首页|芝麻热胁迫响应基因SiHsfB-2b的克隆及表达特性分析

芝麻热胁迫响应基因SiHsfB-2b的克隆及表达特性分析

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为探究B类热激转录因子(HSFs)在芝麻耐热性中的分子功能，基于前期转录组分析鉴定到的编码B类HSF家族蛋白HsfB2b的基因序列，以郑太芝3 号耐热白芝麻品种为材料，采用同源克隆方法克隆了芝麻SiHsfB-2b基因，对其进行生物信息学分析，同时分析其在芝麻不同器官和热胁迫下的表达特性，并对其进行亚细胞定位。结果表明:SiHsfB-2b基因CDS序列全长为 936 bp，编码 311 个氨基酸;系统进化树分析表明SiHsfB-2b与松蒿、独脚金和锈鳞木樨榄的同源蛋白关系较近，含保守DNA结合结构域;亚细胞定位于细胞核内;SiHsfB-2b基因启动子区域含有 13 类顺式作用元件，其中与胁迫响应和激素响应相关的各有 3 类。RT-qPCR分析结果显示，热胁迫可诱导SiHsfB-2b在芝麻根、茎和叶中上调表达，叶中相对表达量最高;在持续高温胁迫下，SiHsfB-2b在芝麻茎和叶中的表达量呈上升趋势，且在胁迫后期的叶中表现出显著上调，而根中的表达量呈现出先上升后下降趋势。综上推测，SiHsfB-2b基因可能通过介导胁迫和激素信号等通路积极响应热胁迫，这可为后期深入研究芝麻耐热性的分子机制奠定理论基础。
Cloning and Expression Characteristics of Sesame SiHsfB-2b Gene Response to Heat Stress
In order to investigate the molecular function of B class heat shock transcription factors(HS-Fs)in heat tolerance of sesame,using the heat-tolerant cultivar Zhengtaizhi 3 as material,the SiHsfB-2b gene was cloned by homologous cloning method based on the gene sequence encoding the B class HSF protein Hs-fB2b identified by previous transcriptome analysis.Meanwhile,its bioinformatics analysis was conducted,its expression characteristics in different organs of sesame and under heat stress were analyzed,and also its sub-cellular localization.The results showed that the full length of SiHsfB-2b CDS was 936 bp,encoding 311 amino acids.The phylogenetic tree analysis showed that SiHsfB-2b was closely related to the homologous proteins of Phtheirospermum japonicum(Thunb.)Kanitz,Striga asiatica L.and Olea europaea subsp.cuspidata,and all they contained the conserved DNA-binding domains.Subcellular localization analysis revealed that the protein was located in nucleus.Furthermore,the promoter region of SiHsfB-2b gene contained 13 types of cis-acting elements,among which,3 types related to stress response and 3 types related to hormone response.The real-time quantitative PCR analysis showed that heat stress could induce the up-regulated expression of SiHsfB-2b in sesame roots,stems and leaves,and the highest relative expression level was observed in leaves.In re-sponse to continuous heat stress,the expression level of SiHsfB-2b in sesame stems and leaves exhibited an in-creasing trend,and the significant up-regulated expression was observed in leaves at the late stage of stress;but in roots,its expression exhibited increasing first and then decreasing.In conclusion,it was speculated that the SiHsfB-2b gene might positively respond to heat stress by mediating stress and hormone signaling pathways.These results could provide a theoretical basis for subsequent in-depth research on molecular mechanism of sesame resistant to heat.

SesameHeat toleranceSiHsfB-2b geneGene cloningQuantitative expression analysisSubcellular localization

田媛、苏小雨、王东勇、李丰、张鹏钰、付锦州、戎亚思、王龙、高桐梅、吴寅

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河南省农业科学院芝麻研究中心,河南 郑州 450002

神农种业实验室,河南 郑州 450002

河南省农业科学院中药材研究所,河南 郑州 450002

芝麻 耐热性 SiHsfB-2b基因 基因克隆 定量表达分析 亚细胞定位

国家自然科学基金青年基金项目国家特色油料产业技术体系项目2022年度河南省重点研发与推广专项河南省农业科学院优秀青年科技基金项目河南省农业科学院自主创新项目中国博士后科学基金资助项目河南省科技攻关项目

32301952CARS-14-1-142221021100842022YQ012023ZC0822020M672239222102110084

2024

10.14083/j.issn.1001-4942.2024.07.002

山东农业科学
山东省农业科学院,山东农学会,山东农业大学

山东农业科学

CSTPCD北大核心
影响因子：0.578
ISSN：1001-4942
年,卷(期)：2024.56(7)