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| 3种甘薯病毒多重RT-PCR检测方法的建立及应用 | |
| 引用本文： | 姜珊珊,冯佳,吴斌,张眉,王升吉,辛志梅,任建雯,陈琳琳,辛相启.3种甘薯病毒多重RT-PCR检测方法的建立及应用[J].植物病理学报,2018,48(6):847-851. |
| 作者姓名： | 姜珊珊 冯佳 吴斌 张眉 王升吉 辛志梅 任建雯 陈琳琳 辛相启 |
| 作者单位： | 山东省农业科学院植物保护研究所,济南250100; 河南农业大学植物保护学院,郑州450002 |
| 基金项目： | 山东省重点研发计划项目(2016GNC111003);山东省农业科学院农业科技创新工程项目(CXGC2016B11) |
| 摘 要： | 正病毒病是引起甘薯品质降低和减产的重要原因之一,现已报道30多种能侵染甘薯的病毒~(1,2])。山东省是甘薯种植大省,病毒种类近10种~(3,4])。甘薯羽状斑驳病毒(Sweet potato feathery mottle virus,SPFM V)、甘薯潜隐病毒(Sweet potato latent virus,SPLV)是为害甘薯的主要病毒,在全国甘薯种植区广泛分布~(5,6])。甘薯病毒2(Sweet potato virus 2,SPV2)为Potyvirus的一个暂定种,多与同属的其他病毒混合侵染~(7])。多重PCR技术由Chamberian等~(8])1988年首次提出,可实现多基因的同时扩增,具有节省时间、提高效率的优点,已初 |
Development and application of a multiplex RT-PCR detection assay for three sweet potato viruses |
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| JIANG Shan-shan,FENG Jia,WU Bin,ZHANG Mei,WANG Sheng-Ji,XIN Zhi-mei,REN Jian-wen,CHEN Lin-lin,XIN Xiang-qi.Development and application of a multiplex RT-PCR detection assay for three sweet potato viruses[J].Acta Phytopathologica Sinica,2018,48(6):847-851. | |
| Authors: | JIANG Shan-shan FENG Jia WU Bin ZHANG Mei WANG Sheng-Ji XIN Zhi-mei REN Jian-wen CHEN Lin-lin XIN Xiang-qi |
| Institution: | Institute of Plant Protection, Shandong Academy of Agricultural Sciences, Jinan 250100, China; College of Plant Protection, Henan Agricultural University, Zhengzhou 450002, China |
| Abstract: | To effectively monitor the occurrence of major sweet potato viruses, the multiplex RT-PCR assay was established for detection of Sweet potato feathery mottle virus (SPFMV), Sweet potato latent virus (SPLV) and Sweet potato virus 2 (SPV2). Three pairs of specific primers were designed based on the conserved region sequences of the coat protein (CP) gene published in GenBank. The mainly factors including the concentration of primer and polymerase, reaction cycles and annealing temperature were optimized. Three specific fragments could be simultaneously amplified by the multiplex RT-PCR assay with the lengths of 317 bp, 569 bp and 994 bp, respectively. The obtained sequences shared at least 99% homology with isolates published in GenBank. Sensibility test showed that the three viruses could be simultaneously detected out in 278.8 ng·L-1 total RNA. The detection rates of SPFMV, SPLV and SPV2 were 100%, 89.6% and 27.1% in 48 samples. Therefore, the multiplex RT-PCR was proven as an efficient method to synchronously detect SPFMV, SPLV and SPV2 in field samples. |
| Keywords: | Sweet potato virus multiplex RT-PCR Sweet potato feathery mottle virus Sweet potato latent virus Sweet potato virus 2 |
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