Research in Plant Disease 2009;15(2):83-87.
Published online August 30, 2009.
콩 종자에서 Xanthomonas axonopodis pv. glycines의 검출을 위한 Direct PCR 방법 개발
이용주, 강미형, 노태환, 이두구, 이건휘, 김시주
Direct PCR Detection of the Causal Agents, Soybean Bacterial Pustule, Xanthomonas axonopodis pv. glycines in Soybean Seeds
Yong Ju Lee, Mi Hyung Kang, Tae Hwan Noh, Du Ku Lee, Geon Hwi Lee, Si Ju Kim
Direct Polymerase Chain Reaction (PCR) method that combines biological and enzymatic amplification of PCR targets was developed for the detection of Xanthomonas axonopodis pv. glycines on soybeen seeds without DNA isolation. Primers Xag F1 and Xag R1 were designed to specifically amplify a 401 bp fragment of the glycinecin A gene of X. axonopodis pv. glycines. Xag F1 and Xag R1 were used to carry out the PCR analysis with genomic DNA from 45 different bacterial strains including phylogenetically related bacteria with X. axonopodis pv. glycines, and other bacterial strains of different genus and species. The PCR assay using this set of primers were able to detect X. axonopodis pv. glycines with DNA concentration as low as 200 fg and 1.8×103 cfu/ml. The Xag was detected from the seed samples incubated for 2 hrs with shaking and the intensity of the band was increase with the incubation time of seeds. The Direct PCR assay method without DNA isolation makes detection of X. axonopodis pv. glycines on soybean seeds easier and more sensitive than other conventional methods. The developed seed assay using direct PCR method will be useful for the specific detection of X. axonopodis pv. glycines in soybean seed samples.
Key Words: Detection, PCR, Soybean seeds, Xanthomonas axonopodis pv. glycines

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