甜橙成花关键基因CsFT的克隆分析与表达载体构建  

Cloning Analysis and Expression Vector Construction of the Key Gene CsFT for Citrus Flowering

作　　者：郭鑫跃 陈伟[1] 杨小慧 刘勇[1] 杨莉[1] Guo Xinyue;Chen Wei;Yang Xiaohui;Liu Yong;Yang Li(School of Agriculture Sciences Jiangxi Agricultural University,Nanchang,330045)

机构地区：[1]江西农业大学农学院,南昌330045

出　　处：《分子植物育种》2019年第2期478-486,共9页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31460506)资助

摘　　要：FT被认为是汇集各个开花途径的关键因子,其超量表达可促进植物提早开花。本研究通过RT-PCR方法,以纽荷尔脐橙(Citrus sinensis (L.) Osbeck)叶片为材料,成功地克隆到了2个FT基因的cDNA序列,分别为534 bp和537 bp,各自编码177个和178个氨基酸,并对其进行了生物信息学相关分析。荧光定量分析了2个CsFT基因在脐橙不同器官的表达情况,结果表明二者在果皮、果肉、茎、叶、花中均有表达,但CsFT1基因在果肉中的表达量最高,CsFT2基因在叶中的表达量最高;这2个基因在不同花器官中均有表达,CsFT1基因在花瓣中表达最高,CsFT2基因雌蕊中表达最高,并都在开花后第7天的花器官中达到最大,随后下降。将2个基因连接到植物表达载体pCAMBIA1301上,得到了过量表达载体pCAMBIA1301-35S-CsFT1和pCAMBIA1301-35S-CsFT2,并成功转化农杆菌,为下一步转化柑橘获得转基因植株提供了技术参考。The FT is c onsidered to be a key factor in the convergence of various flowering pathways, and its over-expression can promote early flowering of plants. In this study, the leaves of Citrus sinensis(L.) Osbeck were used as materials, and the cDNA sequence of two FT genes were cloned through RT-PCR technology. They were534 bp and 537 bp, respectively, encoding 177 and 178 amino acids, and the bioinformatics correlation analysis of them was carried out. The expression of two CsFT genes in different organs of Citrus sinensis was analyzed using real-time PCR, of which the results showed that the two genes were expressed in peel, pulp, stem, leaf and flower.However, the expression of CsFT1 gene was the highest in the pulp, while that of CsFT2 gene was in the leaves.Both of them were expressed in different floral organs, and Cs FT1 gene expressed the highest in petals, and the expression of CsFT2 gene was the highest in pistil. The expression of these two genes both reached the highest in the flower organ in the seventh day after flowering and then decreased. The over-expression vector pCAMBIA1301-35S-CsFT1 and pCAMBIA1301-35S-CsFT2 were obtained by connecting these two genes to the plant expression vector pCAMBIA1301, and Agrobacterium was successfully transformed, which could provide scientific reference for the further transformation of citrus to obtain transgenic plants.

关 键 词：柑橘 FT基因 基因克隆 表达 载体构建 

分 类 号：S666.4[农业科学—果树学]