|Table of Contents|

Qualitative PCR Detection Method for Clavibacter michiganensis subsp.Michiganensis(PDF)

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Issue:
2015年05
Page:
128-131
Research Field:
Publishing date:

Info

Title:
Qualitative PCR Detection Method for Clavibacter michiganensis subsp.Michiganensis
Author(s):
MAO Fu-rong1; LI Fei-wu2; LIU Yan-ni1; LIU Jing-li1; PAN Bo1
(1.Jilin Academy of Vegetables and Flowers，Changchun，Jilin 130033；2.Jilin Academy of Agricultural Sciences，Changchun，Jilin 130033)
Keywords:
phytopathogen; Clavibacter michiganensis subsp.michiganensis; PCR; molecular detection
PACS:
-
DOI:
10.11937/bfyy.201505037
Abstract:
Using two Clavibacter michiganensis subsp.michiganensis strains and six kinds of plant pathogen as experimental materials，through primers design，specificity and sensitivity testing，a PCR method for detection of Clavibacter michiganensis subsp.michiganensis by employing three specific genes of micA，cytC，and TomA were established.The results showed that this developed PCR method resulted in a specific and accurate detection of Clavibacter michiganensis subsp.michiganensis.In particular，the sensitivity of the detection method using TomA gene as test target could reach 5 copy/μL or 103 cfu/mL，which was more sensitive than the other two methods by employing micA and cytC gene，respectively.This study provided a fast and precise technological means for the early diagnosis of Clavibacter michiganensis subsp.michiganensis.

References:

 

[1] Wittmann J，Gartemann K H，Eichenlaub R，et al.Genomic and molecular analysis of phage CMP1 from Clavibacter michiganensis subspecies michiganensis[J].Bacteriophage，2011，1(1)：6-14.

[2] 罗来鑫，赵廷昌，李建强，等.番茄细菌性溃疡病研究进展[J].中国农业科学，2004，37(8):1144-1150.

[3] 赵文军，夏明星，陈红运，等.番茄溃疡病菌PCR快速检测技术[J].植物检疫，2007，21(2):75-77.

[4] 王蕊，罗来鑫，李健强.不同来源番茄溃疡病菌致病力差异研究[J].植物保护，2010，36(1):73-77.

[5] 李江利，欧阳林杰，郭淼淼，等.河南省番茄溃疡病的发现与分子确诊[J].河南农业大学学报，2011，45(3):349-352.

[6] 王念武，王婷，沈建国，等.基于锁式探针的番茄溃疡病菌实时荧光PCR快速检测[J].中国农业科学，2014，47(5):903-911.

[7] 闵现华，刘菁，韩跃武，等.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.

[8] 闵现华，韩跃武，刘菁，等.种传番茄溃疡病菌直接PCR和免疫捕捉PCR检测方法之比较[J].植物检疫，2010，24(4):12-16.

[9] 封立平，尼秀媚，魏晓棠，等.番茄溃疡病菌的环介导等温核酸扩增技术检测方法[J].食品安全质量检测学报，2013，4(4):1207-1212.

[10]Yasuhara-Bell J，Kubota R，Jenkins D M，et al.Loop-mediated amplification of the Clavibacter michiganensis subsp. michiganensis micA gene is highly specific[J].Phytopathology，2013，103(12)：1220-1226.

[11]Cho M S，Lee J H，Her N H，et al.A quantitative and direct PCR assay for the subspecies-specific detection of Clavibacter michiganensis subsp.michiganensis based on a ferredoxin reductase gene[J].Journal of Microbiology，2012，50(3)：496-501.

[12]Gartemann K H，Abt B，Bekel T，et al.The genome sequence of the tomato-pathogenic actinomycete Clavibacter michiganensis subsp. michiganensis NCPPB382 reveals a large island involved in pathogenicity[J].Journal of Bacteriology，2008，190(6)：2138-2149.

[13]王念武，王婷，沈建国，等.超分支滚环扩增技术快速检测番茄溃疡病菌[J].植物检疫，2013，27(5):66-70.

[14]Bach H J，Jessen I，Schloter M，et al.A TaqMan-PCR protocol for quantification and differentiation of the phytopathogenic Clavibacter michiganensis subsp.sepedonicus[J].Journal of Microbiological Methods，2003，52(1)：85-91.

[15]Holtsmark I，Mantzilas D，Eijsink V G，et al.Purification，characterization，and gene sequence of michiganin A，an actagardine-like lantibiotic produced by the tomato pathogen Clavibacter michiganensis subsp.michiganensis[J].Applied and Environmental Microbiology，2006，72(9)：5814-5821.

[16]吴兴海，邵秀玲，厉艳，等.应用分子生物学方法快速检测番茄溃疡病菌的研究[J].江西农业学报，2006，18(2):5-7.

[17]付鹏，郭亚辉，张晓梅，等.番茄溃疡病菌分子检测技术[J].江苏农业学报，2005，21(2):118-122.

Memo

Memo:
-
Last Update: 2015-06-24