摘要
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。
In order to investigate the molecular function of B class heat shock transcription factors(HSFs)in heat tolerance of sesame,using the heattolerant 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 HsfB2b 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 subcellular 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 DNAbinding domains.Subcellular localization analysis revealed that the protein was located in nucleus.Furthermore,the promoter region of SiHsfB-2b gene contained 13 types of cisacting elements,among which,3 types related to stress response and 3 types related to hormone response.The realtime quantitative PCR analysis showed that heat stress could induce the upregulated expression of SiHsfB-2b in sesame roots,stems and leaves,and the highest relative expression level was observed in leaves.In response to continuous heat stress,the expression level of SiHsfB-2b in sesame stems and leaves exhibited an increasing trend,and the significant upregulated 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 indepth research on molecular mechanism of sesame resistant to heat.
作者
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
Tian Yuan;Su Xiaoyu;Wang Dongyong;Li Feng;Zhang Pengyu;Fu Jinzhou;Rong Yasi;Wang Long;Gao Tongmei;Wu Yin(Sesame Research Center,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China;Shennong Seed Industry Laboratory,Zhengzhou 450002,China;Institute of Chinese Herbal Medicines,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China)
出处
《山东农业科学》
北大核心
2024年第7期8-15,共8页
Shandong Agricultural Sciences
基金
国家自然科学基金青年基金项目(32301952)
国家特色油料产业技术体系项目(CARS-14-1-14)
2022年度河南省重点研发与推广专项(222102110084)
河南省农业科学院优秀青年科技基金项目(2022YQ01)
河南省农业科学院自主创新项目(2023ZC082)
中国博士后科学基金资助项目(2020M672239)
河南省科技攻关项目(222102110084)。
