Consensus assessment of the contamination level of publicly available cyanobacterial genomes

PLoS One. 2018 Jul 25;13(7):e0200323. doi: 10.1371/journal.pone.0200323. eCollection 2018.

Abstract

Publicly available genomes are crucial for phylogenetic and metagenomic studies, in which contaminating sequences can be the cause of major problems. This issue is expected to be especially important for Cyanobacteria because axenic strains are notoriously difficult to obtain and keep in culture. Yet, despite their great scientific interest, no data are currently available concerning the quality of publicly available cyanobacterial genomes. As reliably detecting contaminants is a complex task, we designed a pipeline combining six methods in a consensus strategy to assess the contamination level of 440 genome assemblies of Cyanobacteria. Two methods are based on published reference databases of ribosomal genes (SSU rRNA 16S and ribosomal proteins), one is indirectly based on a reference database of marker genes (CheckM), and three are based on complete genome analysis. Among those genome-wide methods, Kraken and DIAMOND blastx share the same reference database that we derived from Ensembl Bacteria, whereas CONCOCT does not require any reference database, instead relying on differences in DNA tetramer frequencies. Given that all the six methods appear to have their own strengths and limitations, we used the consensus of their rankings to infer that >5% of cyanobacterial genome assemblies are highly contaminated by foreign DNA (i.e., contaminants were detected by 5 or 6 methods). Our results will help researchers to check the quality of publicly available genomic data before use in their own analyses. Moreover, we argue that journals should make mandatory the submission of raw read data along with genome assemblies in order to facilitate the detection of contaminants in sequence databases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Consensus
  • Cyanobacteria / genetics*
  • DNA Contamination*
  • DNA, Bacterial / genetics
  • Genes, rRNA / genetics
  • Genetic Markers / genetics
  • Genome, Bacterial / genetics*

Substances

  • DNA, Bacterial
  • Genetic Markers

Grants and funding

This work was supported by operating funds from F.R.S.-FNRS (National Fund for Scientific Research of Belgium) (AW), the European Research Council Stg ELITE FP7/308074 (EJJ), the BELSPO project CCAMBIO (SD/BA/03A) (AW), the BELSPO Interuniversity Attraction Pole Planet TOPERS (EJJ and LC), and the TULIP Laboratory of Excellence (ANR-10-LABX-41) (HP). LC, LM, MVV, RRL and BD are all FRIA (Fonds pour la formation à la Recherche dans l’Industrie et l’Agriculture) fellows of the FRS-FNRS. AW is a Research Associate of the FRS-FNRS. AM’s stay in University of Liège was partially financed by a bilateral cooperation [ARTHRO-ARN: Functional genomics-based approach for the design of genetic engineering tools in the edible cyanobacterium Arthrospira. Scientific and technical cooperation between Republic of Poland and Belgium (Fédération Wallonie-Bruxelles) (2014-2016)]. Computational resources were provided by the Consortium des Équipements de Calcul Intensif (CÉCI), funded by the F.R.S.-FNRS (2.5020.11), and through two grants to DB (University of Liège “Crédit de démarrage 2012” SFRD-12/04; F.R.S.-FNRS “Crédit de recherche 2014” CDR J.0080.15).