杭菊DFR基因克隆及其在花中表达特征分析

发布时间：2018-05-25 23:32

本文选题：杭菊 + DFR基因　；参考：《中草药》2016年07期

【摘要】：目的克隆杭菊Chrysanthemum morifolium二氢黄酮醇还原酶(dihydroflavonol 4-reductase,DFR)基因,并研究其在花芽分化期及开花期不同时期的表达特征。方法根据已报道的菊花Chrysanthemum morifolium DFR基因序列设计引物,通过荧光定量PCR(RT-PCR)技术从杭菊头状花序中扩增出目的基因片段,连接、转化、挑取阳性克隆测序并拼接;采用基于内参基因的相对定量PCR方法,分析该基因表达量特征。结果得到全长1 152 bp的杭菊DFR基因,最大开放阅读框1 029 bp,共编码342个氨基酸。NCBI上进行Blast比对,该基因与其他植物的DFR基因有很高的同源性。RT-PCR表明该基因在花芽分化时期不同阶段以及开花期不同时期花不同部位中表达量均有差异。花芽分化时期杭菊DFR基因在50 d表达量最高;开花期该基因在时期I的管状花中表达量最高,在时期II的花序托中表达量最低。结论克隆得到杭菊DFR基因c DNA序列,该基因表达存在时间和空间上的差异,表达量在花蕾膨大成熟到舌状花开放30%的时间段内最高。该结果为进一步进行该基因在杭菊中的表达与催化产物的关系和调控机制研究奠定了基础。
[Abstract]:Objective to clone the dihydroflavone reductase (Chrysanthemum morifolium) gene from Chrysanthemum morifolium and study its expression in different stages of flower bud differentiation and flowering. Methods according to the reported Chrysanthemum morifolium DFR gene sequence of chrysanthemum, primers were designed and amplified from the head of Chrysanthemum chrysanthemum by fluorescence quantitative PCRRT-PCRR technique. The target gene fragment was ligated, transformed, sequenced and sequenced. The relative quantitative PCR method based on internal reference gene was used to analyze the expression characteristics of this gene. Results A total length of 1 152 BP DFR gene was obtained, and the maximum open reading frame was 1 029 BP, encoding 342 amino acids. NCBI was used for Blast comparison. RT-PCR showed that the gene was expressed in different stages of flower bud differentiation and in different parts of flower in flowering stage. The expression of DFR gene was the highest in flowering stage I and the lowest in inflorescence peduncle of stage II. Conclusion the c DNA sequence of DFR gene was obtained by cloning. The expression of c DNA gene was different in time and space, and the expression level was the highest in the period of blooming from blooming bud to 30% blooming of tongue flower. The results laid a foundation for the further study of the relationship between the expression of the gene and the catalytic products and the regulation mechanism in Hangzhou Chrysanthemum.
【作者单位】：南京农业大学中药材研究所;
【基金】：国家自然科学基金资助项目(81503180)
【分类号】：S567.239

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