黄丽,苏华楠,唐科志,黄爱军,周常勇,李中安

• 1:西南大学园艺园林学院
• 2:中国农业科学院柑桔研究所

摘要(Abstract)：

【目的】为了建立柑橘黄龙病菌亚洲种LAMP快速检测方法。【方法】根据黄龙病菌亚洲种外膜蛋白基因,在种属特异保守区域的6个位点设计2对特异性引物,对反应条件和体系进行优化,并通过产物沉淀观察和SYBRGreen I荧光染料显色的方法来快速判读检测结果。同时对田间样品进行了特异性和灵敏度分析试验。【结果】该方法仅需63℃反应70 min即可得到结果,特异性强,灵敏度高。对柑橘常见病害样品进行LAMP扩增,仅HLB阳性样品扩增产物电泳呈特征性梯状条带。检测灵敏度比常规PCR高100倍,与实时定量PCR相当。在对105份黄龙病田间疑似样品检测中,LAMP阳性检出率为52.4%,常规PCR为37.1%,2种检测方法符合率为84.7%。【结论】建立的黄龙病菌亚洲种LAMP检测方法,快速简便、特异性强、灵敏度高,为快速检测柑橘黄龙病提供了新方法和新思路。

关键词(KeyWords)： 柑橘黄龙病菌亚洲种;外膜蛋白基因;环介导等温扩增技术

Abstract:

Keywords:

基金项目(Foundation): 公益性行业专项(201003067-02,200903004-06);; 教育部创新团队(IRT0976)

作者(Author): 黄丽,苏华楠,唐科志,黄爱军,周常勇,李中安

DOI: 10.13925/j.cnki.gsxb.2012.06.006

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