FUNCTIONAL PHYLOTYPING APPROACH FOR ASSESSING INTRASPECIFIC DIVERSITY OF RUMINOCOCCUS ALBUS WITHIN THE RUMEN MICROBIOME

Inna Rozman Grinberg 1 Guohua Yin 1 Ilya Borovok 1 Margret E. Berg Miller 2 Carl J. Yeoman 3 Bareket Dassa 7 Itzhak Mizrahi 4 Harry J. Flint 5 Zhongtang Yu 6 Edward A. Bayer 7 Bryan A. White 2 Raphael Lamed 1
1Department of Molecular Microbiology and Biotechnology, Tel Aviv University, Tel-Aviv
2Institute for Genomic Biology, University of Illinois, Urbana, Illinois
3Department of Animal and Range Sciences, Montana State University, Bozeman, Montana
4Department of Ruminant Science, Institute of Animal Sciences, Agricultural Research Organization, Beit-Dagan
5Rowett Institute Institute of Nutrition and Health, University of Aberdeen, Aberdeen
6The MAPLE Research Initiative, Department of Animal Sciences, The Ohio State University, Columbus, Ohio
7Department of Biological Chemistry, Weizmann Institute of Science, Rehovot

Ruminococcus albus, a cellulolytic bacterium, is a critical member of the rumen community. Ruminococcus albus lacks a classical cellulosome complex, but it possesses a unique family 37 carbohydrate-binding module (CBM37), which is integrated into a variety of carbohydrate-active enzymes. We developed a potential molecular tool for functional phylotyping of the R. albus population in the rumen, based on a variable region in the cel48A gene. cel48A encodes a single copy of the CBM37-associated family 48 glycoside hydrolase in all known strains of this bacterium. A segment of the cel48A gene was amplified from rumen metagenomic samples of four bovines, and its abundance and diversity were evaluated. Analysis of the obtained sequences revealed the co-existence of multiple functional phylotypes of cel48A in all four animals. These included sequences identical or similar to those of R. albus isolates (reference strains), as well as several novel sequences. The dominant cel48A type varied among animals. This method can be used for detection of intraspecific diversity of R. albus in metagenomic samples. Together with scaC, a previously reported gene marker for R. flavefaciens, we present a set of two species—specific markers for phylotyping of Ruminococci in the herbivore rumen.









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