以新鲜根段进行AM真菌的分子检测及竞争性侵染研究  

MOLECULAR DETECTION OF COMPETITIVE COLONIZATION OF AM FUNGI WITHIN FRESH ROOTS

作　　者：郑世学[1] 董秀丽[1] 喻子牛[1] 赵斌[1] 

机构地区：[1]华中农业大学生命科学技术学院农业微生物学国家重点实验室,武汉430070

出　　处：《菌物学报》2004年第1期126-132,共7页Mycosystema

基　　金：国家自然科学基金(no.30270051);欧盟项目INCO-DEV Project ICA4-CT-2000-30014资助

摘　　要：运用nested-PCR技术和AM真菌特异性引物,建立了用新鲜植物根段直接检测AM真菌的分子生物学方法。以真核生物通用引物LR1和NDL22对混合接种的西红柿新鲜根段进行第1次扩增,将其产物进行稀释,再分别以Glomus intraradices 和Glomus mosseae的种特异性引物8.22和5.25进行第2次扩增。在琼脂糖凝胶上观察到AM真菌种特异性条带;运用该技术检测出混合接种时同一根段内不同的AM真菌,并探讨了真菌在植物根部的竞争性侵染。用盆栽方式种植西红柿,混合接种G. intraradices 和G. mosseae,在1个月后,前者侵染占优势。Two species of arbuscular mycorrhizal fungi, Glomus intraradices and Glomus mosseae , were detected in fresh plant roots by means of molecular approach-nested PCR. Roughly extracted DNA from tomato fresh root was amplified by using universal eukaryotic primers LR1-NDL22. The PCR products were diluted and used as template for the second PCR amplification using species specific primers 8.22 and 5.25 combined with LR1 or NDL22 respectively. Two Glomus species were identified specifically by this technique. In a glasshouse experiment, tomato seedlings were mixed-inoculated with G. intraradices and G. mosseae. Four weeks after inoculation, both inoculated fungi were detected in the same infected root fragments. The present study opens the possibility for molecular monitoring the persistence and dissemination of introduced efficient species of AM fungi in natural ecosystem and evaluation of their potential in field application.

关 键 词：西红柿 根内球囊霉 摩西球囊霉 NESTED-PCR AM真菌 

分 类 号：Q948[生物学—植物学]