Phylogenetics

Phylogenetics Skills

Expert knowledge for phylogenetic tree analysis, visualization, and annotation management.

ITOL Annotation File Troubleshooting

Common Issue: Species Name Mismatches

Problem: Species in tree file don't match annotation files, causing missing data in ITOL visualization.

Root Causes: 1. Tree processing tools (e.g., TimeTree) may abbreviate species names 2. Capitalization inconsistencies (e.g., Alca_Torda vs Alca_torda) 3. Genus-only names replacing full binomial nomenclature

Solution Workflow:

1. Compare tree versions:

   # Find species that exist in original but are different in processed tree
   grep -o "[A-Z][a-z]*_[a-z]*" Tree.nwk | sort -u > original_names.txt
   grep -o "[A-Z][a-z]*_[a-z]*" Tree_final.nwk | sort -u > processed_names.txt
   comm -3 original_names.txt processed_names.txt

2. Identify incomplete names:

   # Species with genus only (no underscore after first word)
   with open('Tree_final.nwk', 'r') as f:
       tree = f.read()
   # Look for patterns like "Myxine:" instead of "Myxine_glutinosa:"

3. Fix systematically:

• Update tree file with complete names
• Update CSV data source
• Update all ITOL annotation files (colorstrip, labels, branch colors)
• Verify counts match across all files

4. Verification checklist:

• [ ] All files have same species count
• [ ] No "Other" or unknown categories remain
• [ ] Legend counts match actual data counts
• [ ] Test species display correctly

ITOL File Synchronization

Critical: When adding/removing species, update ALL annotation files:

• Tree file (.nwk)
• Data source (.csv)
• itol_*_colorstrip_final.txt
• itol_*_labels_final.txt
• itol_branch_colors_final.txt

Verification script:

def verify_itol_sync():
    files = [
        'Tree_final.nwk',
        'itol_taxonomic_colorstrip_final.txt',
        'itol_taxonomic_labels_final.txt',
        'itol_branch_colors_final.txt'
    ]

    counts = {}
    for f in files:
        # Extract species list from each file
        species = extract_species(f)
        counts[f] = len(species)

    if len(set(counts.values())) == 1:
        print(f"✓ All files synchronized: {counts[files[0]]} species")
    else:
        print("✗ Files out of sync:")
        for f, count in counts.items():
            print(f"  {f}: {count}")

Fish Taxonomy Simplification for Visualization

User Preference vs Scientific Detail

Scientific accuracy often requires detailed fish categories:

• Jawless fishes (Agnatha) - hagfish, lampreys
• Cartilaginous fishes (Chondrichthyes) - sharks, rays
• Lobe-finned fishes (Sarcopterygii) - coelacanths, lungfishes
• Ray-finned fishes (Actinopterygii) - most bony fishes

For visualization clarity, users may prefer simplified categories:

• Cartilaginous fishes (includes jawless)
• Bony fishes (includes lobe-finned)

Implementation approach: 1. Start with scientifically accurate categories 2. Present to user for feedback 3. Be ready to simplify based on user preference 4. Document the choice made

Key insight: Users may prioritize:

• Visual simplicity over taxonomic precision
• Fewer categories for cleaner figures
• Practical grouping for their specific use case

Always confirm categorization preferences when creating phylogenetic visualizations, especially for:

• Fish classifications
• Bacterial/archaeal groups
• Plant lineages
• Any domain with complex subdivisions

Bulk Editing ITOL Annotation Files

Safe Update Pattern

When updating ITOL annotation files, use this pattern to avoid data corruption:

def update_itol_file(input_file, species_updates):
    """
    Safely update ITOL annotation file.

    Args:
        input_file: Path to ITOL file
        species_updates: Dict mapping species -> (category, color)
    """
    with open(input_file, 'r') as f:
        lines = f.readlines()

    # Find critical line indices
    data_start = None
    legend_labels_idx = None
    legend_colors_idx = None

    for i, line in enumerate(lines):
        if line.strip() == 'DATA':
            data_start = i
        if line.startswith('LEGEND_LABELS'):
            legend_labels_idx = i
        if line.startswith('LEGEND_COLORS'):
            legend_colors_idx = i

    # Update data section
    for i in range(data_start + 1, len(lines)):
        if not lines[i].strip():
            continue
        parts = lines[i].strip().split('\t')
        if len(parts) >= 3:
            species = parts[0]
            if species in species_updates:
                new_cat, new_color = species_updates[species]
                lines[i] = f"{species}\t{new_color}\t{new_cat}\n"

    # Recalculate category counts
    category_counts = {}
    for i in range(data_start + 1, len(lines)):
        if not lines[i].strip():
            continue
        parts = lines[i].strip().split('\t')
        if len(parts) >= 3:
            category = parts[2]
            category_counts[category] = category_counts.get(category, 0) + 1

    # Update legend with accurate counts
    # [Build new legend line with actual counts]

    # Write atomically
    with open(input_file, 'w') as f:
        f.writelines(lines)

    return category_counts

Key principles: 1. Always recalculate counts after changes 2. Update legend to match actual data 3. Handle all three file types (colorstrip, labels, branch colors) 4. Verify changes with separate verification script

Related Skills

• Analysis/Visualization: Color selection strategies for phylogenetic trees
• VGP Pipeline: Species list management and quality control