芝麻八氢番茄红素脱氢酶基因SiPDS的克隆与序列分析  

Cloning and Sequence Analysis of Si PDS Gene in Sesame indicum L.

作　　者：于沐[1,2] 文艺[1,2] 赵辉[1,2] 苗红梅[3] 刘红彦[1,2] 

机构地区：[1]河南省农业科学院植物保护研究所,郑州450002 [2]农业部华北南部农作物有害生物综合治理重点实验室,郑州450002 [3]河南省农业科学院芝麻研究中心,郑州450002

出　　处：《分子植物育种》2015年第3期589-594,共6页Molecular Plant Breeding

基　　金：现代农业产业技术体系建设专项资金(CARS-15-1-05);河南省创新型科技人才队伍建设工程共同资助

摘　　要：来自芝麻的八氢番茄红素脱氢酶基因(phytoene desaturase gene,PDS)的分离鉴定尚未见报道。本研究根据已报道植物PDS基因的氨基酸保守区,设计简并引物,扩增出编码芝麻新蔡选抗PDS基因c DNA的中间片段。利用电子克隆技术,从芝麻EST数据库中比对得到一条高度同源的序列,并通过RT-PCR方法,从新蔡选抗成功克隆了八氢番茄红素脱氢酶基因的c DNA,命名为Si PDS(Gen Bank登录号:KJ191121)。c DNA序列全长2 273 bp,开放阅读框长度为1 743 bp,编码580个氨基酸。根据克隆获得的全长c DNA序列设计引物,从新蔡选抗DNA中克隆获得Si PDS基因的全长DNA,长度为5 592 bp,转录区包括14个外显子和13个内含子。序列分析表明,Si PDS与其他植物的PDS蛋白序列高度相似;系统进化树分析表明,芝麻Si PDS与黄芩的亲缘关系最近。Si PDS基因可以用作植物病毒诱导的基因沉默载体的内源检测基因,在利用基因工程技术改良芝麻农艺性状方面具有较大的应用价值。The Phytoene desaturase gene（Si PDS） in Sesame indicum L. has not been obtained. In order to clone Si PDS, one pair of degenerated primers was designed based on the conserved regions of plant PDS gene in other plant species in NCBI. The partial sequence of Si PDS was amplified in a sesame cultivar Xincaixuankang. We aligned a highly homologous gene sequence to the amplicon from the sesame EST database by silico cloning method. By reverse transcription PCR（RT-PCR）, a putative c DNA with 2 273 bp in length was successfully cloned,which included a 1 743 bp ORF coding a protein with 580 amino acids, named Si PDS（accession number： KJ191121）.Subsequently, the whole sequence of Si PDS gene was cloned based on the partial c DNA sequence. The gene Si PDS contains 5 592 bp in length including 14 exons and 13 introns. Si PDS shared a high similarity to PDS proteins from different plant species in amino acid level. Phylogenetic analysis indicated a close relationship to Scutellaria baicalensis. The gene Si PDS can be used as an endogenous reporter gene of the virus-induced gene silencing vector,which will be useful in improving the agronomic traits of sesame by genetic engineering methods.

关 键 词：芝麻 八氢番茄红素脱氢酶基因(SiPDS) 序列分析 

分 类 号：S565.3[农业科学—作物学]