A REFERENCE SET OF RUMEN MICROBIAL GENOMES: THE HUNGATE1000 PROJECT. William J. Kelly*1 (bill.kelly@agresearch.co.nz), Graeme T. Attwood1, Peter H. Janssen1, Adrian L. Cookson1, Gemma Henderson1, Suzanne C. Lambie1, Rechelle Perry1, Kenneth Teh1, Nikola Palevich1, Samantha Noel1, Lynne A. Goodwin2, Nicole Shapiro2, Tanja Woyke2, Christopher J. Creevey3, Sinead C. Leahy1. 1AgResearch Ltd, Grasslands Research Centre, Palmerston North, New Zealand; 2DOE Joint Genome Institute, Walnut Creek, California, USA; 3Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, UK. Introduction. Acknowledgements. Research is funded by the New Zealand Government to support the objectives of the Livestock Research Group of the Global Research Alliance on Agricultural Greenhouse Gases. The information contained within this poster should not be taken to represent the views of the Alliance as a whole or its partners. Table 1. Comparison of rumen bacteria covered by culture information ,16S rRNA gene-based studies and genome sequencing. Phylum Cultured genera Cultured isolates Meta-analysis Hungate1000 Actinobacteria 11 (13%) 25 (17%) 41 (2%) 13 (6%) Bacteriodetes 6 (7%) 5 (3%) 907 (38%) 15 (7%) Fibrobacteres 1 (1%) 7 (5%) 16 (1%) 1 (0%) Firmicutes 45 (51%) 90 (62%) 1263 (53%) 186 (84%) Fusobacteria 1 (1%) _ 1 (0%) 1 (0%) Proteobacteria 20 (23%) 16 (11%) 64 (3%) 3 (1%) Spirochaetes 1 (1%) 2 (1%) 48 (2%) 2 (1%) Synergistetes 1 (1%) 1 (1%) 11 (0%) _ Tenericutes 2 (2%) _ 21 (1%) _ Development of the host-microbe relationship begins at birth and continues throughout adult life. The rumen is undeveloped at birth and incomplete information exists about the microbial diversity and colonization dynamics that result in a functioning rumen after weaning. It is conceivable that early-life microbial intervention represents an opportunity to manipulate the indigenous microbial populations of the ruminant and influence the lifelong environmental impact and productivity of the animal. Bifidobacteria are a major component of the microbiota of the ruminant digestive tract from birth until weaning. Two different species were sequenced and their genomes reflect different lifestyles. Genomes of six other rumen Bifidobacterium species are in progress. Bifidobacterium longum subsp. infantis AGR2137 encodes genes for the utilization of bovine milk oligosaccharides and also host-derived glucans. This strain resembles those isolated from human infants but lacks the capacity to utilize fucose and sialic acid substituted oligosaccharides. Bifidobacterium pseudolongum AGR2145 encodes genes for the utilization of a range of plant derived oligosaccharides including fructo-oligosaccharides, galacto- oligosaccharides, arabinoxylo-oligosaccharides and raffinose. This strain also encodes genes that specify sortase-dependent pili which may facilitate host-microbe interaction. Early life. B. longum subsp. infantis 2.27 Mb B. pseudolongum 1.99 Mb Diversity. Butyrivibrio is one of the main genera of rumen bacteria able to initiate the breakdown of hemicellulose and pectin found in plant cell walls. We sequenced and compared the genomes of several strains identified as Butyrivibrio on the basis of 16rRNA analysis. Figure 2 shows three main clusters of strains belonging to the genus Butyrivibrio, although these do not correspond to the three currently recognised species. Strains belonging to the genus Pseudobutyrivibrio form a separate cluster while one additional strain (AE2032) presumably represents a novel genus. The genomes of the Butyrivibrio strains range in size from 3.4 to 5.1 Mb and encode between one and seven multi-domain xylanases belonging to the GH10 and GH11 families. Figure 2. Comparison of the complete ORFeomes of Butyrivibrio genomes by functional genome distribution analysis (Altermann 2012, Front. Microbiol. 3, 48). Figure 3. Genome atlas of Ruminococcus flavefaciens AE3010. From outside to the centre: Genes on forward strand; Genes on reverse