小苍兰3种病毒多重RT-PCR检测体系的建立

樊荣辉; 黄敏玲; 吴建设; 罗远华

小苍兰3种病毒多重RT-PCR检测体系的建立

樊荣辉^1,2,3,
黄敏玲^1,2,3, ,,
吴建设^1,2,3,
罗远华^1,2,3

1.
福建省农业科学院作物研究所,福建福州 350013;
2.
福建省农业科学院花卉研究中心,福建福州 350013;
3.
福建省特色花卉工程技术研究中心,福建福州 350013

基金项目:

福建省科技计划重大专项(2010NZ0003);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(CXTD2011-20)

详细信息

通讯作者:
黄敏玲(1960-),女,研究员,主要从事花卉品种选育与生物技术研究(E-mail:pudang12@yahoo.com.cn)

中图分类号: S 682
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- 被引次数: 0
出版历程
- 收稿日期: 2012-02-16
- 修回日期: 2012-03-22
- 刊出日期: 2012-04-30

Multiplex RT-PCR Detection of Viruses in Freesia

FAN Rong-hui^1,2,3,
HUANG Min-ling^{1,2,3
, ,},
WU Jian-she^1,2,3,
LUO Yuan-hua^1,2,3

1.
Institute of Crop Sciences, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian 350013,China;
2.
Flowers Research Center , Fujian Academy of Agricultural Sciences, Fuzhou , Fujian 350013,China;
3.
Fujian Engineering Research Center for Characteristic Floriculture , Fuzhou , Fujian 350013,China

摘要

摘要: 根据GenBank中已发表的小苍兰花叶病毒Freesia mosaic virus(FreMV)、黄瓜花叶病毒Cucumber mosaic virus(CMV)和菜豆黄花叶病毒Bean yellow mosaic virus(BYMV)外壳蛋白(CP)基因序列保守区域分别设计特异性引物,通过优化多重PCR反应条件,建立能同时检测小苍兰3种病毒的多重PCR检测体系。该体系能够一次扩增出FreMV、CMV和BYMV的特异片段,其大小分别是340、628和212 bp。测序结果表明,3种病毒序列与相应的参考序列相似性均达97%以上。灵敏度测定结果表明,从≥10-2mg的感病植物组织中能够检测到这3种病毒。
- 小苍兰 /
- 病毒 /
- 小苍兰花叶病毒 /
- 黄瓜花叶病毒 /
- 菜豆黄花叶病毒 /
- 多重PCR
Abstract: By using the specific primers designed on the basis of conserved sequences of coat protein (CP) gene, a multiplex PCR protocol for the detection of three main viruses (i.e., Freesia mosaic virus, FreMV, Cucumber mosaic virus, CMV and Bean yellow mosaic virus,BYMV) in freesia was developed. Three specific fragments were simultaneously amplified in a single PCR reaction. Their molecular weights were determined to be 340 bp, 628 bp and 212 bp. The sequence analysis indicated that the three viruses shared at least 97 percent homology with other related viruses. The method’s sensitivity was shown to be capable of positively identifying the three viruses with≥10-2 mg of freesia tissue sample.
- Freesia /
- virus /
- Freesia mosaic virus /
- Cucumber mosaic virus /
- Bean yellow mosaic virus /
- multiplex PCR

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参考文献(2)

[1]	CHOI H I, LIM H R, SONG Y S, et al. The complete genome sequence of freesia mosaic virus and its relationship to other potyviruses[J]. Arch Virol,2010,155: 1183-1185.
[2]	BERTOLINI E, OLMOSAl A, MARTINEZ M C, et al. Single-step multiplex RT-PCR for simultaneous and colourimetric detection of sixRNA viruses in olive trees[J]. Journal of Virological Methods,2001,96: 33-41.