD) and peroxidase (POD) in larval midgut. In addition, two antioxidant response elements (AREs) were identified from the CYP6B50 promoter, indicating that Cu-induced CYP6B50 expression may be related to the ROS burst. Application of ROS scavenger N-acetylcysteine (NAC) effectively suppressed CYP6B50 expression, inhibited P450 activities and impaired larval tolerance to xanthotoxin that had been induced by Cu. These results indicate that the increase in CYP6B50 expression regulated by Cu-induced H2O2 generation contributed to the enhancement of larval tolerance to xanthotoxin in S. litura. Ingestion of heavy metals from their host plants can inadvertently boost the counter-defense system of herbivorous insects to protect themselves against plant defensive toxins.',{'entities': [(87,94,'Gene'),(109,126,'Species'),(255,284,'Gene'),(287,292,'Gene'),(595,599,'Gene'),(800,817,'Species'),(860,864,'Gene'),(961,965,'Gene'),(972,979,'Gene'),(1059,1066,'Gene'),(1596,1603,'Gene'),(1641,1648,'Gene'),(1768,1775,'Gene'),(1798,1802,'Gene'),(1932,1939,'Gene'),(2060,2069,'Species')]}),('timeless2 plays an important role in reproduction and circadian rhythms in the cricket Gryllus bimaculatus. The timeless2 (tim2) gene is an insect orthologue of the mammalian clock gene Timeless (mTim). Although its functional role has been extensively studied in mammals, little is known regarding its role in insects. In the present study, we obtained tim2 cDNA (Gb tim2) from the cricket Gryllus bimaculatus and characterized its functional role in embryonic development, egg production, and circadian rhythms. Gb tim2 gave rise to a 1432 amino acid protein, and showed approximately 65% homology to that of Drosophila melanogaster. When treated with parental Gb tim2<sup>RNAi</sup>, less than 2% of the treated eggs hatched. On the other hand, control eggs treated with DsRed2<sup>RNAi</sup>demonstrated a hatching rate of 70%. In most of the Gb tim2<sup>RNAi</sup>treated embryos, development was arrested in early stages. Egg production in ovaries of adult virgin females treated with Gb tim2<sup>RNAi</sup>was significantly reduced. In addition, while Gb tim2<sup>RNAi</sup>crickets exhibited clear locomotor rhythm synchronized with light cycles, their light-on peak was weaker than that of control crickets. Under constant darkness, the activity rhythm of Gb tim2<sup>RNAi</sup>crickets was often split into two components running with different periods. Molecular analysis revealed that Gb tim2<sup>RNAi</sup>treatment downregulated mRNA levels of Gb per and Gb Clk, and enhanced Gb cyc expression rhythm; no distinct effect was found on Gb tim expression levels. The change in Gb per, Gb Clk and Gb cyc levels may underlie the altered behavioral rhythms in Gb tim2<sup>RNAi</sup>crickets. Both Gb Clk<sup>RNAi</sup>and Gb cyc<sup>RNAi</sup>downregulated Gb tim2 expression, which suggested that transcription of Gb tim2 is mediated by Gb CLK and Gb CYC through E-box. These results suggested that Gb tim2 may be involved in both reproduction and circadian rhythm regulation in crickets.',{'entities': [(0,9,'Gene'),(87,106,'Species'),(112,121,'Gene'),(123,127,'Gene'),(165,174,'Species'),(186,194,'Gene'),(196,200,'Gene'),(354,358,'Gene'),(368,372,'Gene'),(391,410,'Species'),(517,521,'Gene'),(611,634,'Species'),(1551,1554,'Gene'),(1768,1772,'Gene'),(1826,1830,'Gene'),(1911,1915,'Gene')]}),('The NompC channel regulates Nilaparvata lugens proprioception and gentle-touch response. NompC channel is a member of the transient receptor potential (TRP) ion channel superfamily. It can regulate gentle-touch, locomotion, hearing and food texture detection in Drosophila. We cloned the NompC gene of Nilaparvata lugens (NlNompC). The full length NlNompC possessed similar structure as DmNompC, which belongs to TRPN subfamily. The expression pattern analysis of different developmental stages and body parts showed that the transcription of NlNompC was more abundant in adult stage and in the abdomen. Injection of double-stranded RNA (dsRNA) of NlNompC in the third-instar nymphs successfully knocked down the target gene with 75% suppression. At nine days after injection, the survival rate of dsRNA injected nymphs was as low as 9.84%. Behavioral observation revealed that the locomotion of the dsRNA injected nymphs was defective with much less movement compared to the negative control. Feeding and honeydew excretion of the dsRNA injected insects also decreased significantly. These results suggested that NlNompC is a classical mechanotransduction channel that plays important roles in proprioception and locomotion, and is essential for the survival of N. lugens. The results also contribute to the understanding of how TRP channels regulate proprioception.',{'entities': [(4,9,'Gene'),(28,46,'Species'),(89,94,'Gene'),(288,293,'Gene'),(302,320,'Species'),(322,329,'Gene'),(348,355,'Gene'),(543,550,'Gene'),(648,655,'Gene'),(1114,1121,'Gene'),(1263,1272,'Species')]}),('Characterization of two copper/zinc superoxide dismutases (Cu/Zn-SODs) from the desert beetle Microdera punctipennis and their activities in protecting E. coli cells against cold. Superoxide dismutases (SODs) are crucial in scavenging reactive oxygen species (ROS); however, studies regarding SOD functions in insects under cold conditions are rare. In this paper, two novel Cu/Zn-SOD genes in the desert beetle Microdera punctipennis, an extracellular copper/zinc SOD (MpecCu/Zn-SOD) and an intracellular copper/zinc SOD (MpicCu/Zn-SOD), were identified and characterized. The results of quantitative real-time PCR showed that MpecCu/Zn-SOD expression was significantly up-regulated by 4   C exposure for 0.5  h, but MpicCu/Zn-SOD was not. Superoxide anion radical (O2  -) content in beetles under 4   C exposure for 0.5  h showed an initial sharp increase and fluctuated during the cold treatment period, which was consistent with the relative mRNA level of MpecCu/Zn-SOD. The total SOD activity in the beetle was negatively correlated to the O2  - content with a correlation coefficient of -0.437. An E. coli system was employed to study the function of each MpCu/Zn-SOD gene. The fusion proteins Trx-His-MpCu/Zn-SODs were over expressed in E. coli BL21 using pET32a vector, and identified by SDS-PAGE and Western blotting. The transformed bacteria BL21(pET32a-MpecCu/Zn-SOD) and BL21(pET32a-MpicCu/Zn-SOD) showed increased cold tolerance to -4   C as well as increased SOD activity compared to the control BL21(pET32a). The relative conductivity and malondialdehyde content in the two MpCu/Zn-SODs transformed bacteria under -4   C were significantly lower than the control BL21(pET32a). Furthermore, BL21(pET32a-MpecCu/Zn-SOD) had significantly higher SOD activity and cold tolerance than BL21(pET32a-MpicCu/Zn-SOD) under -4   C treatment, and had lower conductivity than BL21(pET32a-MpicCu/Zn-SOD). In conclusion, low temperature led to the accumulation of O2  - in M. punctipennis, which stimulated the expression of extracellular MpCu/Zn-SOD gene and the increase of total SOD activity. E. coli overexpressing Trx-His-MpCu/Zn-SODs increased resistance to cold treatment-induced oxidative stress. Our findings will be helpful in further study of Cu/Zn-SOD genes in insect cold-tolerance.',{'entities': [(59,69,'Gene'),(94,116,'Species'),(152,159,'Species'),(180,201,'Gene'),(203,207,'Gene'),(375,384,'Gene'),(412,434,'Species'),(470,483,'Gene'),(523,536,'Gene'),(628,641,'Gene'),(718,731,'Gene'),(960,973,'Gene'),(1104,1111,'Species'),(1162,1173,'Gene'),(1216,1220,'Gene'),(1244,1256,'Species'),(1589,1601,'Gene'),(1972,1987,'Species'),(2095,2102,'Species'),(2134,2138,'Gene')]}),('Functional characterization of ultraspiracle in Leptinotarsa decemlineata using RNA interference assay. A heterodimer of ultraspiracle (USP) and ecdysone receptor (EcR) mediates 20-hydroxyecdysone (20E) signalling cascade to regulate insect moulting and metamorphosis. However, at least two questions remain to be addressed in terms of the molecular importance of USP in insect species. First, is USP involved in both regulation of ecdysteroidogenesis and mediation of 20E signalling in non-drosophilid insects, as in Drosophila melanogaster? Second, does USP play any role in larval metamorphosis except as the partner of heterodimeric receptor to activate the downstream 20E signalling genes? In this paper, we found that RNA interference (RNAi) of LdUSP in the final (fourth) instar larvae reduced the messenger RNA levels of four ecdysteroidogenesis genes (Ldspo, Ldphm, Lddib and Ldsad) and 20E titre, and repressed the expression of five 20E signal genes (EcRA, HR3, HR4, E74 and E75) in Leptinotarsa decemlineata. The LdUSP RNAi larvae remained as prepupae, with developing antennae, legs and discs of forewings and hindwings. Dietary supplement with 20E restored the expression of the five 20E signal genes, but only partially alleviated the decreased pupation rate in LdUSP RNAi beetles. Knockdown of LdUSP at the penultimate (third) instar larvae did not affect third-fourth instar moulting. However, silencing LdUSP caused similar but less severe impairments on pupation. Accordingly, we propose that USP is undoubtedly necessary for ecdysteroidogenesis, for mediation of 20E signalling and for initiation of metamorphosis in L. decemlineata.',{'entities': [(31,44,'Gene'),(48,73,'Species'),(121,134,'Gene'),(136,139,'Gene'),(145,162,'Gene'),(164,167,'Gene'),(364,367,'Gene'),(397,400,'Gene'),(518,541,'Species'),(556,559,'Gene'),(751,756,'Gene'),(861,866,'Gene'),(868,873,'Gene'),(875,880,'Gene'),(885,890,'Gene'),(994,1019,'Species'),(1025,1030,'Gene'),(1277,1282,'Gene'),(1310,1315,'Gene'),(1421,1426,'Gene'),(1512,1515,'Gene'),(1637,1652,'Species')]}),('Metabolism of selected model substrates and insecticides by recombinant CYP6FD encoded by its gene predominately expressed in the brain of Locusta migratoria. The migratory locust, Locusta migartoria, is a major agricultural insect pest and its resistance to insecticides is becoming more prevalent. Cytochrome P450 monooxygenases (CYPs) are important enzymes for biotransformations of various endogenous and xenobiotic substances. These enzymes play a major role in developing insecticide resistance in many insect species. In this study, we heterologously co-expressed a CYP enzyme (CYP6FD1) and cytochrome P450 reductase (CPR) from L.  migartoria in Sf9 insect cells. The recombinant enzymes were assayed for metabolic activity towards six selected model substrates (luciferin-H, luciferin-Me, luciferin-Be, luciferin-PFBE, luciferin-CEE and 7-ethoxycoumarin), and four selected insecticides (deltamethrin, chlorpyrifos, carbaryl and methoprene). Recombinant CYP6FD1 showed activity towards 7-ethoxycoumarin and luciferin-Me, but no detectable activity towards the other luciferin derivatives. Furthermore, the enzyme efficiently oxidized deltamethrin to hydroxydeltamethrin through an aromatic hydroxylation in a time-dependent manner. However, the enzyme did not show any detectable activity towards the other three insecticides. Our results provide direct evidence that CYP6FD1 is capable of metabolizing deltamethrin. This work is a step towards a more complete characterization of the catalytic capabilities of CYP6FD1 and other xenobiotic metabolizing CYP enzymes in L. migratoria.',{'entities': [(139,157,'Species'),(163,179,'Species'),(300,330,'Gene'),(332,336,'Gene'),(573,576,'Gene'),(585,592,'Gene'),(962,969,'Gene'),(1376,1383,'Gene'),(1519,1526,'Gene'),(1561,1564,'Gene'),(1576,1589,'Species')]}),('Expression Patterns, Molecular Characterization, and Response to Host Stress of CYP Genes from Phenacoccus solenopsis (Hemiptera: Pseudococcidae). The quarantine insect pest Phenacoccus solenopsis (Hemiptera: Pseudococcidae) has a broad host range and is distributed worldwide. Each year, P. solenopsis causes significant crop losses. The detoxification of various xenobiotic compounds involves the cytochrome P450 monooxygenase (CYP) superfamily of enzymes. However, the functions of CYPs in P. solenopsis are poorly understood. In the present study, P. solenopsis was reared from the egg to the adult stage on three host plants: Tomato, cotton, and hibiscus. Thirty-seven P. solenopsis CYP genes were identified and their phylogenetic relationships were analyzed. Eleven CYP genes (PsCYP4NT1, PsCYP4G219, PsCYP6PZ1, PsCYP6PZ5, PsCYP301B1, PsCYP302A1, PsCYP305A22, PsCYP315A1, PsCYP353F1, PsCYP3634A1, and PsCYP3635A2) were selected for quantitative real-time PCR analysis. The results demonstrated marked differences in CYP expression levels in P. solenopsis grown on different host plants. The results will aid the molecular characterization of CYPs and will increase our understanding of CYP expression patterns in P. solenopsis during development and growth on different hosts.',{'entities': [(80,83,'Gene'),(95,117,'Species'),(174,196,'Species'),(289,302,'Species'),(399,428,'Gene'),(430,433,'Gene'),(493,506,'Species'),(552,565,'Species'),(631,637,'Species'),(674,687,'Species'),(688,691,'Gene'),(773,776,'Gene'),(784,793,'Gene'),(795,805,'Gene'),(807,816,'Gene'),(818,827,'Gene'),(829,839,'Gene'),(841,851,'Gene'),(853,864,'Gene'),(866,876,'Gene'),(878,888,'Gene'),(890,901,'Gene'),(907,918,'Gene'),(1022,1025,'Gene'),(1047,1060,'Species'),(1192,1195,'Gene'),(1219,1232,'Species')]}),('CRISPR/Cas9-mediated knockout of both the PxABCC2 and PxABCC3 genes confers high-level resistance to Bacillus thuringiensis Cry1Ac toxin in the diamondback moth, Plutella xylostella (L.). Rapid evolution of resistance by insect pests severely jeopardizes the sustainable utilization of biopesticides and transgenic crops that produce insecticidal crystal proteins derived from the entomopathogenic bacterium Bacillus thuringiensis (Bt). Recently, high levels of resistance to Bt Cry1 toxins have been reported to be genetically linked to the mutation or down-regulation of ABC transporter subfamily C genes ABCC2 and ABCC3 in seven lepidopteran insects, including Plutella xylostella (L.). To further determine the causal relationship between alterations in the PxABCC2 and PxABCC3 genes and Cry1Ac resistance in P. xylostella, the novel CRISPR/Cas9 genome engineering system was utilized to successfully construct two knockout strains: the ABCC2KO strain is homozygous for a 4-bp deletion in exon 3 of the PxABCC2 gene, and the ABCC3KO strain is homozygous for a 5-bp deletion in exon 3 of the PxABCC3 gene, both of which can produce only truncated ABCC proteins. Bioassay results indicated that high levels of resistance to the Cry1Ac protoxin were observed in both the ABCC2KO (724-fold) and ABCC3KO (413-fold) strains compared to the original susceptible DBM1Ac-S strain. Subsequently, dominance degree and genetic complementation tests demonstrated that Cry1Ac resistance in both the knockout strains was incompletely recessive, and Cry1Ac resistance alleles were located in the classic BtR-1 resistance locus that harbored the PxABCC2 and PxABCC3 genes, similar to the near-isogenic resistant NIL-R strain. Moreover, qualitative toxin binding assays revealed that the binding of the Cry1Ac toxin to midgut brush border membrane vesicles (BBMVs) in both knockout strains was dramatically reduced compared to that in the susceptible DBM1Ac-S strain. In summary, our CRISPR/Cas9-mediated genome editing study presents, for the first time, in vivo reverse genetics evidence for both the ABCC2 and ABCC3 proteins as midgut functional receptors for Bt Cry1 toxins in insects, which provides new insight into the pivotal roles of both the ABCC2 and ABCC3 proteins in the complex molecular mechanism of insect resistance to Bt Cry1 toxins.',{'entities': [(42,49,'Gene'),(54,61,'Gene'),(101,123,'Species'),(144,160,'Species'),(162,181,'Species'),(408,430,'Species'),(432,434,'Species'),(476,478,'Species'),(607,612,'Gene'),(617,622,'Gene'),(664,683,'Species'),(762,769,'Gene'),(774,781,'Gene'),(813,826,'Species'),(1007,1014,'Gene'),(1095,1102,'Gene'),(1633,1640,'Gene'),(1645,1652,'Gene'),(2089,2094,'Gene'),(2099,2104,'Gene'),(2149,2151,'Species'),(2238,2243,'Gene'),(2248,2253,'Gene'),(2322,2324,'Species')]}),('Molecular characterization, spatial-temporal expression and magnetic response patterns of iron-sulfur cluster assembly1 (IscA1) in the rice planthopper, Nilaparvata lugens. The mechanisms of magnetoreception have been proposed as the magnetite-based, the chemical radical-pair and biocompass model, in which magnetite particles, the cryptochrome (Cry) or iron-sulfur cluster assembly 1 (IscA1) may be involved. However, little is known about the association among the molecules. Here we investigated the molecular characterization and the mRNA expression of IscA1 in different developmental stages, tissues and magnetic fields in the migratory brown planthopper (BPH), Nilaparvata lugens. NlIscA1 contains an open reading frame of 390 bp, encoding amino acids of 129, with the predicted molecular weight of 14.0 kDa and the isoelectric point of 9.10. Well-conserved Fe-S cluster binding sites were observed in the predicted protein. Phylogenetic analysis demonstrated NlIscA1 to be clustered into the insect s IscA1. NlIscA1 showed up-regulated mRNA expression during the period of migration. The mRNA expression of NlIscA1 could be detected in all the three tissues of head, thorax and abdomen, with the highest expression level in the abdomen. For the macropterous migratory Nilaparvata lugens, mRNA expression of NlIscA1 and N. lugens cryptochrome1 (Nlcry1) were up-regulated under the magnetic fields of 5 Gauss and 10 Gauss in strength (vs. local geomagnetic field), while N. lugens cryptochrome2 (Nlcry2) remained stable. For the brachyterous non-migratory Nilaparvata lugens, no significant changes were found in mRNA expression of NlIscA1, Nlcry1 and Nlcry2 among different magnetic fields. These findings preliminarily reveal that the expression of NlIscA1 and Nlcry1 exhibited coordinated responses to the magnetic field. It suggests some potential associations among the putative magneto-sensitive molecules of cryptochrome and iron-sulfur cluster assembly.',{'entities': [(90,119,'Gene'),(121,126,'Gene'),(135,139,'Species'),(153,171,'Species'),(355,385,'Gene'),(387,392,'Gene'),(558,563,'Gene'),(644,661,'Species'),(663,666,'Species'),(669,687,'Species'),(689,696,'Gene'),(968,975,'Gene'),(1010,1015,'Gene'),(1017,1024,'Gene'),(1116,1123,'Gene'),(1277,1295,'Species'),(1316,1323,'Gene'),(1328,1337,'Species'),(1338,1351,'Gene'),(1353,1359,'Gene'),(1478,1487,'Species'),(1488,1501,'Gene'),(1503,1509,'Gene'),(1563,1581,'Species'),(1639,1646,'Gene'),(1648,1654,'Gene'),(1659,1665,'Gene'),(1758,1765,'Gene'),(1770,1776,'Gene')]}),('Identification and functional characterisation of a novel N-cyanoamidine neonicotinoid metabolising cytochrome P450, CYP9Q6, from the buff-tailed bumblebee Bombus terrestris. Recent work has shown that two bumblebee (Bombus terrestris) cytochrome P450s of the CYP9Q subfamily, CYP9Q4 and CYP9Q5, are important biochemical determinants of sensitivity to neonicotinoid insecticides. Here, we report the characterisation of a third P450 gene CYP9Q6, previously mis-annotated in the genome of B. terrestris, encoding an enzyme that metabolises the N-cyanoamidine neonicotinoids thiacloprid and acetamiprid with high efficiency. The genomic location and complete ORF of CYP9Q6 was corroborated by PCR and its metabolic activity characterised in vitro by expression in an insect cell line. CYP9Q6 metabolises both thiacloprid and acetamiprid more rapidly than the previously reported CYP9Q4 and CYP9Q5. We further demonstrate a direct, in vivo correlation between the expression of the CYP9Q6 enzyme in transgenic Drosophila melanogaster and an increased tolerance to thiacloprid and acetamiprid. We conclude that CYP9Q6 is an efficient metaboliser of N-cyanoamidine neonicotinoids and likely plays a key role in the high tolerance of B. terrestris to these insecticides.',{'entities': [(100,115,'Gene'),(117,123,'Gene'),(134,155,'Species'),(156,173,'Species'),(217,234,'Species'),(236,251,'Gene'),(277,283,'Gene'),(288,294,'Gene'),(439,445,'Gene'),(489,502,'Species'),(665,671,'Gene'),(784,790,'Gene'),(878,884,'Gene'),(889,895,'Gene'),(980,986,'Gene'),(1008,1031,'Species'),(1108,1114,'Gene'),(1229,1242,'Species')]}),('G119S ace-1 mutation conferring insecticide resistance detected in the Culex pipiens complex in Morocco. BACKGROUND: Arboviruses are controlled through insecticide control of their mosquito vector. However, inconsiderate use of insecticides often results in the selection of resistance in treated populations, so that monitoring is required to optimize their usage. Here, Culex pipiens (West Nile and Rift Valley Fever virus vector) specimens were collected from four Moroccan cities. Levels of susceptibility to the organophosphate (OP) insecticide malathion were assessed using World Health Organization (WHO)-recommended bioassays. Individual mosquitoes were tested for the presence of the G119S mutation in the ace-1 gene, the main OP-target resistance mutation. RESULTS: Bioassays showed that mosquitoes from Mohammedia were significantly more resistant to malathion than those from Marrakech. Analyzing the ace-1 genotypes in dead and surviving individuals suggested that other resistance mechanisms may be present in Mohammedia. The ace-1 resistance allele frequencies were relatively moderate (< 0.4). Their analyses in three Moroccan cities (Tangier, Casablanca and Marrakech) however showed disparities between two coexisting Cx. pipiens forms and revealed that the G119S mutation tends to be more frequent in urban than in rural collection sites. CONCLUSION: These findings provide a reference assessment of OP resistance in Morocco and should help the health authorities to develop informed and sustainable vector control programs.  2018 Society of Chemical Industry.',{'entities': [(6,11,'Gene'),(71,92,'Species'),(372,385,'Species'),(401,424,'Species'),(715,720,'Gene'),(913,918,'Gene'),(1040,1045,'Gene'),(1236,1247,'Species')]}),('Transcriptional response of ATP-binding cassette (ABC) transporters to insecticides in the cotton bollworm, Helicoverpa armigera. When any living organism is frequently exposed to any drugs or toxic substances, they evolve different detoxification mechanism to confront with toxicants during absorption and metabolism. Likewise, the insects have evolved detoxification mechanisms as they are frequently exposed to different toxic secondary plant metabolites and commercial insecticides. ABC transporter superfamily is one of the largest and ubiquitous group of proteins which play an important role in phase III of the detoxification process. However, knowledge about this gene family remains largely unknown. To help fill this gap, we have identified a total of 54 ABC transporters in the Helicoverpa armigera genome which are classified into eight subfamilies (A-H) by phylogenetic analysis. The temporal and spatial expression profiles of these 54 ABC transporters throughout H. armigera development stages and seven tissues and their responses to five different insecticides, were investigated using RNA-seq analysis. Furthermore, the mRNA expression of eight selected genes in different tissues and six genes responses to insecticides were confirmed by the quantitative real-time PCR (RT-qPCR). Moreover, H. armigera become more sensitive to abamectin and indoxacarb when P-gp was inhibited. These results provide a foundation for further studies of ABCs in H. armigera.',{'entities': [(28,48,'Gene'),(50,53,'Gene'),(91,106,'Species'),(108,128,'Species'),(487,490,'Gene'),(766,769,'Gene'),(790,810,'Species'),(951,954,'Gene'),(979,990,'Species'),(1310,1321,'Species'),(1463,1474,'Species')]}),('CRISPR/Cas9-mediated ebony knockout results in puparium melanism in Spodoptera litura. Insect body pigmentation and coloration are critical to adaption to the environment. To explore the mechanisms that drive pigmentation, we used the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) genome editing system to target the ebony gene in the non-model insect Spodoptera litura. Ebony is crucial to melanin synthesis in insects. By directly injecting Cas9 messenger RNA and ebony-specific guide RNAs into S. litura embryos, we successfully induced a typical ebony-deficient phenotype of deep coloration of the puparium and induction of melanin formation during the pupal stage. Polymerase chain reaction-based genotype analysis demonstrated that various mutations had occurred at the sites targeted in ebony. Our study clearly demonstrates the function of ebony in the puparium coloration and also provides a potentially useful marker gene for functional studies in S. litura as well as other lepidopteran pests.',{'entities': [(21,26,'Gene'),(68,85,'Species'),(373,378,'Gene'),(408,425,'Species'),(427,432,'Gene'),(522,527,'Gene'),(553,562,'Species'),(563,570,'Species'),(850,855,'Gene'),(904,909,'Gene'),(1014,1023,'Species')]}),('Germline expression of the hunchback orthologues in the asexual viviparous aphids: a conserved feature within the Aphididae. In animals, differentiation of germline from soma usually takes place during embryogenesis. Genes and their products that are preferentially expressed in the embryonic germ cells are regarded as candidates for maintaining germline fate or promoting germline identity. In Drosophila, for example, the protein encoded by the germline gene vasa is specifically restricted to the germ cells, while products of the gap gene hunchback (hb), a somatic gene, are preferentially expressed in the neuroblasts. In this study, we report the expression of both messenger RNA and protein encoded by Aphb, an hb orthologue in the asexual viviparous pea aphid Acyrthosiphon pisum, in germ cells as well as in neuroblasts. We infer that expression of Aphb messenger RNA in the germ cells during the formation of germaria is required for the anterior localization of Aphb in the protruding oocytes. Germarial expression and anterior localization of ApKruppel was also identified but, unlike Aphb, its expression was not detected in the migrating germ cells. Very similar patterns of hb expression were also identified in the green peach aphid Myzus persicae, suggesting that germline expression of hb is conserved within the Aphididae. To date, this pattern of hb germline expression has not been reported in other insects.',{'entities': [(27,36,'Gene'),(544,553,'Gene'),(555,557,'Gene'),(710,714,'Gene'),(719,721,'Gene'),(759,768,'Species'),(769,788,'Species'),(859,863,'Gene'),(974,978,'Gene'),(1056,1065,'Gene'),(1098,1102,'Gene'),(1190,1192,'Gene'),(1232,1249,'Species'),(1250,1264,'Species'),(1305,1307,'Gene'),(1368,1370,'Gene')]}),('Characterization of antennal chemosensilla and associated odorant binding as well as chemosensory proteins in the Eupeodes corollae (Diptera: Syrphidae). Aphidophagous syrphids are important for pest control and pollination in various agroecosystems. However, the mechanism underlying olfaction, which is critical for insect  behavioral processes and fitness, has not been well understood in the family Syrphidae. Hence, we performed a systematic identification and characterisation of the antennal sensilla and two groups of soluble proteins, odorant-binding proteins (OBPs) and chemosensory proteins (CSPs), in the hoverfly Eupeodes corollae. (i) With scanning electron microscopy, four major types of sensilla (chaetic sensilla [two subtypes], trichoid sensilla, basiconic sensilla [two subtypes] and coeloconic sensilla), with numerous microtrichia, were first observed along the entire surface of aristate antennae of both sexes of E. corollae. Of these, only chaetic sensillum was found on the first two antennal segments, scape and pedicel, while the other types of sensilla were located on the flagellum. No marked difference was observed in the morphological structure or distributional pattern of any of the sensilla between the two sexes. (ii) By molecular cloning and bioinformatic analysis, 7 EcorCSPs and 28 EcorOBPs (20 classic OBPs, 5 minus-C OBPs, and 3 plus-C OBPs) were directly identified from the species, which all share the characteristic hallmarks of their family, including the presence of a signal peptide and conserved cysteine signature. (iii) RT-qPCR of these chemosensory genes showed predominately tissue-biased expression patterns; 32 of the 35 EcorOBPs/CSPs were uniquely or primarily expressed in the main olfactory organs, either the antennae or head. (iv) Among these, several genes (EcorCSP2 and EcorOBP1, 9, 12, 15-17, 20) appeared to be antenna-biased. In situ hybridization assays indicated that each antenna-biased chemosensory gene was expressed in a different number of cells, suggesting they might play a more vital role in odour recognition and perception and could be potential candidates to study their biological functions in vivo and in vitro. Together, our current findings provide a basis for future studies on how syrphids utilize chemical cues to regulate their behavior during interactions among the natural enemy, its prey, and host plant in agro-ecosystems.',{'entities': [(114,131,'Species'),(544,567,'Gene'),(570,574,'Gene'),(580,600,'Gene'),(603,607,'Gene'),(626,643,'Species'),(937,948,'Species'),(1306,1314,'Gene'),(1322,1330,'Gene'),(1343,1347,'Gene'),(1359,1363,'Gene'),(1378,1382,'Gene'),(1677,1685,'Gene'),(1686,1690,'Gene')]}),('Characterization of the novel role of NinaB orthologs from Bombyx mori and Tribolium castaneum. Carotenoids can be enzymatically converted to apocarotenoids by carotenoid cleavage dioxygenases. Insect genomes encode only one member of this ancestral enzyme family. We cloned and characterized the ninaB genes from the silk worm (Bombyx mori) and the flour beetle (Tribolium castaneum). We expressed BmNinaB and TcNinaB in E. coli and analyzed their biochemical properties. Both enzymes catalyzed a conversion of carotenoids into cis-retinoids. The enzymes catalyzed a combined trans to cis isomerization at the C11, C12 double bond and oxidative cleavage reaction at the C15, C15  bond of the carotenoid carbon backbone. Analyses of the spatial and temporal expression patterns revealed that ninaB genes were differentially expressed during the beetle and moth life cycles with high expression in reproductive organs. In Bombyx mori, ninaB was almost exclusively expressed in female reproductive organs of the pupa and adult. In Tribolium castaneum, low expression was found in reproductive organs of females but high expressions in male reproductive organs of the pupa and imagoes. We performed RNAi experiments to characterize the role of NinaB in insect reproduction. We observed that RNAi treatment significantly decreased the expression levels of BmninaB and TcninaB and reduced the egg laying capacity of both insects. Together, our study revealed that NinaB s unique enzymatic properties are well conserved among insects and implicate NinaB function in insect reproduction.',{'entities': [(38,43,'Gene'),(59,70,'Species'),(75,94,'Species'),(297,302,'Gene'),(329,340,'Species'),(364,383,'Species'),(399,406,'Gene'),(411,418,'Gene'),(422,429,'Species'),(792,797,'Gene'),(921,932,'Species'),(934,939,'Gene'),(1029,1048,'Species'),(1241,1246,'Gene'),(1352,1359,'Gene'),(1364,1371,'Gene'),(1459,1464,'Gene'),(1542,1547,'Gene')]}),('Characterization and functional analysis of hsp18.3 gene in the red flour beetle, Tribolium castaneum. Small heat shock proteins (sHSPs) are diverse and mainly function as molecular chaperones to protect organisms and cells from various stresses. In this study, hsp18.3, one Tribolium castaneum species-specific shsp, has been identified. Quantitative real-time polymerase chain reaction illustrated that Tchsp18.3 is expressed in all developmental stages, and is highly expressed at early pupal and late adult stages, while it is highly expressed in ovary and fat body at the adult period. Moreover, it was up-regulated 4532  396-fold in response to enhanced heat stress but not to cold stress; meanwhile the lifespan of adults in ds-Tchsp18.3 group reduced by 15.8% from control group under starvation. Laval RNA interference (RNAi) of Tchsp18.3 caused 86.1%  4.5% arrested pupal eclosion and revealed that Tchsp18.3 played an important role in insect development. In addition, parental RNAi of Tchsp18.3 reduced the oviposition amount by 94.7%. These results suggest that Tchsp18.3 is not only essential for the resistance to heat and starvation stress, but also is critical for normal development and reproduction in T. castaneum.',{'entities': [(44,51,'Gene'),(64,80,'Species'),(82,101,'Species'),(103,128,'Gene'),(130,135,'Gene'),(262,269,'Gene'),(275,294,'Species'),(405,414,'Gene'),(735,744,'Gene'),(838,847,'Gene'),(909,918,'Gene'),(997,1006,'Gene'),(1075,1084,'Gene'),(1221,1233,'Species')]}),('Olfactory co-receptor Orco stimulated by Rice stripe virus is essential for host seeking behavior in small brown planthopper. BACKGROUND: Laodelphax striatellus, the small brown planthopper (SBPH), is an economically important pest, besides sucking damage, which transmits rice viruses to cause severe damages to rice. In the process of virus transmission, the host orientation behavior of insect is mainly driven by olfaction. In this context, the molecular basis of olfaction in SBPH is of particular interest. RESULTS: Here, we identified the gene that encodes olfactory receptor co-receptor (Orco) and analyzed its expression profiles in Rice stripe virus (RSV)-infected and RSV-free SBPH. It was found that LstrOrco shared high identity with other Orcos from different order insects. LstrOrco was mainly expressed in the head of SBPH, and its expression was significantly stimulated by RSV-infection. The behavioral bioassay revealed that viruliferous SBPH might have a stronger olfactory and seeking ability for rice than RSV-free insect. After silencing of LstrOrco expression, the olfaction and seeking behavior of nymphs for rice seedlings was significantly inhibited, mainly in the increase of the  no response  percent and the prolongation of the response time. CONCLUSION: These results suggested that Orco played an important role in olfactory signaling and seeking behavior of SBPH, which provided a basic for future development of olfactory-based agriculture pest management strategies.  2018 Society of Chemical Industry.',{'entities': [(0,21,'Gene'),(22,26,'Gene'),(41,58,'Species'),(101,124,'Species'),(138,160,'Species'),(166,189,'Species'),(191,195,'Species'),(273,277,'Species'),(313,317,'Species'),(481,485,'Species'),(564,594,'Gene'),(596,600,'Gene'),(642,659,'Species'),(688,692,'Species'),(712,720,'Gene'),(789,797,'Gene'),(834,838,'Species'),(957,961,'Species'),(1018,1022,'Species'),(1064,1072,'Gene'),(1134,1138,'Species'),(1314,1318,'Gene'),(1391,1395,'Species')]}),('Downregulation of dystrophin expression in pupae of the whitefly Bemisia tabaci inhibits the emergence of adults. The whitefly Bemisia tabaci is a major pest to agriculture. Adults are is able to fly for long distances and to colonize staple crops, herbs and ornamentals, and to vector viruses belonging to several important taxonomic groups. During their early development, whiteflies mature from eggs through several nymphal stages (instars I to IV) until adults emerge from pupae. We aim at reducing whitefly populations by inhibiting the emergence of adults from nymphs. Here we targeted dystrophin, a conserved protein essential for the development of the muscle system in human, animals and insects. We have exploited the fact that whitefly nymphs developing on tomato leaves feed from the plant phloem via their stylets. Thus, we delivered dystrophin-silencing dsRNA to nymphs developing on leaves of tomato plantlets with their roots bathing in the silencing solution. Downregulation of dystrophin expression occurred mainly in pupae. Dystrophin silencing induced also the downregulation of the dystrophin-associated protein genes actin and tropomyosin, and disrupted F-actin. Most significant, the treatment inhibited the emergence of adults from pupae, suggesting that targeting dystrophin may help restraining whitefly populations. This study demonstrates for the first time the important role of dystrophin in the development of a major insect pest to agriculture. This article is protected by copyright. All rights reserved.',{'entities': [(18,28,'Gene'),(65,79,'Species'),(127,141,'Species'),(592,602,'Gene'),(678,683,'Species'),(768,774,'Species'),(847,857,'Gene'),(908,914,'Species'),(995,1005,'Gene'),(1043,1053,'Gene'),(1103,1113,'Gene'),(1289,1299,'Gene'),(1408,1418,'Gene')]}),('RNA interference-mediated knockdown of eye coloration genes in the western tarnished plant bug (Lygus hesperus Knight). Insect eye coloration arises from the accumulation of various pigments. A number of genes that function in the biosynthesis (vermilion, cinnabar, and cardinal) and importation (karmoisin, white, scarlet, and brown) of these pigments, and their precursors, have been identified in diverse species and used as markers for transgenesis and gene editing. To examine their suitability as visible markers in Lygus hesperus Knight (western tarnished plant bug), transcriptomic data were screened for sequences exhibiting homology with the Drosophila melanogaster proteins. Complete open reading frames encoding putative homologs for all seven genes were identified. Bioinformatic-based sequence and phylogenetic analyses supported initial annotations as eye coloration genes. Consistent with their proposed role, each of the genes was expressed in adult heads as well as throughout nymphal and adult development. Adult eyes of those injected with double-stranded RNAs (dsRNAs) for karmoisin, vermilion, cinnabar, cardinal, and scarlet were characterized by a red band along the medial margin extending from the rostral terminus to the antenna. In contrast, eyes of insects injected with dsRNAs for both white and brown were a uniform light brown. White knockdown also produced cuticular and behavioral defects. Based on its expression profile and robust visible phenotype, cardinal would likely prove to be the most suitable marker for developing gene editing methods in Lygus species.',{'entities': [(75,94,'Species'),(96,110,'Species'),(245,254,'Gene'),(256,264,'Gene'),(270,278,'Gene'),(297,306,'Gene'),(308,313,'Gene'),(315,322,'Gene'),(328,333,'Gene'),(522,536,'Species'),(553,572,'Species'),(652,675,'Species'),(1094,1103,'Gene'),(1105,1114,'Gene'),(1116,1124,'Gene'),(1126,1134,'Gene'),(1140,1147,'Gene'),(1316,1321,'Gene'),(1326,1331,'Gene'),(1353,1358,'Gene'),(1360,1365,'Gene'),(1486,1494,'Gene')]}),('Complexities in Bombyx germ cell formation process revealed by Bm-nosO (a Bombyx homolog of nanos) knockout. Inheritance (sequestration of a localized determinant: germplasm) and zygotic induction are two modes of metazoan primordial germ cell (PGC) specification. vasa and nanos homologs are evolutionarily conserved germline marker genes that have been used to examine the ontogeny of germ cells in various animals. In the lepidopteran insect Bombyx mori, although the lack of vasa homolog (BmVLG) protein localization as well as microscopic observation suggested the lack of germplasm, classical embryo manipulation studies and the localization pattern of Bm-nosO (one of the four nanos genes in Bombyx) maternal mRNA in the egg raised the possibility that an inheritance mode is operating in Bombyx. Here, we generated Bm-nosO knockouts to examine whether the localized mRNA acts as a localized germ cell determinant. Contrary to our expectations, Bm-nosO knockout lines could be established. However, these lines frequently produced abnormal eggs, which failed to hatch, to various extent depending on the individuals. We also found that Bm-nosO positively regulated BmVLG expression at least during embryonic stage, directly or indirectly, indicating that these genes were on the same developmental pathway for germ cell formation in Bombyx. These results suggest that these conserved genes are concerned with stable germ cell production. On the other hand, from the aspect of BmVLG as a PGC marker, we showed that maternal Bm-nosO product(s) as well as early zygotic Bm-nosO activity were redundantly involved in PGC specification; elimination of both maternal and zygotic gene activities (as in knockout lines) resulted in the apparent lack of PGCs, indicating that an inheritance mechanism indeed operates in Bombyx. This, however, together with the fact that germ cells are produced at all in Bm-nosO knockout lines, also suggests the possibility that, in Bombyx, not only this inheritance mechanism but also an inductive mechanism acts in concert to form germ cells or that loss of early PGCs are compensated for by germline regeneration: mechanisms that could enable the evolution of preformation. Thus, Bombyx could serve as an important organism in understanding the evolution of germ cell formation mechanisms; transition between preformation and inductive modes.',{'entities': [(63,70,'Gene'),(92,97,'Gene'),(265,269,'Gene'),(274,279,'Gene'),(445,456,'Species'),(479,483,'Gene'),(493,498,'Gene'),(659,666,'Gene'),(684,689,'Gene'),(823,830,'Gene'),(952,959,'Gene'),(1143,1150,'Gene'),(1172,1177,'Gene'),(1483,1488,'Gene'),(1530,1537,'Gene'),(1574,1581,'Gene'),(1903,1910,'Gene')]}),('Function and pharmacology of glutamate-gated chloride channel exon 9 splice variants from the diamondback moth Plutella xylostella. Glutamate-gated chloride channels (GluCls) are found only in invertebrates and mediate fast inhibitory neurotransmission. The structural and functional diversity of GluCls are produced through assembly of multiple subunits and via posttranscriptional alternations. Alternative splicing is the most common way to achieve this in insect GluCls and splicing occurs primarily at exons 3 and 9. As expression pattern and pharmacological properties of exon 9 alternative splices in invertebrate GluCls remain poorly understood, the cDNAs encoding three alternative splice variants (9a, 9b and 9c) of the PxGluCl gene from the diamondback moth Plutella xylostella were constructed and their pharmacological characterizations were examined using electrophysiological studies. Alternative splicing of exon 9 had little to no impact on PxGluCl sensitivity towards the agonist glutamate when subunits were singly or co-expressed in Xenopus oocytes. In contrast, the allosteric modulator abamectin and the chloride channel blocker fipronil had differing effects on PxGluCl splice variants. PxGluCl9c channels were more resistant to abamectin and PxGluCl9b channels were more sensitive to fipronil than other homomeric channels. In addition, heteromeric channels containing different splice variants showed similar sensitivity to abamectin (except for 9c) and reduced sensitivity to fipronil than homomeric channels. These findings suggest that functionally indistinguishable but pharmacologically distinct GluCls could be formed in P. xylostella and that the upregulated constitutive expression of the specific variants may contribute to the evolution of insecticide resistance in P. xylostella and other arthropods.',{'entities': [(29,61,'Gene'),(94,110,'Species'),(111,130,'Species'),(132,165,'Gene'),(167,173,'Gene'),(297,303,'Gene'),(467,473,'Gene'),(621,627,'Gene'),(730,737,'Gene'),(752,768,'Species'),(769,788,'Species'),(958,965,'Gene'),(1053,1060,'Species'),(1185,1192,'Gene'),(1210,1219,'Gene'),(1266,1275,'Gene'),(1626,1632,'Gene'),(1652,1665,'Species'),(1801,1814,'Species')]}),('Sublethal effects of chlorantraniliprole on molting hormone levels and mRNA expressions of three Halloween genes in the rice stem borer, Chilo suppressalis. While sublethal effects of insecticide on insect development have been widely studied, the underlying mechanisms remain elusive. Our previous studies revealed that sublethal concentrations of chlorantraniliprole significantly increased the juvenile hormone levels and resulted in both prolonged developmental time and reduced fecundity in Chilo suppressalis. In the present study, we evaluated the sublethal effects of chlorantraniliprole on molting hormone (MH) levels and mRNA expressions of three Halloween genes including CsCYP307A1, CsCYP306A1 and CsCYP314A1 in C. suppressalis. The results showed that the MH levels in different developmental stages of C. suppressalis were decreased after exposure to LC10 and LC30 of chlorantraniliprole. However, analysis of temporal expression profiles revealed that the mRNA levels of three Halloween genes were not closely correlated with the ecdysteroid titers in C. suppressalis. Notably, the transcript levels of CsCYP307A1, CsCYP306A1 and CsCYP314A1 were induced after treatment with sublethal concentrations of chlorantraniliprole in specific developmental stages. These results indicated that chlorantraniliprole had adverse effects on insect MH biosynthesis, and in addition to the involvement in MH biosynthesis, CsCYP307A1, CsCYP306A1 and CsCYP314A1 may also play important roles in the detoxification metabolism of chlorantraniliprole in C. suppressalis.',{'entities': [(97,106,'Gene'),(120,124,'Species'),(137,155,'Species'),(496,514,'Species'),(657,666,'Gene'),(683,693,'Gene'),(695,705,'Gene'),(710,720,'Gene'),(724,739,'Species'),(816,831,'Species'),(992,1001,'Gene'),(1067,1082,'Species'),(1118,1128,'Gene'),(1130,1140,'Gene'),(1145,1155,'Gene'),(1423,1433,'Gene'),(1435,1445,'Gene'),(1450,1460,'Gene'),(1550,1565,'Species')]}),('Downregulation of imidacloprid resistant genes alters the biological parameters in Colorado potato beetle, Leptinotarsa decemlineata Say (chrysomelidae: Coleoptera). Colorado potato beetle, Leptinotarsa decemlineata Say (coleoptera: chrysomelidae), is the important pest of potato all over the world. This insect pest is resistant to more than 50 active compounds belonging to various chemical groups. Potential of RNA interference (RNAi) was explored to knock down transcript levels of imidacloprid resistant genes in Colorado potato beetle (CPB) under laboratory conditions. Three important genes belonging to cuticular protein (CP), cytochrome P450 monoxygenases (P450) and glutathione synthetase (GSS) families encoding imidacloprid resistance were targeted. Feeding bio-assays were conducted on various stages of imidacloprid resistant CPB lab population by applying HT115 expressing dsRNA on potato leaflets. Survival rate of insects exposed to CP-dsRNA decreased to 4.23%, 15.32% and 47.35% in 2nd, 3rd and 4th instar larvae respectively. Larval weight and pre-adult duration were also affected due to dsRNAs feeding. Synergism of RNAi with imidacloprid conducted on the 2nd instar larvae, exhibited 100% mortality of larvae when subjected to reduced doses of GSS and CP dsRNAs along with imidacloprid. Utilization of three different dsRNAs against imidacloprid resistant CPB population reveal that dsRNAs targeting CP, P450 and GSS enzymes could be useful tool in management of imidacloprid resistant CPB populations.',{'entities': [(18,40,'Gene'),(83,105,'Species'),(107,132,'Species'),(166,188,'Species'),(190,215,'Species'),(274,280,'Species'),(487,509,'Gene'),(519,541,'Species'),(543,546,'Species'),(612,629,'Gene'),(631,633,'Gene'),(636,664,'Gene'),(667,671,'Gene'),(677,699,'Gene'),(701,704,'Gene'),(841,844,'Species'),(898,904,'Species'),(951,953,'Gene'),(1267,1270,'Gene'),(1275,1277,'Gene'),(1379,1382,'Species'),(1423,1425,'Gene'),(1427,1431,'Gene'),(1436,1439,'Gene'),(1509,1512,'Species')]}),('Conserved roles of Osiris genes in insect development, polymorphism and protection. Much of the variation among insects is derived from the different ways that chitin has been moulded to form rigid structures, both internal and external. In this study, we identify a highly conserved expression pattern in an insect-only gene family, the Osiris genes, that is essential for development, but also plays a significant role in phenotypic plasticity and in immunity/toxicity responses. The majority of Osiris genes exist in a highly syntenic cluster, and the cluster itself appears to have arisen very early in the evolution of insects. We used developmental gene expression in the fruit fly, Drosophila melanogaster, the bumble bee, Bombus terrestris, the harvester ant, Pogonomyrmex barbatus, and the wood ant, Formica exsecta, to compare patterns of Osiris gene expression both during development and between alternate caste phenotypes in the polymorphic social insects. Developmental gene expression of Osiris genes is highly conserved across species and correlated with gene location and evolutionary history. The social insect castes are highly divergent in pupal Osiris gene expression. Sets of co-expressed genes that include Osiris genes are enriched in gene ontology terms related to chitin/cuticle and peptidase activity. Osiris genes are essential for cuticle formation in both embryos and pupae, and genes co-expressed with Osiris genes affect wing development. Additionally, Osiris genes and those co-expressed seem to play a conserved role in insect toxicology defences and digestion. Given their role in development, plasticity, and protection, we propose that the Osiris genes play a central role in insect adaptive evolution.',{'entities': [(19,25,'Gene'),(338,344,'Gene'),(498,504,'Gene'),(678,687,'Species'),(689,712,'Species'),(730,747,'Species'),(768,789,'Species'),(809,824,'Species'),(849,855,'Gene'),(1003,1009,'Gene'),(1166,1172,'Gene'),(1230,1236,'Gene'),(1329,1335,'Gene'),(1433,1439,'Gene'),(1485,1491,'Gene'),(1677,1683,'Gene')]}),('Hh signaling from de novo organizers drive lgl neoplasia in Drosophila epithelium. The Hedgehog (Hh) morphogen regulates growth and patterning. Since Hh signaling is also implicated in carcinogenesis, it is conceivable that de novo Hh-secreting organizers, if formed in association with oncogenic hit could be tumor-cooperative. Here we validate this hypothesis using the Drosophila model of cooperative epithelial carcinogenesis. We generate somatic clones with simultaneous loss of tumor suppressor, Lgl, and gain of the posterior compartment selector, Engrailed (En), known to induce synthesis of Hh. We show that lgl UAS-en clones in the anterior wing compartment trigger Hh signaling cascade via cross-talk with their Ci-expressing wild type cell neighbors. Hh-Dpp signaling from clone boundaries of such ectopically formed de novo organizers in turn drive lgl carcinogenesis. By contrast, Ci-expressing lgl clones transform by autocrine and/or juxtracine activation of Hh signaling in only the posterior compartment. We further show that sequestration of the Hh ligand or loss of Dpp receptor, Tkv, in these Hh-sending or -receiving lgl clones arrested their carcinogenesis. Our results therefore reveal a hitherto unrecognized mechanism of tumor cooperation by developmental organizers, which are induced fortuitously by oncogenic hits.',{'entities': [(0,2,'Gene'),(43,46,'Gene'),(60,70,'Species'),(87,95,'Gene'),(97,99,'Gene'),(150,152,'Gene'),(232,234,'Gene'),(502,505,'Gene'),(555,564,'Gene'),(566,568,'Gene'),(600,602,'Gene'),(617,620,'Gene'),(625,627,'Gene'),(676,678,'Gene'),(763,765,'Gene'),(862,865,'Gene'),(909,912,'Gene'),(975,977,'Gene'),(1065,1067,'Gene'),(1086,1098,'Gene'),(1100,1103,'Gene'),(1114,1116,'Gene'),(1139,1142,'Gene')]}),('Genome-wide gene expression profiling reveals that cuticle alterations and P450 detoxification are associated with deltamethrin and DDT resistance in Anopheles arabiensis populations from Ethiopia. BACKGROUND: Vector control is the main intervention in malaria control and elimination strategies. However, the development of insecticide resistance is one of the major challenges for controlling malaria vectors. Anopheles arabiensis populations in Ethiopia showed resistance against both DDT and the pyrethroid deltamethrin. Although an L1014F target-site resistance mutation was present in the voltage gated sodium channel of investigated populations, the levels of resistance indicated the presence of additional resistance mechanisms. In this study, we used genome-wide transcriptome profiling by RNAseq to assess differentially expressed genes between three deltamethrin and DDT resistant An. arabiensis field populations - Asendabo, Chewaka and Tolay - and two susceptible strains - Sekoru and Mozambique. RESULTS: Both RNAseq analysis and RT-qPCR showed that a glutathione-S-transferase, gstd3, and a cytochrome P450 monooxygenase, cyp6p4, were significantly overexpressed in the group of resistant populations compared to the susceptible strains, suggesting that the enzymes they encode play a key role in metabolic resistance against deltamethrin or DDT. Furthermore, a gene ontology enrichment analysis showed that expression changes of cuticle related genes were strongly associated with insecticide resistance. Although this did not translate in increased thickness of the procuticle, a higher cuticular hydrocarbon content was observed in a resistant population. CONCLUSION: Our transcriptome sequencing of deltamethrin and DDT resistant An. arabiensis populations from Ethiopia suggests non-target site resistance mechanisms and paves the way for further investigation of the role of cuticle composition in insecticide resistance of malaria vectors.  2019 Society of Chemical Industry.',{'entities': [(75,79,'Gene'),(150,170,'Species'),(412,432,'Species'),(893,907,'Species'),(1067,1092,'Gene'),(1094,1099,'Gene'),(1107,1136,'Gene'),(1138,1144,'Gene'),(1750,1764,'Species')]}),('Overexpression of Gloverin2 in the Bombyx mori silk gland enhances cocoon/silk antimicrobial activity. The Bombyx mori cocoon/silk possesses many immune-related components, including protease inhibitors, seroins, and antimicrobial peptides, which likely help to protect the pupating larva from infection. However, the natural antimicrobial activity of the B. mori cocoon/silk is still too weak for biomedical applications. With the goal of enhancing this natural activity, we constructed a transgenic vector to overexpress the B. mori antimicrobial peptide Gloverin2 (BmGlv2) under control of the silk gland-specific Serion1 promoter. Transgenic silkworms were generated via embryo microinjection. A low level of BmGlv2 was expressed in the non-transgenic silk gland, but BmGlv2 was efficiently overexpressed and proteolytically activated in the transgenic line. Overexpressed BmGlv2 was secreted and incorporated into the silk during spanning without affecting cocoon/silk formation. Moreover, the transgenic cocoon/silk had significantly greater inhibitory activity against bacteria and fungi than the non-transgenic cocoon/silk. This strategy could help enhance the antimicrobial performance and biomedical application of silk.',{'entities': [(18,27,'Gene'),(35,46,'Species'),(107,118,'Species'),(356,363,'Species'),(527,534,'Species'),(557,566,'Gene'),(568,574,'Gene'),(646,655,'Species'),(713,719,'Gene'),(772,778,'Gene'),(877,883,'Gene')]}),('RNAi-Mediated Knockdown of Tssk1 and Tektin1 Genes Impair Male Fertility in Bactrocera dorsalis. The genetic-based sterile insect technique (SIT) is an effective and environmentally safe strategy to diminish populations of agricultural and horticultural insect pests. Functional characterization of genes related to male fertility can enhance the genetic-based SIT. Tssk1 has been involved to control male fertility in both mammals and insects. Moreover, Tektin1 has also been revealed to influence male fertility in both human and mammals. These findings suggested that Tssk1 and Tektin1 identified from Bactrocera dorsalis could be required for male fertility in B. dorsalis. In this study, expression profiles of these two genes were studied at different developmental stages and in various tissues of adult males. Remarkably, it was found that Tssk1 and Tektin1 were highly expressed in the testis of mature adult males of B. dorsalis. Furthermore, Tssk1 and Tektin1 genes were downregulated by using the RNA interference (RNAi) method. Fertility assays including egg laying, hatching, and spermatozoa count were also performed to investigate male fertility of B. dorsalis. Results showed that knockdown of Tssk1 and Tektin1 caused male sterility up to 58.99% and 64.49%, respectively. As expected, the total numbers of spermatozoa were also significantly reduced by 65.83% and 73.9%, respectively. These results suggested that male sterility was happened wing to the low number of spermatozoa. In conclusion, we demonstrate that Tssk1 and Tektin1 are the novel agents that could be used to enhance the genetic-based SIT, or their double-stranded RNA (dsRNA) can be used as biopesticides to control the population of B. dorsalis.',{'entities': [(27,32,'Gene'),(37,44,'Gene'),(76,95,'Species'),(366,371,'Gene'),(455,462,'Gene'),(522,527,'Species'),(571,576,'Gene'),(581,588,'Gene'),(605,624,'Species'),(665,676,'Species'),(848,853,'Gene'),(858,865,'Gene'),(927,938,'Species'),(953,958,'Gene'),(963,970,'Gene'),(1165,1176,'Species'),(1211,1216,'Gene'),(1221,1228,'Gene'),(1534,1539,'Gene'),(1544,1551,'Gene'),(1721,1732,'Species')]}),('RNA interference in Nilaparvata lugens (Homoptera: Delphacidae) based on dsRNA ingestion. BACKGROUND: An efficient and convenient RNA interference (RNAi) technique involving double-stranded RNA (dsRNA) ingestion is useful for gene function studies of non-model insects. RESULTS: Three dsRNAs targeting different sites within a gene encoding vacuolar ATP synthase subunit E (V-ATPase-E, 21E01) were synthesised for RNAi in Nilaparvata lugens. dsRNA was found to be stable in 0.1 g mL(-1) sucrose solution, but unstable in artificial fodder. Therefore, dsRNAs were orally delivered into N. lugens in 0.1 g mL(-1) sucrose solution. RNAi was induced by all three of the dsRNAs at 0.05  g  L(-1) in N. lugens. Time dynamics analysis of gene silencing indicated that significant suppression of the target gene began as early as 2 days after ingestion of ds2-21E01 and ds3-21E01. However, significant repressive effects were recorded up to 10 days after exposure to ds1-21E01. The maximum reduction in target gene mRNA was observed after 10 days of treatment, with suppression ratios induced by ds1-21E01, ds2-21E01 and ds3-21E01 of 41, 55 and 48% respectively. CONCLUSION: An efficient and convenient RNAi technique involving dsRNA ingestion has been successfully developed for N. lugens. This will be a useful tool for further functional genomic investigation in this organism.',{'entities': [(20,38,'Species'),(341,372,'Gene'),(374,384,'Gene'),(422,440,'Species'),(585,594,'Species'),(694,703,'Species'),(1272,1281,'Species')]}),('Feeding-based RNA interference of a trehalose phosphate synthase gene in the brown planthopper, Nilaparvata lugens. The brown planthopper, Nilaparvata lugens, is the most devastating rice insect pest to have given rise to an outbreak in recent years. RNA interference (RNAi) is a technological breakthrough that has been developed as a powerful tool for studying gene function and for the highly targeted control of insect pests. Here, we examined the effects of using a feeding-based RNAi technique to target the gene trehalose phosphate synthase (TPS) in N. lugens. The full-length cDNA of N. lugens TPS (NlTPS) is 3235 bp and has an open reading frame of 2424 bp, encoding a protein of 807 amino acids. NlTPS was expressed in the fat body, midgut and ovary. Quantitative real-time PCR (qRT-PCR) analysis revealed that NlTPS mRNA is expressed continuously with little change during the life of the insect. Efficient silencing of the TPS gene through double-stranded RNA (dsRNA) feeding led to rapid and significant reduction levels of TPS mRNA and enzymatic activity. Additionally, the development of N. lugens larvae that had been fed with the dsRNA was disturbed, resulting in lethality, and the cumulative survival rates dropped to 75.56, 64.44, 55.56 and 40.00% after continuous ingestion of 0.5  g/ l dsRNA for 2, 4, 7 and 10 days, respectively. These values were significantly lower than those of the insects in the control group, suggesting that NlTPS dsRNA may be useful as a means of insect pest control.',{'entities': [(36,64,'Gene'),(77,94,'Species'),(96,114,'Species'),(120,137,'Species'),(139,157,'Species'),(183,187,'Species'),(519,547,'Gene'),(549,552,'Gene'),(557,566,'Species'),(592,601,'Species'),(602,605,'Gene'),(607,612,'Gene'),(706,711,'Gene'),(821,826,'Gene'),(935,938,'Gene'),(1037,1040,'Gene'),(1103,1112,'Species'),(1455,1460,'Gene')]}),('Native subunit composition of two insect nicotinic receptor subtypes with differing affinities for the insecticide imidacloprid. Neonicotinoid insecticides, such as imidacloprid, are selective agonists of insect nicotinic acetylcholine receptors (nAChRs) and are used extensively to control a variety of insect pest species. The brown planthopper (Nilaparvata lugens), an insect pest of rice crops throughout Asia, is an important target species for control with neonicotinoid insecticides such as imidacloprid. Studies with nAChRs purified from N. lugens have identified two [(3)H]imidacloprid binding sites with different affinities (K(d) = 3.5 +/- 0.6 pM and 1.5 +/- 0.2 nM). Co-immunoprecipitation studies with native preparations of N. lugens nAChRs, using subunit-selective antisera, have demonstrated the co-assembly of Nlalpha1, Nlalpha2 and Nlbeta1 subunits into one receptor complex and of Nlalpha3, Nlalpha8 and Nlbeta1 into another. Immunodepletion of Nlalpha1 or Nlalpha2 subunits resulted in the selective loss of the lower affinity imidacloprid binding site, whereas immunodepletion of Nlalpha3 or Nlalpha8 caused the selective loss of the high-affinity site. Immunodepletion of Nlbeta1 resulted in a complete absence of specific imidacloprid binding. In contrast, immunodepletion with antibodies selective for other N. lugens nAChR subunits (Nlalpha4, Nlalpha6, Nlalpha7 and Nlbeta2) had no significant effect on imidacloprid binding. Taken together, these data suggest that nAChRs containing Nlalpha1, Nlalpha2 and Nlbeta1 constitute the lower affinity binding site, whereas nAChRs containing Nlalpha3, Nlalpha8 and Nlbeta1 constitute the higher affinity binding site for imidacloprid in N. lugens.',{'entities': [(41,59,'Gene'),(212,245,'Gene'),(247,253,'Gene'),(329,346,'Species'),(348,366,'Species'),(387,391,'Species'),(525,531,'Gene'),(546,555,'Species'),(738,747,'Species'),(748,754,'Gene'),(1332,1341,'Species'),(1342,1347,'Gene'),(1491,1497,'Gene'),(1592,1598,'Gene'),(1705,1714,'Species')]}),('Heteromeric co-assembly of two insect nicotinic acetylcholine receptor alpha subunits: influence on sensitivity to neonicotinoid insecticides. Neonicotinoid insecticides, such as imidacloprid, are selective agonists of insect nicotinic acetylcholine receptors (nAChRs) and are used extensively in areas of crop protection and animal health to control a variety of insect pest species. Here, we describe studies performed with nAChR subunits Nlalpha1 and Nlalpha2 cloned from the brown planthopper Nilaparvata lugens, a major insect pest of rice crops in many parts of Asia. The influence of Nlalpha1 and Nlalpha2 subunits upon the functional properties of recombinant nAChRs has been examined by expression in Xenopus oocytes. In addition, the influence of a Nlalpha1 mutation (Y151S), which has been linked to neonicotinoid lab generated resistance in N. lugens, has been examined. As in previous studies of insect alpha subunits, functional expression has been achieved by co-expression with the mammalian beta2 subunit. This approach has revealed a significantly higher apparent affinity of imidacloprid for Nlalpha1/beta2 than for Nlalpha2/beta2 nAChRs. In addition, evidence has been obtained for the co-assembly of Nlalpha1 and Nlalpha2 subunits into  triplet  nAChRs of subunit composition Nlalpha1/Nlalpha2/beta2. Evidence has also been obtained which demonstrates that the resistance-associated Y151S mutation has a significantly reduced effect on neonicotinoid agonist activity when Nlalpha1 is co-assembled with Nlalpha2 than when expressed as the sole alpha subunit in a heteromeric nAChR. These findings may be of importance in assessing the likely impact of the target-site mutations such as Y151S upon neonicotinoid insecticide resistance in insect field populations.',{'entities': [(38,70,'Gene'),(226,259,'Gene'),(261,267,'Gene'),(426,431,'Gene'),(479,496,'Species'),(497,515,'Species'),(540,544,'Species'),(668,674,'Gene'),(710,717,'Species'),(853,862,'Species'),(998,1007,'Species'),(1008,1013,'Gene'),(1150,1156,'Gene'),(1267,1273,'Gene'),(1595,1600,'Gene')]}),('Disruption of sex-specific doublesex exons results in male- and female-specific defects in the black cutworm, Agrotis ipsilon. BACKGROUND: Doublesex (dsx), the downstream gene in the insect sex-determination pathway, is a key regulator of sexually dimorphic development and behavior across a variety of insects. Manipulating expression of dsx could be useful in the genetic control of insects. However, information on the sex-specific function of dsx in non-model insects is lacking. RESULTS: In this work, we isolated a dsx homolog, which is alternatively spliced into six female-specific and one male-specific isoforms, from an important agricultural pest, the black cutworm, Agrotis ipsilon. Studies on the expression of sex-specific Aidsx mRNA during embryonic development showed that the sixth hour post oviposition is the key stage for sex determination in A. ipsilon. Functional analysis of Aidsx was conducted using a CRISPR/Cas9 system targeting female- and male-specific Aidsx exons. Disruptions of sex-specific Aidsx exons resulted in sex-specific, sexually dimorphic defects in external genitals, gonads and antennae, and expression of sex-specific genes as well as production of offspring in both sexes. CONCLUSION: Our results not only demonstrate that dsx is a key player determining A. ipsilon sexually dimorphic traits, but also provide a potential method for the genetic control of this pest.  2018 Society of Chemical Industry.',{'entities': [(27,36,'Gene'),(95,108,'Species'),(110,125,'Species'),(139,148,'Gene'),(150,153,'Gene'),(339,342,'Gene'),(447,450,'Gene'),(521,524,'Gene'),(663,676,'Species'),(678,693,'Species'),(737,742,'Gene'),(863,873,'Species'),(898,903,'Gene'),(981,986,'Gene'),(1022,1027,'Gene'),(1267,1270,'Gene')]}),('Downregulation of female doublesex expression by oral-mediated RNA interference reduces number and fitness of Anopheles gambiae adult females. BACKGROUND: Mosquito-borne diseases affect millions worldwide, with malaria alone killing over 400 thousand people per year and affecting hundreds of millions. To date, the best strategy to prevent the disease remains insecticide-based mosquito control. However, insecticide resistance as well as economic and social factors reduce the effectiveness of the current methodologies. Alternative control technologies are in development, including genetic control such as the sterile insect technique (SIT). The SIT is a pivotal tool in integrated agricultural pest management and could be used to improve malaria vector control. To apply the SIT and most other newer technologies against disease transmitting mosquitoes, it is essential that releases are composed of males with minimal female contamination. The removal of females is an essential requirement because released females can themselves contribute towards nuisance biting and disease transmission. Thus, females need to be eliminated from the cohorts prior to release. Manual separation of Anopheles gambiae pupae or adult mosquitoes based on morphology is time consuming, is not feasible on a large scale and has limited the implementation of the SIT technique. The doublesex (dsx) gene is one of the effector switches of sex determination in the process of sex differentiation in insects. Both males and females have specific splicing variants that are expressed across the different life stages. Using RNA interference (RNAi) to reduce expression of the female specific (dsxF) variant of this gene has proven to have detrimental effects to the females in other mosquito species, such as Aedes aegypti. We tested oral RNAi on dsx (AgdsxF) in An. gambiae. METHODS: We studied the expression pattern of the dsx gene in the An. gambiae G3 strain. We knocked down AgdsxF expression in larvae through oral delivery of double stranded RNA (dsRNA) produced by bacteria and observed its effects in adults. RESULTS: Our results show that feeding of AgdsxF dsRNA can effectively reduce (>  66%) the mRNA of female dsx transcript and that there is a concomitant reduction in the number of female larvae that achieve adulthood. Control groups produced 52% (   3.9% SE) of adult males and 48% (   4.0% SE) females, while AgdsxF dsRNA treated groups had 72.1% (   4.0% SE) males vs 27.8% females (   3.3% SE). In addition, the female adults produce fewer progeny, 37.1% (   8.2% SE) less than the controls. The knockdown was sex-specific and had no impact on total numbers of viable male adults, in the male dsx transcripts or male fitness parameters such as longevity or body size. CONCLUSIONS: These findings indicate that RNAi could be used to improve novel mosquito control strategies that require efficient sex separation and male-only release of An. gambiae by targeting sex determination genes such as AgdsxF. The advantages of using RNAi in a controlled setting for mosquito rearing are numerous, as the dose and time of exposure are controlled, and the possibility of off-target effects and the waste of female production would be significantly reduced.',{'entities': [(25,34,'Gene'),(110,127,'Species'),(1191,1208,'Species'),(1368,1377,'Gene'),(1379,1382,'Gene'),(1791,1804,'Species'),(1829,1832,'Gene'),(1834,1840,'Gene'),(1845,1856,'Species'),(1908,1911,'Gene'),(1924,1935,'Species'),(1963,1969,'Gene'),(2056,2064,'Species'),(2143,2149,'Gene'),(2207,2210,'Gene'),(2288,2294,'Species'),(2369,2374,'Species'),(2411,2417,'Gene'),(2462,2467,'Species'),(2672,2676,'Species'),(2692,2696,'Species'),(2697,2700,'Gene'),(2716,2720,'Species'),(2920,2924,'Species'),(2941,2952,'Species'),(2998,3004,'Gene')]}),('Knockdown of the aminopeptidase N genes decreases susceptibility of Chilo suppressalis larvae to Cry1Ab/Cry1Ac and Cry1Ca. Bacillus thuringiensis (Bt) insecticide is currently the most widely used bioinsecticide. Bt expressing cry genes are some of the most successful foreign-genome-inserting genes used in transgenic insect-resistant crop development. Cry toxins are resistant to lepidopteran pests, such as Chilo suppressalis, a major insect pest of rice worldwide. Since Cry toxins exert their activity by binding to specific receptors in the midgut of target insects, identification of functional Cry toxin receptors in the midgut of C. suppressalis larvae is crucial to evaluate potential resistance mechanisms and develop effective strategies for inhibiting insect resistance. In this study, we isolated two aminopeptidase N genes (APN6 and APN8) from C. suppressalis and determined that they were expressed in the foregut. APN6 was highly expressed at the fourth instar, and APN8 was highly expressed in adult and pupa. Knockdown of CsAPN6 and CsAPN8 by RNA interference resulted in significantly decreased susceptibility of larvae to Bt rice varieties TT51 (expressing cry1Ac/cry1Ab fusion genes) and T1C-19 (expressing cry1Ca), but not T2A-1 (expressing cry2Aa). These findings suggest that both APN6 and APN8 are involved in the toxicity of Cry1Ac/Cry1Ab and Cry1Ca toxins.',{'entities': [(17,33,'Gene'),(68,86,'Species'),(123,145,'Species'),(147,149,'Species'),(213,215,'Species'),(410,428,'Species'),(453,457,'Species'),(639,654,'Species'),(815,831,'Gene'),(839,843,'Gene'),(848,852,'Gene'),(859,874,'Species'),(931,935,'Gene'),(983,987,'Gene'),(1041,1047,'Gene'),(1052,1058,'Gene'),(1143,1145,'Species'),(1146,1150,'Species'),(1306,1310,'Gene'),(1315,1319,'Gene')]}),('Multiple functions of miR-8-3p in the development and metamorphosis of the red flour beetle, Tribolium castaneum. The microRNA miR-8-3p is conserved among insects and closely involved in development and immunity, but its functions in vivo are unexplored in the red flour beetle, Tribolium castaneum. Here, we show that miR-8-3p was highly expressed in late larva and early adult stages, as determined by quantitative real-time PCR. It was enriched in the fat body and cuticle in late larval tissues and abundant in the head and cuticle in early adult tissues, indicating this microRNA plays important roles during T. castaneum development. Specific inhibition of miR-8-3p in late larvae led to metamorphosis defects in the development of wings, eyes, legs and embryo. Moreover, a series of genes related to organism development were identified as miR-8-3p targets by computational prediction and microRNA-messenger RNA interaction validation, including Wingless, Eyg, Fpps and Sema-1a. These genes were critical for the regulation of the larva-to-adult transition. Eyg, as a functional target of miR-8-3p, participates in eye development, which was further confirmed by luciferase assay and loss-of-function analyses. In brief, miR-8-3p is broadly involved in the development of wings, eyes and legs through its target genes and has extensive regulatory roles during T. castaneum development.',{'entities': [(22,30,'Gene'),(75,91,'Species'),(93,112,'Species'),(127,135,'Gene'),(261,277,'Species'),(279,298,'Species'),(319,327,'Gene'),(614,626,'Species'),(663,671,'Gene'),(847,855,'Gene'),(953,961,'Gene'),(963,966,'Gene'),(968,972,'Gene'),(977,984,'Gene'),(1065,1068,'Gene'),(1096,1104,'Gene'),(1228,1236,'Gene'),(1367,1379,'Species')]}),('Multiple functions of CREB-binding protein during postembryonic development: identification of target genes. BACKGROUND: Juvenile hormones (JH) and ecdysteroids control postembryonic development in insects. They serve as valuable targets for pest management. Hence, understanding the molecular mechanisms of their action is of crucial importance. CREB-binding protein (CBP) is a universal transcriptional co-regulator. It controls the expression of several genes including those from hormone signaling pathways through co-activation of many transcription factors. However, the role of CBP during postembryonic development in insects is not well understood. Therefore, we have studied the role of CBP in postembryonic development in Tribolium, a model coleopteran insect. RESULTS: CBP is ubiquitously expressed in the red flour beetle, Tribolium castaneum. RNA interference (RNAi) mediated knockdown of CBP resulted in a decrease in JH induction of Kr-h1 gene expression in Tribolium larvae and led to a block in their development. Moreover, the injection of CBP double-stranded RNA (dsRNA) showed lethal phenotypes within 8 days of injection. RNA-seq and subsequent differential gene expression analysis identified CBP target genes in Tribolium. Knockdown of CBP caused a decrease in the expression of 1306 genes coding for transcription factors and other proteins associated with growth and development. Depletion of CBP impaired the expression of several JH response genes (e.g., Kr-h1, Hairy, early trypsin) and ecdysone response genes (EcR, E74, E75, and broad complex). Further, GO enrichment analyses of the downregulated genes showed enrichment in different functions including developmental processes, pigmentation, anatomical structure development, regulation of biological and cellular processes, etc. CONCLUSION: These data suggest diverse but crucial roles for CBP during postembryonic development in the coleopteran model insect, Tribolium. It can serve as a target for RNAi mediated pest management of this stored product pest.',{'entities': [(22,42,'Gene'),(347,367,'Gene'),(369,372,'Gene'),(585,588,'Gene'),(696,699,'Gene'),(732,741,'Species'),(780,783,'Gene'),(817,833,'Species'),(835,854,'Species'),(902,905,'Gene'),(948,953,'Gene'),(973,982,'Species'),(1058,1061,'Gene'),(1215,1218,'Gene'),(1235,1244,'Species'),(1259,1262,'Gene'),(1418,1421,'Gene'),(1482,1487,'Gene'),(1489,1494,'Gene'),(1496,1509,'Gene'),(1873,1876,'Gene'),(1943,1952,'Species')]}),('Post-embryonic functions of HSP90 in Tribolium castaneum include the regulation of compound eye development. Heat shock protein 90 (HSP90) belongs to a family of conserved chaperons with multiple roles in stress adaptation and development, including spermatogenesis, oogenesis and embryogenesis in insects. In the red flour beetle, Tribolium castaneum, we found that HSP90 is transiently upregulated during larval development, in prepupae, in female pupae and in adults, suggesting multiple post-embryonic roles. We found that silencing HSP90 expression by RNA interference was lethal within 10 days at all developmental stages. Titration experiments revealed that larvae were more susceptible than pupae or beetles. Interestingly, HSP90 silencing in final instar larvae resulted in abnormal pupal phenotypes lacking compound eyes and exhibiting prepupal features, suggesting developmental arrest at the prepupal stage. Our results suggest that HSP90 functions can be expanded beyond the known ones in insect embryogenesis to include roles in post-embryonic development such as the regulation of compound eye development.',{'entities': [(28,33,'Gene'),(37,56,'Species'),(109,130,'Gene'),(132,137,'Gene'),(314,330,'Species'),(332,351,'Species'),(367,372,'Gene'),(537,542,'Gene'),(732,737,'Gene'),(945,950,'Gene')]}),('Functional specialization among insect chitinase family genes revealed by RNA interference. The biological functions of individual members of the large family of chitinase-like proteins from the red flour beetle, Tribolium castaneum (Tc), were examined by using gene-specific RNAi. One chitinase, TcCHT5, was found to be required for pupal-adult molting only. A lethal phenotype was observed when the transcript level of TcCHT5 was down-regulated by injection of TcCHT5-specific dsRNA into larvae. The larvae had metamorphosed into pupae and then to pharate adults but did not complete adult eclosion. Specific knockdown of transcripts for another chitinase, TcCHT10, which has multiple catalytic domains, prevented embryo hatch, larval molting, pupation, and adult metamorphosis, indicating a vital role for TcCHT10 during each of these processes. A third chitinase-like protein, TcCHT7, was required for abdominal contraction and wing/elytra extension immediately after pupation but was dispensable for larval-larval molting, pupation, and adult eclosion. The wing/elytra abnormalities found in TcCHT7-silenced pupae were also manifest in the ensuing adults. A fourth chitinase-like protein, TcIDGF4, exhibited no chitinolytic activity but contributed to adult eclosion. No phenotypic effects were observed after knockdown of transcripts for several other chitinase-like proteins, including imaginal disk growth factor IDGF2. These data indicate functional specialization among insect chitinase family genes, primarily during the molting process, and provide a biological rationale for the presence of a large assortment of chitinase-like proteins.',{'entities': [(32,48,'Gene'),(162,185,'Gene'),(195,211,'Species'),(213,232,'Species'),(234,236,'Species'),(297,303,'Gene'),(421,427,'Gene'),(463,469,'Gene'),(659,666,'Gene'),(809,816,'Gene'),(857,879,'Gene'),(881,887,'Gene'),(1097,1103,'Gene'),(1170,1192,'Gene'),(1194,1201,'Gene'),(1358,1381,'Gene'),(1421,1426,'Gene'),(1480,1496,'Gene'),(1626,1649,'Gene')]}),('Molecular characterization of an NADPH cytochrome P450 reductase from Bemisia tabaci Q: Potential involvement in susceptibility to imidacloprid. NADPH cytochrome P450 reductase (CPR) is an integral component of cytochrome P450-mediated biological reactions, such as the metabolism of xenobiotics, including insecticides. CPR has been reported to be associated with insecticide tolerance in several insects. However, the biochemical characteristics and biological function of CPR in Bemisia tabaci Q (BtCPR) remain undefined. In this study, BtCPR was cloned, and bioinformatic analysis showed that BtCPR is a transmembrane protein with a molecular weight (MW) of 76.73  kDa and contains conserved binding domains (FMN, FAD, and NADPH). Tissue- and developmental stage-dependent expression indicated that the highest expression levels of BtCPR occurred in head tissue and in male adults. Transcripts of BtCPR in the field B. tabaci Q strain were 1.62-fold higher than those of the laboratory B. tabaci Q strain. In both field and laboratory adults, the susceptibility of BtCPR-knockdown B. tabaci Q to imidacloprid substantially increased compared to that of the B. tabaci Q control group. Furthermore, the heterologous expression of BtCPR in Sf9 cells exhibited catalytic activity for cytochrome c reduction, following Michaelis-Menten kinetics. Sf9 cells overexpressing BtCPR had greater viability than the control cells when treated with imidacloprid. The results suggest that BtCPR could affect the susceptibility of B. tabaci Q to imidacloprid and could also be considered a novel target for pest control.',{'entities': [(33,64,'Gene'),(145,176,'Gene'),(178,181,'Gene'),(321,324,'Gene'),(475,478,'Gene'),(500,505,'Gene'),(540,545,'Gene'),(597,602,'Gene'),(836,841,'Gene'),(901,906,'Gene'),(920,929,'Species'),(990,999,'Species'),(1069,1074,'Gene'),(1085,1094,'Species'),(1161,1170,'Species'),(1232,1237,'Gene'),(1370,1375,'Gene'),(1478,1483,'Gene'),(1519,1528,'Species')]}),('Identification and Expression Profiling of Odorant Receptor Protein Genes in Sitophilus zeamais (Coleoptera: Curculionoidea) Using RT-qPCR. This study aimed to identify ORs (odorant receptors) and Orco (odorant receptor coreceptor) genes in Sitophilus zeamais Motschulsky (Coleoptera: Curculionoidea), to explore the relative expression levels of these genes in different adult tissues and obtain information on highly expressed receptor proteins. Putative OR and Orco genes were identified from transcriptomic data previously obtained for S. zeamais using bioinformatics methods. Quantitative real-time PCR was used to compare the differences in expression in seven adult tissues (male antennae, female antennae, heads, thoraxes, abdomens, wings, and legs). The candidate OR and Orco gene sequences were analyzed, and the protein physicochemical properties were predicted. We identified 64 OR genes including the Orco gene. Forty-seven OR genes, including Orco, were over expressed in male or female antennae. Seventeen OR genes appeared to be expressed at elevated levels in male antennae. Twenty-nine genes were expressed at significantly elevated levels in female antennae. In total, 11 OR genes were selected for further sequence analysis. The selected proteins were structurally characterized, and bioinformatics analysis was performed. Overall, in this study, candidate ORs of S. zeamais have been identified for the first time, and these ORs could be molecular targets for interference in the insect olfactory system.',{'entities': [(43,67,'Gene'),(77,95,'Species'),(169,172,'Gene'),(174,191,'Gene'),(197,201,'Gene'),(203,230,'Gene'),(241,259,'Species'),(457,459,'Gene'),(464,468,'Gene'),(540,550,'Species'),(773,775,'Gene'),(780,784,'Gene'),(891,893,'Gene'),(914,918,'Gene'),(937,939,'Gene'),(957,961,'Gene'),(991,993,'Gene'),(1021,1023,'Gene'),(1191,1193,'Gene'),(1377,1380,'Gene'),(1384,1394,'Species'),(1446,1449,'Gene')]}),('Latrophilin mediates insecticides susceptibility and fecundity through two carboxylesterases, esterase4 and esterase6, in Tribolium castaneum. Latrophilin (LPH) is known as an adhesion G-protein-coupled receptor which involved in multiple physiological processes in organisms. Previous studies showed that lph not only involved the susceptibility to anticholinesterase insecticides but also affected fecundity in Tribolium castaneum. However, its regulatory mechanisms in these biological processes are still not clear. Here, we identified two potential downstream carboxylesterase (cce) genes of Tclph, esterase4 and esterase6, and further characterized their interactions with Tclph. After treatment of T. castaneum larvae with carbofuran or dichlorvos insecticides, the transcript levels of Tcest4 and Tcest6 were significantly induced from 12 to 72 h. RNAi against Tcest4 or Tcest6 led to the higher mortality compared with the controls after the insecticides treatment, suggesting that these two genes play a vital role in detoxification of insecticides in T. castaneum. Furthermore, with insecticides exposure to Tclph knockdown beetles, the expression of Tcest4 was upregulated but Tcest6 was downregulated, indicating that beetles existed a compensatory response against the insecticides. Additionally, RNAi of Tcest6 resulted in 43% reductions in female egg laying and completely inhibited egg hatching, which showed the similar phenotype as that of Tclph knockdown. These results indicated that Tclph affected fecundity by positively regulating Tcest6 expression. Our findings will provide a new insight into the molecular mechanisms of Tclph involved in physiological functions in T. castaneum.',{'entities': [(0,11,'Gene'),(94,103,'Gene'),(108,117,'Gene'),(122,141,'Species'),(143,154,'Gene'),(156,159,'Gene'),(185,211,'Gene'),(306,309,'Gene'),(413,432,'Species'),(565,581,'Gene'),(583,586,'Gene'),(597,602,'Gene'),(604,613,'Gene'),(618,627,'Gene'),(679,684,'Gene'),(705,717,'Species'),(718,724,'Species'),(794,800,'Gene'),(805,811,'Gene'),(869,875,'Gene'),(879,885,'Gene'),(1062,1074,'Species'),(1119,1124,'Gene'),(1162,1168,'Gene'),(1189,1195,'Gene'),(1319,1325,'Gene'),(1459,1464,'Gene'),(1505,1510,'Gene'),(1555,1561,'Gene'),(1647,1652,'Gene'),(1692,1704,'Species')]}),('Evolution of the Torso activation cassette, a pathway required for terminal patterning and moulting. Embryonic terminal patterning and moulting are critical developmental processes in insects. In Drosophila and Tribolium both of these processes are regulated by the Torso-activation cassette (TAC). The TAC consists of a common receptor, Torso, ligands Trunk and prothoracicotropic hormone (PTTH), and the spatially restricted protein Torso-like, with combinations of these elements acting mechanistically to activate the receptor in different developmental contexts. In order to trace the evolutionary history of the TAC we determined the presence or absence of TAC components in the genomes of arthropods. Our analyses reveal that Torso, Trunk and PTTH are evolutionarily labile components of the TAC with multiple individual or combined losses occurring in the arthropod lineages leading to and within the insects. These losses are often correlated, with both ligands and receptor missing from the genome of the same species. We determine that the PTTH gene evolved in the common ancestor of Hemiptera and Holometabola, and is missing from the genomes of a number of species with experimentally demonstrated PTTH activity, implying another molecule may be involved in ecdysis in these species. In contrast, the torso-like gene is a common component of pancrustacean genomes.',{'entities': [(17,22,'Gene'),(196,206,'Species'),(211,220,'Species'),(266,271,'Gene'),(338,343,'Gene'),(353,358,'Gene'),(391,395,'Gene'),(435,440,'Gene'),(733,738,'Gene'),(740,745,'Gene'),(750,754,'Gene'),(1051,1055,'Gene'),(1211,1215,'Gene'),(1314,1319,'Gene')]}),('Influence of Metarhizium anisopliae (IMI330189) and Mad1 protein on enzymatic activities and Toll-related genes of migratory locust. Efficacy of Metarhizium anisopliae strain (IMI330189) and Mad1 protein alone or in combination by feeding method to overcome immune-related enzymes and Toll-like pathway genes was investigated in migratory locust. M. anisopliae (IMI330189) is a potent and entomopathogenic fungal strain could be effectively used against insect pests. Similarly, Mad1 protein adheres to insect cuticle, causing virulence to insects. We confirmed maximum 55% of mortality when M. anisopliae (IMI330189) and Mad1 was applied in combination. Similarly, increased PO activity was observed in locust with combined dose of Mad1  +  IMI330189 whereas PO, POD, and SOD activities reduced using Mad1 independently. Four Toll-like signaling pathway genes (MyD88, Cactus, Pelle, and CaN) were investigated from midgut and body of the migratory locust after 72 h of treatments. Subsequently, the expression of MyD88 in the midgut and body significantly decreased with the application of Mad1 and Mad1  +  IMI330189. Performance of these treatments was absolutely non-consistent in both parts of insects. Meanwhile, IMI330189 significantly raised the expression of Cactus in both midgut and body. However, the combined treatment (Mad1  +  IMI330189) significantly reduced the Cactus expression in both body parts. Pelle expression was significantly increased in the midgut with the application of independent treatment of Mad1 and IMI330189 whereas the combined treatment (Mad1  +  IMI330189) suppressed the Pelle expression in midgut. Its expression level was absolutely higher in body with the application of IMI330189 and Mad1  +  IMI330189 only. On the other hand, Mad1 significantly increased the expression of CaN in midgut. However, all three treatments significantly affected and suppressed the expression of CaN gene in body of locust. This shows that the applications of M. anisopliae and Mad1 protein significantly affected Toll signaling pathway genes, which ultimately increased level of susceptibility of locust. However, their effect was significantly different in both parts of locust which recommends that the Toll-related genes are conserved in midgut instead of locust body.',{'entities': [(13,35,'Species'),(115,131,'Species'),(145,167,'Species'),(329,345,'Species'),(347,360,'Species'),(592,605,'Species'),(862,867,'Gene'),(869,875,'Gene'),(877,882,'Gene'),(888,891,'Gene'),(939,955,'Species'),(1014,1019,'Gene'),(1268,1274,'Gene'),(1379,1385,'Gene'),(1417,1422,'Gene'),(1611,1616,'Gene'),(1984,1997,'Species')]}),('Insecticides induce the co-expression of glutathione S-transferases through ROS/CncC pathway in Spodoptera exigua. Glutathione S-transferases (GSTs) are a family of multifunctional enzymes that are involved in detoxification of electrophilic toxic compounds. Although the co-induced expression of GST genes by insecticides in insects has been documented in recent years, the underlying regulatory mechanisms are not understood. In this study, a total of thirty-one cytosolic S. exigua GSTs (SeGSTs) was cloned and identified. The bioinformatics and gene expression patterns were also analyzed. Out of them, SeGSTe9, SeGSTs6, SeGSTe1, SeGSTe6, SeGSTe8, SeGSTe14, and SeGSTd1 were significantly co-expressed following exposure to three insecticides (lambda-cyhalothrin, chlorpyrifos and chlorantraniliprole). The analysis of upstream sequences revealed that all of these seven SeGSTs harbored CncC/Maf binding site. The luciferase reporter assay showed that the pGL3-SeGST promoter construct exhibited a significant increase in luciferase activities after exposure to insecticides, and mutation of CncC/Maf binding site diminish the induction effect. These data indicate that CncC/Maf pathway regulates the co-expression of GST genes in response to different insecticides in S. exigua. Insecticides significantly enhanced the ROS content and treatment with the ROS inhibitor N-acetylcysteine (NAC) decreased the insecticide-induced luciferase activities of the PGL3-GSTe6 promoter construct, but not the CncC-mutated construct. These results indicate that ROS mediates GST gene expression after exposure to insecticides through CncC/Maf pathway. Overall, these data show that insecticides induce the co-expression of glutathione S-transferases through the ROS/CncC pathway in S. exigua.',{'entities': [(41,67,'Gene'),(96,113,'Species'),(115,141,'Gene'),(143,147,'Gene'),(297,300,'Gene'),(475,484,'Species'),(485,489,'Gene'),(491,497,'Gene'),(607,614,'Gene'),(616,623,'Gene'),(625,632,'Gene'),(634,641,'Gene'),(643,650,'Gene'),(652,660,'Gene'),(666,673,'Gene'),(875,881,'Gene'),(1222,1225,'Gene'),(1273,1282,'Species'),(1567,1570,'Gene'),(1774,1783,'Species')]}),('Constitutive overexpression of cytochrome P450 monooxygenase genes contributes to chlorantraniliprole resistance in Chilo suppressalis (Walker). BACKGROUND: The rice striped stem borer (SSB), Chilo suppressalis (Walker), which is one of the most economically important phytophagous pests, has developed resistance to multiple insecticides. The resistance of SSB against chlorantraniliprole has been investigated in detail. However, the mechanism of its metabolic resistance has rarely been studied. RESULTS: A field population from Wuhu City, China was used to establish chlorantraniliprole resistant and susceptible strains (WHR and WHS) by laboratory continuous selection. Enzyme activities data suggested the potential involvement of cytochrome P450 monooxygenase in WHR. CYP6CV5, CYP9A68, CYP321F3, and CYP324A12 were significantly overexpressed in WHR (from 4.48 to 44.88-fold). These four P450 genes were expressed in the late developmental stages of WHR; however, they were almost absent during the egg stage. In addition, their expressions were much more sensitive to chlorantraniliprole induction in WHR than in WHS. Injection of individual and mixture dsRNAs reduced the expression of the four target genes (55.2-73.2% and 43.2-50.2%, respectively) and caused significant larvae mortality (55.1-65.1% and 88.2%, respectively). CONCLUSION: Multiple overexpressed P450 genes were potentially associated with chlorantraniliprole resistance, as confirmed by the RNA interference (RNAi) assay. Our findings suggested that metabolic resistance to chlorantraniliprole might be mediated by P450s.  2018 Society of Chemical Industry.',{'entities': [(31,60,'Gene'),(116,134,'Species'),(161,165,'Species'),(192,210,'Species'),(737,766,'Gene'),(775,782,'Gene'),(784,791,'Gene'),(793,801,'Gene'),(807,816,'Gene')]}),('Modular cis-regulatory logic of yellow gene expression in silkmoth larvae. Colour patterns in butterflies and moths are crucial traits for adaptation. Previous investigations have highlighted genes responsible for pigmentation (ie yellow and ebony). However, the mechanisms by which these genes are regulated in lepidopteran insects remain poorly understood. To elucidate this, molecular studies involving dipterans have largely analysed the cis-regulatory regions of pigmentation genes and have revealed cis-regulatory modularity. Here, we used well-developed transgenic techniques in Bombyx mori and demonstrated that cis-regulatory modularity controls tissue-specific expression of the yellow gene. We first identified which body parts are regulated by the yellow gene via black pigmentation. We then isolated three discrete regulatory elements driving tissue-specific gene expression in three regions of B. mori larvae. Finally, we found that there is no apparent sequence conservation of cis-regulatory regions between B. mori and Drosophila melanogaster, and no expression driven by the regulatory regions of one species when introduced into the other species. Therefore, the trans-regulatory landscapes of the yellow gene differ significantly between the two taxa. The results of this study confirm that lepidopteran species use cis-regulatory modules to control gene expression related to pigmentation, and represent a powerful cadre of transgenic tools for studying evolutionary developmental mechanisms.',{'entities': [(32,38,'Gene'),(58,66,'Species'),(231,237,'Gene'),(242,247,'Gene'),(586,597,'Species'),(689,695,'Gene'),(760,766,'Gene'),(908,915,'Species'),(1024,1031,'Species'),(1036,1059,'Species'),(1217,1223,'Gene')]}),('Targeting the potassium ion channel genes SK and SH as a novel approach for control of insect pests: efficacy and biosafety. BACKGROUND: Potassium ion channels play a critical role in the generation of electrical signals and thus provide potential targets for control of insect pests by RNA interference. RESULTS: Genes encoding the small conductance calcium-activated potassium channel (SK) and the voltage-gated potassium channel (SH) were knocked down in Tribolium castaneum by injection and oral delivery of dsRNA (dsTcSK and dsTcSH, respectively). Irrespective of the delivery mechanism a dose-dependent effect was observed for knockdown (KD) of gene expression and insect mortality for both genes. Larvae fed a 400  ng dsRNA  mg-1 diet showed significant gene (P  <  0.05) knockdown (98% and 83%) for SK and SH, respectively, with corresponding mortalities of 100% and 98% after 7 days. When injected (248.4 ng  larva-1 ), gene KD was 99% and 98% for SK and SH, causing 100% and 73.4% mortality, respectively. All developmental stages tested (larvae, early- and late-stage pupae and adults) showed an RNAi-sensitive response for both genes. LC50 values were lower for SK than SH, irrespective of delivery method, demonstrating that the knockdown of SK had a greater effect on larval mortality. Biosafety studies using adult honeybee Apis mellifera showed that there were no significant differences either in expression levels or mortality of honeybees orally dosed with dsTcSK and dsTcSH compared to control-fed bees. Similarly, there was no significant difference in the titre of deformed wing virus, used as a measure of immune suppression, between experimental and control bees. CONCLUSION: This study demonstrates the potential of using RNAi targeting neural receptors as a technology for the control of T. castaneum.  2019 The Authors. Pest Management Science published by John Wiley _ Sons Ltd on behalf of Society of Chemical Industry.',{'entities': [(42,44,'Gene'),(49,51,'Gene'),(333,386,'Gene'),(388,390,'Gene'),(400,431,'Gene'),(433,435,'Gene'),(458,477,'Species'),(807,809,'Gene'),(814,816,'Gene'),(957,959,'Gene'),(964,966,'Gene'),(1174,1176,'Gene'),(1182,1184,'Gene'),(1255,1257,'Gene'),(1330,1338,'Species'),(1339,1353,'Species'),(1448,1457,'Species'),(1518,1522,'Species'),(1587,1606,'Species'),(1682,1686,'Species'),(1814,1826,'Species')]}),('Transcription factor E93 regulates wing development by directly promoting Dpp signaling in Drosophila. Transcription factor E93 is a steroid hormone ecdysone early response gene and plays crucial roles in both the degradation of larval tissues and the formation of adult organs during insect metamorphosis with the prepupal-pupal-adult transition. However, the molecular mechanism underlying E93 regulation is poorly understood. In this study, we found that specific knockdown of the E93 gene in the Drosophila wing disrupted wing development. Analyzing ChIP-seq signals for E93 in Drosophila wing identified that the decapentaplegic (Dpp) gene was a potential downstream target of E93. ChIP-PCR analysis and dual-luciferase reporter assay confirmed that E93 could bind to the Dpp promoter and enhanced its activity. Furthermore, the expressions of Dpp and other components in the Dpp signaling pathway were upregulated following E93 overexpression in Drosophila S2 cells but were decreased after E93 knockdown in the wing. Moreover, the impairment of the Dpp signaling pathway phenocopied the defects of E93 knockdown on wing development. Taken together, our results suggest that E93 modulates the Dpp signaling pathway to regulate wing development during Drosophila metamorphosis.',{'entities': [(21,24,'Gene'),(74,77,'Gene'),(124,127,'Gene'),(392,395,'Gene'),(484,487,'Gene'),(575,578,'Gene'),(618,633,'Gene'),(635,638,'Gene'),(682,685,'Gene'),(755,758,'Gene'),(777,780,'Gene'),(849,852,'Gene'),(881,884,'Gene'),(930,933,'Gene'),(997,1000,'Gene'),(1056,1059,'Gene'),(1105,1108,'Gene'),(1181,1184,'Gene'),(1199,1202,'Gene')]}),('Orcokinins regulate the expression of neuropeptide precursor genes related to ecdysis in the hemimetabolous insect Rhodnius prolixus. Ecdysis is a vital process for insects, during which they shed the old cuticle in order to emerge as the following developmental stage. Given its relevance for survival and reproduction, ecdysis is tightly regulated by peptidic hormones that conform an interrelated neuromodulatory network. This network was studied in species that undergo a complete metamorphosis, but not in hemimetabola. In a recent work, we demonstrated that orcokinin neuropeptides are essential for ecdysis to occur in the kissing bug Rhodnius prolixus. Here we performed gene silencing, quantitative PCR and in vitro treatments in order to study the interrelationships between RhoprOKs and hormones such as ecdysis triggering hormone, corazonin, eclosion hormone, crustacean cardioactive peptide and ecdysone. Our results suggest that RhoprOKs directly or indirectly regulate the expression of other genes. Whereas RhoprOKA is centrally involved in the regulation of gene expression, RhoprOKB is implicated in processes related to midgut physiology. Therefore, we propose that the different transcripts encoded in RhoprOK gene could integrate signaling cues, in order to coordinate the nutritional state with development and ecdysis. Given the emerging data that point to OKs as important factors for survival and reproduction, they could be candidates in the search for new insect management strategies based on neuroendocrine targets.',{'entities': [(0,10,'Gene'),(115,132,'Species'),(630,641,'Species'),(642,659,'Species'),(785,793,'Gene'),(943,951,'Gene'),(1023,1031,'Gene'),(1092,1100,'Gene'),(1222,1229,'Gene'),(1380,1383,'Gene')]}),('Use of tyrosine hydroxylase RNAi to study Megoura viciae (Hemiptera: Aphididae) sequestration of its host s l-DOPA for body melanism. Melanism in insects is important for their physical protection, immunoreactions, and sclerotization. The vetch aphid, Megoura viciae (Buckton), has relatively strong tanning in its prothorax, head, antennae, cornicles, and legs. It was hypothesized that M. viciae may sequester the high level of l-DOPA in its host Vicia faba to help in its melanization process for ecological adaptation. To confirm this hypothesis, the amount of l-DOPA in M. viciae was modified and quantified. We first generated a Trifolium repens (clover, low l-DOPA containing) host to cut off the extra l-DOPA intake by M. viciae. The rate-limiting tyrosine hydroxylase gene of M. viciae (MV-TH) was then cloned and analyzed. To further reduce the l-DOPA level in the insect, RNAi was used to downregulate the transcriptional level of MV-TH. Our results confirmed that M. viciae could indeed sequester l-DOPA in its body, and its ample storage of this amino acid could be the reason for the strong tanning of its body. M. viciae reared on T. repens could upregulate its MV-TH to enhance l-DOPA biosynthesis and thus maintain a high level of l-DOPA. The MV-TH repression by RNAi lasted for about 3  days, successfully decreasing the l-DOPA level. Aside from a slight decrease in exuvia tanning, no other obvious change in body appearance was detected in the RNAi-treated insect. Although M. viciae can obtain most of its l-DOPA directly from its original host, its internal l-DOPA synthetase is still functional, especially when extra l-DOPA is removed from the diet. This capability to enhance its shield ensures the ecological adaptation of this insect.',{'entities': [(42,56,'Species'),(239,250,'Species'),(252,266,'Species'),(388,397,'Species'),(449,459,'Species'),(575,584,'Species'),(727,736,'Species'),(756,781,'Gene'),(785,794,'Species'),(796,801,'Gene'),(942,947,'Gene'),(976,985,'Species'),(1126,1135,'Species'),(1146,1155,'Species'),(1177,1182,'Gene'),(1260,1265,'Gene'),(1494,1503,'Species')]}),('Bmo-miR-79 downregulates the expression of BmEm4 in the silkworm, Bombyx mori. MicroRNA is an important regulation factor in insect development and metamorphosis. It has been reported that E(spl)m4 is a miRNA-targeted gene, as well as the target of the Notch signaling pathway in Drosophila. The expression of E(spl)m4 can be regulated by microRNA and further affect the neural development of Drosophila. Here, we found that BmEm4, an ortholog of E(spl)m4 from Bombyx mori, was the target gene of bmo-miR-79, with target sites containing the Brd and K boxes of the BmEm4_3 UTR, which was validated by the dual luciferase reporter (DLR) assay. Furthermore, bmo-miR-79 mimics can inhibit the expression of BmEm4 in BmN cells after transfection, and bmo-miR-79 can also inhibit the expression of BmEm4 in different developmental stages of Bombyx mori at a posttranscriptional level, to different degrees. The EMSA test further showed that bmo-miR-79 could bind to BmAGO2, which is the Bombyx mori argonaute2 protein, suggesting that bmo-miR-79 might regulate the expression of BmEm4 by forming miRISC complexes with BmAGO2. Taken together, bmo-miR-79 could regulate the expression of BmEm4 mediated by BmAGO2 and further affect its function in the silkworm Bombyx mori.',{'entities': [(0,10,'Gene'),(56,64,'Species'),(66,77,'Species'),(189,197,'Gene'),(310,318,'Gene'),(447,455,'Gene'),(461,472,'Species'),(497,507,'Gene'),(656,666,'Gene'),(747,757,'Gene'),(836,847,'Species'),(936,946,'Gene'),(961,967,'Gene'),(982,993,'Species'),(994,1004,'Gene'),(1030,1040,'Gene'),(1113,1119,'Gene'),(1137,1147,'Gene'),(1199,1205,'Gene'),(1245,1253,'Species'),(1254,1265,'Species')]}),('Identification and Expression Analysis of Four Small Heat Shock Protein Genes in Cigarette Beetle, Lasioderma serricorne (Fabricius). Small heat shock proteins (sHsps) are molecular chaperones that play crucial roles in the stress adaption of insects. In this study, we identified and characterized four sHsp genes (LsHsp19.4, 20.2, 20.3, and 22.2) from the cigarette beetle, Lasioderma serricorne (Fabricius). The four cDNAs encoded proteins of 169, 180, 181, and 194 amino acids with molecular weights of 19.4, 20.2, 20.3, and 22.2 kDa, respectively. The four LsHsp sequences possessed a typical sHsp domain structure. Quantitative real-time PCR analyses revealed that LsHsp19.4 and 20.3 transcripts were most abundant in pupae, whereas the transcript levels of LsHsp20.2 and 22.2 were highest in adults. Transcripts of three LsHsp genes were highly expressed in the larval fat body, whereas LsHsp20.2 displayed an extremely high expression level in the gut. Expression of the four LsHsp genes was dramatically upregulated in larvae exposed to 20-hydroxyecdysone. The majority of the LsHsp genes were significantly upregulated in response to heat and cold treatments, while LsHsp19.4 was insensitive to cold stress. The four genes were upregulated when challenged by immune triggers (peptidoglycan isolated from Staphylococcus aureus and from Escherichia coli 0111:B4). Exposure to CO2 increased LsHsp20.2 and 20.3 transcript levels, but the LsHsp19.4 transcript level declined. The results suggest that different LsHsp genes play important and distinct regulatory roles in L. serricorne development and in response to diverse stresses.',{'entities': [(53,71,'Gene'),(81,97,'Species'),(99,120,'Species'),(161,166,'Gene'),(304,308,'Gene'),(316,325,'Gene'),(358,374,'Species'),(376,397,'Species'),(562,567,'Gene'),(598,602,'Gene'),(671,680,'Gene'),(764,773,'Gene'),(828,833,'Gene'),(894,903,'Gene'),(984,989,'Gene'),(1086,1091,'Gene'),(1176,1185,'Gene'),(1314,1335,'Species'),(1345,1361,'Species'),(1398,1407,'Gene'),(1444,1453,'Gene'),(1516,1521,'Gene'),(1576,1589,'Species')]}),('Domain structure and expression along the midgut and carcass of peritrophins and cuticle proteins analogous to peritrophins in insects with and without peritrophic membrane. Most insects have a peritrophic membrane (matrix) (PM) surrounding the food bolus. This structure, similarly to the cuticle, is mainly composed of chitin and proteins. The main proteins forming PM are known as peritrophins (PMP), whereas some of the cuticle proteins are the cuticle proteins analogous to peritrophins (CPAP). Both proteins are composed of one or more chitin binding peritrophin-A domain (CBD) and no other recognized domain. Furthermore, insects containing PM usually have two chitin synthase (CS) genes, one mainly expressed in carcass and the other in midgut. In this work we identified PMP, CPAP and CS genes in the genome of insects from the Polyneoptera, Paraneoptera and Holometabola cohorts and analyzed their expression profile in different species from each group. In agreement with the absence of PM, we observed less CBD-containing proteins and only one CS gene in the genome of Paraneoptera species, except for the Phthiraptera Pediculus humanus. The lack of PM in Paraneoptera species was also confirmed by the micrographs of the midgut of two Hemiptera species, Dysdercus peruvianus and Mahanarva fimbriolata which agreed with the RNA-seq data of both species. Our analyses also highlighted a higher number of CBD-containing proteins in Holometabola in relation to the earlier divergent Polyneoptera group, especially regarding the genes composed of more than three CBDs, which are usually associated to PM formation. Finally, we observed a high number of CBD-containing proteins being expressed in both midgut and carcass tissues of several species, which we named as ubiquitous-CBD-containing proteins (UCBP), as their function is unclear. We hypothesized that these proteins can be involved in both cuticle and PM formation or that they can be involved in immune response and/or tracheolae formation.',{'entities': [(64,76,'Gene'),(81,97,'Gene'),(111,123,'Gene'),(384,396,'Gene'),(424,440,'Gene'),(449,465,'Gene'),(479,491,'Gene'),(668,683,'Gene'),(685,687,'Gene'),(794,796,'Gene'),(1019,1042,'Gene'),(1056,1058,'Gene'),(1131,1148,'Species'),(1267,1287,'Species'),(1292,1313,'Species'),(1415,1438,'Gene'),(1661,1684,'Gene'),(1785,1808,'Gene'),(1810,1814,'Gene')]}),('The function of spineless in antenna and wing development of the brown planthopper, Nilaparvata lugens. A wide array of sensilla are distributed on insect antennae, and they play a variety of important roles. Rice planthoppers, destructive pests on rice, have a unique antenna sensilla structure called the  sensory plaque organ . The spineless (ss) gene encodes a bHLH-PAS transcription factor and plays a key role in antenna development. In the current study, a 3029 bp full-length cDNA of the Nilaparvata lugens ss gene (Nlss) was cloned, and it encodes 654 amino acid residues. The highest level of Nlss expression was detected in the thorax of fourth-instar nymphs. Knockdown of Nlss in nymphs led to a decrease in the number and size of plaque organs. Moreover, the flagella of the treated insects were poorly developed, wilted, and even dropped off from the pedicel. Nlss-knockdown also resulted in twisted wings in both long-winged and short-winged brown planthoppers. Y-type olfactometer analyses indicated that antenna defects originating from Nlss depletion resulted in less sensitivity to host volatiles. This study represents the first report of the characteristics and functions of Nlss in N. lugens antenna and wing development and illuminates the function of the plaque organ of N. lugens in host volatile perception.',{'entities': [(16,25,'Gene'),(65,82,'Species'),(84,102,'Species'),(209,213,'Species'),(249,253,'Species'),(335,344,'Gene'),(346,348,'Gene'),(496,514,'Species'),(515,517,'Gene'),(524,528,'Gene'),(603,607,'Gene'),(684,688,'Gene'),(874,878,'Gene'),(957,975,'Species'),(1054,1058,'Gene'),(1196,1200,'Gene'),(1204,1213,'Species'),(1295,1304,'Species')]}),('The limpet transcription factors of Triatoma infestans regulate the response to fungal infection and modulate the expression pattern of defensin genes. As part of the innate humoral response to microbial attack, insects activate the expression of antimicrobial peptides (AMP). Understanding the regulatory mechanisms of this response in the Chagas disease vector Triatoma infestans is important since biological control strategies against pyrethroid-resistant insect populations were recently addressed by using the entomopathogenic fungus Beauveria bassiana. By bioinformatics, gene expression, and silencing techniques in T. infestans nymphs, we achieved sequence and functional characterization of two variants of the limpet transcription factor (Tilimpet) and studied their role as regulators of the AMP expression, particularly defensins, in fungus-infected insects. We found that Tilimpet variants may act differentially since they have divergent sequences and different relative expression ratios, suggesting that Tilimpet-2 could be the main regulator of the higher expressed defensins and Tilimpet-1 might play a complementary or more general role. Also, the six defensins (Tidef-1 to Tidef-6) exhibited different expression levels in fungus-infected nymphs, consistent with their phylogenetic clustering. This study aims to contribute to a better understanding of T. infestans immune response in which limpet is involved, after challenge by B. bassiana infection.',{'entities': [(4,10,'Species'),(36,54,'Species'),(136,144,'Gene'),(363,381,'Species'),(540,558,'Species'),(624,636,'Species'),(721,727,'Gene'),(750,758,'Gene'),(833,842,'Gene'),(886,894,'Gene'),(1021,1029,'Gene'),(1098,1106,'Gene'),(1172,1181,'Gene'),(1183,1190,'Gene'),(1194,1201,'Gene'),(1374,1386,'Species'),(1412,1418,'Gene'),(1451,1462,'Species')]}),('Hormonal control and target genes of ftz-f1 expression in the honeybee Apis mellifera: a positive loop linking juvenile hormone, ftz-f1, and vitellogenin. Ftz-f1 is an orphan member of the nuclear hormone receptor superfamily. A 20-hydroxyecdysone pulse allows ftz-f1 gene expression, which then regulates the activity of downstream genes involved in major developmental progression events. In honeybees, the expression of genes like vitellogenin (vg), prophenoloxidase and juvenile hormone-esterase during late pharate-adult development is known to be hormonally controlled in both queens and workers by increasing juvenile hormone (JH) titres in the presence of declining levels of ecdysteroids. Since Ftz-f1 is known for mediating intracellular JH signalling, we hypothesized that ftz-f1 could mediate JH action during the pharate-adult development of honeybees, thus controlling the expression of these genes. Here, we show that ftz-f1 has caste-specific transcription profiles during this developmental period, with a peak coinciding with the increase in JH titre, and that its expression is upregulated by JH and downregulated by ecdysteroids. RNAi-mediated knock down of ftz-f1 showed that the expression of genes essential for adult development (e.g. vg and cuticular genes) depends on ftz-f1 expression. Finally, a double-repressor hypothesis-inspired vg gene knock-down experiment suggests the existence of a positive molecular loop between JH, ftz-f1 and vg.',{'entities': [(37,43,'Gene'),(62,70,'Species'),(71,85,'Species'),(129,135,'Gene'),(141,153,'Gene'),(155,161,'Gene'),(261,267,'Gene'),(394,403,'Species'),(434,446,'Gene'),(448,450,'Gene'),(453,469,'Gene'),(474,499,'Gene'),(704,710,'Gene'),(784,790,'Gene'),(855,864,'Species'),(933,939,'Gene'),(1178,1184,'Gene'),(1259,1261,'Gene'),(1266,1281,'Gene'),(1294,1300,'Gene'),(1361,1363,'Gene'),(1455,1461,'Gene'),(1466,1468,'Gene')]}),('UDP-Glycosyltransferases are involved in imidacloprid resistance in the Asian citrus psyllid, Diaphorina citri (Hemiptera: Lividae). UDP-glycosyltransferases (UGTs), as phase II detoxification enzymes, are widely distributed within living organisms and play vital roles in the biotransformation of endobiotics and xenobiotics in insects. Insects increase the expression of detoxification enzymes to cope with the stress of xenobiotics, including insecticides. However, the roles of UGTs in insecticide resistance are still seldom reported. In this study, two UGT inhibitors, namely, 5-nitrouracil and sulfinpyrazone, were found to synergistically increase the toxicity of imidacloprid in the resistant population of Diaphorina citri. Based on transcriptome data, a total of 17 putative UGTs were identified. Quantitative real-time PCR showed that fourteen of the 17 UGT genes were overexpressed in the resistant population relative to the susceptible population. Using RNA interference technology to knockdown six UGT genes, the results suggested that silencing the selected UGT375A1, UGT383A1, UGT383B1, and UGT384A1 genes dramatically increased the toxicity of imidacloprid in the resistant population. However, silencing the UGT362B1 and UGT379A1 genes did not result in a significant increase in the toxicity of imidacloprid in the resistant population. These findings revealed that some upregulated UGT genes were involved in imidacloprid resistance in D. citri. These results shed some light upon and further our understanding of the mechanisms of insecticide resistance in insects.',{'entities': [(0,24,'Gene'),(72,92,'Species'),(94,110,'Species'),(133,157,'Gene'),(159,163,'Gene'),(482,486,'Gene'),(559,562,'Gene'),(716,732,'Species'),(786,790,'Gene'),(866,869,'Gene'),(1014,1017,'Gene'),(1075,1083,'Gene'),(1085,1093,'Gene'),(1095,1103,'Gene'),(1109,1117,'Gene'),(1228,1236,'Gene'),(1241,1249,'Gene'),(1404,1407,'Gene'),(1458,1466,'Species')]}),('Microplitis bicoloratus bracovirus modulates innate immune suppression through the eIF4E-eIF4A axis in the insect Spodoptera litura. Eukaryotic initiation factor 4E (eIF4E) is regulated during the innate immune response. However, its translational regulation under innate immune suppression remains largely unexplored. Microplitis bicoloratus bracovirus (MbBV), a symbiotic virus harbored by the parasitoid wasp, Microplitis bicoloratus, suppresses innate immunity in parasitized Spodoptera litura. Here, we generated eIF4E dsRNA and used it to silence the eIF4E gene of S. litura, resulting in a hallmark immunosuppressive phenotype characterized by increased apoptosis of hemocytes and retardation of head capsule width development. In response to natural parasitism, loss of eIF4E function was associated with similar immunosuppression, and we detected no significant differences between the response to parasitism and treatment with eIF4E RNAi. Under MbBV infection, eIF4E overexpression significantly suppressed MbBV-induced increase in apoptosis and suppressed apoptosis to the same extent as co-expression of both eIF4E and eIF4A. There were no significant differences between MbBV-infected and uninfected larvae in which eIF4E was overexpressed. More importantly, in the eIF4E RNAi strain, eIF4A RNAi did not increase apoptosis. Collectively, our results indicate that eIF4E plays a nodal role in the MbBV-suppressed innate immune response via the eIF4E-eIF4A axis.',{'entities': [(0,34,'Species'),(83,88,'Gene'),(89,94,'Gene'),(114,131,'Species'),(133,164,'Gene'),(166,171,'Gene'),(319,353,'Species'),(355,359,'Species'),(413,436,'Species'),(480,497,'Species'),(518,523,'Gene'),(557,562,'Gene'),(571,580,'Species'),(778,783,'Gene'),(937,942,'Gene'),(955,959,'Species'),(971,976,'Gene'),(1017,1021,'Species'),(1121,1126,'Gene'),(1131,1136,'Gene'),(1184,1188,'Species'),(1229,1234,'Gene'),(1279,1284,'Gene'),(1298,1303,'Gene'),(1377,1382,'Gene'),(1409,1413,'Species'),(1456,1461,'Gene'),(1462,1467,'Gene')]}),('Characterisation, analysis of expression and localisation of the opsin gene repertoire from the perspective of photoperiodism in the aphid Acyrthosiphon pisum. Organisms exhibit a wide range of seasonal responses as adaptions to predictable annual changes in their environment. These changes are originally caused by the effect of the Earth s cycles around the sun and its axial tilt. Examples of seasonal responses include floration, migration, reproduction and diapause. In temperate climate zones, the most robust variable to predict seasons is the length of the day (i.e. the photoperiod). The first step to trigger photoperiodic driven responses involves measuring the duration of the light-dark phases, but the molecular clockwork performing this task is poorly characterized. Photopigments such as opsins are known to participate in light perception, being part of the machinery in charge of providing information about the luminous state of the surroundings. Aphids (Hemiptera: Aphididae) are paradigmatic photoperiodic insects, exhibiting a strong induction to diapause when the light regime mimics autumn conditions. The availability of the pea aphid (Acyrthosiphon pisum) genome has facilitated molecular approaches to understand the effect of light stimulus in the photoperiodic induction process. We have identified, experimentally validated and characterized the expression of the full opsin gene repertoire in the pea aphid. Among identified opsin genes in A. pisum, arthropsin is absent in most insects sequenced to date (except for dragonflies and two other hemipterans) but also present in a crustacean, an onychophoran and chelicerates. We have quantified the expression of these genes in aphids exposed to different photoperiodic conditions and at different times of the day and localized their transcripts in the aphid brain. Clear differences in expression patterns were found, thus relating opsin expression with the photoperiodic response.',{'entities': [(65,70,'Gene'),(139,158,'Species'),(805,811,'Gene'),(1151,1160,'Species'),(1162,1181,'Species'),(1400,1405,'Gene'),(1429,1438,'Species'),(1457,1462,'Gene'),(1472,1480,'Species'),(1482,1492,'Gene'),(1914,1919,'Gene')]}),('karmoisin and cardinal ortholog genes participate in the ommochrome synthesis of Nilaparvata lugens (Hemiptera: Delphacidae). Ommochrome is the major source for eye coloration of all insect species so far examined. Phenoxazinone synthetase (PHS) has always been regarded as the terminal step enzyme for ommochrome formation, which is encoded by cardinal or karmoisin genes. Our previous study indicated that the karmoisin ortholog gene (Nl-karmoisin) product in the brown planthopper (BPH) was a monocarboxylate transporter, while not a PHS. Here, based on full-length complementary DNA, the cardinal ortholog gene in BPH (Nl-cardinal) product was predicted to be a haem peroxidase rather than a PHS. We suggest for the first time that neither karmoisin nor cardinal encodes the PHS, but whether PHS participates in BPH eye pigmentation needs further research. Nymphal RNA interference (RNAi) experiments showed that knockdown Nl-cardinal transcript led the BPH ocelli and compound eye to color change from brown to red, while knockdown Nl-karmoisin only made the ocelli present the red phenotype. Notably, not only the Nl-cardinal transcript, dscd injection (Nl-cardinal targeting double-stranded DNA (dsRNA)) also significantly reduced the Nl-karmoisin transcript by 33.7%, while dska (Nl-karmoisin targeting dsRNA) injection did not significantly change the Nl-cardinal transcript. Considering the above RNAi and quantitative real-time polymerase chain reaction results, we propose that Nl-cardinal plays a more important role in ommochrome synthesis than Nl-karmoisin, and it may be an upstream gene of Nl-karmoisin. The present study suggested that both karmoisin and cardinal ortholog genes play a role in ommochrome synthesis in a hemimetabolous insect.',{'entities': [(0,9,'Gene'),(14,22,'Gene'),(81,99,'Species'),(345,353,'Gene'),(357,366,'Gene'),(412,421,'Gene'),(437,449,'Gene'),(466,483,'Species'),(485,488,'Species'),(592,600,'Gene'),(618,621,'Species'),(623,634,'Gene'),(744,753,'Gene'),(758,766,'Gene'),(927,938,'Gene'),(958,961,'Species'),(1037,1049,'Gene'),(1120,1131,'Gene'),(1160,1171,'Gene'),(1242,1254,'Gene'),(1288,1300,'Gene'),(1361,1372,'Gene'),(1493,1501,'Gene'),(1559,1571,'Gene'),(1607,1619,'Gene'),(1659,1668,'Gene'),(1673,1681,'Gene')]}),('A trypsin inhibitor-like protein secreted by Cotesia vestalis teratocytes inhibits hemolymph prophenoloxidase activation of Plutella xylostella. To establish successful infections, endoparasitoid wasps must develop strategies to evade immune responses of the host. Here, we identified and characterized a teratocytes-expressed gene encoding a trypsin inhibitor-like protein containing a cysteine-rich domain from Cotesia vestalis, CvT-TIL. CvT-TIL had a high expression level during the later developmental stage of teratocytes and was secreted into host hemolymph. Further experiments showed CvT-TIL strongly suppressed the prophenoloxidase activation of host hemolymph in a dose-dependent manner by interacting with PxPAP3 of PO cascade. Our results not only provide evidence for an inhibition between CvT-TIL gene and the host s melanization activity, but also expand our knowledge about the mechanisms by which parasitoids regulate humoral immunity of the host.',{'entities': [(2,32,'Gene'),(45,61,'Species'),(124,143,'Species'),(343,373,'Gene'),(413,429,'Species'),(431,438,'Gene'),(440,447,'Gene'),(593,600,'Gene'),(804,811,'Gene')]}),('Insulin-like peptides of the legume pod borer, Maruca vitrata, and their mediation effects on hemolymph trehalose level, larval development, and adult reproduction. Insulin-like peptides (ILPs) of insects mediate various physiological processes including hemolymph sugar level, immature growth, female reproduction, and lifespan. In target cells of ILPs, insulin/insulin-like growth factor signaling (IIS) is highly conserved in animals. IIS in the legume pod borer, Maruca vitrata (Lepidoptera: Crambidae), is known to be involved in maintaining hemolymph trehalose levels and promoting larval growth. However, ILPs in M. vitrata have not been reported yet. This study predicted two ILP genes of Mv-ILP1 and Mv-ILP2 from transcriptome of M. vitrata. Mv-ILP1 and Mv-ILP2 shared high sequence homologies and domain architecture with Drosophila ILPs. Both ILPs exhibited similar expression patterns in most developmental stages, showing high expression levels in adult stage. In the larval stage, Mv-ILP1 and Mv-IlP2 were expressed mostly in the brain and fat body. However, in the adult stage, both ILP genes were expressed more in the abdomen than those in the head containing the brain. RNA interference (RNAi) of either Mv-ILP1 or Mv-ILP2 during larval stage resulted in significant malfunctioning in regulating hemolymph trehalose titers. RNAi-treated larvae also exhibited significant retardation of larval growth. RNAi treatment in adult stage interfered with the ovarian development of females. These results suggest that Mv-ILP1 and Mv-ILP2 play crucial roles in mediating larval growth and adult reproduction.',{'entities': [(0,20,'Gene'),(47,61,'Species'),(188,191,'Gene'),(467,481,'Species'),(620,630,'Species'),(684,687,'Gene'),(700,704,'Gene'),(712,716,'Gene'),(739,749,'Species'),(754,758,'Gene'),(766,770,'Gene'),(998,1002,'Gene'),(1010,1014,'Gene'),(1098,1101,'Gene'),(1225,1229,'Gene'),(1236,1240,'Gene'),(1531,1535,'Gene'),(1543,1547,'Gene')]}),('Identification and functional analysis of a novel chorion protein essential for egg maturation in the brown planthopper. In insect eggs, the chorion has the essential function of protecting the embryo from external agents during development while allowing gas exchange for respiration. In this study, we found a novel gene, Nilaparvata lugens chorion protein (NlChP), that is involved in chorion formation in the brown planthopper, Nilaparvata lugens. NlChP was highly expressed in the follicular cells of female adult brown planthoppers. Knockdown of NlChP resulted in oocyte malformation and the inability to perform oviposition, and electron microscopy showed that the malformed oocytes had thin and rough endochorion layers compared to the control group. Liquid chromatography with tandem mass spectrometry analysis of the eggshell components revealed four unique peptides that were matched to NlChP. Our results demonstrate that NlChP is a novel chorion protein essential for egg maturation in N. lugens, a hemipteran insect with telotrophic meroistic ovaries. NlChP may be a potential target in RNA interference-based insect pest management.',{'entities': [(50,65,'Gene'),(102,119,'Species'),(324,342,'Species'),(343,358,'Gene'),(360,365,'Gene'),(413,430,'Species'),(432,450,'Species'),(452,457,'Gene'),(519,537,'Species'),(552,557,'Gene'),(898,903,'Gene'),(934,939,'Gene'),(951,966,'Gene'),(999,1008,'Species'),(1066,1071,'Gene')]}),('The regulation of three new members of the cytochrome P450 CYP6 family and their promoters in the cotton aphid Aphis gossypii by plant allelochemicals. BACKGROUND: The expression of P450 genes in insects can be induced by plant allelochemicals. To understand the induction mechanisms, we measured the expression profiles of three P450 genes and their promoter activities under the induction of plant allelochemicals. RESULTS: The inducible expression of CYP6CY19 was the highest among three genes, followed by those of CYP6CY22 and CYP6DA1. The regions from -687 to +586 bp of CYP6DA1, from -666 to +140 bp of CYP6CY19 and from -530 to +218 bp of CYP6CY22 were essential for basal transcriptional activity. The cis-elements for plant allelochemicals induction were identified between -193 and +56 bp of CYP6DA1, between -157 and +140 bp of CYP6CY19 and between -108 and +218 bp of CYP6CY22. These promoter regions were found to contain a potential aryl hydrocarbon receptor element binding site with a conservative sequence motif 5 -C/TAC/ANCA/CA-3 . All these four plant allelochemicals were able to induce the expression of these P450 genes. Tannic acid had a better inductive effect than other three plant allelochemicals. CONCLUSIONS: Our study identified the plant allelochemical responsive cis-elements. This provides further research targets aimed at understanding the regulatory mechanisms of P450 genes expression and their interactions with plant allelochemicals in insect pests.  2018 Society of Chemical Industry.',{'entities': [(43,58,'Gene'),(59,63,'Gene'),(98,110,'Species'),(111,125,'Species'),(182,186,'Gene'),(330,334,'Gene'),(454,462,'Gene'),(519,527,'Gene'),(532,539,'Gene'),(577,584,'Gene'),(610,618,'Gene'),(647,655,'Gene'),(803,810,'Gene'),(840,848,'Gene'),(881,889,'Gene'),(1132,1136,'Gene'),(1401,1405,'Gene')]}),('Identification and functional characterization of an odorant receptor in pea aphid, Acyrthosiphon pisum. The sensitive olfactory system is necessary for survival of insects. Odorant receptors (ORs) are located on the dendrites of olfactory receptor neurons and play a critical role in odor detection. Insect ORs are functionally analyzed via heterologous expression in a Xenopus oocyte system using a two-electrode voltage-clamp (TEVC) electrophysiological recording. Here, we have identified a novel OR in the pea aphid, Acyrthosiphon pisum, then we cloned and named it ApisOR4. We analyzed the ApisOR4 tissue expression patterns and found expression only in antennae tissues. Further functional analysis using TEVC revealed that ApisOR4 is broadly tuned to eight volatiles, which elicit electrophysiological response in pea aphid antennae. This study provides an initial functional analysis of aphid ORs and identifies candidate volatiles to be used in developing new strategies for aphid control.',{'entities': [(53,69,'Gene'),(73,82,'Species'),(84,103,'Species'),(174,191,'Gene'),(193,195,'Gene'),(308,311,'Gene'),(371,378,'Species'),(501,503,'Gene'),(511,520,'Species'),(522,541,'Species'),(571,578,'Gene'),(596,603,'Gene'),(731,738,'Gene'),(822,831,'Species'),(902,905,'Gene')]}),('Large gene family expansions and adaptive evolution for odorant and gustatory receptors in the pea aphid, Acyrthosiphon pisum. Gaining insight into the mechanisms of chemoreception in aphids is of primary importance for both integrative studies on the evolution of host plant specialization and applied research in pest control management because aphids rely on their sense of smell and taste to locate and assess their host plants. We made use of the recent genome sequence of the pea aphid, Acyrthosiphon pisum, to address the molecular characterization and evolution of key molecular components of chemoreception: the odorant (Or) and gustatory (Gr) receptor genes. We identified 79 Or and 77 Gr genes in the pea aphid genome and showed that most of them are aphid-specific genes that have undergone recent and rapid expansion in the genome. By addressing selection within sets of paralogous Or and Gr expansions, for the first time in an insect species, we show that the most recently duplicated loci have evolved under positive selection, which might be related to the high degree of ecological specialization of this species. Although more functional studies are still needed for insect chemoreceptors, we provide evidence that Grs and Ors have different sets of positively selected sites, suggesting the possibility that these two gene families might have different binding pockets and bind structurally distinct classes of ligand. The pea aphid is the most basal insect species with a completely sequenced genome to date. The identification of chemoreceptor genes in this species is a key step toward further exploring insect comparative genetics, the genomics of ecological specialization and speciation, and new pest control strategies.',{'entities': [(68,87,'Gene'),(95,104,'Species'),(106,125,'Species'),(482,491,'Species'),(493,512,'Species'),(630,632,'Gene'),(649,651,'Gene'),(686,688,'Gene'),(696,698,'Gene'),(712,721,'Species'),(895,897,'Gene'),(902,904,'Gene'),(1443,1452,'Species')]}),('Over expression of bmo-miR-2819 suppresses BmNPV replication by regulating the BmNPV ie-1 gene in Bombyx mori. Bombyx mori nucleopolyhedrovirus (BmNPV) is a major pathogen that threatens the growth and sustainability of the sericulture industry. Accumulating studies in recent years suggest that insect viruses infection can change the host microRNAs (miRNAs) expression profile and both cellular and viral miRNAs play roles in host-pathogen interactions. Until now, the functional analysis of miRNA encoded by silkworm for host-virus interaction is limited. In this study, we validate the down-regulation of bmo-miR-2819 upon BmNPV infection by qRT-PCR and confirm the BmNPV immediately early 1 gene, ie-1 is one of the targets for bmo-miR-2819 based on the results of dual luciferase report assay. Overexpression of bmo-miR-2819 can significantly decline the abundance of IE-1 protein level in BmNPV-infected silkworm larvae. Further, the expression level of polyhedrin gene and VP39 protein of BmNPV in the infected larvae after applying bmo-miR-2819 mimics was significantly decreased comparing with that of larvae with mimic control. Our results suggest that overexpression of bmo-miR-2819 could suppress BmNPV replication by down-regulating the expression of BmNPV ie-1 gene, which demonstrate that cellular miRNAs could affect virus infection by regulating the expression of virus genes.',{'entities': [(19,31,'Gene'),(43,48,'Species'),(79,84,'Species'),(85,89,'Gene'),(98,109,'Species'),(111,143,'Species'),(145,150,'Species'),(511,519,'Species'),(609,621,'Gene'),(627,632,'Species'),(670,675,'Species'),(702,706,'Gene'),(733,745,'Gene'),(818,830,'Gene'),(874,878,'Gene'),(896,901,'Species'),(911,919,'Species'),(981,985,'Gene'),(997,1002,'Species'),(1041,1053,'Gene'),(1182,1194,'Gene'),(1210,1215,'Species'),(1265,1270,'Species'),(1271,1275,'Gene')]}),('The expression of cockroach insulin-like peptides is differentially regulated by physiological conditions and affected by compensatory regulation. In insects, the insulin receptor (InR) pathway is involved in regulating key physiological processes, including juvenile hormone (JH) synthesis, vitellogenin production, and oocyte growth. This raises the question about which ligand (or ligands) binds to InR to trigger the above effects. We have cloned seven insulin-like peptides (BgILP1 to 7) from female Blattella germanica cockroaches and found that the brain expresses BgILP1 to 6, the fat body BgILP7, and the ovary BgILP2. Starvation induces the reduction of BgILP3, 5, and 6 mRNA levels in the brain, and the various BgILPs are differentially expressed during the gonadotrophic cycle. In addition, by knocking down the BgILPs we were able to identify compensatory regulation at transcriptional level between the different BgILPs, although none of the BgILP knockdown assays, including the knockdown of the seven BgILPs, produced the same phenotypes that we achieved by depleting InR. Taken together, the results indicate that B. germanica ILPs are differentially expressed in tissues and in response to physiological conditions, and that they are affected by compensatory regulation.',{'entities': [(18,27,'Species'),(28,49,'Gene'),(457,478,'Gene'),(480,486,'Gene'),(487,491,'Species'),(505,524,'Species'),(572,578,'Gene'),(598,604,'Gene'),(620,626,'Gene'),(664,670,'Gene'),(723,728,'Gene'),(957,962,'Gene'),(1132,1144,'Species')]}),('Identification, characterization and expression analysis of Anopheles stephensi double peroxidase. Peroxidases catalyze the reduction of peroxides and that, in turn, oxidize various substrates. They have been widely reported to play an important role in mosquito innate immunity against various pathogens. Here, we have characterized double heme peroxidase (AsDBLOX) gene from the Indian malaria vector Anopheles stephensi. It is a true ortholog of An. gambiae DBLOX. This 4209 bp AsDBLOX gene encodes for a protein of 1402 amino acids that has two duplicated peroxidase domains, domain I (from amino acid 61 to 527) and domain II (from amino acid 714 to 1252). The first domain has only substrate binding sites and lacks all other motifs of a functional heme peroxidase (e.g. heme binding site, calcium binding site and homodimer interface). Instead, it has two integrin binding motifs-LDV (Leu-Asp-Val) and RGD (Arg-Gly-Asp). The second peroxidase domain, however, has all the features of a complete heme peroxidase along with an integrin binding motif LDI (Leu-Asp-Ile). Thus, AsDBLOX gene is a unique type of peroxinectin as these groups of proteins are characterized by integrin binding motifs along with a heme peroxidase domain. We also observed that the AsDBLOX gene is expressed in all the life cycle stages of mosquito and is highly induced in the pupal stage of development which indicates its possible role in development.',{'entities': [(60,79,'Species'),(334,356,'Gene'),(358,365,'Gene'),(403,422,'Species'),(449,460,'Species'),(461,466,'Gene'),(481,488,'Gene'),(1080,1087,'Gene'),(1262,1269,'Gene')]}),('Identification and RNAi-based function analysis of chitinase family genes in diamondback moth, Plutella xylostella. BACKGROUND: Insect chitinases play a vital part in chitin degradation in exoskeletons and gut linings during the molting process, therefore are considered as the potential targets for new insecticide design or RNA interference (RNAi) based pest management. The systematic function analysis of chitinase genes have already been well conducted in several insect pests, but still absent in Plutella xylostella. RESULTS: In the present study, 13 full-length chitinase transcripts were obtained in P. xylostella. Developmental and tissue-specific expression pattern analysis revealed that seven chitinase transcripts were periodically expressed during molting stage and mainly expressed in the integument or midgut, including PxCht3, PxCht5, PxCht6-2, PxCht7, PxCht8, PxCht10 and PxCht-h. RNAi-mediated knockdown of these specific expressed genes revealed that PxCht5 and PxCht10 were essential in larval molting, pupation and eclosion, and PxCht7 was indispensable only in eclosion; No significant effects were observed on insect survival or normal development when the rest chitinase transcripts were suppressed by RNAi. CONCLUSION: Our results indicated the function of P. xylostella chitinase family genes during the molting process, and may provide potential targets for RNAi-based management of P. xylostella. This article is protected by copyright. All rights reserved.',{'entities': [(51,60,'Gene'),(77,93,'Species'),(95,114,'Species'),(409,418,'Gene'),(503,522,'Species'),(570,579,'Gene'),(609,622,'Species'),(706,715,'Gene'),(837,843,'Gene'),(845,851,'Gene'),(853,861,'Gene'),(863,869,'Gene'),(871,877,'Gene'),(879,886,'Gene'),(972,978,'Gene'),(1052,1058,'Gene'),(1187,1196,'Gene'),(1284,1297,'Species'),(1298,1307,'Gene'),(1412,1425,'Species')]}),('Identification of the chitinase genes from the diamondback moth, Plutella xylostella. UNASSIGNED: Chitinases have an indispensable function in chitin metabolism and are well characterized in numerous insect species. Although the diamondback moth (DBM) Plutella xylostella, which has a high reproductive potential, short generation time, and characteristic adaptation to adverse environments, has become one of the most serious pests of cruciferous plants worldwide, the information on the chitinases of the moth is presently limited. In the present study, using degenerated polymerase chain reaction (PCR) and rapid amplification of cDNA ends-PCR strategies, four chitinase genes of P. xylostella were cloned, and an exhaustive search was conducted for chitinase-like sequences from the P. xylostella genome and transcriptomic database. Based on the domain analysis of the deduced amino acid sequences and the phylogenetic analysis of the catalytic domain sequences, we identified 15 chitinase genes from P. xylostella. Two of the gut-specific chitinases did not cluster with any of the known phylogenetic groups of chitinases and might be in a new group of the chitinase family. Moreover, in our study, group VIII chitinase was not identified. The structures, classifications and expression patterns of the chitinases of P. xylostella were further delineated, and with this information, further investigations on the functions of chitinase genes in DBM could be facilitated.',{'entities': [(22,31,'Gene'),(47,63,'Species'),(65,84,'Species'),(229,245,'Species'),(252,271,'Species'),(664,673,'Gene'),(683,696,'Species'),(753,762,'Gene'),(787,800,'Species'),(984,993,'Gene'),(1005,1018,'Species'),(1162,1171,'Gene'),(1215,1224,'Gene'),(1322,1335,'Species'),(1431,1440,'Gene')]}),('Molecular characterization of prophenoloxidase-1 (PPO1) and the inhibitory effect of kojic acid on phenoloxidase (PO) activity and on the development of Zeugodacus tau (Walker) (Diptera: Tephritidae). Phenoloxidase (PO) plays a key role in melanin biosynthesis during insect development. Here, we isolated the 2310-bp full-length cDNA of PPO1 from Zeugodacus tau, a destructive horticultural pest. qRT-polymerase chain reaction showed that the ZtPPO1 transcripts were highly expressed during larval-prepupal transition and in the haemolymph. When the larvae were fed a 1.66% kojic acid (KA)-containing diet, the levels of the ZtPPO1 transcripts significantly increased by 2.79- and 3.39-fold in the whole larvae and cuticles, respectively, while the corresponding PO activity was significantly reduced; in addition, the larval and pupal durations were significantly prolonged; pupal weights were lowered; and abnormal phenotypes were observed. An in vitro inhibition experiment indicated that KA was an effective competitive inhibitor of PO in Z. tau. Additionally, the functional analysis showed that 20E could significantly up-regulate the expression of ZtPPO1, induce lower pupal weight, and advance pupation. Knockdown of the ZtPPO1 gene by RNAi significantly decreased mRNA levels after 24 h and led to low pupation rates and incomplete pupae with abnormal phenotypes during the larval-pupal interim period. These results proved that PO is important for the normal growth of Z. tau and that KA can disrupt the development of this pest insect.',{'entities': [(30,48,'Gene'),(50,54,'Gene'),(153,167,'Species'),(338,342,'Gene'),(348,362,'Species'),(444,450,'Gene'),(626,632,'Gene'),(1044,1050,'Species'),(1156,1162,'Gene'),(1230,1236,'Gene'),(1480,1486,'Species')]}),('Egf-like gene is essential for cuticle metabolism in the brown planthopper. Using the mass spectrometry analysis of cuticle casts of brown planthopper (BPH, Nilaparvata lugens) and transcriptome analysis of BPH tissues, we identified a gigantic gene (50,922  bp, 16,973 aa) tentatively called Nlegf-like. Multiple transcripts were found. Nlegf-like encodes an integral membrane protein of 16,973 amino acid residues with 260 EGF-like repeats and 16 Ca2+-binding EGF repeats type (cbEGFs) in the extracellular portion. Nlegf-like was highly expressed in the integument and tended to peak at the middle stage or late stage of each nymph instar. Phylogenetic analysis showed this gene is conserved in many other insects. Different double-stranded RNA-mediated RNA interference targeting eight different regions of the Nlegf-like gene resulted in abnormal cuticle formation or molting and lethal phenotypes. Transmission electron microscopy revealed that the newly formed endocuticle was significantly thinner for RNAi-treated BPHs with phenotype of contracted abdomen, or the old cuticle could not be digested sufficiently for those with phenotype of slender body shape or died with molting difficulty when compared with the control group. We suggest that the Nlegf-like is crucial for metabolism of the cuticle in BPH molting.',{'entities': [(0,8,'Gene'),(57,74,'Species'),(133,150,'Species'),(152,155,'Species'),(157,175,'Species'),(207,210,'Species'),(293,303,'Gene'),(338,348,'Gene'),(425,433,'Gene'),(518,528,'Gene'),(815,825,'Gene'),(1257,1267,'Gene'),(1312,1315,'Species')]}),('MicroRNA Let-7 targets the ecdysone signaling pathway E75 gene to control larval-pupal development in Bactrocera dorsalis. MicroRNAs (miRNAs) regulate various biological processes during insect development; however, their role in larval-pupal development in oriental fruit fly, Bactrocera dorsalis (Hendel) remains unknown. In the current study, we address the biological function of a conserved miRNA, Bdo-Let-7 in the regulation of BdE75 gene, which belongs to the ecdysone signaling pathway and participates in the larval-pupal development in B. dorsalis. Using dual luciferase reporter assay in HEK293T cells we show that Bdo-Let-7 miRNA interacts with the 3  untranslated region of BdE75 gene and suppresses its expression. The Bdo-Let-7 and BdE75 are also co-expressed in the larval-pupal stages and in different tissues of B. dorsalis. In in vivo experiments, the injection of Bdo-Let-7 agomir and antagomir in third instar larvae down- and up-regulated the expression of BdE75, respectively. The 20-hydroxyecdysone (20E) injection assay shows that 20E up-regulated the expression of Bdo-Let-7 on the 5th day of the larvae. Moreover, abnormal pupation and eclosion were observed after larval Bdo-Let-7 antagomir injection. Based on these results, we show that Bdo-Let-7 regulates the ecdysone signaling pathway through the exact dose of BdE75 gene, and is indispensable for normal larval-pupal development in B. dorsalis.',{'entities': [(9,14,'Gene'),(102,121,'Species'),(258,276,'Species'),(278,297,'Species'),(407,412,'Gene'),(434,439,'Gene'),(546,557,'Species'),(630,635,'Gene'),(687,692,'Gene'),(737,742,'Gene'),(747,752,'Gene'),(830,841,'Species'),(888,893,'Gene'),(979,984,'Gene'),(1095,1100,'Gene'),(1203,1208,'Gene'),(1271,1276,'Gene'),(1344,1349,'Gene'),(1416,1427,'Species')]}),('Identification, expression and function of myosin heavy chain family genes in Tribolium castaneum. Functions of myosin heavy chain (myosin) family genes are poorly understood in some insects. To address this, we determined the expression and function of myosin family genes in Tribolium castaneum. TcMyo15 is predominantly expressed in early embryos, late larvae and early adults, but TcMyo7B transcripts significantly increased in late larvae. TcMyo20 transcripts are abundant in pre-adults, whereas TcMhc1 is highly expressed in post-embryonic stages. TcMhc2 shows peak expression in late pupae. TcMyo9 transcripts reached their highest levels in late pupae. TcMyo15, TcMyo7B and TcMhc2 are abundantly expressed in the adult epidermis, gut and testis, respectively. TcMyo9 and TcMyo20 are highly expressed in the epidermis, fat body and ovary, and TcMhc1 exhibits high mRNA levels in the epidermis and accessory gland. TcMyo20 RNAi reduced wing and leg size, fertility and egg hatchability. TcMyo9 knockdown completely inhibited eclosion and fecundity, but TcMyo15 or TcMhc1 silencing only impaired eclosion. TcMhc2 RNAi affected pupation and wing development. This study suggests myosin genes as potential targets of pesticides due to their critical roles in insect development and reproduction.',{'entities': [(43,61,'Gene'),(78,97,'Species'),(112,130,'Gene'),(277,296,'Species'),(298,305,'Gene'),(385,392,'Gene'),(445,452,'Gene'),(501,507,'Gene'),(554,560,'Gene'),(598,604,'Gene'),(661,668,'Gene'),(670,677,'Gene'),(682,688,'Gene'),(768,774,'Gene'),(779,786,'Gene'),(850,856,'Gene'),(921,928,'Gene'),(993,999,'Gene'),(1059,1066,'Gene'),(1070,1076,'Gene'),(1111,1117,'Gene')]}),('Circadian clock genes link photoperiodic signals to lipid accumulation during diapause preparation in the diapause-destined female cabbage beetles Colaphellus bowringi. Many organisms have evolved a series of adaptions, such as dormancy or diapause in insects that enable them to withstand seasonally adverse conditions. In insects, photoperiodic signals received during the diapause induction phase have irreversible effect on diapause initiation. Insects continue to be exposed to diapause-inducing photoperiod after the diapause induction phase during diapause preparation before they enter diapause. However, how photoperiodic signals experienced during the diapause preparation phase (DPP) regulate diapause remains largely unclear. In this paper, we investigate this in the cabbage beetle Colaphellus bowringi. The cabbage beetle is in many respects an ideal experimental model in which to investigate the effect of photoperiodic signals on the DPP because it has facultative reproductive diapause induced by long-day length and has differentiable diapause induction and preparation phases. We found that the lipid content of female cabbage beetles decreased after diapause-destined (DD) individuals were exposed to a diapause-inhibiting photoperiod during the DPP. Two circadian clock negative regulators, per and tim, were probably involved in the photoperiodic response of beetles during the DPP. Per and tim presented obvious oscillation of circadian rhythm and photoperiodic response during the DPP in DD females and knock-down of these genes in DD females caused their lipid content to decrease. Per and tim probably promote lipid accumulation by regulating the expression of genes that regulate lipogenesis and lipolysis. Moreover, decreased lipid accumulation caused by exposure to different photoperiods during the DPP was independent of juvenile hormone. In summary, these results suggest that photoperiodic signals received during the DPP influence lipid accumulation in DD insects. DD insects still have some ability to monitor photoperiodic changes during the DPP and per and tim are probably involved in regulating physiological responses to photoperiodic signals during diapause preparation. These results shed light on the relationship between photoperiodic signals and diapause preparation, and may provide new insights on both how to better utilize insects as resources and for pest management.',{'entities': [(131,145,'Species'),(147,167,'Species'),(780,787,'Species'),(795,815,'Species'),(821,828,'Species'),(1139,1146,'Species'),(1313,1316,'Gene'),(1321,1324,'Gene'),(1406,1409,'Gene'),(1414,1417,'Gene'),(1608,1611,'Gene'),(1616,1619,'Gene'),(2087,2090,'Gene'),(2095,2098,'Gene')]}),('Transcription factors CncC/Maf and AhR/ARNT coordinately regulate the expression of multiple GSTs conferring resistance to chlorpyrifos and cypermethrin in Spodoptera exigua. BACKGROUND: Glutathione S-transferases (GSTs) are a superfamily of multifunctional dimeric proteins existing in both prokaryotic and eukaryotic organisms. They are involved in the detoxification of both endogenous and exogenous electrophiles, including insecticides. However, the molecular mechanisms underlying the regulation of GST genes in insects are poorly understood. RESULTS: We first identified at least three GST genes involved in resistance to the insecticides chlorpyrifos and cypermethrin. Analysis of upstream sequences revealed that three GSTs (SeGSTo2, SeGSTe6 and SeGSTd3) harbor the same cap  n  collar C/muscle aponeurosis fibromatosis (CncC/Maf) binding site, and SeGSTo2 and SeGSTe6 contain the aryl hydrocarbon receptor/aryl hydrocarbon receptor nuclear translocator (AhR/ARNT) binding site. Luciferase reporter assay showed co-transfection of reporter plasmid containing the SeGSTe6 promoter with CncC and/or Maf expressing constructs significantly boosted transcription. Similarly, AhR and/or ARNT expressing constructs also significantly increased the promoter activities. The co-transfection of mutated reporter plasmid with CncC/Maf or AhR/ARNT did not increase transcription activity anymore. Constitutive over-expression of CncC, Maf and AhR was also found in the HZ16 strain, which might be the molecular mechanism for up-regulated expression of multiple detoxification genes conferring resistance to insecticides. CONCLUSION: These results suggest that CncC/Maf and AhR/ARNT coordinately regulate the expression of multiple GST genes involved in insecticide resistance in Spodoptera exigua.  2019 Society of Chemical Industry.',{'entities': [(22,30,'Gene'),(35,43,'Gene'),(93,97,'Gene'),(156,173,'Species'),(187,213,'Gene'),(215,219,'Gene'),(728,732,'Gene'),(734,741,'Gene'),(743,750,'Gene'),(755,762,'Gene'),(830,838,'Gene'),(858,865,'Gene'),(870,877,'Gene'),(964,972,'Gene'),(1072,1079,'Gene'),(1325,1333,'Gene'),(1337,1345,'Gene'),(1658,1666,'Gene'),(1671,1679,'Gene'),(1777,1794,'Species')]}),('Genome-wide identification of chitin-binding proteins and characterization of BmCBP1 in the silkworm, Bombyx mori. The insect cuticle plays important roles in numerous physiological functions to protect the body from invasion of pathogens, physical injury and dehydration. In this report, we conducted a comprehensive genome-wide search for genes encoding proteins with peritrophin A-type (ChtBD2) chitin-binding domain (CBD) in the silkworm, Bombyx mori. One of these genes, which encodes the cuticle protein BmCBP1, was additionally cloned, and its expression and location during the process of development and molting in B. mori were investigated. In total, 46 protein-coding genes were identified in the silkworm genome, including those encoding 15 cuticle proteins analogous to peritrophins with one CBD (CPAP1s), nine cuticle proteins analogous to peritrophins with three CBD (CPAP3s), 15 peritrophic membrane proteins (PMPs), four chitinases, and three chitin deacetylases, which contained at least one ChtBD2 domain. Microarray analysis indicated that CPAP-encoding genes were widely expressed in various tissues, whereas PMP genes were highly expressed in the midgut. Quantitative polymerase chain reaction and western blotting showed that the cuticle protein BmCBP1 was highly expressed in the epidermis and head, particularly during molting and metamorphosis. An immunofluorescence study revealed that chitin co-localized with BmCBP1 at the epidermal surface during molting. Additionally, BmCBP1 was notably up-regulated by 20-hydroxyecdysone treatment. These results provide a genome-level view of the chitin-binding protein in silkworm and suggest that BmCBP1 participates in the formation of the new cuticle during molting.',{'entities': [(30,53,'Gene'),(78,84,'Gene'),(92,100,'Species'),(102,113,'Species'),(433,441,'Species'),(443,454,'Species'),(494,509,'Gene'),(510,516,'Gene'),(624,631,'Species'),(708,716,'Species'),(753,768,'Gene'),(1253,1268,'Gene'),(1269,1275,'Gene'),(1438,1444,'Gene'),(1500,1506,'Gene'),(1614,1636,'Gene'),(1640,1648,'Species'),(1666,1672,'Gene')]}),('RNA Interference in the Tobacco Hornworm, Manduca sexta, Using Plastid-Encoded Long Double-Stranded RNA. RNA interference (RNAi) is a promising method for controlling pest insects by silencing the expression of vital insect genes to interfere with development and physiology; however, certain insect Orders are resistant to this process. In this study, we set out to test the ability of in planta-expressed dsRNA synthesized within the plastids to silence gene expression in an insect recalcitrant to RNAi, the lepidopteran species, Manduca sexta (tobacco hornworm). Using the Manduca vacuolar-type H+ ATPase subunit A (v-ATPaseA) gene as the target, we first evaluated RNAi efficiency of two dsRNA products of different lengths by directly feeding the in vitro-synthesized dsRNAs to M. sexta larvae. We found that a long dsRNA of 2222 bp was the most effective in inducing lethality and silencing the v-ATPaseA gene, when delivered orally in a water droplet. We further transformed the plastid genome of the M. sexta host plant, Nicotiana tabacum, to produce this long dsRNA in its plastids and performed bioassays with M. sexta larvae on the transplastomic plants. In the tested insects, the plastid-derived dsRNA had no effect on larval survival and no statistically significant effect on expression of the v-ATPaseA gene was observed. Comparison of the absolute quantities of the dsRNA present in transplastomic leaf tissue for v-ATPaseA and a control gene, GFP, of a shorter size, revealed a lower concentration for the long dsRNA product compared to the short control product. We suggest that stability and length of the dsRNA may have influenced the quantities produced in the plastids, resulting in inefficient RNAi in the tested insects. Our results imply that many factors dictate the effectiveness of in planta RNAi, including a likely trade-off effect as increasing the dsRNA product length may be countered by a reduction in the amount of dsRNA produced and accumulated in the plastids.',{'entities': [(24,40,'Species'),(42,55,'Species'),(533,546,'Species'),(548,564,'Species'),(577,584,'Species'),(585,618,'Gene'),(620,629,'Gene'),(784,792,'Species'),(1009,1017,'Species'),(1030,1047,'Species'),(1121,1129,'Species')]}),('Transgenic soybean plants expressing Spb18S dsRNA exhibit enhanced resistance to the soybean pod borer Leguminivora glycinivorella (Lepidoptera: Olethreutidae). The soybean pod borer [SPB; Leguminivora glycinivorella (Mats.) Obraztsov] is a major soybean pest in northeastern Asia. A useful method for addressing this problem is the generation of transgenic plants producing double-stranded RNA (dsRNA) that target essential insect genes. In this study, we confirmed that 18S ribosomal RNA is critical for SPB development. Downregulated Spb18S expression induced by dsRNA injection increased larval mortality rates and resulted in early pupation. We also assessed whether Spb18S is silenced in SPB larvae fed on transgenic soybean expressing Spb18S dsRNA. Transgenic plants downregulated Spb18S expression levels and second-instar larval survival rates. Moreover, such plants were less damaged by SPB larvae than control plants under field conditions.',{'entities': [(11,18,'Species'),(85,92,'Species'),(103,130,'Species'),(165,172,'Species'),(189,216,'Species'),(247,254,'Species'),(472,489,'Gene'),(537,543,'Gene'),(672,678,'Gene'),(723,730,'Species'),(745,748,'Gene'),(788,794,'Gene')]}),('Host-mediated RNA interference targeting a cuticular protein gene impaired fecundity in the green peach aphid Myzus persicae. BACKGROUND: The green peach aphid (Myzus persicae) is a devastating sap-sucking insect pest that damages many host plants worldwide and causes billions of dollars of crop losses. Induction of RNA interference (RNAi) through oral feeding of small interfering RNA (siRNA) has been demonstrated in aphids. Therefore, host-mediated delivery of double-stranded RNA (dsRNA) specific to vital structural genes of aphids has been envisaged as a tool for the development of resistance against this aphid species. RESULTS: Cuticular protein (CP) senses seasonal photoperiodism and drives a shift from clonal to sexual generation in aphids. Thus, attenuation of CP gene expression is likely to result in a different reproductive orientation in aphids and thereby affect their fecundity. A gene encoding CP in M. persicae has been targeted for RNAi-mediated knockdown. Transgenic Arabidopsis expressing dsRNA homologous to the MyCP gene was developed. The dsRNA-transgenics produced gene-specific siRNAs fed by aphids infesting the transgenics. A reverse transcription-quantitative polymerase chain reaction (RT-qPCR) study revealed an attenuated level of transcripts of the CP gene in aphid nymphs reared on the transgenic plants. Decreased expression of the CP gene resulted in a noticeable decline in aphid fecundity on the transgenic Arabidopsis plants. CONCLUSION: Increasing genetic resistance is the only sustainable way of minimizing the use of toxic agrochemicals to protect plants. Host-mediated RNAi of important insect genes has been proposed as a potential avenue for developing crop resistance against insect pests. This study demonstrated the potential of MyCP dsRNA in developing RNAi-based resistance to M. persicae. RNAi-mediated resistance is expected to be more durable compared with other transgenic strategies.  2018 Society of Chemical Industry.',{'entities': [(43,60,'Gene'),(142,159,'Species'),(161,175,'Species'),(639,656,'Gene'),(658,660,'Gene'),(777,779,'Gene'),(918,920,'Gene'),(924,935,'Species'),(994,1005,'Species'),(1041,1045,'Gene'),(1289,1291,'Gene'),(1374,1376,'Gene'),(1452,1463,'Species'),(1785,1789,'Gene'),(1835,1846,'Species')]}),('Identification of chemosensory genes from the antennal transcriptome of Indian meal moth Plodia interpunctella. Olfaction plays an indispensable role in mediating insect behavior, such as locating host plants, mating partners, and avoidance of toxins and predators. Olfactory-related proteins are required for olfactory perception of insects. However, very few olfactory-related genes have been reported in Plodia interpunctella up to now. In the present study, we sequenced the antennae transcriptome of P. interpunctella using the next-generation sequencing technology, and identified 117 candidate olfactory-related genes, including 29 odorant-binding proteins (OBPs), 15 chemosensory proteins (CSPs), three sensory neuron membrane proteins (SNMPs), 47 odorant receptors (ORs), 14 ionotropic receptors (IRs) and nine gustatory receptors (GRs). Further analysis of qRT-PCR revealed that nine OBPs, three CSPs, two SNMPs, nine ORs and two GRs were specifically expressed in the male antennae, whereas eight OBPs, six CSPs, one SNMP, 16 ORs, two GRs and seven IRs significantly expressed in the female antennae. Taken together, our results provided useful information for further functional studies on insect genes related to recognition of pheromone and odorant, which might be meaningful targets for pest management.',{'entities': [(72,88,'Species'),(89,110,'Species'),(407,428,'Species'),(505,522,'Species'),(639,663,'Gene'),(665,669,'Gene'),(675,696,'Gene'),(698,702,'Gene'),(711,743,'Gene'),(745,750,'Gene'),(756,772,'Gene'),(775,778,'Gene'),(784,803,'Gene'),(806,809,'Gene'),(820,838,'Gene'),(841,844,'Gene'),(894,898,'Gene'),(906,910,'Gene'),(916,921,'Gene'),(928,931,'Gene'),(940,943,'Gene'),(1008,1012,'Gene'),(1018,1022,'Gene'),(1037,1040,'Gene'),(1046,1049,'Gene'),(1060,1063,'Gene')]}),('Functional analysis by RNAi of an glutaredoxin gene in Helicoverpa armigera. Glutaredoxins play crucial roles in maintaining intracellular redox homeostasis via scavenging of excess reactive oxygen species. In this study, a glutaredoxin domain-containing cysteine-rich gene from Helicoverpa armigera, named HaGdccr, was characterized. Sequence analysis revealed that it contains a glutaredoxin domain and a conserved cysteine and shares high sequence identity with other insect genes. HaGdccr mRNA expression was highest in molting larvae of the 3rd instar and was mainly detected in the central nervous system of larvae and the wings of adults. Quantitative real-time PCR results revealed that the expression of HaGdccr was suppressed at 1 and 6 h and increased at 24 h after the larvae were treated with 4  C and hydrogen peroxide. When the larvae were exposed to 20  C, HaGdccr decreased at 1 h and was induced at 12 and 24 h. HaGdccr transcription level was downregulated at 2 and 12 h and upregulated at 24 h after the adults were exposed to 0  C. However, transcript levels were increased by high temperature in both larvae and adults. After knockdown of HaGdccr by RNA interference, the expression of antioxidant genes, including thioredoxin-like (Trx-like), catalase (CAT), glutathione-S-transferase (GST), thioredoxin reductase (TrxR), and thioredoxin (Trx), was increased, whereas that of thioredoxin peroxidase (Tpx) was decreased. In addition, we found that HaGdccr knockdown enhanced the enzymatic activity of superoxide dismutase and the contents of hydrogen peroxide and ascorbate. Taken together, these results indicate that HaGdccr may play significant roles in protecting organisms against oxidative damage.',{'entities': [(34,46,'Gene'),(55,75,'Species'),(77,89,'Gene'),(224,273,'Gene'),(279,299,'Species'),(307,314,'Gene'),(485,492,'Gene'),(713,720,'Gene'),(873,880,'Gene'),(930,937,'Gene'),(1161,1168,'Gene'),(1237,1253,'Gene'),(1255,1263,'Gene'),(1266,1274,'Gene'),(1276,1279,'Gene'),(1282,1307,'Gene'),(1309,1312,'Gene'),(1315,1336,'Gene'),(1338,1342,'Gene'),(1349,1360,'Gene'),(1362,1365,'Gene'),(1399,1421,'Gene'),(1423,1426,'Gene'),(1470,1477,'Gene'),(1641,1648,'Gene')]}),('Evaluation of potential RNA-interference-target genes to control cotton mealybug, Phenacoccus solenopsis (Hemiptera: Pseudococcuidae). RNA interference (RNAi) of vital insect genes is a potential tool for targeted pest control. However, selection of the right target genes is a challenge because the RNAi efficacy is known to vary among insect species. Cotton mealybug, Phenacoccus solenopsis, is a phloem-feeding economically important crop pest. We evaluated the RNAi of 2 vital genes, Bursicon (PsBur) and V-ATPase (PsV-ATPase) as potential targets in P. solenopsis for its control. PCR fragments of PsBur and PsV-ATPase were amplified using cDNA synthesized from the total RNA. The PCR amplicons were cloned into Potato virus X (PVX) to develop recombinant PVX for the inoculation of Nicotiana tabacum plants for bioassays with healthy P. solenopsis. Reverse-transcription-polymerase chain reaction (RT-PCR) was used to validate the expression of transgenes in the recombinant-PVX-inoculated plants (treated), and suppression of the target genes in the mealybugs exposed to them. The RT-PCR confirmed the expression of transgenes in the treated plants. Mealybug individuals on treated plants either died or showed physical deformities. Further, the population of mealybug was significantly reduced by feeding on N. tabacum expressing RNAi triggers against PsBur and PsV-ATPase. The results conclude that RNAi is activated in P. solenopsis by feeding on N. tabacum expressing RNAi triggering elements of PsBur and PsV-ATPase genes through recombinant PVX vector. Further, V-ATPase and Bursicon genes are potential targets for RNAi-mediated control of P. solenopsis.',{'entities': [(82,104,'Species'),(370,392,'Species'),(488,496,'Gene'),(498,503,'Gene'),(509,517,'Gene'),(519,529,'Gene'),(555,568,'Species'),(603,608,'Gene'),(613,623,'Gene'),(717,731,'Species'),(733,736,'Species'),(761,764,'Species'),(788,805,'Species'),(840,853,'Species'),(981,984,'Species'),(1316,1326,'Species'),(1360,1365,'Gene'),(1370,1380,'Gene'),(1429,1442,'Species'),(1457,1467,'Species'),(1507,1512,'Gene'),(1517,1527,'Gene'),(1554,1557,'Species'),(1575,1583,'Gene'),(1588,1596,'Gene'),(1654,1667,'Species')]}),('Silencing of ecdysone receptor, insect intestinal mucin and sericotropin genes by bacterially produced double-stranded RNA affects larval growth and development in Plutella xylostella and Helicoverpa armigera. RNA interference mediated gene silencing, which is triggered by double-stranded RNA (dsRNA), has become a important tool for functional genomics studies in various systems, including insects. Bacterially produced dsRNA employs the use of a bacterial strain lacking in RNaseIII activity and harbouring a vector with dual T7 promoter sites, which allow the production of intact dsRNA molecules. Here, we report an assessment of the functional relevance of the ecdysone receptor, insect intestinal mucin and sericotropin genes through silencing by dsRNA in two lepidopteran insect pests, Helicoverpa armigera and Plutella xylostella, both of which cause serious crop losses. Oral feeding of dsRNA led to significant reduction in transcripts of the target insect genes, which caused significant larval mortality with various moulting anomalies and an overall developmental delay. We also found a significant decrease in reproductive potential in female moths, with a drop in egg laying and compromised egg hatching from treated larvae as compared to controls. dsRNA was stable in the insect gut and was efficiently processed into small interfering RNAs (siRNAs), thus accounting for the phenotypes observed in the present work. The study revealed the importance of these genes in core insect processes, which are essential for insect development and survival.',{'entities': [(13,30,'Gene'),(50,55,'Gene'),(60,72,'Gene'),(164,183,'Species'),(188,208,'Species'),(668,685,'Gene'),(705,710,'Gene'),(715,727,'Gene'),(795,815,'Species'),(820,839,'Species')]}),('Silencing of CYP6 and APN Genes Affects the Growth and Development of Rice Yellow Stem Borer, Scirpophaga incertulas. UNASSIGNED: RNAi is a powerful tool to target the insect genes involved in host-pest interactions. Key insect genes are the choice for silencing to achieve pest derived resistance where resistance genes are not available in gene pool of host plant. In this study, an attempt was made to determine the effect of dsRNA designed from two genes Cytochrome P450 derivative (CYP6) and Aminopeptidase N (APN) of rice yellow stem borer (YSB) on growth and development of insect. The bioassays involved injection of chemically synthesized 5  FAM labeled 21-nt dsRNA into rice cut stems and allowing the larvae to feed on these stems which resulted in increased mortality and observed growth and development changes in larval length and weight compared with its untreated control at 12-15 days after treatment. These results were further supported by observing the reduction in transcripts expression of these genes in treated larvae. Fluorescence detection in treated larvae also proved that dsRNA was readily taken by larvae when fed on dsRNA treated stems. These results from the present study clearly show that YSB larvae fed on dsRNA designed from Cytochrome P450 and Aminopeptidase N has detrimental effect on larval growth and development. These genes can be deployed to develop YSB resistance in rice using RNAi approach.',{'entities': [(13,17,'Gene'),(22,25,'Gene'),(70,92,'Species'),(94,116,'Species'),(459,474,'Gene'),(487,491,'Gene'),(497,513,'Gene'),(515,518,'Gene'),(523,545,'Species'),(1261,1276,'Gene'),(1281,1297,'Gene')]}),('Expression and characterization of a recombinant i-type lysozyme from the harlequin ladybird beetle Harmonia axyridis. UNASSIGNED: Lysozymes are enzymes that destroy bacterial cell walls by hydrolysing the polysaccharide component of peptidoglycan. In insects, there are two classes of lysozymes, the c-type with muramidase activity and the i-type whose prototypical members from annelids and molluscs possess both muramidase and isopeptidase activities. Many insect genes encoding c-type and i-type lysozymes have been identified during genome and transcriptome analyses, but only c-type lysozymes have been functionally characterized at the protein level. Here we produced one of five i-type lysozymes represented in the immunity-related transcriptome of the invasive harlequin ladybird beetle Harmonia axyridis as recombinant protein. This was the only one containing the serine and histidine residues that are thought to be required for isopeptidase activity. This i-type lysozyme was recombinantly expressed in the yeast Pichia pastoris, but the purified protein was inactive in both muramidase and isopeptidase assays. Transcription and immunofluorescence analysis revealed that this i-type lysozyme is produced in the fat body but is not inducible by immune challenge. These data suggest that i-type lysozymes in insects may have acquired novel and as yet undetermined functions in the course of evolution.',{'entities': [(49,64,'Gene'),(74,99,'Species'),(100,117,'Species'),(493,509,'Gene'),(582,598,'Gene'),(687,703,'Gene'),(770,795,'Species'),(796,813,'Species'),(1300,1316,'Gene')]}),('Silencing the HaAK gene by transgenic plant-mediated RNAi impairs larval growth of Helicoverpa armigera. Insect pests have caused noticeable economic losses in agriculture, and the heavy use of insecticide to control pests not only brings the threats of insecticide resistance but also causes the great pollution to foods and the environment. Transgenic plants producing double-stranded RNA (dsRNA) directed against insect genes have been is currently developed for protection against insect pests. In this study, we used this technology to silence the arginine kinase (AK) gene of Helicoverpa armigera (HaAK), encoding a phosphotransferase that plays a critical role in cellular energy metabolism in invertebrate. Transgenic Arabidopsis plants producing HaAK dsRNA were generated by Agrobacterium-mediated transformation. The maximal mortality rate of 55% was reached when H. armigera first-instar larvae were fed with transgenic plant leaves for 3 days, which was dramatically higher than the 18% mortality recorded in the control group. Moreover, the ingestion of transgenic plants significantly retarded larval growth, and the transcript levels of HaAK were also knocked down by up to 52%. The feeding bioassays further indicated that the inhibition efficiency was correlated with the integrity and concentration of the produced HaAK dsRNA in transgenic plants. These results strongly show that the resistance to H. armigera was improved in transgenic Arabidopsis plants, suggesting that the RNAi targeting of AK has the potential for the control of insect pests.',{'entities': [(14,18,'Gene'),(83,103,'Species'),(553,568,'Gene'),(570,572,'Gene'),(582,602,'Species'),(604,608,'Gene'),(726,737,'Species'),(755,759,'Gene'),(874,885,'Species'),(1152,1156,'Gene'),(1333,1337,'Gene'),(1417,1428,'Species'),(1456,1467,'Species'),(1514,1516,'Gene')]}),('The expression analysis of silk gland-enriched intermediate-size non-coding RNAs in silkworm Bombyx mori. Small non-protein coding RNAs (ncRNAs) play important roles in development, stress response and other cellular processes. Silkworm is an important model for studies on insect genetics and control of Lepidopterous pests. We have previously identified 189 novel intermediate-size ncRNAs in silkworm Bombyx mori, including 40 ncRNAs that showed altered expression in different developmental stages. Here we characterized the functions of these 40 ncRNAs by measuring their expressions in six tissues of the fifth instar larvae using Northern blot and real-time polymerase chain reaction assays. We identified nine ncRNAs (four small nucleolar RNAs and five unclassified ncRNAs) that were enriched in silk gland, including four ncRNAs that showed silk gland-specific expression. We further showed that three of nine silk gland-enriched ncRNAs were predominantly expressed in the anterior silk gland, whereas another three ncRNAs were highly accumulated in the posterior silk gland, suggesting that they may play different roles in fibroin synthesis. Furthermore, an unclassified ncRNA, Bm-152, exhibited converse expression pattern with its antisense host gene gartenzwerg in diverse tissues, and might regulate the expression of gartenzwerg through RNA-protein complex. In addition, two silk gland-enriched ncRNAs Bm-102 and Bm-159 can be found in histone modification complex, which indicated that they might play roles through epigenetic modifications. Taken together, we provided the first expression and preliminary functional analysis of silk gland-enriched ncRNAs, which will help understand the molecular mechanism of silk gland-development and fibroin synthesis.',{'entities': [(84,92,'Species'),(93,104,'Species'),(228,236,'Species'),(394,402,'Species'),(403,414,'Species'),(1263,1274,'Gene'),(1332,1343,'Gene')]}),('Molecular cloning and functional expression of a Drosophila receptor for the neuropeptides capa-1 and -2. The Drosophila Genome Project website contains an annotated gene (CG14575) for a G protein-coupled receptor. We cloned this receptor and found that the cloned cDNA did not correspond to the annotated gene; it partly contained different exons and additional exons located at the 5( )-end of the annotated gene. We expressed the coding part of the cloned cDNA in Chinese hamster ovary cells and found that the receptor was activated by two neuropeptides, capa-1 and -2, encoded by the Drosophila capability gene. Database searches led to the identification of a similar receptor in the genome from the malaria mosquito Anopheles gambiae (58% amino acid residue identities; 76% conserved residues; and 5 introns at identical positions within the two insect genes). Because capa-1 and -2 and related insect neuropeptides stimulate fluid secretion in insect Malpighian (renal) tubules, the identification of this first insect capa receptor will advance our knowledge on insect renal function.',{'entities': [(49,59,'Species'),(91,104,'Gene'),(110,120,'Species'),(172,179,'Gene'),(467,482,'Species'),(559,572,'Gene'),(589,599,'Species'),(706,722,'Species'),(723,740,'Species'),(876,889,'Gene')]}),('Cloning and sequence of the gene encoding the muscle fatty acid binding protein from the desert locust, Schistocerca gregaria. Muscle fatty acid binding protein (FABP) is a major cytosolic protein in flight muscle of the desert locust, Schistocerca gregaria. FABP expression varies greatly during development and periods of increased fatty acid utilization, but the molecular mechanisms that regulate its expression are not known. In this study, the gene coding for locust muscle FABP was amplified by PCR and cloned, together with 1.2 kb of upstream sequence. The sequence coding for the 607 bp cDNA is interrupted by two introns of 12.7 and 2.9 kb, inserted in analogous positions as the first and third intron of the mammalian homologues. Both introns contain repetitive sequences also found in other locust genes, and the second intron contains a GT-microsatellite. The promoter sequence includes a canonical TATA box 24 bp upstream of the transcription start site. The upstream sequence contains various potential myocyte enhancer sequences and a 160 bp segment that is repeated three times. In database searches in the genome database of Drosophila melanogaster, a gene with the same gene organization and promoter structure was identified, likely the dipteran homologue of muscle FABP. Upstream of both insect genes, a conserved 19 bp inverted repeat sequence was detected. A similar but reverse palindrome is also present upstream of all mammalian heart FABP genes, possibly representing a novel element involved in muscle FABP expression.',{'entities': [(89,102,'Species'),(104,125,'Species'),(134,160,'Gene'),(162,166,'Gene'),(221,234,'Species'),(236,257,'Species'),(259,263,'Gene'),(480,484,'Gene'),(720,729,'Species'),(1144,1167,'Species'),(1287,1291,'Gene'),(1446,1455,'Species'),(1462,1466,'Gene'),(1531,1535,'Gene')]}),('Anopheles stephensi Dox-A2 shares common ancestry with genes from distant groups of eukaryotes encoding a 26S proteasome subunit and is in a conserved gene cluster. The sequence of a cloned Anopheles stephensi gene showed 72% inferred amino acid identity with Drosophila melanogaster Dox-A2 and 93% with its putative ortholog in Anopheles gambiae. Dox-A2 is the reported but herein disputed structural locus for diphenol oxidase A2. Database searches identified Dox-A2 related gene sequences from 15 non-insect species from diverse groups. Phylogenetic trees based on alignments of inferred protein sequences, DNA, and protein motif searches and protein secondary structure predictions produced results consistent with expectations for genes that are orthologous. The only inconsistency was that the C-terminus appears to be more primitive in the yeasts than in plants. In mammals, plants, and yeast these genes have been shown to code for a non-ATPase subunit of the PA700 (19S) regulatory complex of 26S proteasome. The analyses indicated that the insect genes contain no divergent structural features, which taken within an appraisal of all available data, makes the reported alternative function highly improbable. A plausible additional role, in which the 26S proteasome is implicated in regulation of phenol oxidase, would also apply to at least the mammalian genes. No function has yet been reported for the other included sequences. These were from genome projects and included Caenorhabiditus elegans, Arabidopsis thaliana, Fugu rubripes, and Toxoplasma gondii. A consensus of the results predicts a protein containing exceptionally long stretches of helix with a hydrophilic C-terminus. Phosphorylation site motifs were identified at two conserved positions. Possible SRY and GATA-1 binding motifs were found at conserved positions upstream of the mosquito genes. The location of A. stephensi Dox-A2 was determined by in situ hybridization at 34D on chromosome arm 3R. It is in a conserved gene cluster with respect to the other insects. However, the A. stephensi cluster contains a gene showing significant sequence identity to human and pigeon carnitine acetyltransferase genes, therefore showing divergence with the distal end of the D. melanogaster cluster.',{'entities': [(0,19,'Species'),(20,26,'Gene'),(190,209,'Species'),(260,283,'Species'),(284,290,'Gene'),(329,346,'Species'),(348,354,'Gene'),(462,468,'Gene'),(847,853,'Species'),(894,899,'Species'),(1307,1321,'Gene'),(1356,1365,'Species'),(1486,1509,'Species'),(1511,1531,'Species'),(1533,1546,'Species'),(1552,1569,'Species'),(1890,1902,'Species'),(1903,1909,'Gene'),(2061,2073,'Species'),(2139,2144,'Species'),(2156,2183,'Gene'),(2247,2262,'Species')]}),('Conserved promoter elements in the CYP6B gene family suggest common ancestry for cytochrome P450 monooxygenases mediating furanocoumarin detoxification. Despite the fact that Papilio glaucus and Papilio polyxenes share no single hostplant species, both species feed to varying extents on hostplants that contain furanocoumarins. P. glaucus contains two nearly identical genes, CYP6B4v2 and CYP6B5v1, and P. polyxenes contains two related genes, CYP6B1v3 and CYP6B3v2. Except for CYP6B3v2, the substrate specificity of which has not yet been defined, each of the encoded cytochrome P450 monooxygenases (P450s) metabolizes an array of linear furanocoumarins. All four genes are transcriptionally induced in larvae by exposure to the furanocoumarin xanthotoxin; several are also induced by other furanocoumarins. Comparisons of the organizational structures of these genes indicate that all have the same intron/exon arrangement. Sequences in the promoter regions of the P. glaucus CYP6B4v2/CYP6B5v1 genes and the P. polyxenes CYP6B3v2 gene are similar but not identical to the -146 to -97 region of CYP6B1v3 gene, which contains a xanthotoxin-responsive element (XRE-xan) important for basal and xanthotoxin-inducible transcription of CYP6B1v3. Complements of the xenobiotic-responsive element (XRE-AhR) in the dioxin-inducible human and rat CYP1A1 genes also exist in all four promoters, suggesting that these genes may be regulated by dioxin. Antioxidant-responsive elements (AREs) in mouse and rat glutathione S-transferase genes and the Barbie box element (Bar) in the bacterial CYP102 gene exist in the CYP6B1v3, CYP6B4v2, and CYP6B5v1 promoters. Similarities in the protein sequences, intron positions, and xanthotoxin- and xenobiotic-responsive promoter elements indicate that these insect CYP6B genes are derived from a common ancestral gene. Evolutionary comparisons between these P450 genes are the first available for a group of insect genes transcriptionally regulated by hostplant allelochemicals and provide insights into the process by which insects evolve specialized feeding habits.',{'entities': [(35,40,'Gene'),(81,111,'Gene'),(175,190,'Species'),(195,212,'Species'),(329,339,'Species'),(377,385,'Gene'),(390,398,'Gene'),(404,416,'Species'),(445,453,'Gene'),(458,466,'Gene'),(479,487,'Gene'),(570,600,'Gene'),(968,978,'Species'),(979,987,'Gene'),(988,996,'Gene'),(1011,1023,'Species'),(1024,1032,'Gene'),(1097,1105,'Gene'),(1233,1241,'Gene'),(1297,1300,'Gene'),(1326,1331,'Species'),(1336,1339,'Species'),(1340,1346,'Gene'),(1485,1490,'Species'),(1495,1498,'Species'),(1606,1614,'Gene'),(1616,1624,'Gene'),(1630,1638,'Gene'),(1795,1800,'Gene')]}),('Primary structure of apolipophorin-III from the migratory locust, Locusta migratoria. Potential amphipathic structures and molecular evolution of an insect apolipoprotein. The amino acid sequence of an insect apolipoprotein, apolipophorin-III from Locusta migratoria, has been deduced from the sequence of its cloned cDNA. The mature hemolymph protein consists of 161 amino acids. Optimized alignments of this protein with apolipophorin-III from the tobacco hornworm, Manduca sexta, disclosed an overall sequence identity of only 29%, even though the two proteins are functionally equivalent. The L. migratoria sequence is composed of 12 repeating peptides that are variable in length. Six amphipathic helical segments of varying length were identified in each protein using a newly described algorithm for detecting such secondary structures. The degree of sequence identity between the two insect apoproteins is considerably less than that observed among orthologous mammalian apolipoproteins. However, calculation of the rates of synonymous and nonsynonymous nucleotide substitutions indicates that the insect genes may be evolving at rates similar to the mammalian apolipoprotein genes. Further comparative analyses of insect and mammalian apolipoproteins should provide insights about the limits of sequence diversity tolerated by their predicted amphipathic helical domains.',{'entities': [(21,38,'Gene'),(48,64,'Species'),(66,84,'Species'),(225,242,'Gene'),(248,266,'Species'),(423,440,'Gene'),(450,466,'Species'),(468,481,'Species'),(597,610,'Species'),(969,978,'Species'),(1159,1168,'Species'),(1234,1243,'Species')]}),('Translational and transcriptional control elements in the untranslated leader of the heat-shock gene hsp22. Downstream of the transcription start site in the Drosophila heat-shock gene hsp22, we have identified a region that is necessary for efficient transcription, and also for selective translation during heat shock. We assayed the expression of mutated genes after P-factor-mediated insertion into the genome. Deletions within the first 26 nucleotides block the preferential translation of hsp22 mRNA at high temperature. The rate of transcription is also decreased, though transcription is still heat-inducible. Leaving this region intact, up to 86% of the leader can be deleted without affecting translation or transcription. The functional region coincides with a region of sequence homology between the heat-shock genes. Only partial homology is found to a conserved sequence, ATCAGTTCT, found at the very 5  end of other insect genes.',{'entities': [(101,106,'Gene'),(158,168,'Species'),(185,190,'Gene'),(495,500,'Gene')]}),('Functional characterization of cis-acting elements mediating flavone-inducible expression of CYP321A1. How plant allelochemicals elicit herbivore counterdefense genes remains largely unknown. To define the cis-acting elements for flavone inducibility of the allelochemical-metabolizing CYP321A1 from Helicoverpa zea, functions of varying length of CYP321A1 promoter are examined in H. zea fatbody cells. Progressive 3  deletions reveal presence of positive elements in the 5  untranslated region (UTR). Progressive 5  deletions map out regions of one essential element, four enhancers, and two silencers. Further progressive 5 deletions localize the essential element to a 36-bp region from -109 to -74. This essential element, designated as xenobiotic response element to flavone (XRE-Fla), contains a 5  AT-only TAAT inverted repeat, a GCT mirror repeat and a 3  antioxidant response element-like element. Internal deletions and substitution mutations show that the TAAT repeat is only necessary for the maximal flavone inducibility, whereas the other two components are necessary for the basal and flavone-induced expression of CYP321A1. Electrophoresis mobility shift assays demonstrate that XRE-Fla specifically binds to H. zea fatbody cell nuclear extracts and flavone treatment increases the nuclear concentrations of the yet-to-be characterized transcription factors binding to XRE-Fla. Taken together, CYP321A1 expression is regulated primarily by XRE-Fla and secondarily by other cis elements scattered in its promoter and 5  UTR.',{'entities': [(93,101,'Gene'),(286,294,'Gene'),(300,315,'Species'),(348,356,'Gene'),(1131,1139,'Gene'),(1411,1419,'Gene')]}),('Molecular cloning of a multidomain cysteine protease and protease inhibitor precursor gene from the tobacco hornworm (Manduca sexta) and functional expression of the cathepsin F-like cysteine protease domain. A Manduca sexta (tobacco hornworm) cysteine protease inhibitor, MsCPI, purified from larval hemolymph has an apparent molecular mass of 11.5 kDa, whereas the size of the mRNA is very large (  9 kilobases). MsCPI cDNA consists of a 9,273 nucleotides that encode a polypeptide of 2,676 amino acids, which includes nine tandemly repeated MsCPI domains, four cystatin-like domains and one procathepsin F-like domain. The procathepsin F-like domain protein was expressed in Escherichia coli and processed to its active mature form by incubation with pepsin. The mature enzyme hydrolyzed Z-Leu-Arg-MCA, Z-Phe-Arg-MCA and Boc-Val-Leu-Lys-MCA rapidly, whereas hydrolysis of Suc-Leu-Tyr-MCA and Z-Arg-Arg-MCA was very slow. The protease was strongly inhibited by MsCPI, egg-white cystatin and sunflower cystatin with K(i) values in the nanomolar range. When the MsCPI tandem protein linked to two MsCPI domains was treated with proteases, it was degraded by the cathepsin F-like protease. However, tryptic digestion converted the MsCPI tandem protein to an active inhibitory form. These data support the hypothesis that the mature MsCPI protein is produced from the MsCPI precursor protein by trypsin-like proteases. The resulting mature MsCPI protein probably plays a role in the regulation of the activity of endogenous cysteine proteases.',{'entities': [(35,52,'Gene'),(100,116,'Species'),(118,131,'Species'),(211,224,'Species'),(226,242,'Species'),(244,271,'Gene'),(273,278,'Gene'),(415,420,'Gene'),(544,549,'Gene'),(963,968,'Gene'),(1062,1067,'Gene'),(1097,1102,'Gene'),(1230,1235,'Gene'),(1331,1336,'Gene'),(1366,1371,'Gene'),(1438,1443,'Gene')]}),('Molecular diversity of PBAN family peptides from fire ants. The PBAN/Pyrokinin peptide family is a major neuropeptide family characterized with a common FXPRLamide in the C-termini. These peptides are ubiquitously distributed in the Insecta and are involved in many essential endocrinal functions, e.g., pheromone production. Previous work demonstrated the localization of PBAN in the fire ant central nervous system, and identified a new family of PBAN from the red imported fire ant, Solenopsis invicta. In this study, we identified five more PBAN/Pyrokinin genes from S. geminata, S. richteri, S. pergandii, S. carolinensis, and a hybrid of S. invicta and S. richteri. The gene sequences were used to determine the phylogenetic relationships of these species and hybrid, which compared well to the morphologically defined fire ant subgroup complexes. The putative PBAN and other peptides were determined from the amino acid sequences of the PBAN/pyrokinin genes. We summarized all known insect PBAN family neuropeptides, and for the first time constructed a phylogenetic tree based on the full amino acid sequences translated from representative PBAN cDNAs. The PBAN/pyrokinin gene is well conserved in Insecta and probably extends into the Arthropod phylum; however, translated pre-propeptides may vary and functional diversity may be retained, lost, or modified during the evolutionary process.',{'entities': [(23,27,'Gene'),(64,68,'Gene'),(69,78,'Gene'),(373,377,'Gene'),(449,453,'Gene'),(463,484,'Species'),(486,504,'Species'),(545,549,'Gene'),(550,559,'Gene'),(571,582,'Species'),(584,595,'Species'),(597,609,'Species'),(611,626,'Species'),(644,654,'Species'),(659,670,'Species'),(867,871,'Gene'),(944,948,'Gene'),(949,958,'Gene'),(997,1001,'Gene'),(1149,1153,'Gene'),(1165,1169,'Gene'),(1170,1179,'Gene')]}),('The cys-loop ligand-gated ion channel gene superfamily of the parasitoid wasp, Nasonia vitripennis. Members of the cys-loop ligand-gated ion channel (cysLGIC) superfamily mediate chemical neurotransmission and are studied extensively as potential targets of drugs used to treat neurological disorders, such as Alzheimer s disease. Insect cys-loop LGICs also have central roles in the nervous system and are targets of highly successful insecticides. Here, we describe the cysLGIC superfamily of the parasitoid wasp, Nasonia vitripennis, which is emerging as a highly useful model organism and is deployed as a biological control of insect pests. The wasp superfamily consists of 26 genes, which is the largest insect cysLGIC superfamily characterized, whereas Drosophila melanogaster, Apis mellifera and Tribolium castaneum have 23, 21 and 24, respectively. As with Apis, Drosophila and Tribolium, Nasonia possesses ion channels predicted to be gated by acetylcholine, gamma-amino butyric acid, glutamate and histamine, as well as orthologues of the Drosophila pH-sensitive chloride channel (pHCl), CG8916 and CG12344. Similar to other insects, wasp cysLGIC diversity is broadened by alternative splicing and RNA A-to-I editing, which may also serve to generate species-specific receptor isoforms. These findings on N. vitripennis enhance our understanding of cysLGIC functional genomics and provide a useful basis for the study of their function in the wasp model, as well as for the development of improved insecticides that spare a major beneficial insect species.',{'entities': [(4,37,'Gene'),(79,98,'Species'),(115,148,'Gene'),(150,157,'Gene'),(338,351,'Gene'),(472,479,'Gene'),(516,535,'Species'),(717,724,'Gene'),(760,783,'Species'),(785,799,'Species'),(804,823,'Species'),(872,882,'Species'),(887,896,'Species'),(898,905,'Species'),(1050,1060,'Species'),(1061,1090,'Gene'),(1092,1096,'Gene'),(1099,1105,'Gene'),(1110,1117,'Gene'),(1150,1157,'Gene'),(1316,1330,'Species'),(1360,1367,'Gene')]}),('Expression of a novel member of the odorant-binding protein gene family in Culex nigripalpus (Diptera: Culicidae). We previously showed that gene expression in the midgut of female Culex nigripalpus Theobald (Diptera: Culicidae) was altered after ingestion of a bloodmeal and some of the expressed cDNAs showed stage-, sex-, and tissue-specific expression. One of these expressed cDNAs, CN G11B, is here shown to be similar to members of a gene family that encodes odorant-binding proteins. CN G11B.1 is a cDNA fragment of 721 bp, is incomplete at the 5 -end, and contains a canonical polyadenylic acid tail signal sequence in the 103-bp 3 -untranslated region. Translation of CN G11B.1 provides a putative protein product of 207 amino acids. GenBank tblastx, VectorBase, and Pfam database searches showed identity to hypothetical proteins from Culex pipiens quinquefasciatus Say (86%) and Aedes aegypti (L.) (55%). These proteins have structural motifs similar to those found in the gene family that includes odorant-binding proteins. CN G11B.1 putative protein has two of the six cysteines that are highly conserved in most invertebrate odorant-binding proteins. CN G11B.1 expression increased in thoraces (6-12 h) and in abdomens (3-48 h) of blood fed female Cx. nigripalpus but was not detected in RNA from heads, indicating a possible role in both chemoreception and blood feeding.',{'entities': [(36,59,'Gene'),(75,92,'Species'),(181,198,'Species'),(845,875,'Species'),(890,903,'Species'),(1010,1033,'Gene'),(1139,1162,'Gene'),(1262,1277,'Species')]}),('The beta2-tubulin gene from three tephritid fruit fly species and use of its promoter for sperm marking. To isolate testis-specific regulatory DNA that could be used in genetically transformed insect pest species to improve their biological control, beta2-tubulin genes and their proximal genomic DNA were isolated from three economically important tephritid pest species, Anastrepha suspensa, Anastrepha ludens, and Bactrocera dorsalis. Gene isolation was first attempted by degenerate PCR on an A. suspensa adult male testes cDNA library, which fortuitously isolated the 2.85 kb beta1-tubulin gene that encodes a 447 amino acid polypeptide. Subsequent PCR using 5  and 3  RACE generated the 1.4 kb Asbeta2-tubulin gene that encodes a 446 amino acid polypeptide. Using primers to conserved sequences, the highly similar A. ludens and B. dorsalis beta2-tubulin genes, encoding identical amino acid sequences, were then isolated. To verify Asbeta2-tubulin gene identification based on gene expression, qRT-PCR showed that Asbeta2-tubulin transcript was most abundant in pupal and adult males, and specific to the testes. This was further tested in transformants having the DsRed.T3 reporter gene regulated by the Asbeta2-tubulin 1.3 kb promoter region. Fluorescent protein was specifically expressed in testes from third instar larvae to adults, and fluorescent sperm could be detected in the spermathecae of non-transgenic females mated to transgenic males.To confirm these matings, a PCR protocol was developed specific to the transgenic sperm DNA.',{'entities': [(4,17,'Gene'),(44,53,'Species'),(250,263,'Gene'),(373,392,'Species'),(394,411,'Species'),(417,436,'Species'),(497,508,'Species'),(700,715,'Gene'),(821,830,'Species'),(835,846,'Species'),(847,860,'Gene'),(939,954,'Gene'),(1021,1036,'Gene'),(1212,1227,'Gene')]}),('Drosophila asterless and vertebrate Cep152 Are orthologs essential for centriole duplication. The centriole is the core structure of centrosome and cilium. Failure to restrict centriole duplication to once per cell cycle has serious consequences and is commonly observed in cancer. Despite its medical importance, the mechanism of centriole formation is poorly understood. Asl was previously reported to be a centrosomal protein essential for centrosome function. Here we identify mecD, a severe loss-of-function allele of the asl gene, and demonstrate that it is required for centriole and cilia formation. Similarly, Cep152, the Asl ortholog in vertebrates, is essential for cilia formation and its function can be partially rescued by the Drosophila Asl. The study of Asl localization suggests that it is closely associated with the centriole wall, but is not part of the centriole structure. By analyzing the biogenesis of centrosomes in cells depleted of Asl, we found that, while pericentriolar material (PCM) function is mildly affected, Asl is essential for daughter centriole formation. The clear absence of several centriolar markers in mecD mutants suggests that Asl is critical early in centriole duplication.',{'entities': [(0,10,'Species'),(11,20,'Gene'),(36,42,'Gene'),(373,376,'Gene'),(527,530,'Gene'),(619,625,'Gene'),(631,634,'Gene'),(753,756,'Gene'),(771,774,'Gene'),(960,963,'Gene'),(1045,1048,'Gene'),(1174,1177,'Gene')]}),('New candidate genes for sex-comb divergence between Drosophila mauritiana and Drosophila simulans. A large-effect QTL for divergence in sex-comb tooth number between Drosophila simulans and D. mauritiana was previously mapped to 73A-84AB. Here we identify genes that are likely contributors to this divergence. We first improved the mapping resolution in the 73A-84AB region using 12 introgression lines and 62 recombinant nearly isogenic lines. To further narrow the list of candidate genes, we assayed leg-specific expression and identified genes with transcript-level evolution consistent with a potential role in sex-comb divergence. Sex combs are formed on the prothoracic (front) legs, but not on the mesothoracic (middle) legs of Drosophila males. We extracted RNA from the prothoracic and mesothoracic pupal legs of two species to determine which of the genes expressed differently between leg types were also divergent for gene expression. Two good functional candidate genes, Scr and dsx, are located in one of our fine-scale QTL regions. In addition, three previously uncharacterized genes (CG15186, CG2016, and CG2791) emerged as new candidates. These genes are located in regions strongly associated with sex-comb tooth number differences and are expressed differently between leg tissues and between species. Further supporting the potential involvement of these genes in sex-comb divergence, we found a significant difference in sex-comb tooth number between co-isogenic D. melanogaster lines with and without P-element insertions at CG2791.',{'entities': [(78,97,'Species'),(166,185,'Species'),(190,203,'Species'),(986,989,'Gene'),(994,997,'Gene'),(1102,1109,'Gene'),(1111,1117,'Gene'),(1123,1129,'Gene'),(1486,1501,'Species'),(1549,1555,'Gene')]}),('Analysis of the insect os-d-like gene family. Insect OS-D-like proteins, also known as chemosensory (CSP) or sensory appendage proteins (SAP), are broadly expressed in various insect tissues, where they are thought to bind short to medium chain length fatty acids and their derivatives. Although their specific function remains uncertain, OS-D-like members have been isolated from sensory organs (including the sensillum lymph in some cases), and a role in olfaction similar to that of the insect odorant binding proteins (OBP) has been suggested for some. We have identified 15 new OS-D-like sequences: four from cDNA clones described herein and 11 from sequence databases. The os-d-like genes from the Anopheles gambiae, Apis mellifera, Drosophila melanogaster, and Drosophila pseudoobscura genomes typically have single, small introns with a conserved splice site. Together with all family members entered on GenBank, a total of 70 OS-D-like proteins, representing the insect orders Diptera, Dictyoptera, Hymenoptera, Lepidoptera, Orthoptera, and Phasmatodea, were analyzed. A neighbor joining distance phenogram identified several protein similarity classes that were characterized by highly conserved sequence motifs, including (A) N-terminal YTTKYDN(V/I)(N/D)(L/V)DEIL, (B) central DGKELKXX(I/L)PDAL, and (C) C-terminal KYDP. In contrast, three similarity classes were characterized by their diversion from these conserved motifs. The functional importance of conserved amino acid residues is discussed in relation to the crystal and NMR structures of MbraCSPA6.',{'entities': [(23,27,'Gene'),(53,57,'Gene'),(109,134,'Gene'),(137,140,'Gene'),(339,343,'Gene'),(497,520,'Gene'),(523,526,'Gene'),(583,587,'Gene'),(679,683,'Gene'),(704,721,'Species'),(723,737,'Species'),(739,762,'Species'),(768,792,'Species'),(935,939,'Gene'),(995,1006,'Species')]}),('Cloning of a putative Bombyx mori TFIIB-related factor (BRF). To identify the protein domains responsible for its conserved and specialized functions, putative TFIIB-Related Factor (BRF) from the silkworm (Bombyx mori) was compared with BRFs from other organisms. The Bombyx BRF coding region was assembled from three separate and overlapping cDNA fragments. Fragments encoding the middle portion and the 3  end were discovered in the Bombyx mori Genome Project "Silkbase" collection through sequence homology with human BRF1, and the fragment encoding the N-terminus was isolated in our laboratory using the 5  RACE method. Southern analysis showed that silkworm BRF is encoded by a single-copy gene. Bombyx BRF contains the following domains that have been noted in all other BRFs, and that are likely, therefore, to provide highly conserved functions: a zinc finger domain, an imperfect repeat, three "BRF Homology" domains, and an acidic domain at the C-terminus. As expected from the evolutionary relationships among insects and mammals, Bombyx BRF is more similar overall to Drosophila BRF (55% identical) than to human BRF1 (42% identical). Detailed examination of individual domains reveals a remarkable exception, however. Domain II of Bombyx BRF is more similar to its human counterpart than to Drosophila Domain II. This result indicates that Domain II has undergone unusual divergence in Drosophila, and suggests a structural basis for Drosophila BRF s unique pattern of interaction with other transcription factors.',{'entities': [(22,54,'Gene'),(56,59,'Gene'),(182,185,'Gene'),(196,204,'Species'),(206,217,'Species'),(275,278,'Gene'),(435,446,'Species'),(515,520,'Species'),(521,525,'Gene'),(655,663,'Species'),(664,667,'Gene'),(709,712,'Gene'),(905,908,'Gene'),(1050,1053,'Gene'),(1081,1091,'Species'),(1092,1095,'Gene'),(1120,1125,'Species'),(1126,1130,'Gene'),(1252,1255,'Gene'),(1279,1284,'Species'),(1305,1315,'Species'),(1400,1410,'Species'),(1448,1458,'Species'),(1459,1462,'Gene')]}),('Structure and evolution of the luciferin-regenerating enzyme (LRE) gene from the firefly Photinus pyralis. To study the structural features of genes for the luciferin-regenerating enzyme (LRE), the entire gene along with 524 bp of upstream sequence was determined from Photinus pyralis (Coleoptera: Lampyridae). The LRE gene revealed an open reading frame composed of five exons divided by four introns ranging in size from 47 to 904 bp. The deduced LRE amino acid sequence showed identity to senescence marker protein-30 (SMP30) from a number of insects and mammals including four putative SMP30 sequences from Anopheles gambiae. Gene structure comparisons showed some intron/exon site conservation with A. gambiae and mammalian SMP30 proteins but not Drosophila. LRE and luciferase sequence comparisons revealed two conserved putative luciferin-binding sites. The evolution of LRE was discussed in relation to its function.',{'entities': [(31,60,'Gene'),(89,105,'Species'),(157,186,'Gene'),(269,285,'Species'),(493,521,'Gene'),(523,528,'Gene'),(591,596,'Gene'),(612,629,'Species'),(720,729,'Species'),(730,735,'Gene'),(753,763,'Species')]}),('The AmCREB gene is an ortholog of the mammalian CREB/CREM family of transcription factors and encodes several splice variants in the honeybee brain. The transcription factor CREB (cAMP response element binding protein) is required for the switch from short-term to long-term synaptic plasticity and from short-term to long-term memory. Its activity is regulated by the cAMP-dependent signalling cascade, which has been shown to play a crucial role in the honeybee s long-term memory formation. To elucidate the role of the CREB in honeybee memory formation we analysed a CREB-homologous gene, AmCREB, which is expressed as several transcripts in the honeybee brain. Eight transcripts have been identified (AmCREB 1-8) that are generated by alternate splicing. One antibody generated against a subset of these variants reveals a cytosolic localization in the mushroom body alpha-lobes, the glomeruli of the antennal lobes, the protocerebral lobes, the central complex and in the optical lobes.',{'entities': [(4,10,'Gene'),(38,47,'Species'),(48,52,'Gene'),(133,141,'Species'),(174,178,'Gene'),(180,217,'Gene'),(455,463,'Species'),(523,527,'Gene'),(531,539,'Species'),(571,575,'Gene'),(593,599,'Gene'),(650,658,'Species'),(706,712,'Gene')]}),('B96Bom encodes a Bombyx mori tyramine receptor negatively coupled to adenylate cyclase. A cDNA encoding a biogenic amine receptor (B96Bom) was isolated from silkworm (Bombyx mori) larvae, and the ligand response of the receptor stably expressed in HEK-293 cells was examined. Tyramine (TA) at 0.1-100 micro m reduced forskolin (10 micro m)-stimulated intracellular cAMP levels by approximately 40%. The inhibitory effect of TA at 1 micro m was abolished by yohimbine and chlorpromazine (each 10 micro m). Although octopamine (OA) also reduced the cAMP levels, the potency was at least two orders of magnitude lower than that of TA. Furthermore, unlabelled TA (IC50 = 5.2 nm) inhibited specific [3H]TA binding to the membranes of B96Bom-transfected HEK-293 cells more potently than did OA (IC50 = 1.4 micro m) and dopamine (IC50 = 1.7 micro m). Taken together with the result of phylogenetic analysis, these findings indicate that the B96Bom receptor is a B. mori TA receptor, which is negatively coupled to adenylate cyclase. The use of this expression system should facilitate physiological studies of TA receptors as well as structure-activity studies of TA receptor ligands.',{'entities': [(17,28,'Species'),(29,46,'Gene'),(106,129,'Gene'),(157,165,'Species'),(167,178,'Species'),(955,962,'Species')]}),('Cloning and characterization of a 70 kDa heat shock cognate gene (HSC70) from two species of Chironomus. In the present study we carried out the isolation and characterization of an HSC70 gene from two midges, Chironomus tentans and C. yoshimatsui. The HSC70 cDNAs are approximately 2424 (C. tentans) and 2464 bp (C. yoshimatsui) long, and contain 1950 and 1956 bp open reading frames, respectively. Analysis of genomic DNA revealed the presence of two introns in these genes. The 5  untranslated regions of the HSC70 genes are adenosine-rich, a feature found in inducible HSP70 genes. The nucleotide and amino acid sequences exhibit high identity with cytosolic HSC70s from other Dipterans. Northern hybridization indicated that HSC70 is expressed at all developmental stages, from embryo to adult, and Southern hybridization confirmed the presence of multiple HSP70 genes in Chironomus.',{'entities': [(34,59,'Gene'),(66,71,'Gene'),(93,103,'Species'),(182,187,'Gene'),(210,228,'Species'),(233,247,'Species'),(253,258,'Gene'),(289,299,'Species'),(314,328,'Species'),(512,517,'Gene'),(573,578,'Gene'),(663,668,'Gene'),(730,735,'Gene'),(862,867,'Gene'),(877,887,'Species')]}),('CYP6B cytochrome p450 monooxygenases from Papilio canadensis and Papilio glaucus: potential contributions of sequence divergence to host plant associations. Two groups of furanocoumarin-inducible cytochrome p450 genes, the CYP6B4 group and the CYP6B17 group, characterized in two closely related tiger swallowtails, Papilio glaucus and Papilio canadensis, are induced to different extents, with generally higher levels of CYP6B transcripts in P. glaucus. To investigate the evolutionary history of these CYP6B genes in the context of their association with furanocoumarin detoxification, we isolated thirteen CYP6B genes from these species. Each of these genes contains an intron at a conserved position (1334 nucleotides from the translation start site), which varies in length due to three insertion/deletions. The proximal 5  end flanking sequence from the transcription initiation site is highly conserved (91-98% nt identity). The sequence 5  to -640 is significantly variable due largely to the presence of three insertion/deletions. The sequence at the 3  end of this region contains a putative xenobiotic response element to xanthotoxin (XRE-xan), important for basal and xanthotoxin-inducible transcription of the P. polyxenes CYP6B1v3 gene, and multiple elements known to regulate vertebrate phase I and II promoters, including an XRE-AhR (Xenobiotic Response Element to Aryl hydrocarbon Receptor), an OCT-1 element (octamer protein binding site), an ARE (Antioxidant Response Element), an EcRE (Ecdysone Response Element), and an imperfect PXR (Pregnane X Receptor) responsive element (PRE). Our analyses of CYP6B genes in these two species indicate that these genes are in an early stage of divergence and that differential exposure of these two species to chemically distinct host plants resulting from geographical isolation has had functional impacts not only on the coding regions of these genes but also on their promoter regions. Thus, changes in p450 regulation as well as catalytic activity may play a role in the evolution of host plant associations in herbivorous insects.',{'entities': [(0,5,'Gene'),(6,21,'Gene'),(42,60,'Species'),(65,80,'Species'),(196,211,'Gene'),(223,229,'Gene'),(244,251,'Gene'),(296,314,'Species'),(316,331,'Species'),(336,354,'Species'),(422,427,'Gene'),(443,453,'Species'),(504,509,'Gene'),(609,614,'Gene'),(1236,1244,'Gene'),(1619,1624,'Gene')]}),('Genomic organization and regulation of three cecropin genes in Anopheles gambiae. Three cecropin genes (AgCecA-C) were identified from Anopheles gambiae, a major vector for malaria in sub-Saharan Africa. These genes form a cluster with AgCecA and AgCecB positioned in opposite orientation, while AgCecC is downstream of AgCecA in the same direction. One intron is present in each of these three genes. Motif searches of promoter regions revealed elements that could be regulated by the NF-kappaB family of transcriptional regulators. The divergent promoter (1186 nucleotides in length) between CecA and CecB and the promoter for CecC were analysed by transfection in An. gambiae cell lines. Results showed that these promoters were up-regulated by lipopolysaccharide. The activity was further elevated when heat-inactivated microbes were used to challenge the cell line. At least one NF-kappaB site was required for inducible expression of both CecA and CecB.',{'entities': [(45,53,'Gene'),(63,80,'Species'),(88,96,'Gene'),(104,112,'Gene'),(135,152,'Species'),(236,242,'Gene'),(247,253,'Gene'),(296,302,'Gene'),(320,326,'Gene'),(667,678,'Species')]}),('In situ hybridization to the Rdl locus on polytene chromosome 3L of Anopheles stephensi. We are interested in generating a Y-autosome translocation of the Resistance to dieldrin (Rdl) locus in the malaria vector mosquito Anopheles stephensi Liston (Diptera: Culicidae), for use in sterile insect release. To ensure stability of the system, a recombination suppressing inversion can also be induced which encompasses the Rdl locus. As a first step, here we report the cloning of fragments of the Rdl gene from both An. stephensi and An. gambiae Giles using degenerate primers in the polymerase chain reaction. These fragments encode the second membrane-spanning region of the gamma-aminobutyric acid receptor and show high levels of both nucleotide and predicted amino acid identity to other Rdl-like receptors. They confirm that, as in all other arthropod species examined, dieldrin resistance in An. stephensi is associated with replacement of alanine302, in this case with a serine. In situ hybridization of the Rdl probe to polytene chromosomes of An. stephensi localizes the gene to the left arm of chromosome 3 (3L) in region 45C. Rdl localization will enable us to identify chromosomal rearrangements encompassing the Rdl locus and help anchor the genome sequence of An. gambiae to the polytene map.',{'entities': [(68,87,'Species'),(155,177,'Gene'),(221,240,'Species'),(514,527,'Species'),(532,549,'Species'),(897,910,'Species'),(1051,1064,'Species'),(1273,1284,'Species')]}),('Isolation and molecular characterization of Musca domestica delta-9 desaturase sequences. We have isolated fatty acyl-CoA desaturase cDNA (Mdomd9) and genomic sequences from the housefly, Musca domestica. Two approximately 1.66 kb cDNAs were recovered. They had identical coding regions and 3  untranslated regions (UTRs), but differed in their 5  UTRs. The open reading frame encodes a 380 amino acid (aa) protein with 82% identity to Drosophila melanogaster desat1, and significant (> 50%) identity with other insect delta-9 desaturases. Functional analyses in a yeast expression system confirmed the cDNA encodes a delta9 desaturase. Northern analysis indicated two transcripts of 1.7 and 2.9 kb that hybridized specifically to the open reading frame. PCR amplification of genomic templates revealed three intron sites that are conserved among other insect species. Southern analysis of genomic DNA indicated at least two desaturase gene copies per haploid genome. There is a high degree of polymorphism, most of which appears to be due to variable intron sequences; curiously, individual flies had varying morphs of intron II and intron III. Together, the data suggest that there are more delta9 desaturase alleles within the population studied than there are loci within the genome, and support other studies suggesting that insect fatty acyl-CoA desaturases are a dynamically evolving gene family.',{'entities': [(44,59,'Species'),(60,78,'Gene'),(107,132,'Gene'),(188,203,'Species'),(436,459,'Species'),(460,466,'Gene'),(565,570,'Species')]}),('Maelstrom is required to position the MTOC in stage 2-6 Drosophila oocytes. The factors that determine intracellular polarity are largely unknown. In Drosophila oocytes one of the earliest polar events is the positioning of the microtubule-organizing center (MTOC). Here we present data that are consistent with the hypothesis that maelstrom is required for posterior positioning of the MTOC.',{'entities': [(0,9,'Gene'),(56,66,'Species'),(150,160,'Species'),(332,341,'Gene')]}),('Expression of a doublesex homologue is altered in sexual mosaics of Ostrinia scapulalis moths infected with Wolbachia. A homologue of the sex-determining gene doublesex, Osdsx, was identified in the adzuki bean borer Ostrinia scapulalis. Three isoforms of the Osdsx transcript (Osdsx(M), Osdsx(FL) and Osdsx(FS)) differing in length were found. Osdsx(M) was specifically found in males, and contained an 852-bp ORF encoding a protein of 284 amino acids. Osdsx(FL) and Osdsx(FS) were found in females, and had the same 813-bp ORF encoding a protein of 271 amino acids. The Osdsx gene was inferred to have six exons and five introns. The variation in the transcript could be explained by the alternative splicing of Osdsx: Osdsx(M) was formed by the splicing of exons 1, 2, 5 and 6, Osdsx(FS) by the splicing of exons 1-4 and 6, and Osdsx(FL) by the splicing of exons 1-6. RT-PCR analysis indicated that Osdsx was transcribed in a sex-specific manner in all somatic tissues examined, regardless of developmental stage. In Wolbachia-induced sexual mosaics of O. scapulalis, which are genetically male, the female-specific isoform of Osdsx (Osdsx(FL)) was shown to be expressed in addition to the male-specific isoform (Osdsx(M)). This finding provides the first evidence that Wolbachia manipulates the sex of its host by interfering either with the sex-specific splicing of Osdsx itself or with another upstream sex determination process.',{'entities': [(16,25,'Gene'),(68,87,'Species'),(108,117,'Species'),(159,168,'Gene'),(199,210,'Species'),(217,236,'Species'),(1020,1029,'Species'),(1273,1282,'Species')]}),('RNA interference of timeless gene does not disrupt circadian locomotor rhythms in the cricket Gryllus bimaculatus. Molecular studies revealed that autoregulatory negative feedback loops consisting of so-called "clock genes" constitute the circadian clock in Drosophila. However, this hypothesis is not fully supported in other insects and is thus to be examined. In the cricket Gryllus bimaculatus, we have previously shown that period (per) plays an essential role in the rhythm generation. In the present study, we cloned cDNA of the clock gene timeless (tim) and investigated its role in the cricket circadian oscillatory mechanism using RNA interference. Molecular structure of the cricket tim has rather high similarity to those of other insect species. Real-time RT-PCR analysis revealed that tim mRNA showed rhythmic expression in both LD and DD similar to that of per, peaking during the (subjective) night. When injected with tim double-stranded RNA (dstim), tim mRNA levels were significantly reduced and its circadian expression rhythm was eliminated. After the dstim treatment, however, adult crickets showed a clear locomotor rhythm in DD, with a free-running period significantly shorter than that of control crickets injected with Discosoma sp. Red2 (DsRed2) dsRNA. These results suggest that in the cricket, tim plays some role in fine-tuning of the free-running period but may not be essential for oscillation of the circadian clock.',{'entities': [(20,28,'Gene'),(94,113,'Species'),(258,268,'Species'),(378,397,'Species'),(429,435,'Gene'),(547,555,'Gene'),(557,560,'Gene'),(694,697,'Gene'),(799,802,'Gene'),(935,938,'Gene'),(968,971,'Gene'),(1173,1179,'Gene'),(1324,1327,'Gene'),(1379,1385,'Gene')]}),('Cloning and characterization of a Gasp homolog from the spruce budworm, Choristoneura fumiferana, and its putative role in cuticle formation. Proteins that are capable of binding chitin play essential roles in the synthesis and structural integrity of the insect cuticle and peritrophic matrix. In the course of developing expressed sequence tag (EST) libraries for the eastern spruce budworm, Choristoneura fumiferana, we identified an abundant cDNA encoding a homolog of the Drosophila "gasp" gene (Gene Analogous to Small Peritrophins). For the present work, we undertook the characterization of this new homolog, CfGasp, in an effort to identify its role during larval development. As shown for DmGasp, the C. fumiferana homolog was found to contain three type-2 chitin-binding domains (CBDs), which were also found in Gasp orthologs retrieved from GenBank. In a phylogenetic analysis, these Gasp proteins formed a tight cluster, distinct from the midgut-specific peritrophins with which they share the cysteine-containing CBDs so far considered absent from cuticular proteins. However, unlike what has been shown for peritrophins, CfGasp transcript levels were low in larval midguts and most abundant in epidermis, while they were low in trachea and ovaries. Transcript levels increased during larval molts in a pattern similar to that observed for exocuticular proteins in other insects. In addition, the recombinant protein was shown to be capable of binding chitin. Altogether, these results suggest a structural role for CfGasp in exocuticle formation.',{'entities': [(34,38,'Gene'),(72,96,'Species'),(394,418,'Species'),(477,487,'Species'),(489,493,'Gene'),(501,537,'Gene'),(711,724,'Species'),(823,827,'Gene'),(896,900,'Gene')]}),('Evolutionary divergence of the paralogs Methoprene tolerant (Met) and germ cell expressed (gce) within the genus Drosophila. Juvenile hormone (JH) signaling underpins both regulatory and developmental pathways in insects. However, the JH receptor is poorly understood. Methoprene tolerant (Met) and germ cell expressed (gce) have been implicated in JH signaling in Drosophila. We investigated the evolution of Met and gce across 12 Drosophila species and found that these paralogs are conserved across at least 63 million years of dipteran evolution. Distinct patterns of selection found using estimates of dN/dS ratios across Drosophila Met and gce coding sequences, along with their incongruent temporal expression profiles in embryonic Drosophila melanogaster, illustrate avenues through which these genes have diverged within the Diptera. Additionally, we demonstrate that the annotated gene CG15032 is the 5  terminus of gce. In mosquitoes and beetles, a single Met-like homolog displays structural similarity to both Met and gce, and the intron locations are conserved with those of gce. We found that Tribolium and mosquito Met orthologs are assembled from Met- and gce-specific domains in a modular fashion. Our results suggest that Drosophila Met and gce experienced divergent evolutionary pressures following the duplication of an ancestral gce-like gene found in less derived holometabolous insects.',{'entities': [(40,59,'Gene'),(61,64,'Gene'),(70,89,'Gene'),(91,94,'Gene'),(113,123,'Species'),(235,246,'Gene'),(269,288,'Gene'),(290,293,'Gene'),(299,318,'Gene'),(320,323,'Gene'),(365,375,'Species'),(410,413,'Gene'),(418,421,'Gene'),(432,442,'Species'),(627,637,'Species'),(638,641,'Gene'),(646,649,'Gene'),(739,762,'Species'),(926,929,'Gene'),(967,970,'Gene'),(1023,1026,'Gene'),(1031,1034,'Gene'),(1089,1092,'Gene'),(1131,1134,'Gene'),(1164,1167,'Gene'),(1173,1176,'Gene'),(1241,1251,'Species'),(1252,1255,'Gene'),(1260,1263,'Gene'),(1351,1354,'Gene')]}),('A gut-specific chitinase gene essential for regulation of chitin content of peritrophic matrix and growth of Ostrinia nubilalis larvae. Chitinases belong to a large and diverse family of hydrolytic enzymes that break down glycosidic bonds of chitin. However, very little is known about the function of chitinase genes in regulating the chitin content in peritrophic matrix (PM) of the midgut in insects. We identified a cDNA putatively encoding a chitinase (OnCht) in European corn borer (ECB; Ostrinia nubilalis). The OnCht transcript was predominately found in larval midgut but undetectable in eggs, pupae, or adults. When the larvae were fed on an artificial diet, the OnCht transcript level increased by 4.4-fold but the transcript level of a gut-specific chitin synthase (OnCHS2) gene decreased by 2.5-fold as compared with those of unfed larvae. In contrast, when the larvae were fed with the food and then starved for 24h, the OnCht transcript level decreased by 1.8-fold but the transcript level of OnCHS2 increased by 1.8-fold. Furthermore, there was a negative relationship between OnCht transcript level and chitin content in the midgut. By using a feeding-based RNAi technique, we were able to reduce the OnCht transcript level by 63-64% in the larval midgut. Consequently, these larvae showed significantly increased chitin content (26%) in the PM but decreased larval body weight (54%) as compared with the control larvae fed on the diet containing GFP dsRNA. Therefore, for the first time, we provide strong evidence that OnCht plays an important role in regulating chitin content of the PM and subsequently affecting the growth and development of the ECB larvae.',{'entities': [(15,24,'Gene'),(109,127,'Species'),(302,311,'Gene'),(447,456,'Gene'),(468,487,'Species'),(494,512,'Species')]}),('All-trans retinoic acid affects subcellular localization of a novel BmNIF3l protein: functional deduce and tissue distribution of NIF3l gene from silkworm (Bombyx mori). A novel cDNA sequence encoding a predicted protein of 271 amino acids containing a conserved NIF3 domain was found from a pupal cDNA library of silkworm. The corresponding gene was named BmNIF3l (Bombyx mori NGG1p interacting factor 3-like). It was found by bioinformatics that BmNIF3l gene consisted of five exons and four introns and BmNIF3l had a high degree of homology to other NIF3-like proteins, especially in the N-terminal and C-terminal regions. A His-tagged BmNIF3l fusion protein with a molecular weight of approximately 33.6 kDa was expressed and purified to homogeneity. We have used the purified fusion protein to produce polyclonal antibodies against BmNIF3l for histochemical analysis. Subcellular localization revealed that BmNIF3l is a cytoplasmic protein that responds to all-trans retinoic acid (ATRA). Western blotting and real-time reverse transcription polymerase chain reaction showed that the expression level of BmNIF3l is higher in tissues undergoing differentiation. Taken together, the results suggest that BmNIF3l functions in transcription.',{'entities': [(130,135,'Gene'),(146,154,'Species'),(156,167,'Species'),(314,322,'Species'),(366,377,'Species')]}),('Life-span phenotypes of elav and Rbp9 in Drosophila suggest functional cooperation of the two ELAV-family protein genes. The ELAV family of RNA-binding proteins is involved in various aspects of the post-transcriptional regulation of gene expression, from alternative splicing to translation. The members of this family have been shown to interact with each other and have been suggested to function as homo- and/or hetero-multimers. However, the functional interactions among them have not been demonstrated in vivo. In this study, we examined the genetic interaction between elav and Rbp9, two of the three genes encoding ELAV-family proteins in Drosophila. Mutants of both elav and Rbp9 showed shorter life spans than the control, with elav showing a shorter life span than Rbp9. The survival curve of elav-Rbp9 double-mutant flies was indistinguishable from that of elav single-mutant flies, suggesting that both mutations affect longevity through the same pathway. Considering the fact that both genes are co-expressed in adult neurons, we hypothesize that ELAV and Rbp9 cooperate to maintain the functional integrity of the adult nervous system.',{'entities': [(24,28,'Gene'),(33,37,'Gene'),(41,51,'Species'),(94,98,'Gene'),(125,129,'Gene'),(577,581,'Gene'),(586,590,'Gene'),(624,628,'Gene'),(648,658,'Species'),(676,680,'Gene'),(685,689,'Gene'),(739,743,'Gene'),(777,781,'Gene'),(805,809,'Gene'),(810,814,'Gene'),(870,874,'Gene'),(1062,1066,'Gene'),(1071,1075,'Gene')]}),('Characterization of a trehalose-6-phosphate synthase gene from Spodoptera exigua and its function identification through RNA interference. Trehalose is an important disaccharide and a key regulation factor for the development of many organisms, including plants, bacteria, fungi and insects. In order to study the trehalose synthesis pathway, a cDNA for a trehalose-6-phosphate synthase from Spodoptera exigua (SeTPS) was cloned which contained an open reading frame of 2481 nucleotides encoding a protein of 826 amino acids with a predicted molecular weight of 92.65kDa. The SeTPS genome has 12 exons and 11 introns. Northern blot and RT-PCR analyses showed that SeTPS mRNA was expressed in the fat body and in the ovary. Competitive RT-PCR revealed that SeTPS mRNA was expressed in the fat body at different developmental stages and was present at a high level in day 1 S. exigua pupae. The concentrations of trehalose and glucose in the hemolymph were determined by HPLC and showed that they varied at different developmental stages and were negatively correlated to each other. The survival rates of the insects injected with dsRNA corresponding to SeTPS gene reached 53.95%, 49.06%, 34.86% and 33.24% for 36, 48, 60 and 204h post-injection respectively which were significantly lower than those of the insects in three control groups. These findings provide new data on the tissue distribution, expression patterns and potential function of the trehalose-6-phosphate synthase gene.',{'entities': [(63,80,'Species'),(392,409,'Species'),(872,881,'Species')]}),('Study of the aminopeptidase N gene family in the lepidopterans Ostrinia nubilalis (H bner) and Bombyx mori (L.): sequences, mapping and expression. Aminopeptidases N (APNs) are a class of ectoenzymes present in lepidopteran larvae midguts, involved in the Bacillus thuringiensis (Bt) toxins mode of action. In the present work, seven aminopeptidases have been cloned from the midgut of Ostrinia nubilalis, the major Lepidopteran corn pest in the temperate climates. Six sequences were identified as APNs because of the presence of the HEXXH(X)18E and GAMEN motifs, as well as the signal peptide and the GPI-anchor sequences. The remaining sequence did not contain the two cellular targeting signals, indicating it belonged to the puromycin-sensitive aminopeptidase (PSA) family. An in silico analysis allowed us to find orthologous sequences in Bombyx mori. A phylogenetic study of lepidopteran aminopeptidase sequences resulted in their clustering into nine classes. Linkage analysis revealed that the onapn genes as well as all bmapn genes clustered in a single linkage group. O. nubilalis aminopeptidases were expressed in all larval instars. In 5th instar larvae tissues, apns transcripts were found mainly in midguts while apn8 was also highly expressed in Malpighian tubules, and psa showed an ubiquitous expression pattern in O. nubilalis and B. mori. The sequence homology and gene organization of apns suggest a single origin from an ancestral lepidopteran apn gene.',{'entities': [(13,29,'Gene'),(63,81,'Species'),(95,106,'Species'),(148,165,'Gene'),(256,278,'Species'),(386,404,'Species'),(429,433,'Species'),(845,856,'Species'),(1079,1091,'Species'),(1333,1345,'Species'),(1350,1357,'Species')]}),('Identification of two piwi genes and their expression profile in honeybee, Apis mellifera. Piwi genes play an important role in regulating spermatogenesis and oogenesis because they participate in the biogenesis of piRNAs, a new class of noncoding RNAs. However, these genes are not well understood in most insects. To understand the function of piwi genes in honeybee reproduction, we amplified two full-length piwi-like genes, Am-aub and Am-ago3. Both the cloned Am-aub and Am-ago3 genes contained typical PAZ and PIWI domains and active catalytic motifs "Asp-Asp-Asp/His/Glu/Lys," suggesting that the two piwi-like genes possessed slicer activity. We examined the expression levels of Am-aub and Am-ago3 in workers, queens, drones, and female larvae by quantitative PCR. Am-aub was more abundant than Am-ago3 in all the tested samples. Both Am-aub and Am-ago3 were highly expressed in drones but not in workers and queens. The significant finding was that the larval food stream influenced the expression of Piwi genes in adult honeybees. This helps to understand the nutritional control of reproductive status in honeybees at the molecular level.',{'entities': [(22,26,'Gene'),(65,73,'Species'),(75,89,'Species'),(91,95,'Gene'),(346,350,'Gene'),(360,368,'Species'),(412,416,'Gene'),(516,520,'Gene'),(608,612,'Gene'),(1011,1015,'Gene'),(1031,1040,'Species'),(1117,1126,'Species')]}),('Isolation of diapause-regulated genes from the flesh fly, Sarcophaga crassipalpis by suppressive subtractive hybridization. Subtractive suppressive hybridization (SSH) was used to characterize the diapause transcriptome of the flesh fly Sarcophaga crassipalpis. Through these efforts, we isolated 97 unique clones which were used as probes in northern hybridization to assess their expression during diapause. Of these, 17 were confirmed to be diapause upregulated and 1 was diapause downregulated, while 12 were shown to be unaffected by diapause in this species. The diapause upregulated genes fall into several broad categories including heat shock proteins, heavy metal responsive genes, neuropeptides, structural genes, regulatory elements, and several genes of unknown function. In combination with other large-scale analyses of gene expression during diapause, this study assists in the characterization of the S. crassipalpis diapause transcriptome, and begins to identify common elements involved in diapause across diverse taxa.',{'entities': [(58,81,'Species'),(237,260,'Species'),(641,659,'Gene'),(918,933,'Species')]}),('Molecular cloning, genomic structure, and genetic mapping of two Rdl-orthologous genes of GABA receptors in the diamondback moth, Plutella xylostella. The Resistance to dieldrin (Rdl) gene encodes a subunit of the insect gamma-aminobutyric acid (GABA) receptor. Cyclodiene resistance in many insects is associated with replacement of a single amino acid (alanine at position 302) with either a serine or a glycine in the Rdl gene. Two Rdl-orthologous genes of GABA receptors (PxGABARalpha1 and PxGABARalpha2) were cloned and sequenced from a susceptible strain (Roth) of Plutella xylostella. PxGABARalpha1 and PxGABARalpha2 showed 84% and 77% identity with the Rdl gene of Drosophila melanogaster at an amino acid level, respectively. The coding regions of PxGABARalpha1 and PxGABARalpha2 both comprise ten exons, with two alternative RNA-splicing forms in exon 3 of both genes. At the orthologous position of alanine-302 in D. melanogaster Rdl, PxGABARalpha1 has a conserved alanine at position 282. PxGABARalpha2 has a serine instead of an alanine at the equivalent position. With two informative DNA markers, both PxGABARalpha1 and PxGABARalpha2 were mapped onto the Z chromosome of P. xylostella.',{'entities': [(65,68,'Gene'),(112,128,'Species'),(130,149,'Species'),(179,182,'Gene'),(421,424,'Gene'),(435,438,'Gene'),(571,590,'Species'),(661,664,'Gene'),(673,696,'Species'),(925,940,'Species'),(941,944,'Gene'),(1186,1199,'Species')]}),('Odorant reception in the malaria mosquito Anopheles gambiae. The mosquito Anopheles gambiae is the major vector of malaria in sub-Saharan Africa. It locates its human hosts primarily through olfaction, but little is known about the molecular basis of this process. Here we functionally characterize the Anopheles gambiae odorant receptor (AgOr) repertoire. We identify receptors that respond strongly to components of human odour and that may act in the process of human recognition. Some of these receptors are narrowly tuned, and some salient odorants elicit strong responses from only one or a few receptors, suggesting a central role for specific transmission channels in human host-seeking behaviour. This analysis of the Anopheles gambiae receptors permits a comparison with the corresponding Drosophila melanogaster odorant receptor repertoire. We find that odorants are differentially encoded by the two species in ways consistent with their ecological needs. Our analysis of the Anopheles gambiae repertoire identifies receptors that may be useful targets for controlling the transmission of malaria.',{'entities': [(42,59,'Species'),(74,91,'Species'),(161,166,'Species'),(303,320,'Species'),(321,337,'Gene'),(418,423,'Species'),(465,470,'Species'),(676,681,'Species'),(727,744,'Species'),(799,822,'Species'),(823,839,'Gene'),(988,1005,'Species')]}),('Molecular characterization of heat shock protein 90, 70 and 70 cognate cDNAs and their expression patterns during thermal stress and pupal diapause in the corn earworm. Three heat shock protein transcripts, hsp90, hsp70, hsc70, isolated from the corn earworm, Helicoverpa zea, were evaluated for their responsiveness to diapause and thermal stress. These Hsps showed high homology to their counterparts in other species. A phylogenetic analysis of the Hsp90 sequence was consistent with the known classification of insects. Northern blot hybridization indicated the presence of hsp90 transcripts in all tissues, but expression in the brain-subesophageal complex was especially pronounced. The genomic organization of hsp90 examined by Southern blot suggested the presence of a single copy of hsp90 in the H. zea genome. The expression patterns after heat shock indicated that hsp70 and hsp90 were heat-inducible, although hsp70 was more strongly induced than hsp90, and hsc70 was indeed a constitutively expressed member of the hsp70 family. Expression of hsp70 and hsc70 were not altered by the diapause program, but hsp90 was down-regulated at this time. Low temperatures (0-4 degrees C) and recovery from low temperature elicited hsp70 and hsp90 responses, but not an hsc70 response. Thus, unlike several other species, H. zea does not up-regulate hsp70 during pupal diapause, but the down-regulation of hsp90 is consistent with the pattern observed in several other species during diapause. Our results also indicate that hsp90 and hsp70 are responsive to low temperature in both diapausing and nondiapausing pupae.',{'entities': [(30,48,'Gene'),(155,167,'Species'),(175,193,'Gene'),(207,212,'Gene'),(214,219,'Gene'),(221,226,'Gene'),(246,258,'Species'),(260,275,'Species'),(452,457,'Gene'),(578,583,'Gene'),(717,722,'Gene'),(792,797,'Gene'),(876,881,'Gene'),(886,891,'Gene'),(922,927,'Gene'),(959,964,'Gene'),(970,975,'Gene'),(1028,1033,'Gene'),(1056,1061,'Gene'),(1066,1071,'Gene'),(1118,1123,'Gene'),(1233,1238,'Gene'),(1243,1248,'Gene'),(1271,1276,'Gene'),(1351,1356,'Gene'),(1407,1412,'Gene'),(1526,1531,'Gene'),(1536,1541,'Gene')]}),('BmCAP, a silkmoth gene encoding multiple protein isoforms characterized by SoHo and SH3 domains: expression analysis during ovarian follicular development. CAP/ArgBP2/vinexin family proteins, adaptor proteins characterized by three SH3 domains at their C-termini and a SoHo domain towards their N-termini, are known to regulate cell adhesion, cytoskeletal organization, and growth factor signaling. Here we present the isolation and ovarian expression of the BmCAP gene which encodes CAP/ArgBP2/vinexin family proteins in the silkmoth, Bombyx mori. Screening for full-length cDNA clones identified three mRNA isoforms, BmCAP-A1, BmCAP-A2 and BmCAP-B, which show expression throughout ovarian follicular development. Using an antibody raised against a unique region between the SoHo and SH3 domains, BmCAP-A protein isoforms were identified that show specific expression in different compartments of the ovarian follicles. Immunofluorescence staining of the cells of the follicular epithelium establishes a dynamic pattern of BmCAP-A protein localization during choriogenesis. During early choriogenesis, BmCAP-A has a diffuse localization in the cytoplasm but could also be found concentrated at the apical and basal sides at the cell-cell junctions. During late choriogenesis, the diffuse cytoplasmic staining of BmCAP-A disappears while the staining pattern at the apical side resembles a blueprint for the eggshell surface structure. We suggest that BmCAP-A isoforms have important functions during ovarian development, which involve not only the traditional roles in actin organization or cell-cell adhesion but also the regulation of secretion of chorion proteins and the sculpting of the chorion surface.',{'entities': [(0,5,'Gene'),(459,464,'Gene'),(536,547,'Species'),(619,624,'Gene'),(629,634,'Gene'),(799,806,'Gene'),(1025,1032,'Gene'),(1104,1111,'Gene'),(1314,1321,'Gene'),(1453,1460,'Gene')]}),('Brownie, a gene involved in building complex respiratory devices in insect eggshells. BACKGROUND: Insect eggshells must combine protection for the yolk and embryo with provisions for respiration and for the entry of sperm, which are ensured by aeropyles and micropyles, respectively. Insects which oviposit the eggs in an egg-case have a double problem of respiration as gas exchange then involves two barriers. An example of this situation is found in the cockroach Blattella germanica, where the aeropyle and the micropyle are combined in a complex structure called the sponge-like body. The sponge-like body has been well described morphologically, but nothing is known about how it is built up. METHODOLOGY/PRINCIPAL FINDINGS: In a library designed to find genes expressed during late chorion formation in B. germanica, we isolated the novel sequence Bg30009 (now called Brownie), which was outstanding due to its high copy number. In the present work, we show that Brownie is expressed in the follicle cells localized in the anterior pole of the oocyte in late choriogenesis. RNA interference (RNAi) of Brownie impaired correct formation of the sponge-like body and, as a result, the egg-case was also ill-formed and the eggs were not viable. CONCLUSIONS/SIGNIFICANCE: Results indicate that the novel gene Brownie plays a pivotal role in building up the sponge-like body. Brownie is the first reported gene involved in the construction of complex eggshell respiratory structures.',{'entities': [(0,7,'Gene'),(467,486,'Species'),(810,822,'Species'),(875,882,'Gene'),(970,977,'Gene'),(1108,1115,'Gene'),(1311,1318,'Gene'),(1377,1384,'Gene')]}),('Digestive beta-glucosidases from the wood-feeding higher termite, Nasutitermes takasagoensis: intestinal distribution, molecular characterization, and alteration in sites of expression. beta-Glucosidase [EC 3.2.1.21] hydrolyzes cellobiose or cello-oligosaccharides into glucose during cellulose digestion in termites. SDS-PAGE and zymogram analyses of the digestive system in the higher termite Nasutitermes takasagoensis revealed that beta-glucosidase activity is localized in the salivary glands and midgut as dimeric glycoproteins. Degenerate PCR using primers based on the N-terminal amino acid sequences of the salivary beta-glucosidase resulted in cDNA fragments of 1.7 kb, encoding 489 amino acids with a sequence similar to glycosyl hydrolase family 1. Moreover, these primers amplified cDNA fragments from the midgut, and the deduced amino acid sequences are 87-91% identical to those of the salivary beta-glucosidases. Successful expression of the cDNAs in Escherichia coli implies that these sequences also encode functional beta-glucosidases. These results indicate that beta-glucosidases that primarily contribute to the digestive process of N. takasagoensis are produced in the midgut. Reverse transcription-PCR analysis indicated the site-specific expression of beta-glucosidase mRNAs in the salivary glands and midgut. These results suggest that termites have developed the ability to produce beta-glucosidases in the midgut, as is the case for endo-beta-1,4-glucanase, in which the site of expression has shifted from the salivary glands of lower termites to the midgut of higher termites.',{'entities': [(66,92,'Species'),(186,202,'Gene'),(395,421,'Species'),(436,452,'Gene'),(625,641,'Gene'),(732,750,'Gene'),(967,983,'Species'),(1036,1052,'Gene'),(1155,1171,'Species'),(1277,1293,'Gene')]}),('RNAi of ace1 and ace2 in Blattella germanica reveals their differential contribution to acetylcholinesterase activity and sensitivity to insecticides. Cyclorrhapha insect genomes contain a single acetylcholinesterase (AChE) gene while other insects contain at least two ace genes (ace1 and ace2). In this study we tested the hypothesis that the two ace paralogous from Blattella germanica have different contributions to AChE activity, using RNA interference (RNAi) to knockdown each one individually. Paralogous-specific depletion of Bgace transcripts was evident in ganglia of injected cockroaches, although the effects at the protein level were less pronounced. Using spectrophotometric and zymogram measurements, we obtained evidence that BgAChE1 represents 65-75% of the total AChE activity in nerve tissue demonstrating that ace1 encodes a predominant AChE. A significant increase in sensitivity of Bgace1-interfered cockroaches was observed after 48 h of exposure to chlorpyrifos. In contrast, Bgace2 knockdown had a negligible effect on mortality to this organophosphate. These results point out a key role, qualitative and/or quantitative, of AChE1 as target of organophosphate insecticides in this species. Silencing the expression of Bgace1 but not Bgace2 also produced an increased mortality in insects when synergized with lambda-cyhalothrin, a situation which resembles the synergistic effects observed between organophosphates and pyrethroids. Gene silencing of ace genes by RNAi offers an exciting approach for examining a possible functional differentiation in ace paralogous.',{'entities': [(8,12,'Gene'),(17,21,'Gene'),(25,44,'Species'),(281,285,'Gene'),(290,294,'Gene'),(369,388,'Species'),(831,835,'Gene')]}),('Structure and expression of the silk adhesive protein Ser2 in Bombyx mori. Sericins are soluble silk components encoded in Bombyx mori by three genes, of which Ser1 and Ser3 have been characterized. The Ser1 and Ser3 proteins were shown to appear later in the last larval instar as the major sericins of cocoon silk. These proteins are, however, virtually absent in the highly adhesive silk spun prior to cocoon spinning, when the larvae construct a loose scaffold for cocoon attachment. We show here that the silk-gland lumen of the feeding last instar larvae contains two abundant adhesive proteins of 230 kDa and 120 kDa that were identified as products of the Ser2 gene. We also describe the sequence, exon-intron structure, alternative splicing and deduced translation products of this gene in the Daizo p50 strain of B. mori. Two mRNAs of 5.7 and 3.1 kb are generated by alternative splicing of the largest exon. The predicted mature proteins contain 1740 and 882 amino acid residues. The repetitive amino acid sequence encoded by exons 9a and 9b is apparently responsible for the adhesiveness of Ser2 products. It has a similar periodic arrangement of motifs containing lysine and proline as a highly adhesive protein of the mussel Mytilus edulis.',{'entities': [(54,58,'Gene'),(62,73,'Species'),(123,134,'Species'),(160,164,'Gene'),(169,173,'Gene'),(203,207,'Gene'),(212,216,'Gene'),(664,668,'Gene'),(823,830,'Species'),(1103,1107,'Gene'),(1239,1253,'Species')]}),('Wing defects in Drosophila xenicid mutant clones are caused by C-terminal deletion of additional sex combs (Asx). BACKGROUND: The coordinated action of genes that control patterning, cell fate determination, cell size, and cell adhesion is required for proper wing formation in Drosophila. Defects in any of these basic processes can lead to wing aberrations, including blisters. The xenicid mutation was originally identified in a screen designed to uncover regulators of adhesion between wing surfaces [1]. PRINCIPAL FINDINGS: Here, we demonstrate that expression of the betaPS integrin or the patterning protein Engrailed are not affected in developing wing imaginal discs in xenicid mutants. Instead, expression of the homeotic protein Ultrabithorax (Ubx) is strongly increased in xenicid mutant cells. CONCLUSION: Our results suggest that upregulation of Ubx transforms cells from a wing blade fate to a haltere fate, and that the presence of haltere cells within the wing blade is the primary defect leading to the adult wing phenotypes observed.',{'entities': [(16,26,'Species'),(27,34,'Gene'),(86,106,'Gene'),(108,111,'Gene'),(278,288,'Species'),(384,391,'Gene'),(573,588,'Gene'),(615,624,'Gene'),(679,686,'Gene'),(740,753,'Gene'),(755,758,'Gene'),(785,792,'Gene'),(860,863,'Gene')]}),('Functional analysis of saxophone, the Drosophila gene encoding the BMP type I receptor ortholog of human ALK1/ACVRL1 and ACVR1/ALK2. In metazoans, bone morphogenetic proteins (BMPs) direct a myriad of developmental and adult homeostatic events through their heterotetrameric type I and type II receptor complexes. We examined 3 existing and 12 newly generated mutations in the Drosophila type I receptor gene, saxophone (sax), the ortholog of the human Activin Receptor-Like Kinase1 and -2 (ALK1/ACVRL1 and ALK2/ACVR1) genes. Our genetic analyses identified two distinct classes of sax alleles. The first class consists of homozygous viable gain-of-function (GOF) alleles that exhibit (1) synthetic lethality in combination with mutations in BMP pathway components, and (2) significant maternal effect lethality that can be rescued by an increased dosage of the BMP encoding gene, dpp+. In contrast, the second class consists of alleles that are recessive lethal and do not exhibit lethality in combination with mutations in other BMP pathway components. The alleles in this second class are clearly loss-of-function (LOF) with both complete and partial loss-of-function mutations represented. We find that one allele in the second class of recessive lethals exhibits dominant-negative behavior, albeit distinct from the GOF activity of the first class of viable alleles. On the basis of the fact that the first class of viable alleles can be reverted to lethality and on our ability to independently generate recessive lethal sax mutations, our analysis demonstrates that sax is an essential gene. Consistent with this conclusion, we find that a normal sax transcript is produced by saxP, a viable allele previously reported to be null, and that this allele can be reverted to lethality. Interestingly, we determine that two mutations in the first class of sax alleles show the same amino acid substitutions as mutations in the human receptors ALK1/ACVRl-1 and ACVR1/ALK2, responsible for cases of hereditary hemorrhagic telangiectasia type 2 (HHT2) and fibrodysplasia ossificans progressiva (FOP), respectively. Finally, the data presented here identify different functional requirements for the Sax receptor, support the proposal that Sax participates in a heteromeric receptor complex, and provide a mechanistic framework for future investigations into disease states that arise from defects in BMP/TGF-beta signaling.',{'entities': [(23,32,'Gene'),(38,48,'Species'),(67,70,'Gene'),(99,104,'Species'),(105,109,'Gene'),(110,116,'Gene'),(121,126,'Gene'),(127,131,'Gene'),(147,173,'Gene'),(377,387,'Species'),(410,419,'Gene'),(421,424,'Gene'),(447,452,'Species'),(453,489,'Gene'),(491,495,'Gene'),(496,502,'Gene'),(507,511,'Gene'),(512,517,'Gene'),(582,585,'Gene'),(742,745,'Gene'),(862,865,'Gene'),(1031,1034,'Gene'),(1527,1530,'Gene'),(1573,1576,'Gene'),(1654,1657,'Gene'),(1858,1861,'Gene'),(1929,1934,'Species'),(1945,1949,'Gene'),(1950,1957,'Gene'),(1962,1967,'Gene'),(1968,1972,'Gene'),(1999,2043,'Gene'),(2045,2049,'Gene'),(2198,2201,'Gene'),(2238,2241,'Gene'),(2399,2402,'Gene')]}),('Genome-wide analysis of Notch signalling in Drosophila by transgenic RNAi. Genome-wide RNA interference (RNAi) screens have identified near-complete sets of genes involved in cellular processes. However, this methodology has not yet been used to study complex developmental processes in a tissue-specific manner. Here we report the use of a library of Drosophila strains expressing inducible hairpin RNAi constructs to study the Notch signalling pathway during external sensory organ development. We assigned putative loss-of-function phenotypes to 21.2% of the protein-coding Drosophila genes. Using secondary assays, we identified 6 new genes involved in asymmetric cell division and 23 novel genes regulating the Notch signalling pathway. By integrating our phenotypic results with protein interaction data, we constructed a genome-wide, functionally validated interaction network governing Notch signalling and asymmetric cell division. We used clustering algorithms to identify nuclear import pathways and the COP9 signallosome as Notch regulators. Our results show that complex developmental processes can be analysed on a genome-wide level and provide a unique resource for functional annotation of the Drosophila genome.',{'entities': [(44,54,'Species'),(352,362,'Species'),(577,587,'Species'),(1015,1019,'Gene'),(1210,1220,'Species')]}),('CNAct-1 gene is differentially expressed in the subtropical mosquito Culex nigripalpus (Diptera: Culicidae), the primary West Nile Virus vector in Florida. Analysis of differentially expressed genes is a common molecular biological tool to investigate changes in mosquito genes after a bloodmeal or parasite exposure. We report here the characterization of a differentially expressed actin gene, CNAct-1, from the subtropical mosquito, Culex nigripalpus Theobald (Diptera: Culicidae). The CNAct-1 genomic clone is 1.525 kb, includes one 66-bp intron, and a 328-bp 3 -untranslated region. The 376-amino acid putative translation product shares high similarity with muscle-specific actin proteins from other insects, including Culex pipiens pipiens L., Aedes aegypti (L.), Anopheles gambiae Giles and Drosophila melanogaster (Meigen). CNAct-1 is expressed in second and third instars, late pupae, and adult females and males. Interestingly, Cx. nigripalpus actin was highly expressed in female mosquito midgut tissue isolated 6-12 h after ingestion of a bloodmeal. This expression profile indicates a unique function for CNAct-1 in midgut processes that are initiated after blood ingestion.',{'entities': [(0,7,'Gene'),(69,86,'Species'),(121,136,'Species'),(384,389,'Gene'),(396,403,'Gene'),(436,453,'Species'),(489,496,'Gene'),(680,685,'Gene'),(725,748,'Species'),(751,764,'Species'),(771,788,'Species'),(799,822,'Species'),(833,840,'Gene'),(955,960,'Gene'),(1119,1126,'Gene')]}),('Population analysis using the nuclear white gene detects Pliocene/Pleistocene lineage divergence within Anopheles nuneztovari in South America. Anopheles (Nyssorhynchus) nuneztovari Gabald n (Diptera: Culicidae), a locally important malaria vector in some regions of South America, has been hypothesized to consist of at least two cryptic incipient species. We investigated its phylogeographic structure in several South American localities to determine the number of lineages and levels of divergence using the nuclear white gene, a marker that detected two recently diverged genotypes in the primary Neotropical malaria vector Anopheles darlingi Root. In An. nuneztovari, five distinct lineages (1-5) were elucidated: (1) populations from northeastern and central Amazonia; (2) populations from Venezuela east and west of the Andes; (3) populations from Colombia and Venezuela west of the Andes; (4) southeastern and western Amazonian Brazil populations, and (5) southeastern and western Amazonian Brazil and Bolivian populations. There was a large amount of genetic differentiation among these lineages. The deepest and earliest divergence was found between lineage 3 and lineages 1, 2 and 4, which probably accounts for the detection of lineage 3 in some earlier studies. The multiple lineages within Amazonia are partially congruent with previous mtDNA and ITS2 data, but were undetected in many earlier studies, probably because of their recent (Pleistocene) divergence and the differential mutation rates of the markers. The estimates for the five lineages, interpreted as recently evolved or incipient species, date to the Pleistocene and Pliocene. We hypothesize that the diversification in An. nuneztovari is the result of an interaction between the Miocene/Pliocene marine incursion and Pleistocene climatic changes leading to refugial isolation. The identification of cryptic lineages in An. nuneztovari could have a significant impact on local vector control measures.',{'entities': [(38,43,'Gene'),(104,125,'Species'),(520,525,'Gene'),(629,647,'Species'),(657,672,'Species'),(1700,1715,'Species'),(1900,1915,'Species')]}),('Isolation and expression analysis of a homolog of the 14-3-3 epsilon gene in the diamondback moth, Plutella xylostella. A full-length 14-3-3 gene homolog (also referred to as the Px14-3-3 epsilon " " or Px14-3-3  gene) was cloned from cDNA of the diamondback moth, Plutella xylostella. The Px14-3-3 transcript is 789 nucleotides in length, and the predicted polypeptide is 263 amino acids in length, with a calculated molecular mass of 29.6  kDa. The Px14-3-3 gene contains the typical and predicted 14-3-3 domains and motifs. The amino acid sequence of the diamondback moth 14-3-3 gene is very similar to that of other insect epsilons ( ) but not to other insect zetas ( ). In particular, the protein sequence of the Px14-3-3 gene shows high identity to the Bombyx mori epsilon (96.2%). Western blot analysis using an antibody against Px14-3-3  verified the expression of 14-3-3  in the larval, pupal, and adult stages. The Px14-3-3  expression patterns in all the different tissue types were examined in the fourth instar larvae. Px14-3-3  was detected in every tissue examined, including the body fat, hemocytes, brain, gut, and cuticle.',{'entities': [(54,68,'Gene'),(81,97,'Species'),(99,118,'Species'),(247,263,'Species'),(265,284,'Species'),(558,574,'Species'),(759,770,'Species')]}),('A caspase-like decoy molecule enhances the activity of a paralogous caspase in the yellow fever mosquito, Aedes aegypti. Caspases are cysteine proteases that play critical roles in apoptosis and other key cellular processes. A mechanism of caspase regulation that has been described in mammals and nematodes involves caspase-like decoy molecules, enzymatically inactive caspase homologs that have arisen by gene duplication and acquired the ability to regulate other caspases. Caspase-like decoy molecules are not found in Drosophila melanogaster, raising the question of whether this type of caspase regulation exists in insects. Phylogenomic analysis of caspase genes from twelve Drosophila and three mosquito species revealed several examples of duplicated caspase homologs lacking critical catalytic residues, making them candidate caspase-like decoy molecules. One of these, CASPS18 from the mosquito Aedes aegypti, is a homolog of the D. melanogaster caspase Decay and contains substitutions in two critical amino acid positions, including the catalytic cysteine residue. As expected, CASPS18 lacked caspase activity, but co-expression of CASPS18 with a paralogous caspase, CASPS19, in mosquito cells or co-incubation of CASPS18 and CASPS19 recombinant proteins resulted in greatly enhanced CASPS19 activity. The discovery of potential caspase-like decoy molecules in several insect species opens new avenues for investigating caspase regulation in insects, particularly in disease vectors such as mosquitoes.',{'entities': [(2,14,'Gene'),(68,75,'Gene'),(83,104,'Species'),(106,119,'Species'),(240,247,'Gene'),(298,307,'Species'),(317,324,'Gene'),(370,377,'Gene'),(477,484,'Gene'),(523,546,'Species'),(593,600,'Gene'),(656,663,'Gene'),(760,767,'Gene'),(836,843,'Gene'),(880,887,'Gene'),(906,919,'Species'),(941,956,'Species'),(957,964,'Gene'),(1091,1098,'Gene'),(1106,1113,'Gene'),(1145,1152,'Gene'),(1171,1178,'Gene'),(1180,1187,'Gene'),(1227,1234,'Gene'),(1239,1246,'Gene'),(1297,1304,'Gene'),(1342,1349,'Gene'),(1433,1440,'Gene')]}),('Heat shock proteins contribute to mosquito dehydration tolerance. This study examines the responses of heat shock protein transcripts, Hsp70 and Hsp90, to dehydration stress in three mosquito species, Aedes aegypti, Anopheles gambiae and Culex pipiens. We first defined the water balance attributes of adult females of each species, monitored expression of the hsp transcripts in response to dehydration, and then knocked down expression of the transcripts using RNA interference (RNAi) to evaluate potential functions of the Hsps in maintenance of water balance. Fully hydrated females of all three species contained nearly the same amount of water (66-68%), but water loss rates differed among the species, with A. aegypti having the lowest water loss rate (2.6%/h), followed by C. pipiens (3.3%/h), and A. gambiae (5.1%/h). In all three species water could be replaced only by drinking water (or blood). Both A. aegypti and C. pipiens tolerated a loss of 36% of their body water, but A. gambiae was more vulnerable to water loss, tolerating a loss of only 29% of its body water. Dehydration elicited expression of hsp70 in all three species, but only C. pipiens continued to express this transcript during rehydration. Hsp90 was constitutively expressed and expression levels remained fairly constant during dehydration and rehydration, except expression was not noted during rehydration of C. pipiens. Injection of dsRNA to knock down expression of hsp70 (83% reduction) and hsp90 (46% reduction) in A. aegypti did not alter water content or water loss rates, but the dehydration tolerance was lower. Instead of surviving a 36% water loss, females were able to survive only a 28% water loss in response to RNAi directed against hsp70 and a 26% water loss when RNAi was directed against hsp90. These results indicate a critical function for these Hsps in mosquito dehydration tolerance.',{'entities': [(103,121,'Gene'),(135,140,'Gene'),(145,150,'Gene'),(201,214,'Species'),(216,233,'Species'),(238,251,'Species'),(781,791,'Species'),(927,937,'Species'),(1117,1122,'Gene'),(1154,1164,'Species'),(1222,1227,'Gene'),(1394,1404,'Species'),(1453,1458,'Gene'),(1479,1484,'Gene'),(1732,1737,'Gene'),(1790,1795,'Gene')]})]


def main(model=None, output_dir=None, n_iter=1):
    """Load the model, set up the pipeline and train the entity recognizer."""
    if model is not None:
        nlp = spacy.load(model)  # load existing spaCy model
        print("Loaded model '%s'" % model)
    else:
        nlp = spacy.blank("en")  # create blank Language class
        print("Created blank 'en' model")

    # create the built-in pipeline components and add them to the pipeline
    # nlp.create_pipe works for built-ins that are registered with spaCy
    if "ner" not in nlp.pipe_names:
        ner = nlp.create_pipe("ner")
        nlp.add_pipe(ner, last=True)
    # otherwise, get it so we can add labels
    else:
        ner = nlp.get_pipe("ner")

    # add labels
    for _, annotations in TRAIN_DATA:
        for ent in annotations.get("entities"):
            ner.add_label(ent[2])

    # get names of other pipes to disable them during training
    pipe_exceptions = ["ner", "trf_wordpiecer", "trf_tok2vec"]
    other_pipes = [pipe for pipe in nlp.pipe_names if pipe not in pipe_exceptions]
    with nlp.disable_pipes(*other_pipes):  # only train NER
        # reset and initialize the weights randomly – but only if we're
        # training a new model
        if model is None:
            nlp.begin_training()
        for itn in range(n_iter):
            random.shuffle(TRAIN_DATA)
            losses = {}
            # batch up the examples using spaCy's minibatch
            batches = minibatch(TRAIN_DATA, size=compounding(4.0, 32.0, 1.001))
            for batch in batches:
                texts, annotations = zip(*batch)
                nlp.update(
                    texts,  # batch of texts
                    annotations,  # batch of annotations
                    drop=0.5,  # dropout - make it harder to memorise data
                    losses=losses,
                )
            print(len(texts))
            print("Losses", losses)

    # test the trained model
    for text, _ in TRAIN_DATA:
        doc = nlp(text)
        print("Entities", [(ent.text, ent.label_) for ent in doc.ents])
        #print("Tokens", [(t.text, t.ent_type_, t.ent_iob) for t in doc])

    # save model to output directory
    if output_dir is not None:
        output_dir = Path(output_dir)
        if not output_dir.exists():
            output_dir.mkdir()
        nlp.to_disk(output_dir)
        print("Saved model to", output_dir)

        # test the saved model
        print("Loading from", output_dir)
        nlp2 = spacy.load(output_dir)
        for text, _ in TRAIN_DATA:
            doc = nlp2(text)
            print("Entities", [(ent.text, ent.label_) for ent in doc.ents])
            #print("Tokens", [(t.text, t.ent_type_, t.ent_iob) for t in doc])


if __name__ == "__main__":
    main()