Preprint now on bioRxiv:
Dsuite - fast D-statistics and related admixture evidence from VCF files
Milan Malinsky
bioRxiv 634477; doi: https://doi.org/10.1101/634477
Malawi cichlid data used in the manuscript:
VCF file; SETS.txt file
Simulated 20-species data used in the manuscript: VCF file; SETS.txt file; TREE_FILE.nwk (input tree)
There is also a very nice and detailed tutorial by @mmatschiner.
To compile you must have a reasonably recent GCC (>=4.9.0) or clang compiler (on mac OS this comes with Command Line Tools) and the zlib compression library (https://www.zlib.net). Both should already be present on most systems.
$ git clone https://github.com/millanek/Dsuite.git
$ cd Dsuite
$ makeThe Dsuite executable will be in the Build folder, so to run it type e.g. ./Build/Dsuite; this will show the available commands. To execute e.g. the Dtrios command, type ./Build/Dsuite Dtrios.
If you want to plot the results of the f-branch calcuation (see below), you will need to install the python script for this using setuptools. You need an internet connection as some python dependencies will be downloaded from pypi.org. It may be necessary to exit python or conda virtual environments for this to work correctly.
$ cd utils
$ python3 setup.py install --user --prefix=The above should work on both mac and linux. Note that there is no text (not even whitespace) after the = above. If you want to use your own virtual environments, you can alternatively not run setup.py and just install the dependencies with pip or conda.
- A VCF file, which can be compressed with gzip or bgzip. It can contain multiallelic loci and indels, but only biallelic loci will be used.
- Population/species map
SETS.txt: a text file with one individual per row and a tab separating the individual’s name from the name of the species/population it belongs to, as shown below:
Ind1 Species1
Ind2 Species1
Ind3 Species2
Ind4 Species2
Ind5 Species3
Ind6 Outgroup
Ind7 Outgroup
Ind8 xxx
... ...
IndN Species_n
At least one individual needs to be specified to be the outgroup by using the Outgroup keyword. If you want some individuals to be ignored entirely, use the xxx keyword instead. Therefore, you don't have to subset your VCF file if you only want to use a subset of the samples in it.
- A tree in Newick format. The tree should have leaf labels corresponding to the species/population names. Branch lengths can be present but are not used.
Valid examples:
(Species2,(Species1,(Species3,Species4)));
(Species2:6.0,(Species1:5.0,(Species3:3.0,Species4:4.0))); - The
test_trios.txtfile forDinvestigate. One trio of populations/species per line, separated by a tab in the orderP1 P2 P3:
Species1 Species2 Species3
Species1 Species4 Species2
... ... ...
Dsuite Dtrios - Calculate the D (ABBA-BABA) and f4-ratio (f_G) statistics for all possible trios of populations/species
Calculate the D (ABBA/BABA) and f4-ratio (f_G) statistics for all trios of species in the dataset (the outgroup being fixed)
the results are as definded in Patterson et al. 2012 (equivalent to Durand et al. 2011 when the Outgroup is fixed for the ancestral allele)
The SETS.txt should have two columns: SAMPLE_ID SPECIES_ID
The outgroup (can be multiple samples) should be specified by using the keywork Outgroup in place of the SPECIES_ID
-h, --help display this help and exit
-k, --JKnum (default=20) the number of Jackknife blocks to divide the dataset into; should be at least 20 for the whole dataset
-j, --JKwindow (default=NA) Jackknife block size in number of informative SNPs (as used in v0.2)
when specified, this is used in place of the --JKnum option
-r, --region=start,length (optional) only process a subset of the VCF file
-t, --tree=TREE_FILE.nwk (optional) a file with a tree in the newick format specifying the relationships between populations/species
D and f4-ratio values for trios arranged according to the tree will be output in a file with _tree.txt suffix
-n, --run-name run-name will be included in the output file name
The output files with suffixes BBAA.txt, Dmin.txt, and optionally tree.txt (if the -t option was used) contain the results: the D statistics, unadjusted p-values, and the f4-ratios (f_G). Please read the manuscript for more details.
The output files with suffixes combine.txt and combine_stderr.txt are used as input to DtriosCombine. If you don't need to use DtriosCombine, you can safely delete these files.
Combine the BBAA, ABBA, and BABA counts from multiple files (e.g per-chromosome) and output the overall D stats,
p-values and f4-ratio values
-h, --help display this help and exit
-n, --run-name run-name will be included in the output file name
-t , --tree=TREE_FILE.nwk (optional) a file with a tree in the newick format specifying the relationships between populations/species
D and f4-ratio values for trios arranged according to the tree will be output in a file with _tree.txt suffix
-s , --subset=start,length (optional) only process a subset of the trios
As for Dtrios, there are output files with suffixes BBAA.txt, Dmin.txt, and optionally tree.txt (if the -t option was used). They contain the overall combined results: the D statistics, unadjusted p-values, and the f4-ratios (f_G).
Dinvestigate - Follow up analyses for trios with significantly elevated D: calculates D, f_d and f_dM in windows along the genome
Outputs D, f_d (Martin et al. 2014 MBE), and f_dM (Malinsky et al., 2015) in genomic windows
The SETS.txt file should have two columns: SAMPLE_ID POPULATION_ID
The test_trios.txt should contain names of three populations for which the statistics will be calculated:
POP1 POP2 POP3
There can be multiple lines and then the program generates multiple ouput files, named like POP1_POP2_POP3_localFstats_SIZE_STEP.txt
-h, --help display this help and exit
-w SIZE,STEP --window=SIZE,STEP (required) D, f_D, and f_dM statistics for windows containing SIZE useable SNPs, moving by STEP (default: 50,25)
-n, --run-name run-name will be included in the output file name
Fbranch - A heuristic approach designed to aid the interpretation of many correlated f4-ratio results
Usage: Dsuite Fbranch [OPTIONS] TREE_FILE.nwk FVALS_tree.txt > fbranch.txt
Implements the 'f-branch' type calculations developed by Hannes Svardal for Malinsky et al., 2018, Nat. Ecol. Evo.
Uses the f4-ratio (f_G) values produced by Dsuite Dtrios (or DtriosCombine) with the --tree option; this is the output of Dtrios with the "_tree.txt" suffix
-h, --help display this help and exit
The f-branch statistic in matrix-like format. Use the plotting function below to display the f-branch statistic.
The output of Dsuite Fbranch can be plotted with ./utils/dtools.py (see installation instructions above).
usage: dtools.py [-h] [-n RUN_NAME] [--outgroup OUTGROUP] [--use_distances]
[--ladderize]
fbranch.txt tree.newick
Plot f-branch statistic as produced by Dsuite. Produces .png and .svg files.
positional arguments:
fbranch Path to file containing f-branch matrix as produced by
Dsuite Fbranch.
tree Path to .newick tree file as given to Dsuite Fbranch.
optional arguments:
-h, --help show this help message and exit
-n RUN_NAME, --run-name RUN_NAME
Base file name for output plots. (default: fbranch)
--outgroup OUTGROUP Outgroup name in newick file. (default: Outgroup)
--use_distances Use actual node distances from newick file when
plotting tree. (default: False)
--ladderize Ladderize the input tree before plotting. (default:
False)
Running dtools.py yields a .png and an .svg file of the f-branch statistic along the input tree. You can edit the .svg file in a vector graphics editor (e.g., inkscape to your liking). See Malinsky et al. 2018 Fig. 3 and the Dsuite preprint biorxiv for examples and interpretation of f-branch plots.
Selected updates (full update history is accessible on gitHub):
v0.3 r21: Automatic estimation of Jackknife window size to get a desired number of blocks
Progress update in %
f4-ratios are calculated by default by Dtrios
Updated documentation
v0.2 r20: Subset option returns to DtriosCombine
v0.2 r19: Fixed a bug in Fbranch where P1 and P2 positions where A in P2 positions and B in P1 positions were not considered
v0.2 r18: Full implementation of D and f4-ratio in line with the Patterson et al. 2012 definitions
(affects only analyses where the outgroup allele is not fixed)
v0.2 r15: Ironed bugs in Dinvestigate, truly useable from this point
v0.2 r6: First Fbranch version
v0.1 r1: First workable Dsuite release 8th May 2019