Abstract

Molecular fingerprinting of 16S rRNA genes using terminal restriction fragment length polymorphism (T-RFLP) and denaturing gradient gel electrophoresis (DGGE) was used to characterize the temporal and spatial variability among sponge-associated bacteria from Mycale hentscheli having distinct bioactive chemotypes. Cluster analysis of T-RFLP and DGGE profiles from M. hentscheli chemotypes largely grouped sponge microbial diversity to their distinct chemotype pattern. Repeat sampling of individual M. hentscheli at one location over a 21-month period showed that the T-RFLP profiles from individual sponges had similarity indices ranging from 60% to 82% and calculated DGGE similarities between 23% and 95%. However, a portion (>35% from DGGE and >19% from T-RFLP) of the microbial community from M. hentscheli appeared to be spatially conserved through all M. hentscheli populations. Sequence analysis of DGGE band fragments showed a similarity among the bands originating from different individuals, different times, and different locations. The sponge-associated relationship of these bands was confirmed, with sequences having similarity to sponge-associated bacteria reported from global locations. This study highlights the spatial and temporal complexity in the distribution of bacterial communities associated with different chemotypes of the marine sponge M. hentscheli.

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