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中国农学通报 ›› 2016, Vol. 32 ›› Issue (4): 150-154.doi: 10.11924/j.issn.1000-6850.casb15100072

• 生物技术科学 • 上一篇    下一篇

砀山梨黑斑病分子检测技术研究

李云飞1,陈雪娇1,张爱芳2,杨 雪2,陈 雨2,姚 剑1   

  1. (1安徽出入境检验检疫局,合肥 230022;2安徽省农业科学院植物保护与农产品质量安全研究所,合肥 230031)
  • 收稿日期:2015-10-19 修回日期:2015-12-07 接受日期:2015-12-11 出版日期:2016-02-01 发布日期:2016-02-01
  • 通讯作者: 姚剑
  • 基金资助:
    国家质量监督检验检疫总局项目“砀山梨黑斑病的生物学特性与快速精确检测技术研究”(2014IK011);安徽省农科院学科建设面上项目“梨黑斑病快速精确检测技术研究与应用”(14A1130);安徽省农科院创新团队“农田有害生物抗药性监测与治理技术创新”(12C1105);国家质量监督检验检疫总局公益性行业科研专项“花卉、水稻、棉花等重大疫病检疫关键技术研究”(201410076)。

Molecular Detection of Alternaria alternata Causing Black Spot of Dangshan Pear

Li Yunfei1, Chen Xuejiao1, Zhang Aifang2, Yang Xue2, Chen Yu2, Yao Jian1   

  1. (1Anhui Entry-Exit Inspection and Quarantine Bureau, Hefei 230022;2Institute of Plant Protection and Aro-Products Safety, Anhui Academy of Agricultural Sciences, Hefei 230031)
  • Received:2015-10-19 Revised:2015-12-07 Accepted:2015-12-11 Online:2016-02-01 Published:2016-02-01

摘要: 黑斑病是砀山梨上最重要的病害之一,建立一套砀山梨黑斑病的分子检测技术对该病害的早期及时防治及口岸检验检疫具有重要的意义。本研究通过分析比较砀山梨黑斑病菌与其他19种不同真菌的ITS序列,设计了1对特异性引物,能够从该病菌中特异性地扩增出1条398 bp的条带,而其他测试真菌中不能扩增出该条带。在灵敏度检测中,该菌的基因组DNA检出限为10 pg/μL,同时在发病的砀山梨果实组织样品和接种梨黑斑病菌的梨果实组织样品中检测出梨黑斑病菌。因此,可以将该对引物用于梨黑斑病早期诊断及病菌的PCR检测,为指导果农及时进行药剂防治提供有效的技术支撑。

关键词: 天然草地, 天然草地, 豆科牧草, 补播, 技术, 草地群落

Abstract: Black spot is one of the most severe pear diseases. Establishing a PCR detection method is important for early diagnosis and port plant quarantine. In this study, the ITS sequence of Alternaria alternata and other 19 different fungi were analyzed and compared, and a pair of specific primers were designed which could specifically amplify a 398 bp fragment from Alternaria alternata, but not from other tested fungi. In the sensitivity test, the detection limit of the bacteria genomic DNA were 10 pg/μL, Alternaria alternata was detected in both diseased pear fruit with symptoms of black spot or inoculated pear fruit by Alternaria alternata. Therefore, this pair of primers could be applied to early diagnosis and PCR detection of pear black spot. The study provided useful technical support for controlling this disease by fungicides for farmers.