Platon: identification and characterization of bacterial plasmid contigs from short-read draft assemblies.
TL;DR Platon detects plasmid contigs within bacterial draft genomes from WGS short-read assemblies. Therefore, Platon analyzes the natural distribution biases of certain protein coding genes between chromosomes and plasmids. This analysis is complemented by comprehensive contig characterizations upon which several heuristics are applied.
Platon conducts three analysis steps:
- It predicts and searches coding sequences against a custom and pre-computed database comprising marker protein sequences (MPS) and related replicon distribution scores (RDS). These scores express the empirically measured frequency biases of protein sequence distributions between plasmids and chromosomes pre-computed on complete NCBI RefSeq replicons. Platon calculates the mean RDS for each contig and either classifies them as chromosome if the RDS is below a sensitivity cutoff determined to 95% sensitivity or as plasmid if the RDS is above a specificity cutoff determined to 99.9% specificity. Exact values for these thresholds have been computed based on Monte Carlo simulations of artifical replicon fragments created from complete RefSeq chromosome and plasmid sequences.
- Contigs passing the sensitivity filter get comprehensivley characterized. Hereby, Platon tries to circularize the contig sequences, searches for rRNA, replication, mobilization and conjugation genes, oriT sequences, incompatibility group DNA probes and finally performs a BLAST+ search against the NCBI plasmid database.
- Finally, to increase the overall sensitivity, Platon classifies all remaining contigs based on the gathered information by several heuristics.
Platon accepts draft assemblies in fasta format. If contigs have been assembled with SPAdes, Platon is able to extract the coverage information from the contig names.
For each contig classified as plasmid sequence the following columns are printed
to STDOUT
as tab separated values:
- Contig ID
- Length
- Coverage
- # ORFs
- RDS
- Circularity
- Incompatibility Type(s)
- # Replication Genes
- # Mobilization Genes
- # OriT Sequences
- # Conjugation Genes
- # rRNA Genes
- # Plasmid Database Hits
In addition, Platon writes the following files into the output directory:
<prefix>
.plasmid.fasta: contigs classified as plasmids or plasmodal origin<prefix>
.chromosome.fasta: contigs classified as chromosomal origin<prefix>
.tsv: dense information as printed to STDOUT (see above)<prefix>
.json: comprehensive results and information on each single plasmid contig. All files are prefixed (<prefix>
) as the input genome fasta file.
Platon can be installed in 2 different ways, though we advise to use Conda/BioConda.
In all cases, the custom database must be downloaded which we provide for download:
Example:
$ conda install -c conda-forge -c bioconda -c defaults platon
$ wget https://zenodo.org/record/3751774/files/db.tar.gz
$ tar -xzf db.tar.gz
$ rm db.tar.gz
$ platon --db ./db genome.fasta
- clone the the repository
- install Platon & Python dependencies per pip
- download & extract the database
Example:
$ git clone [email protected]:oschwengers/platon.git
$ cd platon
$ python3 -m pip install .
$ cd ..
$ wget https://zenodo.org/record/3751774/files/db.tar.gz
$ tar -xzf db.tar.gz
$ rm db.tar.gz
$ platon/bin/platon --db ./db genome.fasta
Info: Just move the extracted database directory into the platon directory. Platon will automatically recognise it and thus, the database path doesn't need to be specified:
$ ...
$ mv db/ platon
$ platon/bin/platon genome.fasta
Usage:
usage: platon [-h] [--db DB] [--threads THREADS] [--verbose] [--characterize]
[--output OUTPUT] [--version]
<genome>
Identification and characterization of bacterial plasmid contigs from short-read draft assemblies.
positional arguments:
<genome> draft genome in fasta format
optional arguments:
-h, --help show this help message and exit
--db DB, -d DB database path (default = <platon_path>/db)
--mode {sensitivity,accuracy,specificity}, -m {sensitivity,accuracy,specificity}
applied filter mode: sensitivity: RDS only (>= 95%
sensitivity); specificity: RDS only (>=99.9%
specificity); accuracy: RDS & characterization
heuristics (highest accuracy)
--threads THREADS, -t THREADS
number of threads to use (default = number of
available CPUs)
--verbose, -v print verbose information
--characterize, -c deactivate filters; characterize all contigs
--output OUTPUT, -o OUTPUT
output directory (default = current working directory)
--version, -V show program's version number and exit
Simple:
$ platon genome.fasta
Expert: writing results to results
directory with verbose output using 8 threads:
$ platon -db ~/db --output results/ --verbose --threads 8 genome.fasta
Platon provides 3 different modi controlling which filters will be used.
Accuracy
mode is the preset default.
In the sensitivity
mode Platon will classifiy all contigs with an RDS
value below
the sensitivity threshold as chromosomal and all remaining contigs as plasmid.
This threshold was defined to account for 95% sensitivity and computed via Monte Carlo
simulations of artifical contigs resulting in an RDS=-7.7.
-> use this mode to exclude chromosomal contigs.
In the specificity
mode Platon will classifiy all contigs with an RDS
value above
the specificity threshold as plasmid and all remaining contigs as chromosomal.
This threshold was defined to account for 99.9% specificity and computed via Monte Carlo
simulations of artifical contigs resulting in an RDS=0.4.
In the accuracy
mode Platon will classifiy all contigs with:
- an
RDS
value below the sensitivity threshold as chromosomal - an
RDS
value above the specificity threshold as plasmid and in addition all contigs as plasmid for which one of the following is true: it - can be circularized
- has an incompatibility group sequence
- has a replication or mobilization HMM hit
- has an oriT hit
- has an RDS above the conservative score (0.1), a RefSeq plasmid hit and no rRNA hit
Platon depends on a custom database based on MPS, RDS, RefSeq Plasmid database, PlasmidFinder db as well as manually curated MOB HMM models from MOBscan, custom conjugation and replication HMM models and oriT sequences from MOB-suite. This database based on UniProt UniRef90 release 2020_01 can be downloaded here: (zipped 1.4 Gb, unzipped 2.4 Gb)
Platon was developed and tested in Python 3.5 and depends on BioPython (>=1.71).
Additionally, it depends on the following 3rd party executables:
- Prodigal (2.6.3) https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2848648 https://github.com/hyattpd/Prodigal
- Diamond (0.9.32) https://pubmed.ncbi.nlm.nih.gov/25402007 http://www.diamondsearch.org
- Blast+ (2.7.1) https://www.ncbi.nlm.nih.gov/pubmed/2231712 https://blast.ncbi.nlm.nih.gov
- MUMmer (4.0.0-beta2) https://www.ncbi.nlm.nih.gov/pmc/articles/PMC395750/ https://github.com/gmarcais/mummer
- HMMER (3.2.1) https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3695513/ http://hmmer.org/
- INFERNAL (1.1.2) https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3810854 http://eddylab.org/infernal
A manuscript is submitted. To temporarily cite our work, please transitionally refer to:
Schwengers O., Barth P., Falgenhauer L., Hain T., Chakraborty T., Goesmann A. (2020) Platon: identification and characterization of bacterial plasmid contigs in short-read draft assemblies exploiting protein-sequence-based replicon distribution scores. bioRxiv 2020.04.21.053082; doi: https://doi.org/10.1101/2020.04.21.053082
As Platon takes advantage of the inc groups, MOB HMMs and oriT sequences of the following databases, please also cite:
-
Carattoli A., Zankari E., Garcia-Fernandez A., Voldby Larsen M., Lund O., Villa L., Aarestrup F.M., Hasman H. (2014) PlasmidFinder and pMLST: in silico detection and typing of plasmids. Antimicrobial Agents and Chemotherapy, https://doi.org/10.1128/AAC.02412-14
-
Garcillán-Barcia M. P., Redondo-Salvo S., Vielva L., de la Cruz F. (2020) MOBscan: Automated Annotation of MOB Relaxases. Methods in Molecular Biology, https://doi.org/10.1007/978-1-4939-9877-7_21
-
Robertson J., Nash J. H. E. (2018) MOB-suite: Software Tools for Clustering, Reconstruction and Typing of Plasmids From Draft Assemblies. Microbial Genomics, https://doi.org/10.1099/mgen.0.000206
If you run into any issues with Platon, we'd be happy to hear about it!
Please, start the pipeline with -v
(verbose) and do not hesitate
to file an issue including as much of the following as possible:
- a detailed description of the issue
- the platon cmd line output
- the
<prefix>.json
file if possible - A reproducible example of the issue with a small dataset that you can share (helps us identify whether the issue is specific to a particular computer, operating system, and/or dataset).