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园艺学报 ›› 2006, Vol. 33 ›› Issue (4): 845-848.

• 研究报告 • 上一篇    下一篇

香蕉两种主要病毒多重PCR检测方法的建立

彭军1;王国芬1;黄俊生1, 2*;代鹏1;谢玉萍1;李伟东3

  

  1. (1 中国热带农业科学院环境与植物保护研究所, 海南儋州571737; 2 中国热带农业科学院热带生物技术研究所热带作物生物技术国家重点实验室, 海南海口571101; 3 中华人民共和国海南出入境检验检疫局, 海南海口570311)
  • 收稿日期:2005-08-16 修回日期:2005-12-01 出版日期:2006-08-25 发布日期:2006-08-25

Development of a Multiplex PCR Protocol for the Detection of Two Virusesof Banana

Peng Jun1;Wang Guofen1;Huang Junsheng1, 2*;Dai Peng1;Xie Yuping1;Li Weidong3

  

  1. (1 Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, Hainan 571737,China; 2 State Key Laboratory of Tropical Crop Biotechnology, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan 571101, China; 3Hainan Entry-exit Inspection and Quarantine Bureau of Peoples Republic of China, Haikou, Hainan 570311, China)
  • Received:2005-08-16 Revised:2005-12-01 Online:2006-08-25 Published:2006-08-25

摘要:

根据香蕉束顶病(Banana bunchy top virus, BBTV) 和香蕉花叶心腐病(Cucumber mosaic virus,CMV) 的复制酶基因、外壳蛋白基因序列分别设计特异性引物对, 在BBTV单一PCR和CMV RT-PCR优化体系的基础上, 建立了可同时检测BBTV、CMV的多重PCR检测方法。此方法可以特异地从感染BBTV和CMV的样品中扩增出2条带, BBTV (748 bp) 和CMV (557 bp) 。扩增产物序列测定结果表明, BBTV扩增产物与GenBank中其他分离物的核苷酸序列同源性为91% ~99% , CMV扩增产物与GenBank中其他分离物的核苷酸序列同源性为93%~98%。

关键词: 香蕉, 香蕉束顶病毒, 黄瓜花叶病毒, 病毒检测, RT-PCR

Abstract: A multiplex RT-PCR was developed for the simultaneous detection of two viruses of banana and plantain (Musa spp. ). Two sets of specific primer pairs were designed according to the gene sequence of Banana bunchy top virus (BBTV) replicase region and Cucumber mosaic virus (CMV) coat protein region, respectively. Based upon the establishment of the optimized PCR and RT-PCR detection of BBTV and CMV, the multiplex RT-PCR which can simultaneously detect the two banana viruses was developed. It was shown that all samples infected with BBT and CMV could be amplified simultaneously by multiplex RT-PCR, and yield two specific bands of BBTV (748 bp) and CMV (557 bp) visualized by agarose gel electrophoresis. Sequence analysis of the amplified products showed high homology of BBTV (91% - 99% ) and CMV (93% - 98% )
with the sequences of BBTV and CMV in GenBank.

Key words: Banana, Banana bunchy top virus (BBTV), Cucumber mosaic virus (CMV), Virus detection, RT-PCR