马铃薯StDHN-2基因的克隆与表达分析  

Cloning and Expression Analysis of the StDHN-2 Gene in Potato(Solanum tuberosum)

作　　者：魏晶晶 马丽丽 张宁[1,2] 司怀军[1,2] WEI Jing-Jing;MA Li-Li;ZHANG Ning;SI Huai-Jun(College of Life Science and Technology,Gansu Agricultural University,Lanzhou 730070,China;State Key Laboratory of Aridland Crop Science,Gansu Agricultural University,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学生命科学技术学院,兰州730070 [2]甘肃农业大学干旱生境作物学国家重点实验室,兰州730070

出　　处：《农业生物技术学报》2025年第4期731-742,共12页Journal of Agricultural Biotechnology

基　　金：国家重点研发计划(2022YFD1602103);甘肃省科技重大专项计划(23ZDNA006)。

摘　　要：植物脱水蛋白(dehydrins,DHN)是一类在植物遭受干旱、盐碱等逆境时表达的特异性蛋白。为探究马铃薯(Solanum tuberosum)脱水蛋白(StDHN-2)基因在植株抗旱方面的功能,本研究以马铃薯品种'Atlantic'为材料,克隆了StDHN-2基因的CDS序列,通过同源重组法构建了融合蛋白表达载体,亚细胞定位结果表明,StDHN-2蛋白可能定位于细胞核上。对StDHN-2基因进行基础的生物信息学分析,发现StDHN-2基因组全长931 bp,其中编码区序列为474 bp,可编码157个氨基酸;蛋白分子量为16659.06 D,理论等电点为7.23,为亲水蛋白,无跨膜结构域。该蛋白二级结构以无规则卷曲为主,没有β-转角结构。通过构建系统发育树,发现StDHN-2在进化关系上和多疣茄(Solanum verrucosum)的亲缘关系最近;顺式作用元件分析表明,StDHN-2基因启动子区包含ABRE、DRE、P-box和LTR等多个与逆境响应相关的元件;利用qRT-PCR技术分析了StDHN-2基因在PEG、NaCl和脱落酸(abscisic acid,ABA)3种胁迫处理下的表达模式发现,该基因在3种胁迫处理下都会出现一定程度的上调表达。研究结果初步说明StDHN-2基因可作为马铃薯抗旱的优良候选基因,参与植物抗旱的生物学过程。本研究为解析马铃薯StDHN-2基因的抗旱响应机制提供了基础资料。Plant dehydrins(dehydrins,DHN)are a class of specific proteins expressed when plants are subjected to adversities such as drought and salinity.To investigate the function of potato(Solanum tuberosum)StDHN-2 gene in plant drought tolerance,in this study,the CDS of StDHN-2 gene was cloned from potato cultivar'Atlantic'plantlets,and the fusion protein expression vector was constructed by homologous recombination,the subcellular localization results of StDHN-2 protein showed that the protein might be located in the nucleus.Based on the bioinformatics analysis of StDHN-2 gene,it was found that the genome had a total length of 931 bp,including a coding region sequence of 474 bp,which encoded 157 amino acids;The molecular weight of StDHN-2 protein was 16659.06 D,and the theoretical isoelectric point was 7.23.The analysis showed that StDHN-2 protein was a hydrophilic protein without transmembrane domain.The secondary structure of the StDHN-2 protein was mainly the random coil,and there was noβ-turn structure.By constructing a phylogenetic tree,it was found that StDHN-2 protein had the closest evolutionary relationship with Solanum verrucosum;The cis-acting element analysis showed that the promoter region of StDHN-2 gene contained ABRE,DRE,P-box,LTR and other elements related to abiotic stress response;The expression patterns of StDHN-2 gene were analyzed under PEG,NaCl,and abscisic acid(ABA)stress treatments by using qRT-PCR analysis,it was found that the gene was up-regulated under all 3 kinds of stress treatments.The results of the study preliminarily indicated that the StDHN-2 gene could be a good candidate gene for drought tolerance in potato and involved in the biological process of drought tolerance in plants.This study provides basic information for analyzing the drought response mechanism of potato StDHN-2 gene.

关 键 词：马铃薯 StDHN-2基因 表达分析 生物信息学分析 亚细胞定位 

分 类 号：S532[农业科学—作物学]