Department of Environmental Science and Engineering, Ewha Womans University, 11-1 Daehyun-dong, Seodaemun-gu, Seoul 120-750, Korea.
J Microbiol Biotechnol. 2012 Mar;22(3):360-70. doi: 10.4014/jmb.1109.09052.
Current ecological knowledge of methanotrophic biofilms is incomplete, although they have been broadly studied in biotechnological processes. Four individual DNA samples were prepared from a methanotrophic biofilm, and a multiplex 16S rDNA pyrosequencing was performed. A complete library (before being de-multiplexed) contained 33,639 sequences (average length, 415 nt). Interestingly, methanotrophs were not dominant, only making up 23% of the community. Methylosinus, Methylomonas, and Methylosarcina were the dominant methanotrophs. Type II methanotrophs were more abundant than type I (56 vs. 44%), but less richer and diverse. Dominant non-methanotrophic genera included Hydrogenophaga, Flavobacterium, and Hyphomicrobium. The library was de-multiplexed into four libraries, with different sequencing efforts (3,915-20,133 sequences). Sørrenson abundance similarity results showed that the four libraries were almost identical (indices > 0.97), and phylogenetic comparisons using UniFrac test and P-test revealed the same results. It was demonstrated that the pyrosequencing was highly reproducible. These survey results can provide an insight into the management and/or manipulation of methanotrophic biofilms.
目前对甲烷营养生物膜的生态知识还不完全了解，尽管它们在生物技术过程中已经得到了广泛的研究。从甲烷营养生物膜中制备了四个单独的 DNA 样本，并进行了多重 16S rDNA 焦磷酸测序。一个完整的文库（未进行解复用之前）包含 33639 条序列（平均长度为 415nt）。有趣的是，甲烷营养菌并不占优势，只占群落的 23%。甲基单胞菌、甲基杆菌和甲基八叠球菌是主要的甲烷营养菌。与 I 型相比，II 型甲烷营养菌更为丰富（56%对 44%），但丰富度和多样性较低。优势非甲烷营养菌属包括氢噬菌属、黄杆菌属和微菌属。该文库被解复用为四个文库，测序工作量不同（3915-20133 条序列）。Sørrenson 丰度相似性结果表明，四个文库几乎相同（指数>0.97），使用 UniFrac 测试和 P 测试进行的系统发育比较也得到了相同的结果。结果表明焦磷酸测序具有高度的可重复性。这些调查结果可以为甲烷营养生物膜的管理和/或操纵提供一些见解。
