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为了进一步研究调控节间长度相关基因的表达模式及为克隆该半矮生性状的基因提供依据,采用cDNA-AFLP分析方法对半矮生型桃节间“生长跃变期”差异表达的基因进行筛选,并对差异的转录衍生片段(TDFs)进行回收、克隆、测序、功能注释及qRT-PCR验证。采用256对引物,共产生9021个TDFs,有明显差异的TDFs共899个,成功回收到497个TDFs,254条序列与已知基因高度相似。差异表达的基因主要涉及到信号转导、生长发育、转录调控、蛋白质代谢、防御和物质与能量代谢等。选取其中26个基因进行实时荧光定量PCR验证,结果表明26个基因的qRT-PCR表达丰度分析与cDNA-AFLP结果一致。分离到植物生长素、细胞分裂素、油菜素内酯、脱落酸、细胞周期等调控途径中生长调控相关的候选基因,可为阐明节间生长调控机制奠定基础。
In order to further study the regulation of the expression pattern of internode length-related genes and provide the basis for cloning the genes of the semidwarf traits, the cDNA-AFLP analysis was used to analyze the differential expression between semi-dwarf peach internode (TDFs) were recovered, cloned, sequenced, functional annotated and verified by qRT-PCR. Using 256 pairs of primers, a total of 9021 TDFs were generated. There were 899 TDFs with significant difference. 497 TDFs were successfully recovered, and 254 sequences were highly similar to the known genes. Differentially expressed genes are mainly involved in signal transduction, growth and development, transcriptional regulation, protein metabolism, defense and material and energy metabolism. Twenty-six genes were selected for real-time PCR. The results showed that the analysis of qRT-PCR expression of 26 genes was consistent with that of cDNA-AFLP. Isolation of auxin, cytokinin, brassinosteroid, abscisic acid, cell cycle and other regulatory pathways related to the regulation of the candidate genes, can lay the foundation for the elucidation of the mechanism of internode growth.