马铃薯StCYP734A1基因克隆、表达模式及生物信息学分析  

Cloning, Expression Pattern and Bioinformation Analysis of StCYP734A1 Gene in Solanum tuberosum

作　　者：张杰[1] 邓孟胜 蔡诚诚 冉爽 余丽萍 杨翠芹[1,2] 王西瑶[1,2] Zhang Jie;Deng Mengsheng;Cai Chengcheng;Ran Shuang;Yu Liping;Yang Cuiqin;Wang Xiyao(Potato Research and Development Center,Sichuan Agricultural University,Chengdu,611130;National Crop Science Experimental TeachingDemonstration Center,Sichuan Agricultural University,Chengdu,611130)

机构地区：[1]四川农业大学,马铃薯研究与开发中心,成都611130 [2]四川农业大学,作物科学国家级实验教学示范中心,成都611130

出　　处：《分子植物育种》2019年第15期4883-4893,共11页Molecular Plant Breeding

基　　金：现代农业产业技术体系四川薯类创新团队项目(川农业函[2014] 91号)资助

摘　　要：CYP734As作为失活油菜素内酯(BRs)的关键基因,调节植物生长发育,可未见马铃薯CYP734As基因的相关报道。为探索CYP734As在马铃薯块茎生长发育中发挥的功能,本试验以RT-PCR技术从马铃薯品种‘费乌瑞它’中克隆到StCYP734A1基因全长序列,并对其进行生物信息学分析;利用Real-time PCR技术分析不同组织及块茎不同贮藏时期基因表达模式。结果显示,该序列开放阅读框(ORF)为1 644 bp,编码547个氨基酸,蛋白质相对分子质量约62.07 kD,理论等电点(pI) 9.31,在7~26位氨基酸存在一个跨膜区,亚细胞定位预测主要在线粒体和细胞核,系统进化分析表明与番茄LeCYP734A8亲缘关系最近。荧光定量分析马铃薯StCYP734A1在不同组织和块茎不同贮藏时期基因表达量,根中StCYP734A1表达量最高,其次是茎、叶、花、果实。随着贮藏时间的延长,马铃薯StCYP734A1表达量升高,打破休眠时,该基因表达量显著降低。本试验结果为进一步研究马铃薯StCYP734A1基因的功能提供了依据。CYP734As as a key gene in inactivating brassinosteroids (BRs), it can regulate the growth and development of plant, but related report of potato CYP734As gene was not found. In order to explore the function of CYP734As in the growth and development of potato tubers, the full-length sequence of StCYP734A1 gene was cloned from the potato variety 'Favorita' by RT-PCR, and the bioinformatics analysis was carried out. Real-time PCR was used to analyze gene expression patterns in different tissues and tubers during different storage periods. The results showed that the open reading frame (ORF) of this sequence was 1 644 bp, encoding 547 amino acids, the relative molecular mass of the protein was about 62.07 kD, the theoretical isoelectric point (pI) was 9.31, and there was a transmembrane region at 7~26 amino acids. The prediction of subcellular localization was mainly in mitochondria and nucleus, and phylogenetic analysis showed that the relationship between tomato and LeCYP- 734A8 was the most recent. Fluorescence quantitative analysis of the expression of potato StCYP734A1 in different tissues and tubers during different storage periods, the expression of StCYP734A1 in roots was the highest, then followed by stems, leaves, flowers and fruits. With the prolongation of storage time, the increasing expression of StCYP734A 1 in potato, and the expression of this gene decreased significantly when breaking dormancy. The results of this experiment lay the foundation for further study of the function of the potato StCYP734A 1 gene.

关 键 词：马铃薯(Solanum TUBEROSUM L.) StCYP734A1 基因克隆 表达分析 生物信息学分析 

分 类 号：S532[农业科学—作物学] Q943.2[生物学—植物学]