Physical isolation of endospores from environmental samples by targeted lysis of vegetative cells

Tina Wunderlin, Thomas Junier, Christophe Paul, Nicole Jeanneret, Pilar Junier

Research output: Contribution to journalArticlepeer-review

4 Citations (Scopus)

Abstract

Endospore formation is a survival strategy found among some bacteria from the phylum Firmicutes. During endospore formation, these bacteria enter a morpho-physiological resting state that enhances survival under adverse environmental conditions. Even though endospore-forming Firmicutes are one of the most frequently enriched and isolated bacterial groups in culturing studies, they are often absent from diversity studies based on molecular methods. The resistance of the spore core is considered one of the factors limiting the recovery of DNA from endospores. We developed a method that takes advantage of the higher resistance of endospores to separate them from other cells in a complex microbial community using physical, enzymatic and chemical lysis methods. The endospore-only preparation thus obtained can be used for re-culturing or to perform downstream analysis such as tailored DNA extraction optimized for endospores and subsequent DNA sequencing. This method, applied to sediment samples, has allowed the enrichment of endospores and after sequencing, has revealed a large diversity of endospore-formers in freshwater lake sediments. We expect that the application of this method to other samples will yield a similar outcome.
Original languageEnglish
Article numbere53411
Pages (from-to)e53411-1-e53411-7
Number of pages7
JournalJournal of visualized experiments : JoVE
Issue number107
DOIs
Publication statusPublished - Jan 2016

Keywords

  • environmental sciences
  • endospores
  • separation
  • vegetative cells
  • endospore-forming bacteria
  • cell lysis
  • diversity
  • microbial ecology
  • Separation
  • Environmental sciences
  • Diversity
  • Microbial ecology
  • Cell lysis
  • Issue 107
  • Endospore-forming bacteria
  • Vegetative cells
  • Endospores

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