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根据水稻细菌性基腐病病菌菌株的16S-23S细菌核糖体DNA ITS区域的特异区间,设计了1对水稻细菌性基腐病病菌特异性扩增引物X1和X2,拟建立水稻细菌性基腐病实时荧光PCR检测方法。试验结果表明,在供试的8种细菌中,只能特异性的扩增出水稻细菌性基腐病病菌的目标条带,实时荧光PCR能够检测出的目标菌DNA量最低浓度为5×10-3ng/μL,比常规PCR的灵敏度高100倍。实时荧光PCR也能够在人工接种的水稻中检测到病菌的存在。
According to the specific interval of bacterial ribosomal DNA ITS of 16S-23S bacterial strain of rice bacterial rot fungi, 1 pair of primers X1 and X2 were designed to amplify the bacterial bacterial pathogens of rice, Real-time fluorescence PCR detection method. The results showed that in the eight kinds of bacteria tested, only the specific bands of bacteria could be amplified. The minimum concentration of target bacteria detected by real-time fluorescence PCR was 5 × 10 -3ng / μL, 100 times more sensitive than conventional PCR. Real-time fluorescence PCR also detects the presence of germs in artificially inoculated rice.