Community Composition of Marine Bacterioplankton Determined by 16S rRNA Gene Clone Libraries and Fluorescence In Situ Hybridization

TLDR

The study aimed to determine the composition of bacterioplankton communities in coastal California surface waters using 16S rRNA gene clone libraries and fluorescence in situ hybridization to compare the community structures inferred from these two culture‑independent approaches. The authors constructed 16S rRNA gene clone libraries and applied fluorescence in situ hybridization with specific probes to assess bacterial community composition. Clone libraries showed α‑proteobacteria dominated (~55%) and γ‑proteobacteria (~28%), whereas FISH revealed the Cytophaga‑Flavobacter group dominated 8 of 11 communities, demonstrating that clone libraries over‑represent α‑proteobacteria and under‑represent Cytophaga‑Flavobacter, which can be numerically dominant in coastal marine bacterioplankton.

Abstract

ABSTRACT We determined the compositions of bacterioplankton communities in surface waters of coastal California using clone libraries of 16S rRNA genes and fluorescence in situ hybridization (FISH) in order to compare the community structures inferred from these two culture-independent approaches. The compositions of two clone libraries were quite similar to those of clone libraries of marine bacterioplankton examined by previous studies. Clones from γ-proteobacteria comprised ca. 28% of the libraries, while approximately 55% of the clones came from α-proteobacteria, which dominated the clone libraries. The Cytophaga-Flavobacter group and three others each comprised 10% or fewer of the clone libraries. The community composition determined by FISH differed substantially from the composition implied by the clone libraries. The Cytophaga-Flavobacter group dominated 8 of the 11 communities assayed by FISH, including the two communities assayed using clone libraries. On average only 10% of DAPI (4′,6′-diamidino-2-phenylindole)-stained bacteria were detected by FISH with a probe for α-proteobacteria, but 30% of DAPI-stained bacteria appeared to be in the Cytophaga-Flavobacter group as determined by FISH. α-Proteobacteria were greatly overrepresented in clone libraries compared to their relative abundance determined by FISH, while the Cytophaga-Flavobacter group was underrepresented in clone libraries. Our data show that the Cytophaga-Flavobacter group can be a numerically dominant component of coastal marine bacterioplankton communities.