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Nested PCR检测种传番茄细菌性溃疡病菌
引用本文:闵现华,刘 箐,韩跃武,文朝慧,王军平,刘姝彤,刘雅莉,宋 蕤,施颖波.Nested PCR检测种传番茄细菌性溃疡病菌[J].西北农业学报,2011,20(7):28-31.
作者姓名:闵现华  刘 箐  韩跃武  文朝慧  王军平  刘姝彤  刘雅莉  宋 蕤  施颖波
作者单位:1. 兰州大学基础医学院医学生物化学与分子生物学研究所,兰州 730000;甘肃出入境检验检疫局,国家质检总局外繁种子检疫重点实验室,兰州730020
2. 甘肃出入境检验检疫局,国家质检总局外繁种子检疫重点实验室,兰州730020
3. 兰州大学基础医学院医学生物化学与分子生物学研究所,兰州,730000
基金项目:甘肃省科技厅中小企业创新基金计划(0802NCCA028); 国家质检总局项目(2007IK259,2009IK276)
摘    要:利用番茄溃疡病菌特异性引物对纯菌液、模拟带菌种子及自然感染种子提取液分别进行Direct-PCR和Nested-PCR检测。结果在纯菌液中,Direct-PCR的检测灵敏度为105cfu/mL,Nested-PCR为102cfu/mL;在模拟带菌种子提取液中,Direct-PCR的检测灵敏度为107cfu/mL,Nested-PCR为104cfu/mL;在自然感染种子提取液中,稀释104倍后Nested-PCR仍然可以检出。建立的番茄种子Nested-PCR检测技术,无需经过核酸提取步骤,可以在8 h内完成整个检测过程,方便快捷,成本低廉,可作为番茄种子带菌的常规检测方法。

关 键 词:番茄溃疡病  番茄种子  Direct-PCR  Nested-PCR  

Nested PCR for Detection of Seed borne Pathogen of Tomato Bacterial Canker, Clavibacter michiganensis subsp. michiganensis
MIN Xianhu,LIU Qing,HAN Yuewu,WEN Zhaohui,WANG Junping,LIU Shutong,LIU Yali,SONG Rui and SHI Yingbo.Nested PCR for Detection of Seed borne Pathogen of Tomato Bacterial Canker, Clavibacter michiganensis subsp. michiganensis[J].Acta Agriculturae Boreali-occidentalis Sinica,2011,20(7):28-31.
Authors:MIN Xianhu  LIU Qing  HAN Yuewu  WEN Zhaohui  WANG Junping  LIU Shutong  LIU Yali  SONG Rui and SHI Yingbo
Institution:MIN Xianhua{1,2},LIU Qing{2},HAN Yuewu1,WEN Zhaohui2,WANG Junping2,LIU Shutong2,LIU Yali2,SONG Rui2 and SHI Yingbo2 (1.Institute of Medical Biochemistry and Molecular Biology,School of Basic Medical Sciences,Lanzhou University,Lanzhou 730000,China,2.Key Laboratory of Seed Quarantine,Gansu Entry-exit Inspection and Quarantine Bureau,Lanzhou 730020,China)
Abstract:Tomato bacterial canker is the most important bacterial diseases which caused substantial economic losses in the worldwide.Seeds were the main pathway of long-distance spread and seed-borne Clavibacter michiganensis subsp.michiganensis(Cmm) were difficult to detect by current methods because of they were low rate of infected.Direct-PCR and Nested-PCR were performed for the detection of Cmm in bacterial suspensions,artificial contaminated seed extracts and naturally infected seed extracts,respectively.The de...
Keywords:Tomato bacterial canker  Tomato seeds  Direct-PCR  Nested-PCR  
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