滇牡丹类黄酮7-O-葡萄糖基转移酶基因的鉴定与表达分析

Characterization and Expression Analysis of A Favonoid 7-O-glucosyltransferase Gene in Paeonia delavayi

以野生滇牡丹为试验材料,以其转录组数据为基础,采用反转录PCR技术克隆得到一个具完整开放阅读框的类黄酮7-O-葡萄糖基转移酶基因(Pd7GT),并通过生物信息学分析手段、基因表达对该基因进行分析。结果显示:该基因全长1 446bp (Gen Bank登录号为KX394687),可编码481个蛋白氨基酸;生物信息学分析发现Pd7GT蛋白C末端含有典型糖基转移酶识别区(WAPQV)和UDP-葡萄糖基配体绑定位点(HCGWNS)的PSPG盒子,其蛋白序列GWAPQVMILEHEAVGGFVTHCGWNSTLEGISAGLPLVTWPIFAEQFYNEK,与可可(XP_007042481)、Herrannia umbratica (XP_021298085)、毛果杨(XP_006379195)、巨桉(XP_010066837)等以葡萄糖为糖基配体的类黄酮7-O-糖基转移酶聚为一类; Pd7GT基因在组织茎中表达量最高、不同花发育时期的花谢期表达量最高、不同颜色花瓣中黄花花瓣表达量最高。本研究为滇牡丹糖基转移酶异源表达、分子育种等研究提供一定的基础,为未来通过基因工程培育具新颖花色和抗性的滇牡丹新品种提供必要材料。

Taken wild Paeonia delavayi as experimental materials,we isolated and cloned a full length cDNA of flavonoid 7-O-glucosyltransferase gene through analyzing the transcriptome of P.delavayi by the methods of RT-PCR,which has a whole opening reading frame,and carried out analysis to Pd7GT with bioinformatics and gene expression.The results showed that the full length of Pd7GT gene 1 446bp(GenBank ID:KX394687),could encode 481 amino acids;the results of bioinformatics analysis revealed there a PSPG box in the C-terminal of Pd7GT,which includes typical identical region of glucosyltransferase(WAPQV)and UDP-glucose binding sites(HCGWNS),the proteinic sequence of Pd7GT GWAPQVMILEHEAVGGFVTHCGWNSTLEGISAGLPLVTWPIFAEQFYNEK;the dendrogram of Pd7GT clarified that Pd7GT and Theobroma cacao(XP_007042481),Herrannia umbratica(XP_021298085),Populus trichocarpa(XP_006379195),Eucalyptus grandis(XP_010066837)clustered one clade,which clade takes UDP-glucose as sugar moiety and be flavonoid 7-O-glucosyltransferase.The present study some basis to heterologous expression of the glucosyltransferase and molecular breeding of P.delavayi,and culturing the new cultivar that has new color and resistance through genetic engineering.