福州和成都竹花叶病毒的RT-PCR法检测  

Detection of Bamboo mosaic virus by RT-PCR amplification in Fuzhou and Chengdu

作　　者：林文武[1] 杨文婷[1] 张洁[1] 刘宇艳[1] 王润语 吴祖建[1] 

机构地区：[1]福建农林大学植物病毒研究所,福建省植物病毒学重点实验室,福州350002 [2]福建省福州第一中学,福州350001

出　　处：《植物病理学报》2016年第4期469-473,共5页Acta Phytopathologica Sinica

基　　金：国家重点基础研究发展计划“973”项目(2014CB138402);国家自然科学基金资助项目(31171821);教育部博士点基金(20113515110001);国家科技支撑计划(2012BAD19B03);福建省教育厅重点项目(JA025226)

摘　　要：竹花叶病毒(Bamboo mosaic virus,BaMV)是马铃薯X病毒属唯一携带有卫星RNA的成员,目前已经在巴西、美国以及中国台湾等地的竹类栽培区产生危害,严重影响竹材的经济价值,而在中国大陆还不曾有相关报道。根据GenBank上报道的BaMV和其伴随卫星RNA(satBaMV)序列以及未公布的序列分别设计了1对特异性引物,在优化RT-PCR条件的基础上,成功建立了BaMV的RT-PCR检测方法。CpC-F/R用于扩增部分BaMV-CP序列(527 bp),SaC-F/R用于扩增satBaMV部分序列(472 bp)。把所建立的方法用于检测分别采自成都和福州的不同品种竹子病叶,结果均检测出了BaMV和satBaMV,而无症样品未得到任何产物。本研究在基因水平上为BaMV的检测提供了快速、准确、灵敏的新体系,为我国竹类栽培过程中竹花叶病毒的检测和防治提供了有效手段。Bamboo mosaic virus （BaMV） is the only member of Potexvirus carrying satellite RNA. BaMV had been reported in the bamboo cultivation areas in Brazil, USA and Taiwan （China） and affected the economic value of bamboos severely. However, there have been few reports on BaMV occurrence in the mainland of China thus far. Two pairs of specific primers were designed according to the sequences of BaMV and its associated sa- tellite RNA （satBaMV） deposited in GenBank or undisclosed sequences, respectively. The RT-PCR methods of detecting BaMV and satBaMV were established by optimizing the RT-PCR conditions. Primer pairs CpC-F/R were used to amplify the partial coat protein gene of BaMV（527 bp）, while primer pairs SaC-F/R were used to amplify the partial sequence of satBaMV （472 bp）. The symptomatic leaves of different bamboo species collected from Chengdu and Fuzhou were analyzed by RT-PCR. Results showed that bamboo plants with disease symptoms contain the BaMV and satBaMV whereas no fragment was detected in the asymptomatic samples. Our study es- tablished a rapid, accurate and sensitive method for detection of BaMV in the field, thus providing an efficient way to monitor and control BaMV infection during bamboo cultivation in China.

关 键 词：竹花叶病毒 卫星RNA RT-PCR 检测 

分 类 号：S432.41[农业科学—植物病理学]