土壤学报2009，Vol.46Issue(6)：1096-1101,6.
培养方法对土壤可培养细菌多样性的影响
EFFECT OF CULTIVATION METHODS ON BACTERIAL DIVERSITY BY PCR RFLP ANALYSIS
摘要
Abstract
Improving the culturability of soil microorganisms and isolating pure strains are the bases of the study of microbial ecology. Three culturing media were used for time interval counting of soil bacteria. Bacterial universal primers were used to amplify 16S rDNA fragments; PCR products digested by restriction endonucleases Hha I and their fingerprints analyzed for influence of culturing methods on soil microbial diversity and culturability. Results indicate that the number of bacteria grown in media LB, CSEA and WSA was 14.84×10~7, 10.27×10~7 and 6.91×10~7 CFU per gram dry soil, respectively, after 192 h of incubation. The diversity index of soil bacteria was the highest in WSA, while the lowest in LB. Certain similarity was discovered in bacteria community between the three media, i.e. the Jaccard index was 57.69% between LB and CSEA, 53.13% between LB and WSA and 66.67% between CSEA and WSA. Phylogenetic analysis suggests that the dominant bacterial groups in the soil belong to β-Proteobacteria,γ-Proteobacteria and Actinobacteria. The sequence of 16S rDNA of some OTUs was found to have high homology with Burkholderiaceae bacterium, Rhodococcus and Mycobacterium, so it is presumed that the cells of these bacteria can secrete Resuscitation Promoting Factors, which may effectively improve culturability of soil microorganisms.关键词
微生物多样性/可培养性/RFLP分析/培养基/培养时间Key words
Microbial diversity/Culturability/RFLP analysis/Culture media/Incubation time分类
农业科技引用本文复制引用
王萍,崔中利,刘标,孙波,曹慧..培养方法对土壤可培养细菌多样性的影响[J].土壤学报,2009,46(6):1096-1101,6.基金项目
国家自然科学基金项目(40871125,40871123)、环保公益性行业科研专项(200709047)和江苏省自然科学基金项目(BK2006501)资助 （40871125,40871123）
