中国大陆科学钻探(CCSD)地下岩心样品中的细菌群落分析

16S rDNA analysis of bacterial community in Donghai terrestrial deep subsurface:Microbiology research of the Chinese Continent Scientific Drilling.

通过建立高效率的微生物未培养DNA提取技术和16SrDNA分析技术,在中国大陆钻探深层岩芯中发现微生物存在的证据。从地下430.00米(S1)与1033.77米(S13)深度岩芯中分别提取到微生物DNA,进行16SrDNAPCR扩增,通过对S13样品DNA中克隆得到的54个16SrDNA的序列测定,初步构建了该深度地下微生物群落的系统进化树,发现了23种不同的微生物序列存在。通过DAPI染色证明在地下1033.77米的微生物数量约为1×104个/克岩石,采用实时定量PCR分析分析,发现了其中两种微生物(CCSD1305与CCSD1307)的数量为6.6×104+/g岩石与2.18×106个/g岩石。

Evidences for the existence of bacteria in the terrestrial deep subsurface of the Chinese Continent Scientific Drilling have been found through independent culture methods and by high efficiency DNA extraction and analysis of ribosomal DNA (16S rDNA) sequences. Crude DNA extracted from 430.00 (SI) and 1033.17 (S13) meter-deep-subsurface rocks, was used as template for polymerase chain reaction (PCR). After sequence analysis of 16S rDNA obtained from the S13 sample a phylogenetic tree was constructed suggesting the existence of at leas 23 different bacteria species. The cell number in the S13 is about 1.0 x 10(4) cells per gram rock as determined by 4, 6-Diamidino-2-phenylindole (DAPT) staining method. Using real time PCR, we determined the numbers of two different bacterial species (CCSD1305 and CCSD1307) to be 6.6 X 10(4) and 2.18 X 10(6) cells, respectively, per gain rock in the S13.