新疆农业科学2018，Vol.55Issue(1)：9-15,7.DOI:10.6048/j.issn.1001-4330.2018.01.002
新疆棉花根际细菌TaqMan探针实时荧光定量PCR的建立及时空动态分析
Establishme nt and Spatiotemporal Dynamic Analysis of Rhizosphere Bacteria of Cotton in Xingjiang using Real-time Fluorescent TaqMa n-quantitative PCR
摘要
Abstract
[Objective] In order to determine the total amount of rhizosphere bacteria of cotton during dif-ferent growth periods and study the spatiotemporal dynamics,a rapid real-time fluorescent TaqMan-quanti-tative PCR technique was established.[Method]A set of primers and TaqMan probe specific for rhizosphere bacteria of cotton were designed according to the conserved region of 16S rDNA gene, and the recombinant plasmid were constructed as a standard curve, and practical soil samples were detected using the technique.[Result]The number of rhizosphere bacteria in the rhizosphere of Xinjiang cotton varied in different growth periods.The number of inter -rhizosphere bacteria in Korla, Alaer and Hami were the same between the seedling stage and the flowering period, while there was a difference between flower stage and boll opening stage;The trend of rhizosphere bacteria in the whole growth period of cotton in Shihezi, Wusu and Tumu Shuker were basically the same;The rhizosphere bacteria began to increase slowly from seedling stage in Jing-he,and there was little change between bud stage and flower stage,but it increased rapidly from boll opening stage.The number of total rhizosphere bacteria of cotton showed different trends in different sampling sites of cotton in Xinjiang.The spatial variation trend of number of rhizosphere bacteria was decreasing in the Eastern, Southern,Northern Xinjiang.Among them,the largest number of rhizosphere bacteria was in Hami in Eastern Xingjiang,and reached 1.7×107copies/g(FRW)in boll opening stage, the least was only 8.5×104cop-ies/g(FRW)in seedling stage in Jinghe in northern Xingjiang.[Conclusion]The method of real-time fluo-rescent TaqMan-quantitative PCR can be used to rapidly detect the number of total rhizosphere bacteria of cotton,and when using this method we find that the number of total rhizosphere bacteria showed different trends in different growth periods and sampling sites of cotton in Xinjiang.The largest number of rhizosphere bacteria was found in Hami in boll opening stage,the least was detected in seedling stage in Jinghe County.关键词
棉花/根际细菌/TaqMan探针/实时荧光定量PCRKey words
otton/rhizosphere bacteria/TaqMan probe/real-time fluorescent quantitative PCR分类
农业科技引用本文复制引用
张涛,欧提库尔,李玉国,娄恺,史应武,李雪艳,杨红梅,楚敏,高雁,曾军,霍向东,林青..新疆棉花根际细菌TaqMan探针实时荧光定量PCR的建立及时空动态分析[J].新疆农业科学,2018,55(1):9-15,7.基金项目
国家自然科学基金面上项目(41471220) （41471220）
新疆维吾尔自治区优秀青年科技人才培养项目(qn2005yx023) （qn2005yx023）
中国博士后科学基金项目(2016M602953XB)The National Natural Science Foundation of China(41471220) （2016M602953XB）
The Outstanding Young Talent Training Program of Xinjiang Uygur Autonomous Region(2015A020209126) （2015A020209126）
China PhD Science Foundation Program(No.2016M602953XB) （No.2016M602953XB）
