利用一步法RT-PCR检测柑橘裂皮病类病毒

园艺学报 ›› 2005, Vol. 32 ›› Issue (3): 408-413.

利用一步法RT-PCR检测柑橘裂皮病类病毒

唐科志¹;彭　军²;王雪峰¹;周　彦¹;周常勇¹;刘科宏¹;刘　英¹;杨方云¹

¹中国农业科学院柑橘研究所, 重庆400712; ²西南农业大学植物保护学院, 重庆400716

收稿日期:2004-10-25 修回日期:2005-02-04 出版日期:2005-06-25 发布日期:2005-06-25

Quick Detection of Citrus exocortis viroid (CEVd) by One-step RT-PCR

Tang Kezhi¹;Peng Jun²;Wang Xuefeng¹;Zhou Yan¹;Zhou Changyong¹;Liu Kehong¹;Liu Ying¹;Yang Fangyun¹

¹Citrus Research Institute of Chinese Academy of Agriculture Sciences, Chongqing 400712, China; ²College of Plant Protection, Southwest Agricultural University, Chongqing 400716, China

Received:2004-10-25 Revised:2005-02-04 Online:2005-06-25 Published:2005-06-25

摘要/Abstract

摘要： 柑橘裂皮病(CEVd) 是限制柑橘产量的一种重要类病毒病害, 国内外已经建立了多种方法检测该病害的病原。本研究初步建立了快速灵敏检测CEVd的一步法RT-PCR技术。经过与生物学鉴定和双向聚丙烯酰胺凝胶电泳(sPAGE) 方法的比较检测, 表明该RT-PCR方法检测周期短, 灵敏度高, 有利于对类病毒在春季和冬季浓度低时进行快速准确的检测。应用该技术对大量田间寄主进行了检测。

关键词: 柑橘裂皮病, 一步法RT-PCR, 生物检测, 双向聚丙烯酰胺凝胶电泳

Abstract: Citrus exocortis viroid (CEVd) causes an important citrus disease throughout the world. To date, variousmethods have been developed for detecting this viroid. One-step reverse transcrip tion and polymerase chain reaction (RT-PCR) has been approached to detect CEVd. Compared with biological indexing and detection by sequential polyacrylamide gel electrophoresis (sPAGE) , detection by One-step RT-PCR was more sensitive and quicker than that by other methods, overcoming the detection difficulty derived from low viroid concentrations in winter and spring. Large number field trees were detected by the One-step RT-PCR method.

Key words: Citrus exocortis viroid, One-step RT-PCR, Biological indexing, sPAGE

中图分类号:

中图分类号: S 666

唐科志;彭　军;王雪峰;周　彦;周常勇;刘科宏;刘　英;杨方云. 利用一步法RT-PCR检测柑橘裂皮病类病毒[J]. 园艺学报, 2005, 32(3): 408-413.

Tang Kezhi;Peng Jun;Wang Xuefeng;Zhou Yan;Zhou Changyong;Liu Kehong;Liu Ying;Yang Fangyun. Quick Detection of Citrus exocortis viroid (CEVd) by One-step RT-PCR[J]. ACTA HORTICULTURAE SINICA, 2005, 32(3): 408-413.