湖北农业科学 ›› 2018, Vol. 57 ›› Issue (2): 67-72.doi: 10.14088/j.cnki.issn0439-8114.2018.02.017

• 植物保护 • 上一篇    下一篇

入境旅客携带苹果中粉红聚端孢菌的检测与鉴定

许萍萍1, 粟寒2, 王振华3, 刘中慧4, 李甜甜4, 李彬2, 吴翠萍2   

  1. 1.南京出入境检验检疫局,南京 210011;
    2.江苏出入境检验检疫局,南京 210019;
    3.湖北出入境检验检疫局,武汉 430050;
    4.徐州出入境检验检疫局,江苏 徐州 221006
  • 收稿日期:2017-08-30 出版日期:2018-01-25 发布日期:2018-01-10
  • 通讯作者: 李 彬,(电话)13815871522(电子信箱)libin@jsciq.gov.cn。
  • 作者简介:许萍萍(1982-),女,江苏阜宁人,农艺师,主要从事植物检疫及研究工作,(电话)18360859806(电子信箱)xupp@jsciq.gov.cn
  • 基金资助:
    国家质量监督检验检疫总局科技计划项目(2015IK157); 江苏出入境检验检疫局科技计划项目(2017KJ08)

Detection and Identification of Trichothecium roseum from Imported Apple Carried by Passenger

XU Ping-ping1, SU Han2, WANG Zhen-hua3, LIU Zhong-hui4, LI Tian-tian4, LI Bin2, WU Cui-ping2   

  1. 1.Nanjing Entry-Exit Inspection and Quarantine Bureau, Nanjing 210011,China;
    2.Jiangsu Entry-Exit Inspection and Quarantine Bureau,Nanjing 210019,China;
    3.Hubei Entry-Exit Inspection and Quarantine Bureau,Wuhan 430050,China;
    4.Xuzhou Entry-Exit Inspection and Quarantine Bureau,Xuzhou 221006,Jiangsu,China
  • Received:2017-08-30 Online:2018-01-25 Published:2018-01-10

摘要: 首次建立了粉红聚端孢菌(Trichothecium roseum)实时荧光定量PCR检测方法,该检测方法扩增效率高、特异性好,灵敏度可达0.1 pg菌丝体DNA量;结合形态学观察及致病性测定,对从机场入境旅客携带的苹果中分离到的1株疑似菌株Tr-1685进行检测和鉴定。Tr-1685实时荧光PCR检测结果为阳性。该菌株在PDA培养基上生长较快,菌落平展,边缘不规则,表面形成淡粉红色的霉层,产孢量丰富。分生孢子梗直立,长短不一,顶端着生分生孢子。分生孢子双细胞、光滑、棍棒状,平均大小为17.5 μm×8.7 μm,基部细胞弯曲状。该病原菌接种苹果4 d后,出现褐色腐烂,8 d后出现淡粉色霉层及分生孢子。分离获得的菌株Tr-1685鉴定为粉红聚端孢菌,与实时荧光PCR结果一致。

关键词: 粉红聚端孢菌(Trichothecium roseum), 入境旅客, 苹果, 鉴定, 实时荧光定量PCR检测方法

Abstract: The real-time PCR method of Trichothecium roseum was developed for the first time, and the method of T. roseum appeared high amplification efficiency and good specificity. Sensitivity of the method reached to 0.1 pg mycelium DNA amount. The isolate Tr-1685 similar to T. roseum which was obtained from imported apple carried by airport incoming passenger had been detected and identified using real-time PCR along with morphology obersevation and pathogenicity test. The real-time PCR result of the isolate Tr-1685 showed positive. The isolate bacterial strain grew fast on PDA media. The colony was flat and irregular in appearance,covered by light pink mould which produced lots of conidia. Conidiophores of the isolate were erect and different in length,which bore conidia at the tip. Conidia of the isolate was 17.5 μm×8.7 μm on average,which was smooth and clavate. Each conidium was two celled with curved basal cell. The isolate caused brown rot on apple 4 d after inoculation. Pinkish mould and conidia appeared 8 d after inoculation. The isolate Tr-1685 was identified as T. roseum,consistent with the result of the real-time PCR.

Key words: Trichothecium roseum, incoming passengers, apple, identification, real-time PCR

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