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
Mares, Jan; Lara, Yannick; Dadakova, Inaet al.
2015 • In Journal of Phycology, 51 (2), p. 288–297
[en] 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 Petalonema and Stigonema. 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 Stigonema from Europe, North and Central America, and Hawaii, and the type species of Petalonema from Slovakia were sequenced. Contrary to some previous studies, which proposed a relationship with heteropolar nostocalean cyanobacteria, Petalonema appeared to belong to the family Scytonemataceae. Analysis of Stigonema 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 Stigonema clustered in a phylogenetic branch derived from uni- to biseriate S. ocellatum 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.
Research center :
CIP - Centre d'Ingénierie des Protéines - ULiège
Disciplines :
Environmental sciences & ecology Microbiology
Author, co-author :
Mares, Jan
Lara, Yannick ; Université de Liège > Centre d'ingénierie des protéines
Dadakova, Ina
Hauer, Tomas
Uher, Bohuslav
Wilmotte, Annick ; Université de Liège > Département des sciences de la vie > Physiologie et génétique bactériennes
Kastovsky, Jan
Language :
English
Title :
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
Alternative titles :
[fr] Analyse phylogénétique des cyanobactéries terrestres résistantes à la mise en culture et avec des larges gaines (Stigonema spp. et Petalonema alatum, Nostocales, Cyanobacterie) en utilisant le séquençage de cellules ou filaments isolés à partir d'échantillons environnementaux
Publication date :
April 2015
Journal title :
Journal of Phycology
ISSN :
0022-3646
eISSN :
1529-8817
Publisher :
Blackwell Publishing, Oxford, United Kingdom
Volume :
51
Issue :
2
Pages :
288–297
Peer reviewed :
Peer Reviewed verified by ORBi
Name of the research project :
Single cell sequencing of cyanobacteria
Funders :
F.R.S.-FNRS - Fonds de la Recherche Scientifique [BE] ULg - Université de Liège [BE]
Funding text :
Université de Liège (short-term research fellowship for foreign doctoral students)
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