【作者】 朱志成；

【导师】 高洁；

【作者基本信息】 吉林农业大学 ， 植物病理学， 2005， 硕士

【摘要】 通过对采自北京、河南、河北、山东、甘肃、四川等省部分麦区的小麦白粉菌菌株做对三唑酮敏感性测定,结果表明:对三唑酮的EC50值,北京的菌株最低为7.7318μg/mL,抗性水平为3.699;山东的菌株最高为698.3μg/mL,抗性水平为334.156。以上结果表明,不同地区小麦白粉菌对三唑酮杀菌剂的敏感性差异大,同一地区的小麦白粉菌对三唑酮杀菌剂的敏感性也存在较大差异。也表明在我国小麦白粉菌对三唑酮已存有抗药性,而且不同地区的抗性之间存在着差异。 用520个10碱基的随机引物对小麦白粉菌对三唑酮不同敏感性的菌株进行RAPD扩增,有478个单引物能扩增出多态性条带,占扩增引物的92%,扩增片段在200-2000bp之间。在对抗性和敏感菌株构成的两个池中的混和菌株DNA扩增,共24条引物能扩增出多态性,分别由引物OPA5、OPA18、OPC10、OPC12、OPE18、OPH8、OPH11、OPI9、OPI12、OPK3、OPK15、OPL12、OPL13、OPP9、OPQ3、OPQ13、OPR5、OPS1、OPS11、OPY14、OPY19、OPZ17、S1170获得。在24条有多态性条带的引物中,寻找到引物OPZ17能扩增出与其不同敏感性状相应的多态性条带,在敏感菌株Es-1-s、01-8-2-（2）中能扩增出一条长度约为500bp的带(即下文中的克隆的标记OPZ17₅₂₄),而在中抗、高抗菌株中Es-2R（中抗）、子99-5-3-（2）（中抗）、E31（高抗）、Es-3R（高抗）却没有相应的条带扩出来,说明该条带可能与病菌对三唑酮敏感性相关,将其进行克隆、测序,根据序列设计特异性引物,转化成SCAR标记。 用转化的特异性引物在经鉴定已知对三唑酮不同敏感性的野生菌体中进行DNA扩增检测,检测到参试的北京、河南、河北、山东、甘肃、四川等省市部分麦区的11株敏感菌株中有8株扩增出特异性条带,而在15株抗性菌株中无此条带,表明可以为抗药性检测提供依据参考。可为进一步小麦白粉茵对三唑酮不同抗性建立起分子鉴定体系奠定基础。更多还原

【Abstract】 The sensitivities of Blumeria graminis f.sp. tritici to triadimefon were identified and the isolates,which were collected from Beijing,Henan , Hebei, Sandong, Gansu and Sichuan were test. The results showed that the lowest EC50 of isolates collected from Beijing was 7.7318μg/mL, which of the resistance was 3. 699; the highest EC50 of isolates collected from Sandong was 698. 39μg/mL, which of the resistance was 334.156. The results present that it was different sensitivities which come from different province in China. There were the difference of sensitive isolates to triadimefon in the same region. The results indicated that there were the resistant isolates to triadimefon in China, and it was different resistance to triadimefon which exsist different province in China.520 random primers were used to amplify some isolates with different sensitive to triadimefon by RAPD analysis of the genome DNA. Twenty-four primers could amplify specific DNA bands in two genome DNA pools, respectively produced by primers OPA5,OPA18,OPC10,OPC12,OPE18,OPH8,OPH11,OPI9,OPI12,OPK3,OPK15,OPL12,OPL 13,OPP9,OPQ3,OPQ13,OPR5,OPS1,OPS11,OPY14,OPY19,OPZ17,S1170.A band of about 500-600bp was obtained with primer OPZ17 amplified the genomic DNA of different sensitive isolates. The band was found from sensitive isolates to triadimenfon, and it was absent in the products from resistant isolates.The fragment OPZ17 was recovered from polyacrylamide gel and cloned in pGEM-T easy vector, then sequenced from two end. According to the sequences, two pairs of specific primers were designed. The RAPD markers was convert to SCAR markers.The different sensitive isolates to triadimefon which were identified from Beijing,Henan, Hebei, Sandong, Gansu and Sichuan in 2004-2005 were amplified with the pair of SCAR primers. The results showed that there were the specific bands could be revealed from 8 of in the 11 sensitive isolates, and no band was detected in the 15 fungicide resistant isolates. This could be convenient tool for monitoring the fungicide resistance of Blumeria graminis f.sp. tritici to triadimefon to detected.更多还原