Adapting MolecularGenetic Methods for Studying
the Taxonomic Diversity of Microbial Communities Associated
with Macrophytes

I. V. Rybakova^a, N. L. Bel’kova^b, N. A. Lapteva^a, and E. V. Sukhanova^a

^aInstitute for the Biology of Inland Waters, Russian Academy of Sciences, Borok, 152742 Russia

^bLimnological Institute, Siberian Division of the Russian Academy of Sciences,
Ulan-Batorskaya ul., 3, PB 278, Irkutsk, 664033 Russia

Abstract—The combination of molecular–genetic techniques used in the study is applied to investigate micro-
organisms associated with macrophytes. The method of enzymatic lysis with phenol–chloroform extraction is
optimal for the total bacterial DNA isolation from both periphyton organisms and enriched cultures. Amplifying
the total DNA on conservative primers at the two-step PCR regime is recommended. An analysis of the taxo-
nomic diversity of the microbial community from biofilm on the reed grass and in the enriched cultures propa-
gated on various growth media has been carried out. The results have revealed a high diversity of periphyton
microorganisms associated with reed grass, including representatives of such phylogenetic lines as proteobac-
teria (, , , and subgroups), Bacteroidetes/Chlorobi, Chlamydiae/Verrucomicrobia, and cyanobacteria. The
low diversity of sequences in enriched cultures is represented by dominating genotypes of Cellvibrio with a
high percentage of homology and uncultivated bacilla.

Key words: molecular–genetic approach, bacterioperiphyton, taxonomy, rRNA gene 16S, phylogeny.

DOI:10.1134/S1995082909010155

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