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采用cDNA-AFLP技术,对成株抗条锈小麦品种兴资9104在成株期受条锈菌生理小种CY32侵染后5d内9个时间点的基因表达谱进行了分析。共筛选64对引物,产生32320个转录本(TDF);用37对引物检测到2201个(6.81%)差异TDF,其中926个TDF诱导表达,1275个下调表达。经大规模克隆、测序分析,最终获得330个差异TDF,聚类分析得到259个EST(unigenes),命名为aTaPST1至aTaPST259(GenBank登录号为FL645754～FL646011和FL646262)。经Blastx比对和功能分类分析,其中96条EST(37.07%)未找到同源性匹配,68条(26.25%)与未知功能蛋白同源性较高;其余95条ESTs主要涉及能量(11.20%)、基础代谢(4.63%)、转录调控(3.86%)、抗病与防御(3.86%)、蛋白质运输和储存(3.09%)、蛋白质合成和细胞生长(各2.32%)以及信号转导(1.54%)等。选取抗病与防御、转录调控及信号转导类等相关的6个差异基因,qRT-PCR分析结果显示其表达模式符合cDNA-AFLP表达谱。小麦成株抗条锈性分子机制涉及植物多方面生理生化反应,包括抗病与防御、转录调控、蛋白质代谢、信号转导、以及非生物胁迫等多种途径相关基因的协同控制。
Using cDNA-AFLP technique, the gene expression profile of the resistant-resistant wheat cultivar Xingzao 9104 was analyzed at 9 time points within 5 days after being infected with the stripe rust CY32 at adult stage. A total of 64 pairs of primers were screened to generate 32,320 transcripts (TDF). Twenty-one (6.81%) differential TDFs were detected with 37 pairs of primers, of which 926 TDFs were induced and 1275 were down-regulated. After a large-scale cloning and sequencing analysis, 330 differential TDFs were finally obtained, and 259 ESTs (unigenes) were obtained by cluster analysis and named aTaPST1 to aTaPST259 (GenBank accession numbers FL645754 to FL646011 and FL646262). There was no homology match between 96 ESTs (37.07%) and 68 (26.25%) with unknown function proteins. The remaining 95 ESTs mainly involved energy (11.20% ), Basal metabolism (4.63%), transcriptional regulation (3.86%), disease resistance and defense (3.86%), protein transport and storage (3.09%), protein synthesis and cell growth %)Wait. Six differential genes related to disease resistance, defense, transcription regulation and signal transduction were selected and analyzed by qRT-PCR. The results of qRT-PCR showed that the expression patterns were in accordance with the cDNA-AFLP expression profile. The molecular mechanism of adult plant stripe rust resistance involves many physiological and biochemical reactions in plants, including synergistic control of genes involved in many pathways including disease resistance and defense, transcriptional regulation, protein metabolism, signal transduction, and abiotic stress.