甘薯羽状斑驳病毒（SPFMV）ELISA鉴定及RT-PCR检测方法的建立

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

doi:10.19303/j.issn.1008-0384.2013.12.016

甘薯羽状斑驳病毒（SPFMV）ELISA鉴定及RT-PCR检测方法的建立

doi: 10.19303/j.issn.1008-0384.2013.12.016

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, 主要从事甘薯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: 122
- HTML全文浏览量: 38
- PDF下载量: 1
- 被引次数: 0
出版历程
- 收稿日期: 2013-10-21
- 刊出日期: 2013-12-18

ELISA Identification and Development of RT-PCR Detection of Sweet Potato Feathery Mottle Virus

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

摘要

摘要: 利用双抗体酶联免疫吸附测定法 (DAS-ELISA) 对采自福建省51份甘薯叶片进行检测, 结果显示51份样品中有24份SPFMV呈阳性, 阳性率为47.06%, 其中泉州样品SPFMV阳性率最高, 为71.40%, 其次为南安和龙岩的样品, 阳性率均为50%。根据GenBank中公布的SPFMV外壳蛋白 (CP) 基因序列保守区域设计了1对特异性引物, 通过RT-PCR反应程序的优化建立了能检测SPFMV的检测方法。该检测方法能够扩增出SPFMV特异性片段, 片段大小为441bp, 测序结果表明, SPFMV序列与参考序列的同源性92%⁹7%。
- 甘薯 /
- 病毒 /
- 甘薯羽状斑驳病毒 /
- ELISA /
- RT-PCR
Abstract: 51samples of sweet potato from Fujian province were detected by the double antibody enzyme-linked immunosorbent method (DAS-ELISA) .The results showed that 24samples were SPFMV-positive, the positive rate were 47.06%;the highest positive rate was 71.40%, detected in the samples from Quanzhou, and the next were 50.00%, detected in samples from Nanan and Longyan.A pair of primer was designed and synthesized according to the published nucleotide sequence of coat protein gene of SPFMV, and a specific RT-PCR detection procedure was established.The amplified SPFMV-specific fragment in PCR contained 441bps, and showed 92%-97%, homology with the SPFMV reference sequence.
- sweet potato /
- virus /
- sweet potato feathery mottle virus /
- ELISA /
- RT-PCR

HTML全文

参考文献(14)

[1]	LUAN Y S, ZHANG J, LIU D M, et al.Molecular characterization of sweet potato leaf curl virus isolate from China (SPLCV-CN) and its phylogenetic relationship with other members of the Geminiviridae[J].Virus Genes, 2007, 35 (2) :379-385.
[2]	LOZANO G, TRENADO H P, VALVERDE R A, et al.Novel begomovirus species of recombinant nature in sweet potato (Ipomoea batatas) and Ipomoea indica:taxonomic and phylogenetic implications[J].Journal of General Virology, 2009, 90 (10) :2550-2562.
[3]	CLARK C A, DAVIS J A, ABAD J A, et al.sweet potato viruses:15years of progress on understanding and managing complex diseases[J].Plant Disease, 2012, 96 (2) :168-185.
[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]	乔奇, 张德胜, 秦艳红, 等.中国甘薯病毒种类的初步鉴定[J].植物病理学报, 2012, 42 (1) :10-16.
[6]	RICCBMANN J L, LAIN S, GARCIAJ A.Highlights and prospects of potyvirus molecular biology[J].Gen Virol, 1992, 73:1-16.
[7]	CHAVI F, ROBERTSON A J, BENEDICTUS J M.Verduin survey and cbaracterization of viruses in sweet potato from Zimbabwe[J].Plant Dis, 1997, 81:1115-1122.
[8]	SHIGUCHI NI, MORI M, SUZUKI F, et al.Specific severe strains of sweet potato feathery mottle virus (SPFMV) by reverse transcription and polymerase chain reaction[J].Ann Pbytopatbol Soc Japan, 1995, (61) :119-192.
[9]	COLINET D, KUMMER J.Identification of a sweet potato feathery mottle virus isolate from China (SPFMV-CH) by the polymerase chain reaction with degenerate primers[J].Journal of Virological Methods, 1993, 45 (2) :149-159.
[10]	UNTIVEROS M, QUISPE D, KREUZE J.Analysis of complete genomic sequences of isolates of the Sweet potato feathery mottle virus strains C and EA:Molecular evidence for two distinct potyvirus species and two P1protein domains[J].Archives of Virology, 2010, 155 (12) :2059-2063.
[11]	COLINET D, NGUYEN M, KUMMERT J, et al.Differentiation among potyviruses infecting sweet potato based on genus-and virus-specific reverse transcription polymerase chain reaction[J].Plant Disease, 1998, 82 (2) :223-229.
[12]	赵冬兰, 唐君, 张允刚, 等.甘薯羽状斑驳病毒RT-PCR检测[J].西北农业学报, 2007, 16 (6) :199-202.
[13]	包改丽, 左瑞娟, 饶维力, 等.云南甘薯病毒的检测及主要病毒的多样性分析[J].微生物学报, 2013, 40 (2) :236-248.
[14]	张业辉, 张振臣, 蒋士君, 等.中国甘薯病毒种类的初步鉴定[J].植物病理学报, 2012, 42 (1) :10-16.