摘要
旨在探讨油茶CoTOC1基因在非生物胁迫和外源激素调控下的功能及表达特性,以揭示其在油茶抗逆性中的潜在作用。试验克隆油茶核心品种‘湘林210’的CoTOC1基因,并通过生物信息学分析预测其蛋白结构和性质,利用亚细胞定位实验确定其细胞内分布,酵母双杂交实验验证其转录自激活活性,最终通过qRT-PCR分析其组织特异性表达及在胁迫处理下的表达特征。结果显示,成功克隆CoTOC1基因的CDS序列,长度为1686 bp,编码561个氨基酸。生物信息学分析表明,CoTOC1蛋白含有REC和CCT保守结构域,属于亲水性不稳定蛋白。亚细胞定位试验表明,CoTOC1主要定位于细胞核;酵母双杂交试验验证其具有转录自激活活性。组织特异性表达分析显示,CoTOC1在花中的表达量最高,其次为茎、根、叶和果实。在非生物胁迫和激素处理下,CoTOC1表达发生显著变化:盐胁迫处理12 h内达到高峰,高温胁迫36 h后表达量升至对照组的2.24倍,干旱胁迫下呈现先升后降的趋势;ABA和GA处理12 h后基因表达显著上调。
To investigate the function and expression characteristics of the CoTOC1 gene in Camellia oleifera under abiotic stress and exogenous hormone regulation,and to reveal its potential role in enhancing the stress resistance of C.oleifera,the CoTOC1 gene was cloned from the core cultivar‘Xianglin 210’,yielding a coding sequence(CDS)of 1686 bp encoding 561 amino acid protein.Bioinformatics analysis indicated that the CoTOC1 protein contains conserved REC and CCT domains,and was classified as hydrophilic and structurally unstable.Subcellular localization experiments demonstrated that CoTOC1 was predominantly localized in the nucleus,and yeast two-hybrid assays confirmed its self-activation ability.Tissue-specific expression analysis showed the highest transcript levels in flowers,followed by stems,roots,leaves,and fruits.Under abiotic stress and hormone treatments,CoTOC1 expression exhibited significant dynamic changes,with salt stress inducing peak expression within 12 hours,and high-temperature stress causing a 2.24-fold increase after 36 hours.Drought stress led to an initial increase followed by a decrease,while ABA and GA treatments significantly upregulated CoTOC1 expression within 12 hours.These findings provide insights into the role of CoTOC1 in abiotic stress response and hormone signal transduction,laying a foundation for further functional analysis in C.oleifera.
作者
万镇滔
王瑞
张英
寻成峰
张震
邓雪梅
何之龙
WAN Zhentao;WANG Rui;ZHANG Ying;XUN Chengfeng;ZHANG Zhen;DENG Xuemei;HE Zhilong(College of Forestry,Central South University of Forestry&Technology,Changsha 410004,China;National Engineering Research Center for Oil-tea Camellia,Hunan Academy of Forestry,Changsha 410004,China)
出处
《西北农业学报》
北大核心
2025年第12期2301-2310,共10页
Acta Agriculturae Boreali-occidentalis Sinica
基金
湖南省十大技术攻关项目(2024NK1020)
湖南省重点领域研发计划项目(2023NK2005)
湖南省自然科学基金面上项目(2022JJ30325)。
关键词
油茶
CoTOC1
非生物胁迫
激素调控
基因表达分析
Camellia oleifera
CoTOC1
Abiotic stress
Hormonal regulation
Gene expression analysis
