黄瓜铜伴侣蛋白基因CsATX1克隆及表达分析  

Cloning and Expression Analysis of Copper Chaperone Gene CsATX1 from Cucumber

作　　者：孙婷婷 孟令波 李淑敏[3] Sun Tingting;Meng Lingbo;Li Shumin(Department of Food Engineering,Harbin University,Harbin,150086;School of Geography and Tourism,Harbin University,Harbin,150086;School of Resources and Environment,Northeast Agricultural University,Harbin,150030)

机构地区：[1]哈尔滨学院食品工程学院,哈尔滨150086 [2]哈尔滨学院地理与旅游学院,哈尔滨150086 [3]东北农业大学资源与环境学院,哈尔滨150030

出　　处：《分子植物育种》2024年第4期1051-1057,共7页Molecular Plant Breeding

基　　金：黑龙江省自然科学基金联合引导项目(LH2019C041);黑龙江省博士后科研启动基金项目(LBH-Q16140)共同资助。

摘　　要：中国黄瓜品种繁多,但黄瓜的抗逆性不强,容易产生病虫害。因此,分析和验证抗病相关的基因,探究黄瓜抗病的分子机理,可为培育黄瓜抗性品种提供理论基础。本研究以黄瓜为试验材料,采用RT-PCR技术克隆了黄瓜铜伴侣蛋白基因CsATX1的cDNA序列全长,并对其进行生物信息学分析;采用荧光定量PCR的方法分析CsATX1基因在细菌性角斑病侵染0~96 h下的表达情况,以期探究其与黄瓜抗病性的关系。结果表明:CsATX1基因序列全长为768 bp,含有一个288 bp的ORF阅读框,可编码95个氨基酸,该基因编码蛋白的相对分子质量约为9.95 kD,理论等电点是5.07,为亲水性蛋白,不具有跨膜结构,不含信号肽序列。系统进化树分析表明Cs ATX1与葫芦科同源蛋白的亲缘关系最近。RT-qPCR分析表明,CsATX1在黄瓜叶中有表达,在黄瓜细菌性角斑病菌侵染下,该基因在黄瓜叶中表达显著增高,受黄瓜细菌性角斑病菌的诱导。该研究结果为深入探究CsATX1基因功能提供科学依据。There are many varieties of cucumbers in China,but the resistance of cucumbers is not strong,and they are prone to pest and disease.Therefore analysis and validation of disease resistance related genes in cucumber and exploring its molecular mechanism can provide a theoretical basis for breeding resistant cucumber varieties.In this study,taking cucumber as experimental materials,the CsATX1 gene cDNA full-length sequence was cloned by RT-PCR technology.Characteristics including the physicochemical properties and conserved domain of the deduced CsATX1 protein were determined by a series of bioinformatics tools.RT-qPCR technology was performed to measure the transcript levels of CsATX1 gene induced by Pseudomonas syringae pv.lachrymans to clone CsATX1 gene and study the relationship between CsATX1 gene and disease resistance of cucumber.The results showed that the full-length nucleotide sequence of CsATX1 was 768 bp,containing a complete open reading frame of 288 bp which encoded a polypeptide of 95 amino acids.Bioinformatics analysis of the amino acid sequence showed that the molecular weight of encoded protein was 9.95 kD,and theoretical isoelectric point was 5.07.This protein was a hydrophilic protein,without transmembrane and signal peptide sequence.Phylogenetic tree analysis indicated that CsATX1 had the closest relationship with the homologous protein of Solanaceae.The expression analysis of the gene by RT-qPCR showed that the CsATX1 expressd in C.sativus leaves.Induced by P.syringae pv.lachrymans,the transcript levels of CsATX1 in cucumber leaves remarkably increased with the extension of induction time.The results of this study provides theoretical basis for further exploring the function of CsATX1 gene.

关 键 词：黄瓜 铜伴侣蛋白 基因克隆 黄瓜细菌性角斑病 荧光定量PCR 

分 类 号：S642.2[农业科学—蔬菜学]