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利用cDNA-AFLP技术,对甜瓜抗白粉病品种‘云甜-930’在接种Podosphaera xanthii生理小种2F.后的基因表达谱进行分析。256对引物共产生188个具良好多态性的转录本(TDF),其中109个上调表达,79个下调表达。经过对差异片段的回收、克隆、测序分析,最终得到60个EST。Blastx比对和功能分类分析表明,参与物质合成与代谢的属第一大类,占48%,其他主要涉及物质运输(12%)、防御系统(12%)、转录调控(8%)、能量代谢(8%)、信号转导(4%)等,7条EST(8%)与未知功能蛋白同源性较高。选取与代谢、抗病防御、信号转导及蛋白转运等相关的4个差异基因TDF12(SEH)、TDF67(SAMDC)、TDF76(CDPK)和TDF82(PDR8)进行qRT-PCR验证,结果显示其表达模式符合cDNA-AFLP表达谱,同时表明这些基因可能参与了甜瓜与白粉病菌的互作过程。
Using cDNA-AFLP technique, we analyzed the gene expression profile of melon-resistant powdery mildew ’Yuntian930’ after inoculation with Podosphaera xanthii physiological race 2F. Totally, 256 pairs of primers generated 188 polymorphic transcripts (TDF), of which 109 were up-regulated and 79 were down-regulated. After the recovery of the different fragments, cloning, sequencing analysis, the final 60 EST. Blastx alignment and functional taxonomic analysis showed that the first major group involved in the synthesis and metabolism of materials accounted for 48%, while the other major involved material transport (12%), defense system (12%), transcriptional regulation (8%), energy Metabolism (8%), signal transduction (4%) and so on. Seven ESTs (8%) have high homology with unknown functional proteins. Four differential genes TDF12 (SEH), TDF67 (SAMDC), TDF76 (CDPK) and TDF82 (PDR8) related to metabolism, disease resistance and defense, signal transduction and protein transport were selected and validated by qRT- The model conformed to the cDNA-AFLP expression profiles, indicating that these genes may be involved in the interaction between melon and B.fungi.