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以玉米自交系黄早四和黄早四Ht2近等基因型系为材料,利用cDNA-AFLP差异显示方法,分析玉米抗大斑病Ht2相关基因受大斑病菌1号小种胁迫后的差异表达情况。用筛选的70对引物在黄早四Ht2中扩增出76个差异显示片段,对差异条带回收、克隆和测序,并在NCBI上进行BLAST分析,有52个在GenBank中发现相似性较高、功能已知的EST序列,按其功能分为以下8类:基础能量代谢、跨膜运输蛋白、抗逆或抗病相关蛋白、蛋白质代谢、染色体组成蛋白及DNA复制和转录中的蛋白、信号传导、生长发育调控因子和转录因子,而其中以参与基础能量代谢的基因和抗逆或抗病相关基因所占比例较大。功能已知的52个差异表达片段在玉米1～9号染色体上均有分布,且在非亲和互作前期(6h)主要是一些信号相关基因的表达,12h表达的基因多属于抗病反应初期阶段的相关基因,与防卫有关的基因主要在48～72h表达,生长发育类基因在所检测的非亲和互作的不同阶段均表现作用,在非亲和互作后期主要是蛋白质代谢基因的表达。
The maize inbred lines Huangzao 4 and Huangzao 4 Ht2 near-isogenic lines were used as materials to analyze the difference of Ht2-related genes in the susceptible maize leaf spot turf by the cDNA-AFLP differential display method Express the situation. A total of 76 differentially expressed fragments were amplified by screening 70 pairs of primers in Huangzhaosi Ht2. The differential bands were recovered, cloned and sequenced. BLAST analysis was performed on NCBI, and 52 of them showed high similarity in GenBank The functionally well-characterized EST sequences fall into the following eight categories: basal energy metabolism, transmembrane transport proteins, anti-resistance or disease-resistance-related proteins, protein metabolism, chromosomal proteins and proteins in DNA replication and transcription, signaling Transduction, growth and development regulatory factors and transcription factors, of which genes involved in basic energy metabolism and anti-resistance or resistance-related genes accounted for a large proportion. The 52 differentially expressed fragments with known functions were distributed on chromosomes 1 to 9 of maize and mainly expressed signal-related genes at the early stage of non-affinity interaction (6h). Most of the genes expressed at 12h were resistant to disease The genes related to defense at the early stage are mainly expressed at 48-72h, while the genes of growth and development are all expressed at different stages of non-affinity interactions detected. In the late stage of non-affinity interaction, the genes of protein metabolism expression.