香蕉炭疽菌rDNA ITS区的分子鉴定与检测  

Rapid molecular identification and detection of Colletotrichum musae with species-specific primers based on the internal transcribed spacer(ITS) region

作　　者：杨腊英[1] 黄华平[1] 唐复润[1] 胡美娇[1] 张世清[1] 黄俊生[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热带作物生物技术国家重点实验室,海口571101

出　　处：《植物病理学报》2006年第3期219-225,共7页Acta Phytopathologica Sinica

基　　金：国家科技部农业微生物资源平台项目(2004DKA30560-8);农业部南亚热作专项(ly2004-5)

摘　　要：香蕉炭疽病菌(Colletotrichum musae)是一种引起香蕉采后病害的最重要病原,本研究用真菌18S^28S间的内转录间隔区(internal transcribed spacer,ITS)通用引物18SF和28SR扩增香蕉炭疽菌和其它外群真菌的基因组DNA,扩增出约510 bp的片段;通过克隆测序香蕉炭疽菌的ITS全序列并与GenBank中炭疽菌属其它种的ITS序列比对,设计出香蕉炭疽菌的特异性引物ColM1和ColM2。用此特异引物可以从香蕉炭疽菌株中扩增出382 bp的特异性片段,而其余20个参试菌株和香蕉组织的PCR反应结果为阴性,灵敏度实验证明可以检测到目标DNA的浓度为0.1 pg。该方法可用于快速、准确和灵敏地检测香蕉炭疽菌,为快速监测组织中有无香蕉炭疽病菌潜伏侵染与及早采取防治措施提供积极的指导意义。A polymerase chain reaction （PCR） protocol was developed to specifically detect C. musae, the causal agent of banana anthracnose. Approximately 510 bp fragment of C. musae and the other outgroup fungal genome DNA were amplified by using the universal primers 18SF and 28SR of internal transcribed spacer （ITS） region of eukaryote 18S -28S ribosomal DNA. The species-specific PCR primes ColM1 and ColM2 for C. musae were designed after multiple sequence alignment of C. musae and the other species of Colletotrichum genus. By which a 382 bp single fragment with DNA from the isolates of C. musae was amplified, no product was amplified with DNA from other fungi and DNA of banana tissue. 0. 1 pg C. musae genomic DNA was detected by species-specific primers. The PCR protocol provides a rapid and reliable tool for routine detection and identification of C. musae. In addition , it is important that we can inspect whether C. musae infect the banana tissues or not and also significative to take control measures timely.

关 键 词：香蕉炭疽菌 内转录间隔区(ITS) 特异性引物 PCR检测 

分 类 号：S436.67[农业科学—农业昆虫与害虫防治]