云南马铃薯青枯病菌的PCR检测

SPECIFIC DETECTION OF RALSTONIA SOLANACEARUM IN POTATO BY PCR

利用针对马铃薯青枯病菌hrp基因族DNA序列设计合成特异性寡核苷酸引物 (引物 1:5’GAAGAGGAACGACG GAAAGC - 3’和引物 2 :5’CGAACAGCCCACAGACAAGA - 3’) ,进行马铃薯青枯病菌PCR特异性扩增试验。该试验合成的引物能从马铃薯青枯病菌总基因组DNA和细菌纯培养 ,以及人工接种和自然发病的马铃薯块茎中特异性扩增青枯病菌hrp基因区段 1993bp的分子片段。该试验结果为马铃薯青枯病菌的鉴定、检测及病害流行学研究提供了新的技术和方法。

A sensitive and specific assay based on a PCR protocol was develop for the detection of Ralstonia solanacearum , the pathogen for potato bacterial blight. Oligonucleotide specific primers (P1: 5'GAAGAGGAACGACGGAAAGC-3' and P2:5'CGAACAGCCCACAGACAAGA-3') were selected within the hrp gene region. Specific amplification of hrp fragments (1993 bp) was obtained for all the R. solanacearum strains studied. No amplification was observed for a wide range of other bacterial species. When applied to either pure culture or infected potato tubers, PCR allowed detection as low as 10 5 cfu/mL.