摘要
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。
This study aimed to select and identify genes related to regulate the length of internode, and provide basis for revealing expression pattern of regulating internode length related genes and cloning the gene of controlling semi-dwarf trait, cDNA-AFLP technique was employed to investigate differentially expressed genes. The transcript-derived fragments （TDFs） was recycled, cloned, sequenced and functionally annotated and then validate by qRT-PCR. A total of 9 021 TDFs were screened by 256 primer combinations, of which 899 TDFs were identified differentially expression. Totally, 497 TDFs were successfully sequenced and 254 sequences were high similar to genes with known functions. The results indicated that these genes were involved in signal transduction, growth and development, transcription regulation, stress tolerance, protein degradation and synthesis and so on. The expression abundance of 26 genes validated by real-time quantitative RT-PCR was consistent with the result of cDNA-AFLP analysis. We generated candidate genes in plant IAA, CK, BR, ABA and cell cycle regulation pathways, which will help to elucidate the molecular regulation mechanism of internode length.
出处
《园艺学报》
CAS
CSCD
北大核心
2013年第5期943-952,共10页
Acta Horticulturae Sinica
基金
国家自然科学基金项目(30871682)
