滇水金凤花距发育相关基因ABP的克隆及表达分析

Cloning and expression analysis of ABP gene related to spur development in Impatiens uliginosa

为探究滇水金凤(Impatiens uliginosa)ABP基因的结构和表达特征,该研究以滇水金凤为材料,采用RT-PCR技术对滇水金凤ABP基因进行克隆,运用DNAMAN和MEGA对其所编码的蛋白序列进行同源性分析和系统进化分析,并利用qRT-PCR分析ABP基因的时空表达模式。结果表明:(1)滇水金凤ABP基因的cDNA全长为627 bp,编码208 aa,命名为IuABP基因,其蛋白具有Cupin超家族蛋白的典型结构。(2)同源性分析表明滇水金凤ABP基因的氨基酸序列与喜马拉雅凤仙花(I.glandulifera)、月季(Rose chinensis)、木薯(Manihot esculenta)等物种的同源性均达71%;系统进化分析表明IuABP与喜马拉雅凤仙花(Impatiens glandulifera)聚为一支,亲缘关系最近。(3)qRT-PCR分析表明IuABP基因在滇水金凤花距发育的3个时期及2个部位均有表达。随着花距的发育,IuABP基因在滇水金凤花距檐部的表达量呈先下降后上升的趋势,在盛花期时达最高,而在花距距部的表达量逐渐下降。以上结果为进一步研究滇水金凤ABP基因在花距发育中的功能及其表达调控机制提供了一定的理论参考。

The purpose of this study was to explore the structural and expression characteristics of ABP gene from Impatiens uliginosa.ABP gene related to spur development of I.uliginosa was cloned by using RT-PCR method,whose homology and phylogenetic analyses of protein sequence were analyzed by using DNAMAN and MEGA softwares.In addition,the spatiotemporal expression patterns of ABP gene were investigated by qRT-PCR method.The results were as follows:(1)ABP gene of I.uliginosa was successfully cloned,whose full-length cDNA sequences was 627 bp,encoding 208 aa,and named IuABP.The protein encoded by ABP gene had the typical structure of Cupin superfamily proteins.(2)According to the result of its homology analysis,it showed that the homology of ABP gene of I.uliginosa reached 71%with those of I.glandulifera,Rose chinensis and Manihot esculenta.Based on phylogenetic analysis,it was found that IuABP and Impatiens glandulifera clustered into a branch,with the most close genetic relationship.(3)qRT-PCR analysis showed that the IuABP were expressed in both three stages and two different parts of spur development of I.uliginosa.With the development of spur,the expression level of IuABP in the blade had a tendency of declining at the beginning and rising up later,and reached the highest in the blooming stage,while the expression level decreased gradually in the spur cup.These results provide a theoretical reference for further studies on the function and the expression regulation mechanism of IuABP gene in spur development.