大豆根际土壤真菌分子生物学鉴定方法  

Molecular Identification of the Fungi in the Rhizospheric Soil of Soybean

作　　者：接伟光[1] 张勇[1] 蔡柏岩[2] 白莉[1] 王丽阳[2] 李二平[3] 

机构地区：[1]黑龙江东方学院食品与环境工程学部,黑龙江哈尔滨150086 [2]黑龙江大学生命科学学院,分子生物学重点实验室,黑龙江哈尔滨150080 [3]东北林业大学生命科学学院,黑龙江哈尔滨150040

出　　处：《大豆科学》2011年第3期384-387,共4页Soybean Science

基　　金：黑龙江省教育厅2010年度科学技术研究(指导)项目(11553080);黑龙江省自然科学基金资助项目(C200918);黑龙江省博士后科研启动金资助项目(LBH-Q09022);黑龙江大学高层次人才支持计划资助项目(生态修复团队Hdtd2010-12)

摘　　要：以大豆根际土壤为研究对象,采用湿筛倾析-蔗糖离心法分离大豆根际土壤真菌孢子。应用Nested-PCR技术扩增其28S rDNA D1/D2区域,并结合DNA测序、系统发育分析,对其进行分类鉴定。结果表明:筛选出的大豆根际土壤真菌孢子DT-1与土壤真菌(AB438763)有较高的序列同源性,为97%;DT-2与Mucor racemosus(Y213713)有较高的序列同源性,为99%。此方法适用于大豆根际土壤真菌的鉴定,并且能够快速准确地得到试验结果。The fungi in the rhizospheric soil of soybean played a vital role in the formation of soil fertility,the spread of soil-borne diseases,the occurrence of allelopathy,etc.In particularly,soybean root rot caused by soil fungi has became a major disease which result in soybean production declined dramatically in China.Therefore,the rapid identification of the fungi in rhizospheric soil of soybean has great significance in preventing and curing fungal diseases of soybean.In this study,the fungal spores were isolated from soybean rhizospheres following a procedure including wet sieving,decanting,and separation in a sucrose gradient.Nested-PCR was conducted to specifically amplify the large-subunit（28S） rDNA D1/D2 domain sequences.They were identified from sequencing,and phylogenetic analysis.It showed that sequence alignment of the DT-1 28S rDNA D1/D2 domain sequence to fungi sequences in the database revealed that DT-1 shared 97%homology with the corresponding DT-1 sequence;DT-2 clusters shared 99%homology with the corresponding Mucor racemosus（Y213713） sequence.Results demonstrate that the method can get results quickly and accurately,and is suitable for identification of fungi in rhizospheric soil of soybean.

关 键 词：土壤 真菌 大豆 NESTED-PCR 

分 类 号：S565.1[农业科学—作物学]