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为明确库尔勒香梨萼片脱落和宿存与kfpMYB基因表达的关系,以其花器官为材料,根据库尔勒香梨kfpMYB基因c DNA序列及梨基因组信息设计引物,通过PCR获得了该基因长度为1 659 bp的基因组DNA序列和2 440 bp的上游调控序列,利用PLACE和Plant Care数据库进行启动子顺式作用元件的预测分析。生物信息学分析表明,kfpMYB启动子序列中存在一些与激素调节相关的元件,如脱落酸响应顺式作用元件ABRERATCAL、DPBFCOREDCDC3和EBOXBNNAPA,赤霉素信号抑制因子WRKY71OS、GARE-motif和TATC-box,生长素诱导有关元件NTBBF1ARROLB和TGA-element。利用实时荧光定量PCR检测了不同生长调节物质处理对kfpMYB转录水平的影响,实时荧光定量PCR分析结果表明ABA、ETH、GA、NAA和果树促控剂(PBO)对kfpMYB的表达均有不同程度的影响,说明这些调控元件参与了基因的表达。
In order to clarify the relationship between the exfoliation and persistence of sepals and the expression of kfpMYB in Korla, the primers were designed according to the krpMYB gene cDNA sequence and pear genomic information of the flower organs of Korla L., and the length of the gene was 1 659 bp genomic DNA sequence and a 2 440 bp upstream regulatory sequence were used to predict the promoter cis-acting elements using PLACE and Plant Care databases. Bioinformatics analysis showed that some elements related to hormone regulation such as ABRERATCAL, DPBFCOREDCDC3 and EBOXBNNAPA, gibberellin inhibitor WRKY71OS, GARE-motif and TATC-box were present in kfpMYB promoter sequence, Auxin-induced elements NTBBF1ARROLB and TGA-element. The effects of different growth regulators on the transcription of kfpMYB were detected by real-time fluorescence quantitative PCR. The results of real-time fluorescence quantitative PCR showed that the expression of kfpMYB was affected by ABA, ETH, GA, NAA and PBO Influence, indicating that these regulatory elements involved in gene expression.