水稻白叶枯病菌对拌种灵抗药性分子机制研究.pdfVIP

中国农业科学 2005,38(1):64-69 Scientia Agricultura Sinica 水稻白叶枯病菌对拌种灵抗药性分子机制研究 张宇君，李 俊，赵 伟，周明国 （南京农业大学植物保护学院，南京 210095） 摘要： 通过紫外诱变获得了抗拌种灵水稻白叶枯病菌（Xanthomonas oryzae pv.oryzae ）的突变体，这些突变 体可以在含 100 μg·ml-1拌种灵平板上生长，而敏感菌株在 10 μg·ml-1 浓度下则不能生长。敏感菌株琥珀酸脱氢酶活 性受拌种灵强烈抑制，而抗药突变体的酶活性较低，且不受药剂抑制。应用 6 对引物，从水稻白叶枯病菌野生敏 感菌株和室内诱导抗药性菌株中扩增到琥珀酸脱氢酶基因全序列。该基因全长 3 616 bp ，编码 1 115 个氨基酸，含 有 2 个内含子。与柑橘溃疡病菌（Xanthomonas axonopodis pv. citri ）琥珀酸脱氢酶核苷酸同源性93% ，氨基酸同 源性 97%；与其它 6 种病原细菌琥珀酸脱氢酶基因编码的氨基酸序列同源性为 43% ～95%。敏感菌株和抗药菌株 的琥珀酸脱氢酶序列分析表明，琥珀酸脱氢酶铁硫蛋白亚基中229 位氨基酸由组氨酸（CAC ）突变为酪氨酸（TAC ） 是导致X. oryzae 对拌种灵产生抗药性的主要原因。 关键词：水稻白叶枯病菌；拌种灵；琥珀酸脱氢酶；作用方式；抗药性 Molecular Mechanism of Amicarthiazol Resistance in Xanthomonas oryzae pv.oryzae ZHANG Yu-jun , LI Jun , ZHAO Wei, ZHOU Ming-guo (College of Plant Protection, Nanjing Agricultural University, Nanjing 210095) Abstract: The resistance mutants of wild-type Xanthomonas oryzae pv.oryzae to amicarthiazol were got by ultraviolet(UV) irradiation in laboratory. These mutants could grow in the medium containing 100 μg·ml-1 of amicarthiaol. However the wild-type sensitive strain could not grow at 10 μg·ml-1. The activity of succinate dehydrogenase(SDH) was strongly inhibited by amicarthiazol while the activity of SDH from resistant mutants was very low and was not inhibited by the chemical. Six pairs of special primer based on the conserved region of Xanthomonas axonopodis pv.citri were designed and used to amplify the gene encoding succinate dehydrogenase from X.oryzae . Whole gene of succinate dehydrogenase has 3 616bp, 1 115 deduced amino acids and two introns. The homology