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

0505474 - MBÚ 2020 RIV JP eng J - Journal Article
Strunecký, O. - Kopejtka, Karel - Goecke, Franz - Tomasch, J. - Lukavský, Jaromír - Neori, A. - Kahl, S. - Pieper, D. H. - Pilarski, P. - Kaftan, David - Koblížek, Michal
High diversity of thermophilic cyanobacteria in Rupite hot spring identified by microscopy, cultivation, single-cell PCR and amplicon sequencing.
Extremophiles. Roč. 23, č. 1 (2019), s. 35-48. ISSN 1431-0651. E-ISSN 1433-4909
R&D Projects: GA ČR GA15-00703S; GA ČR GA15-00113S; GA MŠMT(CZ) LO1416
Institutional support: RVO:61388971 ; RVO:67985939
Keywords : Bulgaria * Cyanobacteria * Extremophile
OECD category: Biochemistry and molecular biology; Plant sciences, botany (BU-J)
Impact factor: 2.462, year: 2019
Method of publishing: Limited access
https://link.springer.com/article/10.1007%2Fs00792-018-1058-z

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 degrees 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 degrees C. Chlorogloeopsis sp. was the most abundant at the highest temperature (68 degrees C), followed by Leptolyngbya boryana, Thermoleptolyngbya albertanoae, Synechococcus bigranulatus, Oculatella sp., and Desertifilum sp. thriving above 60 degrees C, while Leptolyngbya geysericola, Geitlerinema splendidum, and Cyanobacterium aponinum were found above 50 degrees C.
Permanent Link: http://hdl.handle.net/11104/0296964