A multiplex PCR method for detection of Clavibacter michiganensis subsp. michiganensis with co-amplification of its host DNA
A multiplex PCR method for detection of Clavibacter michiganensis subsp. michiganensis with co-amplification of its host DNA
Yan ZHANG, Wenxiang YANG, Yaning LI, Daqun LIU(), Ting ZHANG
College of Plant Protection, Biological Control Center for Plant Disease and Plant Pests of Hebei Province, Agricultural University of Hebei, Baoding 071001, China
A multiplex PCR assay system was developed for the detection of Clavibacter michiganensis subsp. michiganensis (Cmm), which combined two tests in one reaction mixture. Cmm-specific primers PSA-4/PSA-R and Solanum lycopersicum–specific primers NS-7-F/NS-8-R (internal PCR control primer) were combined in one PCR reaction mixture with Cmm and plant DNA as template. The primer sets could amplify the target product successfully. Different combinations and concentrations of primers and annealing temperatures were tested, respectively. The detection level of the optimized multiplex PCR assay was up to 5×102 cfu·mL-1. To verify the applicability of this system, it was employed to detect Cmm in tomato seeds and plantlet samples. Seeds mixed with Cmm and diseased plantlets were detected successfully. The multiplex PCR system will avoid false-negative results and provide a reliable method for the detection of Cmm.
Corresponding Author(s):
LIU Daqun,Email:ldq@hebau.edu.cn
引用本文:
. A multiplex PCR method for detection of Clavibacter michiganensis subsp. michiganensis with co-amplification of its host DNA[J]. Frontiers of Agriculture in China, 2009, 3(2): 140-145.
Yan ZHANG, Wenxiang YANG, Yaning LI, Daqun LIU, Ting ZHANG. A multiplex PCR method for detection of Clavibacter michiganensis subsp. michiganensis with co-amplification of its host DNA. Front Agric Chin, 2009, 3(2): 140-145.
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