水稻黄单胞菌的检测方法分析-analysis of detection methods for xanthomonas oryzae in rice.docxVIP

II II 同时检测水稻白叶枯病菌和细菌性条斑病菌。可以从白叶枯病菌中扩增出164bp及 333bp的两个条带，从水稻细菌性条斑病菌中扩增出333bp及694bp的两个条带，从同 时包含两种菌的菌液中扩增出164bp、333bp及694bp的三个条带，能够有效检测这两 种菌及其混合菌。利用模拟种子样品进行多重PCR检测，也能得到同样的结果，因此 该多重PCR技术能够应用于日常的种子检测工作中。 关键词：水稻黄单胞菌；16S rDNA 序列分析；多重 PCR 体系；检测 PAGE PAGE IV Abstract The research and application of hybrid rice in China is in the leading position internationally, and thus, export of hybrid rice have comparative advantages. Two pathogenic races of Xanthononas oryzae including Xanthomonas oryzae pv. oryzae and Xanthomonas oryzae pv. oryzicola, which were listed as the quarantine pests by many countries, are the most important bacterial diseases on rice. In order to maitan the image of export products and ensure the quality of export of hybrid rice seeds, this study the 16S rDNA of X. oryzae pv. oryzae and X. oryzae pv. Oryzicola were sequenced and analyzed, and the identification of X. oryzae pv. oryzae and X. oryzae pv. Oryzicola were testified by inoculation methods in the field experiments. The specific primers were designed to further identify the two pathogens and multiplex PCR were extablished to detect these two pathogens in rice samples. The main results were as follow: Sequence analysis of 16S rDNA and confirmation by inoculation in the field experiments The universal primers (27f/1492r) were used to amplify the 16S rDNA of 3 and 8 baterial strains from rice leaves and rice seeds, respectively, and the sequence analysis showed that the strains ZFL-13 and YFL-04 were X. oryzae pv. oryzicola, and ZFL-07 and YFL-08 were X. oryzae pv. oryzae. The results above were confirmed by inoculation methods in the field experiments and the results were in consistent with the 16S rDNA analysis. Molecular detection of X. oryzae pv. oryzicola Through sequence analysis, the specific primers (Xocf/Xocr) were designed based on the LPS gene from X. oryzae pv. oryzicola to amplify the fragments of X. oryzae pv. oryzae, X. oryzae pv. oryzicola and the other bacterial pathogens in rice see