铁皮石斛DoTCP2基因克隆及非生物胁迫下的表达分析  

Cloning and expression analysis of DoTCP2 gene in Dendrobium officinale under abiotic stress

作　　者：刘羽佳 卓来凤 马立钦 黄秋妮 张勇 张颖智 李翔 刘博婷[1,2] LIU Yujia;ZHUO Laifeng;MA Liqin;HUANG Qiuni;ZHANG Yong;ZHANG Yingzhi;LI Xiang;LIU Boting(Guangdong Provincial Key Laboratory of Utilization and Conservation of Food and Medicinal Resources in Northerrn Region/Shaoguan University,Shaoguan 512005,China;College of Biology and Agriculture,Shaoguan University,Shaoguan 512005,China;College of Food,Shaoguan University,Shaoguan 512005,China)

机构地区：[1]广东省粵北食药资源利用与保护重点实验室/韶关学院,广东韶关512005 [2]韶关学院生物与农业学院,广东韶关512005 [3]韶关学院食品学院,广东韶关512005

出　　处：《江苏农业学报》2025年第3期588-598,共11页Jiangsu Journal of Agricultural Sciences

基　　金：广东省基础与应用基础研究基金面上项目(2024A1515013167);广东省普通高校重点领域专项(2022ZDZX4044、2024ZDZX2023);韶关学院科技计划项目(240823118037510);韶关学院科研重点项目(SZ2023KJ19)。

摘　　要：TCP转录因子是植物特有的一类调控生长发育和逆境胁迫反应的重要调控因子。为探究铁皮石斛(Dendrobium officinale)TCP2基因(DoTCP2)的表达特性和抗逆响应能力,通过同源克隆技术从广东丹霞铁皮石斛品种的叶组织中成功分离出DoTCP2基因,并对该基因序列进行生物信息学分析及表达模式探究。结果显示,DoTCP2(GenBank登录号:PP534463)cDNA全长1 122 bp,位于第7号染色体,含有4个外显子和3个内含子,编码长度为373个氨基酸的多肽链,与参考序列(序列号:LOC110114709)进行比对分析,发现存在4个碱基差异,其中3个为同义替换。DoTCP2蛋白理论相对分子量为40 600,理论等电点为8.94,为亲水性稳定蛋白质,具有TCP保守结构域,定位于细胞核,无信号肽和跨膜结构域,含有46个磷酸化位点。DoTCP2与鼓槌石斛(Dendrobium chrysotoxum)TCP蛋白序列同源性高达97.87%,且遗传距离最近。DoTCP2基因具有组织表达特异性,在根组织中的相对表达量极显著高于其他组织,可能在根发育和营养物质的吸收以及运输等过程中发挥作用。DoTCP2基因在低温胁迫处理6 h时的相对表达量极显著上调(P<0.01),暗示DoTCP2可能在低温胁迫响应中发挥重要作用;在模拟干旱和外源脱落酸(ABA)处理下,DoTCP2基因的相对表达量呈现下降趋势,且在处理24 h后相对表达量最低,与处理前差异达到极显著水平(P<0.01),推测该基因可能通过ABA信号通路在调控铁皮石斛应答干旱胁迫中发挥负调控作用。本研究结果可为深入探究DoTCP2基因的抗逆功能及其在植物应对非生物胁迫响应中的潜在作用机制提供前期研究基础。TCP transcription factors are a plant-specific group of regulators that play a vital role in governing growth,development,and responses to environmental stress.To investigate the expression characteristics and stress response capabilities of the TCP2 gene(DoTCP2)from Dendrobium officinale,the gene was successfully isolated from leaf tissues of the Guangdong Danxia variety of Dendrobium officinale using homology cloning.Subsequent bioinformatics analysis and expression pattern research were conducted.The results showed that the cDNA of DoTCP2(GenBank accession number was PP534463)was 1122 bp in full length,located on chromosome seven,consisted of four exons and three introns,and encoded a polypeptide chain of 373 amino acids.Compared to the reference sequence(accession number:LOC110114709),there were four nucleotide differences,of which three were synonymous substitutions.The theoretical relative molecular weight of the DoTCP2 protein was 40600,and its theoretical isoelectric point was 8.94.It was a hydrophilic and stable protein with a conserved TCP domain,localized in the nucleus,without signal peptides or transmembrane regions,and contained 46 phosphorylation sites.The DoTCP2 protein shared 97.87%sequence homology with the TCP protein of Dendrobium chrysotoxum and exhibited the closest genetic distance.DoTCP2 exhibited tissue-specific expression,with highly significantly higher relative expression levels in root tissues compared to other tissues,suggesting its potential role in root development,nutrient absorption,and transport processes.The relative expression of DoTCP2 was highly significantly upregulated after six hours of cold stress treatment(P<0.01),indicating that DoTCP2 might play an important role in the response to cold stress.Under simulated drought treatment and exogenous abscisic acid(ABA)treatment,the relative expression of DoTCP2 showed a downward trend.The lowest expression levels were observed after 24 hours of treatment,with highly significant differences compared to the untreated levels(P<0.0

关 键 词：铁皮石斛 TCP转录因子 DoTCP2基因 非生物胁迫 表达模式 

分 类 号：Q785[生物学—分子生物学]