bio-comparative-genomics-whole-genome-duplication

name	bio-comparative-genomics-whole-genome-duplication
description	Detect, date, and contextualize whole-genome duplication (WGD / paleopolyploidy) events using wgd v2 (Chen et al 2024), KsRates (Sensalari 2022 substitution-rate-corrected Ks dating), DupGen_finder (Qiao 2019), MAPS (Li 2018 phylogenomic), POInT (Conant 2008 ordered-block), SLEDGe (2024 ML-based), Whale.jl (Bayesian DL+WGD), and synteny-anchored paranome construction. Use when identifying ancient polyploidy from Ks distributions and synteny block analysis, positioning WGD events relative to speciation, distinguishing tandem from segmental from WGD duplications, dating the 2R/3R vertebrate / fish / salmonid WGDs, building paranome and Ks-age mixture models, applying KsRates substitution-rate correction across lineages, or testing alternative biased-fractionation / dosage-balance models post-WGD.
origin	openai4s
category	bioskills/comparative-genomics
metadata	{"tool_type":"mixed","primary_tool":"wgd","third_party":{"name":"GPTomics/bioSkills","repository":"https://github.com/GPTomics/bioSkills","commit":"d91ed3d563019e649dc854c56ccd62551359488a","license":"MIT"}}

Version Compatibility

Reference examples tested with: wgd v2.0.31+ (heche-psb/wgd; Chen et al 2024 Bioinformatics 40:btae272), KsRates 1.1.3+ (VIB-PSB/ksrates; Sensalari 2022 Bioinformatics 38:530), DupGen_finder (Qiao 2019 Genome Biol 20:38), MAPS 1.0 (Li 2018), POInT (Conant lab), SLEDGe (bioRxiv 2024.01.17.574559), Whale.jl 2.0+, ksrates pip 1.1+, MCScanX 1.0+, PAML 4.10+ (yn00/codeml for Ks), BLAT 36+, DIAMOND 2.1+, R 4.4+, mclust 6.1+ (for mixture models). Python 3.10+ required for wgd v2.

Before using code patterns, verify installed versions match. If versions differ:

• CLI: wgd --version; ksrates --version; wgd ksd --help
• Python: pip show wgd ksrates
• R: packageVersion('mclust')

If code throws wgd ksd: cannot find PAML output, KsRates: insufficient sister species, MAPS: missing tree, these tools have specific input expectations: wgd needs codon-aware MAFFT/MUSCLE alignment; KsRates needs configured config_ksrates.txt; MAPS needs nucleotide tree. The deprecated arzwa/wgd v1 is replaced by heche-psb/wgd v2.

Whole Genome Duplication Analysis

"Are there WGD events in this lineage and when did they occur?" -> WGD detection combines Ks distributions (synonymous-substitution rates between gene paralog pairs, showing peaks at past polyploidy events) and synteny block analysis (parallel collinear blocks within a genome). Modern best practice uses wgd v2 (Chen et al 2024 Bioinformatics 40:btae272) as an integrated pipeline. KsRates (Sensalari 2022 Bioinformatics 38:530) is mandatory for cross-lineage comparison because substitution rates vary across the tree -- ignoring this places WGDs incorrectly relative to speciation events. The fundamental tradeoff: Ks plot peaks are visually obvious but biologically ambiguous between (1) small-scale tandem duplications, (2) segmental duplications, and (3) true WGD; combining Ks with synteny anchors disambiguates these.

• CLI: wgd dmd and wgd ksd -- paranome construction + Ks distribution
• CLI: wgd syn -- synteny-anchored WGD signal extraction
• CLI: ksrates init then ksrates wgd-paralogs ortho -- substitution-rate-corrected positioning
• CLI: MCScanX -h then dupgen_finder -- duplication-class assignment
• CLI: mapsR -- gene-tree-based WGD phylogenetic placement

Algorithmic Taxonomy

Tool	Approach	Output	Strength	Fails when
wgd v2 (Chen et al 2024 Bioinformatics 40:btae272)	Integrated paranome + Ks + synteny + WGD dating	Ks distributions, collinearity plots, GMM/ELMM mixture fits, dating	Standard 2024 pipeline; replaces deprecated arzwa/wgd v1	Saturation at Ks > 2; single-lineage substitution rate variation
KsRates (Sensalari 2022 Bioinformatics 38:530)	Substitution-rate correction via outgroup pairs	Adjusted Ks ages of focal-species paralogs vs orthologs	MANDATORY when comparing WGDs across lineages with different rates	Requires at least 2 outgroups for rate calibration
DupGen_finder (Qiao 2019 Genome Biol 20:38)	Classifies duplications by genomic context	Per-gene class: tandem, proximal, transposed, dispersed, WGD	Disambiguates duplication type	Class assignment depends on intervening-gene-count windows
MAPS (Li 2018)	Phylogenomic placement of WGD via gene-tree topology mapping	WGD position on species tree	Detects WGDs from gene-tree-species-tree discordance	Computationally heavy; requires many gene trees
POInT (Conant & Wolfe 2008 Genetics 179:1681)	Order-aware reconstruction of WGD chromosomes	Reconstructed ancestral WGD genome	Strong inference for syntenic-block ages	Lineage-specific tuning required
SLEDGe (bioRxiv 2024.01.17.574559)	ML classifier on Ks plot features	WGD vs no-WGD binary call + confidence	Reduces visual-peak-fitting subjectivity	Newer; less validated
wgd v1 (arzwa/wgd; DEPRECATED)	Predecessor of v2	--	Historical	Use v2 (heche-psb/wgd)
Whale.jl (Zwaenepoel & Van de Peer 2019 MBE 36:1384)	Bayesian DL + WGD reconciliation	WGD posterior at species-tree nodes	Native WGD modeling; integrates with [[gene-tree-species-tree-reconciliation]]	Julia ecosystem
WGDexploreR (legacy)	Visualize Ks plots	Plots only	Visualization aid	Not for inference
McLuster-WGD (custom workflows)	Mixture-model fitting on Ks	GMM / ELMM components	For custom peak fitting	Not a standard tool

Methodology evolves; verify the current wgd v2 manual and the Chen & Zwaenepoel 2023 review chapter (in Polyploidy: Methods and Protocols) before locking on a single workflow. The 2R vertebrate / 3R teleost / Ss4R salmonid WGDs are well-established; novel WGD claims require concordance across Ks, synteny, and phylogenomic placement.

Decision Tree by Experimental Scenario

Scenario	Recommended approach	Why
Plant comparative genomics; suspect WGD	wgd v2 + KsRates	Modern standard pipeline; plants are WGD-prone
Vertebrate 2R or 3R WGD analysis	wgd v2 + MAPS phylogenomic placement	Vertebrate-specific; combines Ks + tree topology
Salmonid Ss4R WGD	Whale.jl with WGD node in species tree	Native WGD modeling in Bayesian framework
Distinguish WGD from sequential small duplications	Ks GMM fit + synteny block analysis (wgd syn)	Ks peak alone insufficient; synteny confirms
Date WGD relative to a speciation event	KsRates with outgroup speciation calibration	Required when substitution rates differ
Identify duplicates retained vs lost after WGD	DupGen_finder + Ks distribution	Classification + age estimation
Detect novel WGD in non-model organism	wgd dmd -> wgd ksd -> visual GMM/ELMM mixture	Standard discovery workflow
Reconstruct ancestral WGD genome architecture	POInT	Order-aware ancestral reconstruction
Test post-WGD bias in fractionation (dosage-balance)	DupGen_finder + per-gene functional annotation	Compare retained vs lost genes
Distinguish recent vs ancient WGD	KsRates + Ks peak location after correction	Ks peak < 0.1 recent; 0.5-1.5 ancient
WGD in clade with rapid evolution (e.g. fish)	KsRates mandatory	Without rate correction, Ks peaks displaced
ML classification of WGD signal	SLEDGe	Newer ML-based alternative to manual fitting
Integrate WGD with DTL inference	Whale.jl OR ALE with WGD branch	See [[gene-tree-species-tree-reconciliation]]
Pan-clade WGD survey (e.g. all angiosperms)	wgd v2 batch + ksrates aggregated	Standard for sustained phylogenomic surveys
Polyploid genome with subgenomes	DupGen_finder for tandem/transposed + AnchorWave proali for subgenome-aware synteny	Subgenome assignment first
Recently diverged species pair, possible recent WGD	minimap2 -x asm5 + SyRI + Ks distribution	High-resolution recent-WGD detection

Per-Tool Failure Modes

Saturation at Ks > 2

Trigger: Computing Ks for ancient WGD candidates from distantly related taxa.

Mechanism: Synonymous substitutions saturate after Ks ~2; each site has undergone multiple substitutions. Observed Ks underestimates true Ks; the relationship between Ks and time becomes non-monotonic above 1.5. WGD peaks at age 200+ Myr in vertebrates / 100+ Myr in plants are at Ks > 2 and unreliable (Vanneste 2013 MBE 30:177).

Symptom: wgd ksd output shows peak at Ks > 1.5 with broad distribution; KsRates rate-corrected Ks even more uncertain; visual fitting yields ambiguous components.

Fix: Restrict Ks-based WGD inference to Ks < 1.5; for older WGDs, use phylogenomic methods (MAPS, POInT, Whale.jl) which don't rely on Ks alone. Report Ks-based dating with explicit saturation caveat. For 2R vertebrate WGD (~500 Myr), MAPS / Whale.jl is required because Ks is saturated.

Substitution-rate variation across lineages

Trigger: Comparing WGD position in two lineages with different molecular evolutionary rates.

Mechanism: If lineage A evolves twice as fast as lineage B, the same biological time is at twice the Ks in lineage A. A WGD at Ks = 0.5 in lineage A and Ks = 0.25 in lineage B might be the same event biologically.

Symptom: WGD claimed at "different times" in different lineages; speciation-vs-WGD relative timing flips depending on which lineage is the focal species.

Fix: Always use KsRates (Sensalari 2022) when comparing WGDs across lineages. KsRates uses outgroup-species speciation events to calibrate the relative substitution rates, then rescales Ks to a common scale. Single-lineage Ks dating is unreliable for inter-lineage comparison.

Tandem duplication masquerading as WGD peak

Trigger: Ks distribution shows a peak at low Ks (< 0.5); user concludes "recent WGD."

Mechanism: Tandem duplications create paralog pairs with low Ks; these accumulate in clusters (especially in NLR genes in plants, OR genes in mammals). The Ks peak reflects clustered tandem origins, not WGD.

Symptom: "WGD" peak Ks-distribution is dominated by paralogs from tandem clusters; synteny analysis shows few cross-chromosome parallel blocks.

Fix: Use DupGen_finder to classify each paralog pair as tandem / proximal / transposed / dispersed / WGD. The true WGD signal comes from the WGD-classified pairs. Recompute Ks distribution restricted to WGD-classified pairs (or anchor-pair restricted to synteny blocks). wgd v2 integrates this filtering.

Synteny block age inconsistency

Trigger: Different synteny blocks from the same suspected WGD have different Ks distributions.

Mechanism: Real WGD blocks should share a synchronized Ks distribution centered at the WGD age; if blocks have very different ages, the inferred "WGD" was probably a series of segmental duplications.

Symptom: Per-block Ks median varies widely; some blocks show Ks ~ 0.3 and others Ks ~ 1.0; not consistent with single WGD.

Fix: Compute per-block Ks distribution; require synchronization (interquartile range overlapping across blocks). wgd v2 reports per-block age dispersion. If dispersion is high, downgrade to "potentially WGD-like" or attribute to ancient segmental duplication history.

Mixture model under/overfitting components

Trigger: Fitting GMM (gaussian mixture model) or ELMM (exponential-lognormal mixture model) to Ks distribution.

Mechanism: Mixture models with too few components miss real WGD signal; too many components find spurious peaks. BIC-based model selection is standard but sensitive to bin choice in histograms.

Symptom: Different runs with different bin counts give different component numbers; visual peaks don't match component means.

Fix: Use BIC-based component selection with 5-fold cross-validation; report uncertainty in component count. wgd v2 fits both GMM and ELMM and reports BIC for each model up to 5 components. Visually validate against synteny block analysis.

Comparing wgd v1 (deprecated) vs v2 outputs

Trigger: Mixing scripts written for arzwa/wgd v1 with heche-psb/wgd v2.

Mechanism: wgd v1 was deprecated 2023; v2 has different command-line interface, default parameters, and output file structure. Scripts from v1 fail or produce different results in v2.

Symptom: Older scripts using wgd ksd v1 syntax don't work in v2; output paths differ.

Fix: Update all scripts to wgd v2 syntax. The v1 wgd is no longer maintained; v2 is the current standard.

KsRates failing with insufficient species

Trigger: Running KsRates with one focal species and one outgroup.

Mechanism: KsRates calibrates substitution-rate variation via at least two outgroup speciation events; one outgroup species provides only one rate point and cannot resolve rate heterogeneity.

Symptom: KsRates errors or produces unreliable rate-corrected Ks.

Fix: Include >= 2 outgroup species at different phylogenetic distances; for plants, a model angiosperm + gymnosperm pair; for vertebrates, lamprey + invertebrate outgroup. Document outgroup sampling.

Polyploid genome confusion

Trigger: Running wgd v2 on an unassigned-subgenome polyploid (e.g. wheat hexaploid without A/B/D subgenome labels).

Mechanism: WGD signal is muddled when subgenomes are not separated; orthologs across subgenomes appear as paralogs, intermixing tandem / segmental / WGD / homeolog signal.

Symptom: Ks distribution shows multiple overlapping peaks at varying intensities; component fits are unstable.

Fix: Assign subgenomes first using k-mer methods (KMC2 + SubPhaser; Jia 2022 New Phytol 235:801) or synteny-based assignment (GENESPACE; Lovell 2022). Then run wgd v2 on each subgenome separately. Document subgenome assignment.

Confusion of homeologous (WGD) vs orthologous (speciation) pairs

Trigger: Computing Ks across recent WGD species without explicit homeolog labeling.

Mechanism: In recent WGD lineages (e.g. salmonid Ss4R), the "ortholog" between species and the "homeolog within species" can have similar Ks; standard ortholog detection (OrthoFinder) lumps both.

Symptom: wgd v2 reports Ks distributions but doesn't distinguish homeologs from orthologs; downstream dating confused.

Fix: Use synteny-anchored ortholog detection (GENESPACE, ProteinOrtho-synteny) to separate cross-species orthologs from within-species homeologs. Run wgd v2 separately on each class. Report each Ks distribution explicitly.

Quantitative Thresholds

Quantity	Threshold	Source / Rationale
Ks saturation upper limit	Ks < 1.5 for reliable inference; >= 2 saturated	Vanneste 2013 MBE 30:177
Recent WGD Ks range	Ks 0.1-0.5	Standard convention
Ancient WGD Ks range	Ks 0.5-1.5	Standard convention
WGD synteny block minimum	>= 5 anchors per block	wgd v2 / GENESPACE default
Tandem duplicate window for DupGen_finder	5 genes default; species-tunable	Qiao 2019
Proximal duplicate window	5-25 genes	Standard
Number of mixture components to consider	1-5 with BIC selection	wgd v2 default
Cross-block age synchrony for WGD	per-block Ks IQR overlap	Visual + statistical
KsRates minimum outgroups	>= 2 species at different distances	Sensalari 2022
KsRates substitution rate correction valid range	Ks < 1.5 in original; correction can extend slightly	KsRates docs
MAPS minimum gene trees	>= 1000 single-copy ortholog trees	Li 2018
Whale.jl WGD detection power	depends on retention rate; > 0.3 retention typically detectable	Zwaenepoel 2019
Vertebrate 2R Ks	saturated; estimated 500-700 Myr	Dehal 2005
Teleost 3R Ks	saturated; estimated 250-350 Myr	Glasauer 2014
Salmonid Ss4R Ks	80-100 Myr; Ks ~0.1-0.2	Lien 2016
Plant 1R / 2R	varies clade; core-eudicot gamma paleohexaploidy ~120 Myr	Jiao 2012 (dating); Soltis 2009 (placement)
Synonymous codon site count	dS reliable when >= 30 synonymous sites per pair	Yang 2007 PAML
Per-pair gene length minimum	>= 300 bp CDS	wgd v2 default
Codon-aware MSA aligner	MAFFT --auto or MUSCLE; for distant pairs PRANK	wgd v2 supports all
Bootstrap support for MAPS	>= 80 on gene-tree branches	Li 2018

wgd v2 Standard Workflow

Goal: Detect and date WGD events from a focal-species proteome and CDS, with synteny-anchored Ks distribution.

Approach: Build paranome -> compute Ks distribution -> identify synteny anchors -> mixture model fit -> visualize.

# Install (Python 3.10+ required)
pip install wgd

# 1. Build paranome (all-vs-all paralog identification)
wgd dmd cds.fasta -o output/paranome.tsv -t 16

# 2. Compute Ks distribution
# Verify exact flags with `wgd ksd --help` (the wgd v2 CLI evolves; --pairwise / --ks-method spelling differs across versions).
wgd ksd output/paranome.tsv cds.fasta -o output/ksd \
    --aligner mafft --n-threads 16

# 3. Synteny anchors (intra-genome)
wgd syn output/paranome.tsv gff.bed cds.fasta -o output/syn \
    --feature gene --gene-attribute Name --min-block-size 5

# 4. Mixture model on synteny-anchored Ks
wgd mix output/ksd/ks_distributions.tsv -o output/mix \
    --model both --components 1 2 3 4 5

# 5. Visualize
wgd viz output/mix/mix_results.tsv -o output/viz \
    --type histogram --bins 50

'''Parse wgd v2 outputs and identify WGD peak from Ks distribution.'''
import pandas as pd
from scipy import stats


def load_ks_distribution(ksd_file):
    '''Load wgd ksd output: pair  Ks  Ka  ...'''
    df = pd.read_csv(ksd_file, sep='\t')
    # Filter saturated
    df_filt = df[(df['Ks'] > 0) & (df['Ks'] < 2)]
    return df_filt


def fit_mixture(ks_values, n_components=3):
    '''Fit GMM (gaussian mixture model) on Ks via sklearn.'''
    from sklearn.mixture import GaussianMixture
    gmm = GaussianMixture(n_components=n_components, random_state=42)
    gmm.fit(ks_values.reshape(-1, 1))
    return {
        'means': gmm.means_.flatten(),
        'variances': gmm.covariances_.flatten(),
        'weights': gmm.weights_,
        'bic': gmm.bic(ks_values.reshape(-1, 1))
    }


def identify_wgd_peaks(ks_values, min_components=1, max_components=5):
    '''BIC-based model selection.'''
    results = {}
    for n in range(min_components, max_components + 1):
        results[n] = fit_mixture(ks_values, n)
    best_n = min(results.keys(), key= k: results[k][])
     best_n, results[best_n]

KsRates Substitution-Rate-Corrected Workflow

Goal: Position a focal-species WGD relative to speciation events with substitution-rate correction.

Approach: Define focal species + 2+ outgroups -> compute orthologous Ks (focal vs outgroup) -> compute paralogous Ks (focal-internal) -> rescale via outgroup calibration.

# KsRates is best driven through its Nextflow pipeline (which orchestrates ortholog Ks,
# paralog Ks, rate correction, and plotting). Subcommand naming differs across releases;
# always verify with `ksrates --help` against the installed version.

# 1. Generate a working config (subcommand spelling varies by release; e.g. `init` in some,
#    `generate-config` in others). Inspect `ksrates --help`.
ksrates init config_ksrates.txt        # OR: ksrates generate-config config_ksrates.txt

# 2. Edit config_ksrates.txt to set focal_species, outgroups, FASTA + GFF paths, tree.

# 3. Run the full pipeline (Nextflow-driven):
ksrates --config config_ksrates.txt --n-threads 16
# Or invoke individual stages (subcommand names: see `ksrates --help`).

The output plot shows the rate-corrected focal-species paralog Ks distribution with vertical lines indicating outgroup-species speciation events; WGD peaks before vs after speciation events can be distinguished.

DupGen_finder for Duplication Class Assignment

Goal: Classify each paralog pair as tandem / proximal / transposed / dispersed / WGD by genomic context.

Approach: MCScanX collinearity + intervening-gene count -> classify each duplicate by class.

# Input: MCScanX collinearity file + GFF
git clone https://github.com/qiao-xin/DupGen_finder
cd DupGen_finder

# Run with intervening-gene count
./DupGen_finder.pl \
    -i input_collinearity_file \
    -t species_name \
    -c gene_count_file \
    -o output

# Output: tandem, proximal, transposed, dispersed, wgd duplicate lists

'''Aggregate DupGen_finder output for downstream Ks distribution per class.'''
def load_dupgen(class_dir):
    classes = {}
    for cls in ('tandem', 'proximal', 'transposed', 'dispersed', 'wgd'):
        path = f'{class_dir}/{cls}.pairs'
        try:
            classes[cls] = pd.read_csv(path, sep='\t', header=None,
                                       names=['gene1', 'gene2'])
        except FileNotFoundError:
            classes[cls] = pd.DataFrame(columns=['gene1', 'gene2'])
    return classes

MAPS Phylogenomic WGD Placement

Goal: Place a WGD event on a species tree using gene-tree-species-tree mapping.

Approach: Build many single-copy ortholog gene trees -> map each tree's topology to the species tree -> identify branches with topology consistent with a WGD event.

MAPS is heavyweight; requires CRG database setup and significant compute. See https://bitbucket.org/barker-lab/maps/src for the current pipeline.

Reconciliation: When Methods Disagree

Pattern	Likely cause	Action
wgd v2 peak at Ks 0.5, MAPS supports WGD on different branch	Rate variation across lineages	KsRates with outgroups; trust rate-corrected position
Ks peak prominent, synteny anchors sparse	Tandem-driven peak	DupGen_finder classification; verify WGD class
Synteny blocks present, Ks peak diffuse	Old WGD; saturated Ks	Restrict to recent paralogs; expect Ks 0.5-1.5
One outgroup's KsRates says WGD before speciation, other after	Outgroup substitution rate variation	Use multiple outgroups; report ranges; explicit caveat
GMM fits 2 components, ELMM fits 3	Model class differs	Visual inspection; report both; choose by BIC
DupGen_finder calls "WGD" but Ks > 2	Saturation	Ks unreliable; verify via synteny + phylogenomic placement
Two recently sequenced sister species disagree on WGD date	Different annotation pipelines	Re-annotate consistently; verify orthology
wgd v2 detects WGD; Whale.jl posterior doesn't	Different inference frameworks	Whale.jl is Bayesian and gene-tree-based; trust if MAPS / phylogeny supports
Salmonid Ss4R detectable by wgd v2 in young salmonids	Recent WGD; clearer signal	Confirmation, not contradiction

Operational rule for publication: Concordance across Ks distribution (wgd v2), synteny-anchored block analysis (wgd syn / GENESPACE), KsRates rate-corrected positioning, and at least one phylogenomic method (MAPS / Whale.jl / ALE with WGD node) = publication-grade WGD claim. Single-Ks-peak claims should be downgraded to "Ks evidence for possible WGD."

Cohort Gotchas

• Plant WGD legacy: All angiosperms share at least one ancestral WGD (the epsilon event, ~192 Myr); the core-eudicot gamma paleohexaploidy is younger (~120 Myr; Jiao 2012 Genome Biol 13:R3), core-eudicot-specific (Soltis 2009 Am J Bot 96:336); confirm consensus against published plant lineages
• Vertebrate 2R: ~500-600 Myr; Ks saturated; use MAPS phylogenomic placement (Dehal & Boore 2005)
• Teleost 3R: ~320 Myr; in addition to 2R; doubles ohnologs in fish
• Salmonid Ss4R: ~80-100 Myr; recent enough that Ks is informative; ohnologs identifiable
• Plant polyploids (wheat, Brassica): recent; subgenomes assignable; subgenome-stratify before wgd v2
• Allopolyploids vs autopolyploids: allopolyploids have higher inter-subgenome divergence (easier to assign subgenomes); autopolyploids have similar subgenomes (chimeric assembly risk)
• Lineage-specific extra duplications post-WGD: make Ks peak broader; differentiate from sequential WGDs

Anticipated Reviewer Pushback

Pushback	Standard response
"Saturation?"	Ks < 1.5 reported; for older WGDs, MAPS or Whale.jl used
"Rate variation?"	KsRates applied with 2+ outgroups; rate-corrected positioning
"Tandem vs WGD?"	DupGen_finder classification; WGD-class subset reported
"Synteny confirmation?"	wgd syn or GENESPACE; per-block Ks synchrony reported
"Phylogenomic support?"	MAPS, Whale.jl, or ALE-with-WGD agrees
"Mixture model uncertainty?"	BIC-based model selection; 1-5 components tested
"Subgenome assignment (polyploid)?"	k-mer-based or synteny-based subgenome assignment first
"Why wgd v2 over wgd v1?"	v1 deprecated 2023; v2 (heche-psb/wgd) is current
"Comparison to published WGD dates?"	Match published vertebrate 2R / teleost 3R / salmonid Ss4R; cite primary literature

Common Errors

Error / symptom	Cause	Solution
wgd ksd reports few Ks values	CDS sequences not in-frame or short	Verify CDS quality; filter short sequences
KsRates "no orthologs found"	Outgroup species names mismatched	Normalize species labels exactly
GMM fits with 5 components, all similar means	Overfitting	Use BIC; restrict max components to 3-4
Per-block Ks IQR > 0.5	Block age dispersion; likely segmental not WGD	Trust dispersion; reclassify as segmental
wgd v2 ELMM converges to single component	Insufficient pairs	Reduce filtering; include more paralog pairs
MAPS slow / OOM	Many gene trees; large dataset	Cluster; reduce to representative single-copy orthologs
Whale.jl Turing fails to mix	NUTS step size; tree dimension	HMC manual tuning; or reduce species tree
DupGen_finder produces empty tandem set	Default window inappropriate	Adjust window for genome density
Ks distribution has secondary peak at 1.8	Saturation artifact (not WGD)	Recompute with PRANK or PAML codeml; restrict Ks < 1.5
Polyploid "WGD" appears at Ks 0.05	Recent polyploidy is ohnologous WGD, expected	Verify with subgenome assignment
Plant analysis lacks reference outgroup gymnosperm	KsRates needs gymnosperm + angiosperm	Add Selaginella or moss outgroup

Tool Installation Notes

# wgd v2
pip install wgd
# Or: git clone https://github.com/heche-psb/wgd && cd wgd && pip install .

# KsRates
pip install ksrates

# DupGen_finder (Perl + MCScanX dependency)
git clone https://github.com/qiao-xin/DupGen_finder
# Install MCScanX (Wang 2012): git clone https://github.com/wyp1125/MCScanX && cd MCScanX && make

# MAPS (Python; requires CRG, ete3)
git clone https://bitbucket.org/barker-lab/maps

# Whale.jl (Julia)
julia -e 'using Pkg; Pkg.add("Whale")'

# SLEDGe
git clone https://github.com/SLEDGe-team/SLEDGe

# POInT (C++)
git clone https://github.com/gconant0/PoInT
cd PoInT && make

# PAML for Ks computation (yn00 method)
conda install -c bioconda paml

# DIAMOND / BLAST for paranome
conda install -c bioconda diamond blast

# R packages
install.packages(c('mclust', 'rmixmod'))

For new analyses, default to wgd v2 + KsRates as the primary pipeline; MAPS or Whale.jl for confirmation on phylogenomic placement.

References

• Chen H et al 2024 Bioinformatics 40:btae272 (wgd v2)
• Sensalari C et al 2022 Bioinformatics 38:530 (KsRates)
• Qiao X et al 2019 Genome Biol 20:38 (DupGen_finder)
• Li Z et al 2018 PNAS 115:4713 (MAPS phylogenomic WGD placement)
• Conant GC & Wolfe KH 2008 Genetics 179:1681 (POInT)
• Zwaenepoel A & Van de Peer Y 2019 MBE 36:1384 (Whale.jl)
• Sutherland BL et al 2024 bioRxiv 2024.01.17.574559 (SLEDGe ML classifier)
• Vanneste K et al 2013 MBE 30:177 (Ks saturation)
• Holland PWH et al 1994 Development Suppl:125 (2R hypothesis)
• Dehal P & Boore JL 2005 PLoS Biol 3:e314 (vertebrate 2R confirmation)
• Glasauer SMK & Neuhauss SCF 2014 Mol Genet Genomics 289:1045 (teleost 3R)
• Lien S et al 2016 Nature 533:200 (salmonid Ss4R)
• Soltis DE et al 2009 Am J Bot 96:336 (polyploidy and angiosperm diversification)
• Jiao Y et al 2012 Genome Biol 13:R3 (dates the core-eudicot gamma triplication)
• Birchler JA & Veitia RA 2007 Plant Cell 19:395 (gene balance hypothesis)
• Force A et al 1999 Genetics 151:1531 (subfunctionalization)
• Maere S et al 2005 PNAS 102:5454 (post-WGD retention bias)
• Tang H et al 2008 GR 18:1944 (synteny / MCScan)
• Lovell JT et al 2022 eLife 11:e78526 (GENESPACE)
• Jia K-H et al 2022 New Phytol 235:801 (SubPhaser subgenome assignment)
• Yang Z 2007 PAML manual (yn00 codon Ks)
• Chen H & Zwaenepoel A 2023 Methods Mol Biol 2545:3 (inference of ancient polyploidy from genomic data)

Related Skills

• comparative-genomics/synteny-analysis - Synteny-anchored Ks (wgd syn / GENESPACE)
• comparative-genomics/ortholog-inference - Ortholog detection feeds DupGen / wgd
• comparative-genomics/gene-tree-species-tree-reconciliation - Whale.jl native WGD modeling; ALE with WGD branch
• comparative-genomics/gene-family-evolution - CAFE5 birth-death often shows post-WGD retention bias
• comparative-genomics/positive-selection - Selection on retained ohnologs (post-WGD diversification)
• comparative-genomics/ancestral-reconstruction - Ancestral pre-WGD gene state
• phylogenetics/divergence-dating - Time-calibrated WGD positioning
• alignment/multiple-alignment - Codon-aware MSA for Ks computation
• alignment/pairwise-alignment - Pairwise Ks via PAML yn00 / codeml
• genome-assembly/assembly-qc - BUSCO / Compleasm before WGD analysis