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Rationale and background:

BUSCOassessing genome assembly and annotation completeness with single-copy orthologs 

Felipe A. Simão, Robert M. Waterhouse, Panagiotis Ioannidis, Evgenia V. Kriventseva, & Evgeny M. Zdobnov Zdobnov’s Computational Evolutionary Genomics Group

Bioinformatics, published online June 9, 2015 (doi: 10.1093/bioinformatics/btv351)

BUSCO (Benchmarking UniversalSingle-Copy Orthologs) is a tool that provides measures for quantitative assessment of genome assembly, gene set, and transcriptome completeness based on evolutionarily informed expectations of gene content from near-universal single-copy orthologs selected from OrthoDBBUSCO assessments are implemented in open-source software, with comprehensive lineage-specific sets of Benchmarking Universal Single-Copy Orthologs for arthropods, vertebrates, metazoans, fungi, eukaryotes, and bacteria. These conserved orthologs are ideal candidates for large-scale phylogenomics studies, and the annotated BUSCO gene models built during genome assessments provide a comprehensive gene predictor training set for use as part of genome annotation pipelines. BUSCO assessments offer intuitive metrics, based on evolutionarily informed expectations of gene content from hundreds of species, to gauge completeness of rapidly accumulating genomic data and satisfy an Iberian's quest for quality - "Busco calidad/qualidade". The software is freely available to download at (http://busco.ezlab.org/). 


Pre-Requisites

  1. A CyVerse account. (Register for an CyVerse account here - user.cyverse.org)
  2. Mandatory arguments 
    1. Output folder name (name to use for the run and all temporary files (appended))
    2. Input file (genome assembly/gene set/transcript set file in FASTA format)
    3. Lineage data (Location of the BUSCO lineage data to use. You can select the BUSCO profile files for your species of interest from the Data window under Community Data -> iplantcollaborative -> example_data -> BUSCO.sample.data )
    4. Mode of analysis (genome, protein and trans. Default: genome)
  3. Optional arguments
    1. species (If your species is not in the list, selecting a closely-related species usually produces better results).
    2. e-value (Use a custom blast e-value cutoff. Default: 0.03) 
    3. long (Performs full optimization for Augustus gene finding training Default: Off

Test with sample data

Test data for this app appears directly in the Discovery Environment in the Data window under Community Data -> iplantcollaborative -> example_data -> BUSCO.sample.data 

Execute BUSCO with the following input data

  1.  Output folder - run_example
  2. Input file - target.fa 
  3. lineage data - example
  4. mode - genome (default)
  5. species - fly
  6. e-value - 0.03 (default) 

Results 

Successful execution of the BUSCO assessment pipeline will create a directory named run_example along with logs directoryThe directory will contain several files and directories:

1- Files

  1. short_summary_run_sample.txt - Contains a plain text summary of the results in BUSCO notation. Also gives a brief breakdown of the metrics.

    # BUSCO version is: 2.0 
    # The lineage dataset is: sample dataset BUSCO 2.0 (Creation date: 07.10.2016, number of species: 23, number of BUSCOs: 10)
    # To reproduce this run: python /usr/bin/BUSCO.py -i target.fa -o run_sample -l example/ -m genome -c 4 -sp fly -e 0.03
    #
    # Summarized benchmarking in BUSCO notation for file target.fa
    # BUSCO was run in mode: genome

    C:80.0%[S:80.0%,D:0.0%],F:0.0%,M:20.0%,n:10

    8 Complete BUSCOs (C)
    8 Complete and single-copy BUSCOs (S)
    0 Complete and duplicated BUSCOs (D)
    0 Fragmented BUSCOs (F)
    2 Missing BUSCOs (M)
    10 Total BUSCO groups searched

  2.  full_table_run_sample.txt - Contains the complete results in a tabular format with scores and lengths of BUSCO matches, and coordinates (for genome mode) or gene/protein IDs (for transcriptome or proteins mode).
  3. missing_busco_list_run_sample.tsv - Contains a list of missing BUSCOs.

2- Directories

  1. augustus_output - Augustus-predicted genes, only created during genome assessment. File: augustus.log = full details on Augustus jobs File: training_set_XXXX.txt = genes used for Augustus training Folder: predicted_genes = Augustus raw gene output Folder: extracted_proteins = Augustus protein FASTA output Folder: retraining_parameters = Augustus training results Folder: gb = GenBank format complete BUSCOs Folder: gffs = General Feature Format complete BUSCOs
  2. blast_output - tBLASTn results, not created for assessment of proteins. File: tblastn_XXXX.txt = tabular tBLASTn results File: coordinates_XXXX.txt = locations of BUSCO matches (genome mode)
  3. hmmer_output Tabular format HMMER output of searches with BUSCO HMMs
  4. single_copy_busco_sequences - FASTA format file for each complete single-copy BUSCO identified. .faa files contain protein sequences .fna files contain coding sequences (DNA, genome mode only).

More information on BUSCO-v2 inputs, outputs and parameters can be found in this manual

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