中国农业科学院农业信息研究所 中国农学会
Agricultural Information Institute of CAAS China Association of Agricultural Science Societies
En
Ch
中国农业科学院农业信息研究所 中国农学会 主办

AI检索

订阅

返回顶部
当前位置: 首页 > 文章 > 反向斑点杂交法快速检测甘薯羽状斑驳病毒和甘薯G病毒 南方农业学报 2014,45 (1) 43-48
Position: Home > Articles > Detection of SPFMV and SPVG by using reverse dot blot hybridization system Journal of Southern Agriculture 2014,45 (1) 43-48

反向斑点杂交法快速检测甘薯羽状斑驳病毒和甘薯G病毒

作  者：
何海旺;何虎翼;谭冠宁;刘义明;何新民;唐洲萍;李丽淑;王晖
单  位：
广西农业科学院经济作物研究所;防城港市农业技术推广服务中心
关键词：
甘薯;甘薯羽状斑驳病毒;甘薯G病毒毒;病毒检测;反向斑点杂交技术
摘  要：
【目的】以克隆得到的甘薯羽状斑驳病毒(Sweet potato feathery mottle virus,SPFMV)和甘薯G病毒(Sweet potato virus G,SPVG)基因片段为探针建立反向斑点杂交技术体系,应用于甘薯组培脱毒苗带毒情况检测,为生产优质甘薯组培脱毒苗提供保障。【方法】根据已公布的侵染甘薯的SPFMV和SPVG的核苷酸序列设计两对特异引物,以RT-PCR法从染病的甘薯叶片扩增SPFMV和SPVG的两个片段,并以克隆到的两个病毒片段及内参基因Actin片段为探针建立反向斑点杂交体系。【结果】分别克隆出长度约310和500 bp的片段,经BLAST比对,所获得的310 bp片段为SPFMV的外壳蛋白基因片段,同源性为97%,500 bp片段为SPVG的外壳蛋白基因片段,同源性为99%。分别用带有SPFMV和SPVG片段的重组质粒pUC-SPFMV和pUC-SPVG进行反向斑点杂交,发现不同病毒能获得单一信号,无交叉现象。以染病甘薯和脱毒甘薯苗为样品,用该种病毒的探针进行反向斑点杂交,染病植株样品中能获得单一信号,而脱毒苗甘薯样品未见任何信号,杂交结果与RT-PCR检测结果一致。【结论】用建立的反向斑点杂交技术体系能有效检测甘薯中的SPFMV和SPVG,无交叉信号,可用于甘薯组培脱毒苗的前期检测。
译  名：
Detection of SPFMV and SPVG by using reverse dot blot hybridization system
作  者：
HE Hai-wang;HE Hu-yi;TAN Guan-ning;LIU Yi-ming;HE Xin-min;TANG Zhou-ping;LI Li-shu;WANG Hui;Cash Crops Research Institute,Guangxi Academy of Agricultural Sciences;Agricultural Technology Center of Fangchenggang City;
关键词：
sweet potato;;SPFMV;;SPVG;;virus detection;;reverse dot blot hybridization
摘  要：
【Objective】Two sweet potato viruses, feathery mottle virus(SPFMV) and virus G(SPVG), were cloned, and the reverse dot blot hybridization system was built. Then this system was used for detecting the sweet potato virus so as to provide references for producing high quality virus-free seedlings. 【Method】According to the published nucleotide sequences of SPFMV and SPVG, two specific primers were designed and two fragments were cloned by reverse transcription-polymerase chain reaction(RT-PCR) from total RNA extracted from sweet potato leaf which were suspiciously infected with sweet potato virus. Then, the two fragments and Actin gene were used as the probes in this reverse dot blot hybridization system. 【Result】Two fragments, about 310 and 500 bp were obtained. It was found by sequencing and blasting that they were coat proteins of SPFMV and SPVG with the homology of 97% and 99%, respectively. Reverse dot blot hybridization of recombinant plasmids pUC-SPFMV and pUC-SPVG showed that single signals were attained from different viruses. No cross signals were found. Then the infected and virus-free sweet potatoes were detected by reverse dot blot hybridization using probes. The results showed that single signals were found in the affected samples while no signal was observed in virus-free samples, which accorded with RT-PCR results. 【Conclusion】SPFMV and SPVG can be detected by this hybridized system and there was no cross signal from the sample with virus. So this system can be used to detect virus of virus-free sweet potato seedlings in the initial stage.

相似文章

导出引用
计量
文章访问数: 8
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊

友情链接Friendly link
农业信息研究所Agricultural information institute 农业智能知识服务平台Agricultural intelligent knowledge service platform 农业学术服务平台Agricultural Knowledge Service System 中国知网CNKI 维普网VIP 万方WANFANG DATA
联系我们Contact us

电话Tell：010-82109652-808

邮箱E-mail：agrijournal@caas.cn

服务Service
关于我们About us 意见建议Opinions and suggest
版权说明Copyright
单位：中国农业科学院农业信息研究所 中国农学会 © 2017中国农业科学院农业信息研究所 ICP备案号：京ICP备09089781号-46