橡胶树多主棒孢菌rDNA-ITS区的分子鉴定及检测  

Rapid Molecular Identification and Detection of Corynespora cassiicola of Hevea brasiliensis

作　　者：刘先宝[1] 蔡吉苗[1] 潘羡心[1] 高宏华[1] 彭建华[1] 黄贵修[1] 

机构地区：[1]中国热带农业科学院环境与植物保护研究所海南省热带农业有害生物检测监控重点实验室,海南儋州571737

出　　处：《热带作物学报》2008年第4期489-493,共5页Chinese Journal of Tropical Crops

基　　金：科技部科研院所社会公益研究专项基金项目(No.2004DIA4J012);科技部农业成果转化项目(No.2006326018);中央级公益性科研院所基本科研业务费专项及热带作物栽培生理学重点开放实验室开放课题基金项目(KLOF0611)资助

摘　　要：以来自国内外的18个多主棒孢病菌[Corynespora cassiicola(Ber.&Curt.)]菌株为研究材料,提取其基因组DNA进行rDNA–ITS区序列扩增,并进行了5.8S rDNA及其两侧ITS序列分析。序列分析结果表明,种内各菌株间ITS序列同源性高达99.9%以上,部分菌株间出现个别碱基的差异。根据ITS区序列设计的一对特异引物(CorP1/CorP2)可以在供试的多主棒孢病菌中扩增出1条约277 bp的特异谱带,其余18个参试菌株和橡胶树叶片组织未能扩增出该特异谱带,检测灵敏度可达浓度为0.1 pg的目标DNA。The Corynespora lea f fall disease （CLFD） of Hevea brasiliensis caused by the fungus Corynespora cassiicola has emerged as a major constraint for natural rubber production over the last two decades in the world. Eighteen strains from China were used in this study. Its genomic DNAs were isolated, and the ribosomal DNA internal transcribed spacer （ITS） region were amplified and sequenced. The results showed that there was no obvious variance in ITS sequence and just some slight variance in bases. The species-specific PCR primers CorP1 and CorP2 for C. cassiicola were designed after multiple sequence alignment of C. cassiicola and the other species of Corynespora genus. An about 300 bp single fragment with DNA from the isolates of C. cassiicola was amplified, and no product was amplified with DNA from the other 18 fungi and DNA of rubber tree leaf tissue. Then 0.1 pg C. cassiicola genomic DNA was detected by species-specific primers.

关 键 词：橡胶树 多主棒孢菌 内转录间隔区(ITS) 特异性引物 PCR检测 

分 类 号：S763.102[农业科学—森林保护学] S794.1[农业科学—林学]