甘薯羽状斑驳病毒和褪绿矮化病毒双重RT-PCR检测方法的建立

许泳清; 李华伟; 邱思鑫; 刘中华; 邱永祥; 罗文彬; 纪荣昌; 汤浩; 余华

doi:10.19303/j.issn.1008-0384.2014.11.014

甘薯羽状斑驳病毒和褪绿矮化病毒双重RT-PCR检测方法的建立

doi: 10.19303/j.issn.1008-0384.2014.11.014

1. 福建省农业科学院作物研究所,福建福州,350013;
2. 福建省农业科学院中心实验室,福建福州,350003

基金项目:

国家甘薯产业技术体系项目 (CARS-11-B-10-2013)

福建省财政专项——福建省农业科学院科技创新团队建设项目 (CXTD-1-1301)

详细信息

作者简介:
许泳清 (1980-) , 女, 助理研究员, 主要从事薯类作物组织培养与病毒检测研究 (E-mail:qingqing0722@126.com);邱思鑫 (1974-) , 男, 博士, 研究员, 主要从甘薯病虫害防控技术研究 (E-mail:qiusixin@yahoo.com.cn);余华 (1957-) , 女, 研究员, 主要从事甘薯病虫害防控技术研究 (E-mail:yuhua@faas.cn)

中图分类号: S435.31
计量
- 文章访问数: 100
- HTML全文浏览量: 30
- PDF下载量: 1
- 被引次数: 0
出版历程
- 收稿日期: 2014-07-30
- 刊出日期: 2014-11-18

Development of Dutiplex RT-PCR for the Detection of SPFMV and SPCSV in Sweet Potato

1. Institute of Crop Sciences,Fujian Academy of Agricultural Sciences;
2. Central Laboratory of Agricultural Sciences of Fujian Academy

摘要

摘要: 根据GenBank中公布的甘薯羽状斑驳病毒 (Sweet potato feathery mottle virus, SPFMV) 外壳蛋白 (CP) 基因和甘薯褪绿矮化病毒 (Sweet potato chlorotic stunt virus, SPCSV) 热激蛋白 (Hsp70) 基因序列保守区域设计了2对特异性引物, 通过RT-PCR反应程序的优化, 建立了能同时检测SPFMV和SPCSV的双重PCR检测体系。该检测体系能够1次扩增出SPFMV和SPCSV的特异性片段, 片段大小为461bp和304bp, 测序结果表明, 2种病毒序列与参考序列的同源性达93%以上。
- 甘薯 /
- 甘薯羽状斑驳病毒 /
- 甘薯褪绿矮化病毒 /
- 双重RT-PCR
Abstract: A duplex PCR protocol for the detection of two main viruses in sweet potato was developed based on the coat protein (CP) gene of Sweet potato feathery mottle virus (SPFMV) and the (Hsp70) gene of Sweet potato chlorotic stunt virus (SPCSV) .They were able to amplify unique DNA fragments of 461 bp and 304 bp in a single PCR reaction.The sequence analysis suggst that the two viruses shared at least 93 percent homology with other related viruses.
- sweet potato /
- SPCSV /
- SPFMV /
- duplex RT-PCR

HTML全文

参考文献(10)

[1]	GIBSON R W, MPEMBE I, ALICAI T, et al.Symptoms, actiology and serological analysis of sweet potato virus disease in Uganda[J].Plant Pathology, 1998, 47:95-102.
[2]	GUTIERREZ D L, FUENTES S, SALAZAR L F.Sweet potato virus disease (SPVD) :distribution, incidence, and effect on sweet potato yield in Peru[J].Plant Disease, 2003, 87:297-302.
[3]	NJERU R W, MBURU M W K, CHERAMGOI E, et al.Studies on the physiological effects of viruses on sweet potato yield in Kenya[J].Annals of Applied Biology, 2004, 145:71-76.
[4]	张振臣, 乔奇, 秦艳红, 等.我国发现由甘薯褪绿矮化病毒和甘薯羽状斑驳病毒协生共侵染引起的甘薯病毒病害[J].植物病理学报, 2012, 43 (3) :328-333.
[5]	MENZELW, JELKMANN W, MAISS E.Detection of four apple viruses by multiplex RT-PCR assays with coamplification of plant mRNA as internal control[J].Journal of Virological M ethods, 2002, 99:81-92.
[6]	THOMPSON J R, WETZEL S, KLERKS M M, et al.Multiplex RT PCR detection of four aphid-borne strawberry viruses in Fragaria spp in combination with a plant mRNA specific internal control[J].Journal of Virological Methods, 2003, 11 (2) :85-93.
[7]	张业辉, 张振臣, 蒋士君, 等.3种甘薯病毒多重RT-PCR检测方法的建立[J].植物病理学报, 2010, 40 (1) :95-98.
[8]	张盼, 兰新芝, 乔奇, 等.甘薯病毒病害 (SPVD) 的多重RTPCR检测方法及其应用[J].植物保护, 2013, 39 (2) :86-90.
[9]	范旭东, 董雅凤, 张尊平.3种苹果潜隐病毒多重RT-PCR检测体系的建立[J].园艺学报, 2009, 36 (12) :1821-1826.
[10]	KARYEIJA R F, KREUZE J F, GIBSON R W, et al.Synergistic interactions of a potyvirus and a phloem limited crinivirus in sweet potato plants[J].Virology, 2000, 269:26-36.