深港两地工程土壤细菌多样性的T-RFLP分析  

T- RFLP Analysis of Bacterial Diversity in Engineering Soils from Shenzhen and Hongkong

作　　者：龙海[1,2] 章桂明[1] 冯建军[1] 李芳荣[1] 程颖慧[1] 焦懿[1,2] 王颖[1] 李一农[1] 

机构地区：[1]深圳出入境检验检疫局,广东深圳518001 [2]深圳市检验检疫科学研究院深圳市外来有害生物检测技术研发重点实验室,广东深圳518045

出　　处：《生物技术通讯》2016年第1期106-109,共4页Letters in Biotechnology

基　　金：国家质检总局科技计划(2015IK266)

摘　　要：目的:建立一种高效的末端限制性片段长度多态性(T-RFLP)方法,检测工程土壤细菌。方法:以深圳和香港两地不同土层(0.5,10,20和30 m)的工程土壤为研究对象,提取并纯化总DNA,利用细菌16S r DNA基因通用引物8f-FAM/1492r进行PCR扩增,扩增产物分别用HhaⅠ、HaeⅢ和MspⅠ限制性内切酶酶切,酶切产物经毛细管电泳测序,序列上传至数据库进行分析。结果:经过比对分析,香港段土壤中大约存在细菌141属,235种;深圳段大约存在132属,206种;32个细菌种属只在香港土壤中有分布,3个细菌种属只在深圳土壤中有分布;植物病原细菌有14个种属。结论:建立的T-RFLP方法能够高效、快速地分析工程土壤细菌。Objective： To establish a rapid terminal restriction fragment length polymorphism（T-RFLP） method for detecting the bacterial diversity in engineering soils. Methods： The soil sampls from Shenzhen and Hongkong were collected from 0.5, 10, 20 and 30 m depth respectively. The total DNA was extracted and purified from the samples. 16S rDNA was amplified from the total DNA using primers 8f-FAM/1492r and then digested with Hha I, Hae Ⅲ and Msp I respectively. The products of enzyme digestion were sequenced by capillary eleetrophoresis. The sequences were uploaded to the database for comparison. Results： There were about 141 genus（235 species） of bacteria in soils of Hongkong and 132 genus（206 species） of bacteria in soils of Shenzhen. 32 species of bacte- ria were only distributed in soils of Hongkong and 3 species were only distributed in soils of Shenzhen. There were 14 species of plant bacteria. Conclusion： The T-RFLP method can effectively and quickly analyze bacteria in the engineering soil.

关 键 词：工程土壤 细菌多样性 末端限制性片段长度多态性 

分 类 号：Q789[生物学—分子生物学]