Sunday, May 4, 2008
2-58

Microbial and genetic analysis of a microbial community actively decaying poplar biomass

Daniel Van der Lelie1, Safiyh Taghavi1, Shi-You Ding2, and Michael E. Himmel2. (1) Biology, Brookhaven National Laboratory, 50, Bell Avenue, Upton, NY 11973, (2) Chemical and Biosciences Center, National Renewable Energy Laboratory, 1617 Cole Blvd., Golden, CO 80401

As primary decomposers, microbial communities have evolved as competitors and collaborators in deconstructing biomass. To understand and exploit these complex microbial communities and their dynamics for the conversion of recalcitrant plant biomass to useful bioenergy feedstocks, a highly integrated research initiative is required. A prerequisite is to have insight into the composition and metabolic potential of this lignocellulosic biomass degrading community. To achieve this goal several complementary strategies are used to study the microbial community actively decaying poplar woodchips:

Analysis of total community composition: After the isolation of metagenome DNA, 16S rRNA genes were PCR amplified, shotgun cloned and sequenced. In total, 238 clones were sequenced and identified to their closest matching species. The distribution of the species showed that members of the order Clostridiales, many of which are closely related to uncultivatable bacteria, comprise 85% of this community. The presence of the majority of members of this order in the community is expected because of the mesophylic, anaerobic conditions characteristic for the sample.

Isolation and characterization of cultivable microorganisms: the high number of uncultivable microorganisms in this consortium was confirmed by cultivation studies. None of the cultivable bacteria represented the dominant members of the community as determined via 16S RDNA sequencing. Isolated strains are presently screened for their glycosylhydrolase activity.

Metagenome sequencing: in order to obtain a thorough understanding of the diversity, structure, functional interdependence, and metabolic capabilities of this community. This approach should provide unprecedented insights in the diversity of glycosylhydrolases present in plant biomass decomposing microbial communities.


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