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为深入了解玉米抗灰斑病的分子机理,本试验以抗/感玉米灰斑病自交系78599-1、OH43Ht N/掖478、K12为试验材料,利用DDRT-PCR方法筛选玉米6叶期时接种玉米灰斑病菌粗毒素0、5、7和9 d时玉米叶片中差异表达的m RNA并通过反式Northern进行验证。结果发现在抗/感材料78599-1、OH43Ht N/掖478、K12中分别分离到6、5、5和3条差异表达片段;对19条差异表达片段进行测序和blast分析,得到6个候选基因片段,它们与玉米中MTN3、CONSTANS-CO5家族的基因、脱落酸受体基因和酰基辅酶A结合蛋白基因同源性分别为97%、99%、97%和100%;另外获得2个基因片段与未知功能基因同源性均为99%。研究结果将为深入研究抗灰斑病分子机理提供理论依据。
In order to further understand the molecular mechanism of resistance to Cercospora solani in maize, in this study, resistant and susceptible lines were screened by 6-leaf stage of DDRT-PCR using inbred lines 78599-1 and OH43Ht N / t 478 and K12 as test materials. The differentially expressed m RNA in maize leaves at 0, 5, 7, and 9 d inoculation with R. solani crude inoculation was verified by trans Northern analysis. The results showed that 6, 5, 5 and 3 differentially expressed fragments were isolated from 78599-1, OH43Ht N / Ye 478 and K12, respectively. Nineteen differentially expressed fragments were sequenced and blast analyzed to obtain 6 candidates Gene fragments, which share 97%, 99%, 97% and 100% homology with the genes of MTN3 and CONSTANS-CO5 in maize, abscisic acid receptor gene and acyl-CoA binding protein respectively, and additionally obtained two genes The homology between the fragment and unknown function gene is 99%. The results will provide a theoretical basis for further study on the molecular mechanism of resistance to Cercospora sojae.