Phylogenetic analysis of cultivation‐resistant terrestrial cyanobacteria with massive sheaths (Stigonema spp. and Petalonema alatum, Nostocales, Cyanobacteria) using single‐cell and filament sequencing of environmental samples. Issue 2 (12th February 2015)

• Record Type: Journal Article
• Authors: Mareš, Jan; Lara, Yannick; Dadáková, Iva; Hauer, Tomáš; Uher, Bohuslav; Wilmotte, Annick; Kaštovský, Jan; Collier, J. L.
• Is Part Of: Journal of phycology. Volume 51:Issue 2(2015)
• Page Start: 288
• Page End: 297
• Publication Date: 2015-02-12
• Subjects: Algae -- Periodicals; 579.8
• DOI: 10.1111/jpy.12273
• Languages: English
• ISSNs: 0022-3646

Abstract

<abstract abstract-type="main" id="jpy12273-abs-0001"> <title> <x xml:space="preserve">Abstract</x> </title> <p>Molecular assessment of a large portion of traditional cyanobacterial taxa has been hindered by the failure to isolate and grow them in culture. In this study, we developed an optimized protocol for single cell/filament isolation and 16S rRNA gene sequencing of terrestrial cyanobacteria with large mucilaginous sheaths, and applied it to determine the phylogenetic position of typical members of the genera <italic>Petalonema</italic> and <italic>Stigonema</italic>. A methodology based on a glass‐capillary isolation technique and a semi‐nested PCR protocol enabled reliable sequencing of the 16S rRNA gene from all samples analyzed. Ten samples covering seven species of <italic>Stigonema</italic> from Europe, North and Central America, and Hawaii, and the type species of <italic>Petalonema</italic> from Slovakia were sequenced. Contrary to some previous studies, which proposed a relationship with heteropolar nostocalean cyanobacteria, <italic>Petalonema</italic> appeared to belong to the family Scytonemataceae. Analysis of <italic>Stigonema</italic> specimens recovered a unique coherent phylogenetic cluster, substantially broadening our knowledge of the molecular diversity within this genus. Neither the uni‐ to biseriate species nor the multiseriate species formed monophyletic subclusters within the genus. Typical multiseriate species of <italic>Stigonema</italic> clustered in a phylogenetic branch derived from uni‐ to biseriate <italic>S. ocellatum</italic> Thuret ex Bornet & Flahault in our analysis, suggesting that species with more complex thalli may have evolved from the more simple ones. We propose the technique tested in this study as a promising tool for a future revision of the molecular taxonomy in cyanobacteria.</p> </abstract>