rDNA analyses of planktonic heterocystous cyanobacteria, including members of the genera Anabaenopsis and Cyanospira

Unité des Cyanobactéries (CNRS URA 2172), Département de Biochimie et Génétique Moléculaire, Institut Pasteur, 28 rue du Dr Roux, 75724 Paris Cedex 15, France 1 Author for correspondence: Isabelle Iteman. Tel: +33 1 4568 8416. Fax: +33 1 4061 3042. e-mail: iiteman{at}pasteur.fr The taxonomic coherence and phylogenetic relationships of 11 planktonic heterocystous cyanobacterial isolates were examined by investigating two areas of the rRNA operon, the 16S rRNA gene ( rrnS ) and the internal transcribed spacer (ITS) located between the 16S rRNA and 23S rRNA genes. The rrnS sequences were determined for five strains, including representatives of Anabaena flos - aquae , Aphanizomenon flos - aquae , Nodularia sp. and two alkaliphilic planktonic members of the genera Anabaenopsis and Cyanospira , whose phylogenetic position was previously unknown. Comparison of the data with those previously published for individual groups of planktonic heterocystous cyanobacteria showed that, with the exception of members assigned to the genus Cylindrospermopsis , all the planktonic strains form a distinct subclade within the monophyletic clade of heterocystous cyanobacteria. Within this subclade five different phylogenetic clusters were distinguished. The phylogenetic groupings of Anabaena and Aphanizomenon strains within three of these clusters were not always consistent with their generic or specific assignments based on classical morphological definitions, and the high degree of sequence similarity between strains of Anabaenopsis and Cyanospira suggests that they may be assignable to a single genus. Ribotyping and additional studies performed on PCR amplicons of the 16S rDNA or the ITS for the 11 planktonic heterocystous strains demonstrated that they all contain multiple rrn operons and ITS regions of variable size. Finally, evidence is provided for intra-genomic sequence heterogeneity of the 16S rRNA genes within most of the individual isolates. Keywords: 16S rRNA, internal transcribed spacer, phylogeny, sequence heterogeneity, restriction fragment length polymorphism Abbreviations: IGS, intergenic spacer region; ITS, internal transcribed spacer of the rRNA operon; PCC, Pasteur Culture Collection of Cyanobacteria; PPFD, photosynthetic photon flux density; RB, relative binding The GenBank accession numbers of the 16S rDNA gene sequences reported in this paper are AY038032 – AY038037 .