萼脊兰FPPS基因的克隆及表达分析  

The Cloning and Expression Analysis of FPPS Gene from Sedirea japonica

作　　者：蒋素华[1] 王默霏[1] 郝平安 袁秀云[1] 马杰[1] 崔波[1] Jiang Suhua;Wang Mofei;Hao Pingan;Yuan Xiuyun;Ma Jie;Cui Bo(Institute of Bioengineering, Zhengzhou Normal University, Zhengzhou,450044;College of Life Sciences, Henan Agricultural University, Zhengzhou,450002)

机构地区：[1]郑州师范学院生物工程研究所,郑州450044 [2]河南农业大学生命科学学院,郑州450002

出　　处：《分子植物育种》2018年第23期7606-7612,共7页Molecular Plant Breeding

基　　金：河南省科技厅科技发展计划项目(152107000071);郑州市科技发展计划项目(20150448);郑州师范学院科技创新团队支持计划共同资助

摘　　要：为了深入研究兰科植物花香的调控机制,该研究以萼脊兰为材料,利用RACE技术克隆萼脊兰花香物质的关键酶基因FPPS的全长序列并对其进行生物信息学分析和时空表达分析。结果显示:FPPS基因全长为1317bp,开放阅读框(ORF)长度为1047bp,编码348个氨基酸(GenBank登录号为MG018616)。FPPS编码的蛋白质为亲水性蛋白,等电点(pI)为5.11。萼脊兰FPPS氨基酸序列与春兰的相似性为92%,与霍山石斛的相似性为90%。FPPS蛋白分布于线粒体、叶绿体和分泌途径中,且可信度为3。预测FPPS蛋白质结构功能,可能有意义的功能包括翻译、氨基酸的生物合成、脂肪酸代谢和辅酶因子的生物合成等,他们的几率分别是4.666、4.108、3.689、3.040;对其蛋白质二级结构进行预测,α螺旋占39.37%,无规则卷曲占43.10%,延伸链占17.53%;以4kk2.1.A为模板进行蛋白质的三维结构预测,与FPPS蛋白一致性为72.14%。荧光实时定量PCR结果显示,FPPS基因在盛花期的花瓣中表达量最高,时空特异性明显,为研究萼脊兰花香成分和花香分子生物学打下了良好基础。In order to study the regulation mechanism of fragrance in orchid, this research took Sedirea japonica as a material and used rapid amplification of cDNA ends(RACE) to clone full length sequence of key enzyme gene FPPS from aromatic constituent in Sedirea japonica. Bioinformatics analysis and temporal and spatial expression analysis were also carried out. The results showed that the full-length sequence of FPPS gene was 1 317 bp and the opening reading frame(ORF) was 1 047 bp, which encoded 348 amino acids(GenBank Accession No. MG018616).The protein was hydrophilic and the theoretical pI was 5.11. The comparability of the FPPS amino sequences of Sedirea japonica with cymbidium was 92%, and that with Dendrobium huoshanense was 90%. The FPPS protein was distributed in the mitochondria, chloroplast and secretory pathway with a credibility of 3. This research predicted the protein structure function of FPPS, which might include translation, the biosynthesis of amino acid, fatty acid metabolism, biosynthesis of cofactors and so on, of which the odds were 4.666, 4.108, 3.689, 4.108, respectively. The FPPS protein secondary structure components showed that a-helix(h), extended strand(e), and random coil(c) accounted for 39.37%, 17.53%, and 43.10%, respectively. The three-dimensional structure of protein predicted by using 4 kk2.1.A as template had 72.14% consistent with FPPS. The results of real-time RT-PCR showed that FPPS genes expressed highest in full bloom petals with obvious spatial and temporal specificity obvious. This study would lay a good foundation for the study of floral components and floral molecular biology in Sedirea japonica.

关 键 词：萼脊兰 FPPS基因 克隆 表达分析 

分 类 号：S682.31[农业科学—观赏园艺]