High diversity of thermophilic cyanobacteria in Rupite hot spring identified by microscopy, cultivation, single-cell PCR and amplicon sequencing

Otakar Strunecký, Karel Kopejtka, Franz Goecke, Jürgen Tomasch, Jaromír Lukavský, Amir Neori, Silke Kahl, Dietmar H. Pieper, Plamen Pilarski, David Kaftan, Michal Koblížek

Research output: Contribution to journalArticlepeer-review

Abstract

Genotypic and morphological diversity of cyanobacteria in the Rupite hot spring (Bulgaria) was investigated by means of optical microscopy, cultivation, single-cell PCR, and 16S rRNA gene amplicon sequencing. Altogether, 34 sites were investigated along the 71–39 °C temperature gradient. Analysis of samples from eight representative sites shown that Illumina, optical microscopy, and Roche 454 identified 72, 45 and 19% respective occurrences of all cumulatively present taxa. Optical microscopy failed to detect species of minor occurrence; whereas, amplicon sequencing technologies suffered from failed primer annealing and the presence of species with extensive extracellular polysaccharides production. Amplicon sequencing of the 16S rRNA gene V5–V6 region performed by Illumina identified the cyanobacteria most reliably to the generic level. Nevertheless, only the combined use of optical microscopy, cultivation and sequencing methods allowed for reliable estimate of the cyanobacterial diversity. Here, we show that Rupite hot-spring system hosts one of the richest cyanobacterial flora reported from a single site above 50 °C. Chlorogloeopsis sp. was the most abundant at the highest temperature (68 °C), followed by Leptolyngbya boryana, Thermoleptolyngbya albertanoae, Synechococcus bigranulatus, Oculatella sp., and Desertifilum sp. thriving above 60 °C, while Leptolyngbya geysericola, Geitlerinema splendidum, and Cyanobacterium aponinum were found above 50 °C.

Original languageEnglish
Pages (from-to)35-48
Number of pages14
JournalExtremophiles
Volume23
Issue number1
DOIs
StatePublished - 22 Jan 2019
Externally publishedYes

Bibliographical note

Publisher Copyright:
© 2018, Springer Japan KK, part of Springer Nature.

Keywords

  • Bulgaria
  • Cyanobacteria
  • Extremophile
  • Hot spring

ASJC Scopus subject areas

  • Microbiology
  • Molecular Medicine

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