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当前位置: 首页 > 文章 > Nested-PCR检测种传番茄细菌性溃疡病菌 西北农业学报 2011,20 (7) 28-31
Position: Home > Articles > Nested-PCR for Detection of Seed-borne Pathogen of Tomato Bacterial Canker,Clavibacter michiganensis subsp.michiganensis Acta Agriculturae Boreali-occidentalis Sinica 2011,20 (7) 28-31

Nested-PCR检测种传番茄细菌性溃疡病菌

作  者：
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
单  位：
兰州大学基础医学院医学生物化学与分子生物学研究所;甘肃出入境检验检疫局
关键词：
番茄溃疡病;番茄种子;Direct-PCR;Nested-PCR
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。
译  名：
Nested-PCR for Detection of Seed-borne Pathogen of Tomato Bacterial Canker,Clavibacter michiganensis subsp.michiganensis
作  者：
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)
关键词：
Tomato bacterial canker;Tomato seeds;Direct-PCR;Nested-PCR
摘  要：
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 detection sensitivity of Direct-PCR is 105 cfu/mL and 107 cfu/mL in bacterial suspensions and artificial contaminated seed extracts,respectively.But the sensitivity of Nested-PCR was 102 cfu/mL and 104 cfu/mL respectively.And Nested-PCR could provide positive result in 104 fold dilution in naturally infected seed extracts.The Nested-PCR developed in our study was convenient and low-cost,which could be used a routine method for the detection of tomato bacterial canker.

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