香菜CsEF-1α基因克隆与表达分析  

Cloning and Expression Analysis of CsEF-1αGene in Coriander

作　　者：谢芳杰 轩正英 张娟[1] 王新建[1,2,3] 吴翠云[1,2,3] 杜红斌[1] XIE Fangjie;XUAN Zhengying;ZHANG Juan;WANG Xinjian;WU Cuiyun;DU Hongbin(College of Horticulture and Forestry,Tarim University,Alar,Xinjiang 843300,China;National-Local Joint Engineering Laboratory of High Efficiency and Superior-Quality Cultivation and Fruit Deep Processing Technology on Characteristic Fruit Trees,Alar,Xinjiang 843300,China;XPCC Key Laboratory of Production and Utilization of Biological Resources in Tarim Basin,Alar,Xinjiang 843300,China)

机构地区：[1]塔里木大学园艺与林学学院,新疆阿拉尔843300 [2]塔里木大学南疆特色果树高效优质栽培与深加工技术国家地方联合工程实验室,新疆阿拉尔843300 [3]塔里木大学新疆生产建设兵团塔里木盆地生物资源保护利用兵团重点实验室,新疆阿拉尔843300

出　　处：《西北植物学报》2022年第11期1835-1843,共9页Acta Botanica Boreali-Occidentalia Sinica

基　　金：新疆生产建设兵团重大科技计划项目(2016AA002-2);塔里木大学校长基金项目(TDZKSS202245)。

摘　　要：翻译延伸因子EF-1α(elongation factor 1 alpha)是细胞中最丰富的蛋白质之一,其在确保mRNA正确解码以产生细胞蛋白质方面发挥重要作用。该研究采用RT-PCR扩增方法克隆香菜CsEF-1α基因序列,利用生物信息学对CsEF-1α基因结构、序列特征及系统进化等进行分析,并采用qPCR探究CsEF-1α基因在香菜不同生长时期和非生物胁迫下的表达模式,为进一步揭示EF-1α基因调控机制的研究奠定基础。结果显示:(1)成功克隆获得香菜CsEF-1α基因序列;CsEF-1α基因包含1个1344 bp的开放阅读框,编码447个氨基酸,分子式为C 2202 H 3544 N 594 O 644 S 20,蛋白质分子量为49.29 kD,等电点为9.12;氨基酸序列组成中赖氨酸数量最多(49个,占11.0%),色氨酸数量最少(3个,占0.7%);属碱性蛋白。(2)CsEF-1α蛋白主要由无规则卷曲(36.91%)和α-螺旋(30.43%)构成,定位于细胞质;系统进化树分析显示,CsEF-1α与胡萝卜、野生番茄、青蒿素和非洲菊的亲缘关系较接近;启动子分析包括4种植物生长发育元件、3种激素响应元件和3种胁迫响应元件。(3)qRT-PCR结果显示,CsEF-1α基因的表达量随着香菜生长发育时间的延长而升高,并且与转录丰度的变化一致;CsEF-1α基因对4种不同非生物胁迫的响应表达模式有所差异;随着胁迫时间的延长,在盐胁迫下CsEF-1α基因表现出先升高后降低趋势,而在低温、高温和干旱胁迫下表现出先降低再升高的趋势。研究表明,CsEF-1α基因参与了香菜对非生物胁迫的应答,在香菜生长发育和非生物胁迫中具有重要调控作用。Translation elongation factor EF-1α(elongation factor 1 alpha)is one of the most abundant proteins in cells,which plays an important role in ensuring the correct decoding of mRNA to produce cellular proteins.In this study,the gene sequence of CsEF-1αof coriander was cloned by RT-PCR,and the structure,sequence characteristics,and phylogenetic evolution were analyzed by bioinformatics software.Meanwhile,the qRT-PCR was used to detect the expression patterns of CsEF-1αin coriander at different growth stages and different abiotic stress treatments.These provide the foundation for further research on the regulatory mechanism of the EF-1αgene.The results showed that:(1)the CsEF-1αgene sequence of coriander was cloned successfully.CsEF-1αgene contained a 1344 bp open reading frame encoding 447 amino acids.The molecular formula of CsEF-1αwas C 2202 H 3544 N 594 O 644 S 20,the protein molecular weight was 49.29 kD and the isoelectric point was 9.12.CsEF-1αis a basic protein with the highest number of lysine(49,11.0%)and the lowest number of tryptophan(3,0.7%).(2)The protein of CsEF-1αwas mainly composed of random coils(36.91%)andα-helices(30.43%),and localized in the cytoplasm.Phylogenetic tree analysis showed that CsEF-1αin coriander was closely related to carrots,wild tomatoes,artemisinin,and Gerbera.Promoter prediction found that the promoter of CsEF-1αcontained four plant growth developmental elements,three hormone response elements,and three stress response elements.(3)The results of the qRT-PCR showed that the expression profiles of the CsEF-1αincreased with the growth of coriander and were consistent with the transcript abundance.The expression pattern of CsEF-1αdiffered under four different abiotic stresses.With the increase of stress treatment time,the CsEF-1αgene showed a tendency to increase and then decrease under salt stress,while it showed a tendency to decrease and then increase under low temperature,high temperature,and drought stress.These results showed that the CsEF-1αgene in coriander i

关 键 词：香菜 CsEF-1α 生物信息学 基因表达模式 非生物胁迫 

分 类 号：Q785[生物学—分子生物学] Q786[农业科学—蔬菜学] S636.9[农业科学—园艺学]