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园艺学报 ›› 2011, Vol. 38 ›› Issue (11): 2193-2198.

• 研究报告 • 上一篇    下一篇

应用real-time RT-PCR监测柑橘衰退病毒强、弱毒株的时序变化

邹 勤1,2,周 彦1,李中安1,周常勇1,2,*,刘永清2,苏华楠2   

  1. (1中国农业科学院柑桔研究所国家柑桔工程技术研究中心,重庆400712;2西南大学植物保护学院,重庆400716)
  • 收稿日期:2011-07-01 修回日期:2011-10-24 出版日期:2011-11-25 发布日期:2011-11-25
  • 通讯作者: 周常勇

Monitoring the Temporal Change of the Mild and Severe Strain of Citrus tristeza virus by Real-time RT-PCR

ZOU Qin1,2,ZHOU Yan1,LI Zhong-an1,ZHOU Chang-yong1,2,*,LIU Yong-qing2,and SU Hua-nan2   

  1. (1National Citrus Engineering Research Centre,Citrus Research Institute,Chinese Academy of Agricultural Sciences,Chongqing 400712,China;2College of Plant Protection,Southwest University,Chongqing 400716,China)
  • Received:2011-07-01 Revised:2011-10-24 Online:2011-11-25 Published:2011-11-25
  • Contact: ZHOU Chang-yong

摘要: 使用褐色橘蚜在提前7个月预免疫接种了柑橘衰退病毒(Citrus tristeza virus,CTV)弱毒株CT11的锦橙植株上拮抗接种强毒株CT14,应用real-time RT-PCR技术,监测供试植株中CTV强、弱毒株的含量变化,同时采用内参基因18S rRNAF/R及nad5F/R校正检测结果。试验结果表明,在拮抗接种3个月内始终未能在预免疫的拮抗植株中检测出强毒株CT14。在25头蚜虫拮抗接种后45 d和50头蚜虫拮抗接种后10 d,绝大多数预免疫的拮抗植株中弱毒株的含量高于预免疫的负对照植株,此后弱毒株的含量基本下降到负对照植株的水平,而负对照植株内弱毒株的含量变化不明显。

关键词: 柑橘, 柑橘衰退病毒, real-time RT-PCR, 弱毒株交叉保护

Abstract: Jincheng sweet orange seedlings were pre-inoculated with CTV mild isolate CT11 by grafting,and seven months later,challenged by severe CTV isolate CT14 with Toxoptera citricida. To identify and quantitate the mild and severe CTV isolates within the same plant,a real-time RT-PCR technique was modified and housekeeping genes 18S rRNAF/R and nad5F/R were approached for rectification. The results showed that CT14 was not detectable in any challenge-inoculated plants three months after CT14 attacked through aphids. The titer of CT11 in most samples of the plants challenge-inoculated via 25 or 50 aphids per plant was higher than that in the pre-immunized control plants on the 45th or 10th day,respectively,then decreased to the same level of the pre-immunized control,whereas the titer of the mild isolate in the pre-immunized control plants was much stable.

Key words: Citrus, Citrus tristeza virus, real-time RT-PCR, mild strain cross protection

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