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选用耐渍性有明显差异的玉米自交系Mol7和Hz32,对二叶一心期的幼苗进行淹水处理,构建了这两个材料各自正向和反向的SSH(suppression subtractive hybridization)文库.从SSH 文库中筛选到105个阳性克隆;利用处理及对照mRNA与Mol7全生育期cDNA芯片分别杂交, 得到57个3倍以上表达差异的克隆.对所有162个克隆进行序列分析,结果表明:所有差异表达的克隆代表85个TUGs(tentative unique genes),这些TUGs主要参与厌氧代谢、逆境自我保护、信号传导、细胞结构和能量代谢等生理生化途径,同时还包括41个未知功能的cDNA,说明玉米根系对淹水胁迫响应涉及到复杂的代谢反应.对从Mol7和Hz32中筛选到的差异表达克隆进行比较分析发现,在淹水条件下,耐渍性不同的材料具有不同的耐渍相关基因的表达模式.利用Northern及RT-PCR检测了部分阳性克隆的表达谱,并克隆到一个与耐渍相关的新基因(Sicyp51).
The maize inbred lines Mol7 and Hz32 with significant difference in waterlogging resistance were used to inoculate seedlings of one leaf stage with two floats and construct the suppression subtractive hybridization (SSH) library for each of the two materials. SSH clones were screened out 105 positive clones; using processing and control mRNA and Mol7 full-growth cDNA hybridization were 57 times more than three times the expression of the clone were obtained.All 162 clones were sequenced and the results showed that: all the differences The expressed clones represented 85 TUGs (tentative unique genes). These TUGs are mainly involved in physiological and biochemical pathways such as anaerobic metabolism, self-protection during stress, signal transduction, cell structure and energy metabolism, as well as 41 unknown function cDNAs, indicating that corn Root response to waterlogging involved complex metabolic reactions.Compared with differentially expressed clones screened from Mol7 and Hz32, it was found that under different flooding conditions, materials with different waterlogging resistance had different tolerance genes The expression patterns of some positive clones were detected by Northern and RT-PCR, and a new gene related to stain tolerance (Sicyp51) was cloned.