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中国农学通报

中国农学通报 ›› 2015, Vol. 31 ›› Issue (6): 234-240.doi: 10.11924/j.issn.1000-6850.casb14110109

所属专题： 食用菌； 食用菌

• 食品 营养 检测 安全 • 上一篇    下一篇

主栽金针菇品种中病毒的快速检测和鉴定

张俊玲1,章炉军2,谭 琦2,尚晓冬2,张俊玲1,2,章炉军1,谭琦1,2,尚晓冬1   

  1. (1上海市农业科学院食用菌研究所/国家食用菌工程技术研究中心,上海 201403；2南京农业大学生命科学学院,南京 210095）
  • 收稿日期:2014-11-18 修回日期:2015-02-11 接受日期:2014-12-01 出版日期:2015-03-20 发布日期:2015-03-20
  • 通讯作者: 尚晓冬
  • 基金资助:
    国家科技支撑项目“食用菌新品种培育及制种关键技术研究”(2013BAD16B02)；上海种业专项“香菇重要基因资源的收集、评价及应用”[沪农科种字(2012)第6号]；农业科技成果转化资金项目“优质高效金针菇新品种‘金针菇G1’的推广应用”(2014GB2C000086)。

Rapid Detection and Identification of Mycovirus in the Main Cultivated Strains of Flammulina velutipes

Zhang Junling1, Zhang Lujun2, Tan Qi2, Shang Xiaodong2, Zhang Junlin1,2, Zhang Lujun1, Tan Qi1,2, Shang Xiaodong1   

  1. (1Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences/National Engineering Research Center of Edible Fungi, Shanghai 201403;2College of Life Sciences, Nanjing Agricultural University, Nanjing 210095)
  • Received:2014-11-18 Revised:2015-02-11 Accepted:2014-12-01 Online:2015-03-20 Published:2015-03-20

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摘要： 为了探索一种成本低、速度快、易操作、特异性强的dsRNA提取方法来检测金针菇中的病毒，并对目前主栽金针菇品种进行病毒的快速检测和鉴定。以各地主栽的金针菇品种为主要研究材料，采用试剂盒法和Redzol法2种方法检测dsRNA，将这2种方法进行比较，并通过反转录PCR和测序确定病毒种类。结果表明，试剂盒法和Redzol法均在主栽金针菇品种F-4889和F-4891菌丝体中检测到FvBV的病毒组分。与试剂盒法相比，Redzol法检测时间短，样品需求量小，易操作，成本较低，不需要繁琐的前期准备工作，适于实验室和大中型食用菌生产企业使用。Redzol法能从金针菇菌丝体中准确提取dsRNA病毒组分，并在主栽品种F-4889和F-4891菌丝体中成功检测到FvBV的病毒组分，为栽培生产、优化市场种质资源、育种选材等提供了理论依据。

关键词: 达维, 达维, 路径, 暴雨, 双台风

Abstract: In this study, a lower cost, time saving, simple operation and specificity dsRNA extraction method was explored to detect and identify the mycovirus in the main cultivated strains of Flammulina velutipes. We applied two different methods to extract the dsRNA in the main cultivated strains, extraction Kit method and Redzol method, and compared the advantages of these two methods. To identify the virus type, reverse transcription PCR was employed to amplify the dsRNAs and sequence a part of them. Results showed that both Kit and Redzol methods discovered the dsRNA in strain F-4889 and F-4891 of the tested strains. Compared with the Kit method, Redzol method was time saving, simple operation, cost saving and easy pretreatment, besides it just need small amount of samples. So it was more suitable for general laboratory and large to middle-size edible fungi factory. Redzol method can accurately extract dsRNA from the mycelium of F. velutipes, and detect FvBV virus in strain F-4889 and F-4891, which provide the theory basis for cultivation and breeding, and can optimize the germplasm resources of commercial strains.