3种检疫性葡萄黄化植原体的快速检测  

A PCR-based assay for rapid detection and precise differentiation of three quarantine phytoplasmas responsible for yellows diseases in grapevine

作　　者：姜华[1] 王有福[2] 姜一[3] 魏薇[3] Rasa Jomantiene Robert E.Davis 赵衍[3] 

机构地区：[1]辽宁师范大学生命科学学院,大连116029 [2]辽宁出入境检验检疫局,大连116001 [3]美国农业部植物分子病理研究室,美国马里兰州20705

出　　处：《植物病理学报》2011年第4期352-360,共9页Acta Phytopathologica Sinica

基　　金：辽宁省教育厅项目(2008S135);国家质检总局项目(2006IK218)

摘　　要：葡萄上的植原体病害由于引起叶片黄化而被称为葡萄黄化病。由于这一病害极为严重,葡萄黄化植原体被列为我国的植物检疫对象。其中,葡萄金黄化植原体(16SrV)、维吉尼亚葡萄黄化植原体(16SrⅢ)和澳大利亚葡萄黄化植原体(16SrⅫ)是引起葡萄黄化病的主要3个株系,它们导致的病害症状相似,难以区分。本文进行了3个株系16S rRNA基因DNA序列比对,而后根据同源性相对低的序列设计了43条特异性引物、103对引物对组合,对葡萄黄化植原体3个株系各自的DNA及混合DNA进行PCR扩增,从中筛选出来特异性较强的8个引物对组合。这些引物对组合,能够同步、特异、快速地检测3种葡萄黄化植原体。Grapevine yellows（GY） is a destructive disease complex that affects a wide variety of grapevine cultivars in almost all major wine production countries.The causative agents of GY are phloem-restricted cell wall-less bacteria termed phytoplasmas.Thus far,diverse strains belonging to five distinct ＇Candidatus Phytoplasma＇ species and four 16S rDNA RFLP groups have been identified to be responsible for various GY epidemics around the world and some have been listed as quarantine pests in many countries.The disease symptoms induced by different GY phytoplasmas are very similar,making accurate visual diagnosis/quarantine inspection difficult.In the present study,16S rRNA gene sequences from three major GY phytoplasma strains,Flavescence doree phytoplasma（16SrV）,Virginia grapevine yellows phytoplasma（16SrⅢ）,and Australian grapevine yellows phytoplasma（16SrⅫ）,were analyzed and several variable regions were identified.Based on the analysis,43 primers were designed and 103 primer combinations were tested in PCR reactions aimed at precise differentiation of the three GY phytoplasma strains.The study identified reliable molecular markers and primer pairs that enable rapid and specific detection and differentiation of the three GY phytoplasma strains.

关 键 词：葡萄 植原体 PCR检测 

分 类 号：Q939[生物学—微生物学] S41[农业科学—植物保护]