Duplex PCR Detection of Pseudomonas savastanoi pv.phaseolicola and Curtobacterium flaccumfaciens pv,flaccumfaciens in Soybean

The paper aimed to establish a duplex PCR method for simultaneous detection of Pseudomonas savastanoi pv. Phaseolicola （Psp） and Curtobaeterium /accumfadens pv. Flaccumfaciens （Cff）. Based on the argK gene of Psp in GenBank, the primers PSPF1/PSPR2 were designed. The duplex PCR assay was dereloped using the combined primers PSPF1/PSPR2 and CflF1/CffR2, which were specific primers for Cff. The reaction conditions were optimized and specificity md sensitivity of the duplex PCR were tested. The expected DNA fragment was specifically amplified from the genomic DNA of Psp and Cff. Specificity was conirmed in the artificially inoculated soybean samples imparted. Thus, the duplex PCR developed in this study could be used for the simultaneous detection of Psp md Cff from imported soybean.

大豆携带菜豆晕疫病菌PCR检测技术研究

晕疫病是豆科作物上重要的检疫性细菌病害。本研究在传统PCR检测技术的基础上,分析了细菌洗涤液、洗涤时间以及洗涤液容量对一粒带菌大豆检出率的影响,以及有效检出不同带菌率大豆的最佳检测时间。结果表明,洗涤液种类、洗涤时间以及洗涤液容量对检测结果影响不大,但对不同带菌率大豆洗涤时间的长短直接影响病菌的检出率,实验表明振荡洗涤时间8 h,可检测到的大豆种子最低带菌率为0.0625%。本研究所建立的检测方法简便易行,可有效应用于口岸检测进口大豆中的菜豆晕疫病菌。