为了探讨热激蛋白在低温锻炼诱导的耐热性形成过程中的保护功能,进一步分析葡萄幼苗对温度逆境产生交叉适应性的细胞学机制,试验以葡萄(Vitis vinifera L. cv. Jingxiu)幼苗为试材,采用胶体金免疫电镜定位技术,定位观察了小分子热激蛋白...为了探讨热激蛋白在低温锻炼诱导的耐热性形成过程中的保护功能,进一步分析葡萄幼苗对温度逆境产生交叉适应性的细胞学机制,试验以葡萄(Vitis vinifera L. cv. Jingxiu)幼苗为试材,采用胶体金免疫电镜定位技术,定位观察了小分子热激蛋白17.6(sHSP17.6)在葡萄叶片中的亚细胞分布情况,结果表明,低温锻炼预处理可增强其在高温胁迫条件下的表达水平,细胞核和叶绿体中代表sHSP17.6的免疫金颗粒密度比相应的对照细胞明显增多,尤其是胁迫处理3 h时。这一结果为sHSP17.6参与低温锻炼诱导的耐热性提供了更直观的细胞学证据。展开更多
棉铃虫Helicoverpa armigera是一种重要的农业害虫,本文旨在研究小分子热激蛋白在其生长发育过程、抵御高温及Cry1Ac杀虫蛋白中的功能。利用PCR结合RACE技术克隆了棉铃虫sHSP22.0(small heat shock protein 22.0)基因,通过生物信息学软...棉铃虫Helicoverpa armigera是一种重要的农业害虫,本文旨在研究小分子热激蛋白在其生长发育过程、抵御高温及Cry1Ac杀虫蛋白中的功能。利用PCR结合RACE技术克隆了棉铃虫sHSP22.0(small heat shock protein 22.0)基因,通过生物信息学软件分析了棉铃虫sHSP22.0基因序列,利用实时荧光定量qRT-PCR分析了该基因在棉铃虫不同生长发育阶段和组织中的表达模式;并分析了该基因受高温及Cry1Ac全长蛋白的诱导效应。获得了棉铃虫sHSP22.0(GenBank登录号:XP_021196802.1)761 bp cDNA片段,其开放阅读框576 bp,编码191个氨基酸,具有小分子热激蛋白典型的α-晶体结构域(α-crystallin domain,ACD)。sHSP22.0在棉铃虫4龄和5龄期特异性表达,尤其在5龄幼虫的表皮、中肠和后肠内特异性表达。该小分子热激蛋白受温度和Cry1Ac全长蛋白的诱导后显著高表达。sHSP22.0不仅在暴食期及肠道内特异性表达,而且响应Cry1Ac全长蛋白的诱导,表明它在棉铃虫消化吸收及抵御外源物的活动中可能起到重要的作用。展开更多
Exploring the molecular mechanism of soybean response to drought stress,providing a basis for genetic improvement and breeding of heat-resistant varieties,relying on the transcriptome sequencing data of unpollinated o...Exploring the molecular mechanism of soybean response to drought stress,providing a basis for genetic improvement and breeding of heat-resistant varieties,relying on the transcriptome sequencing data of unpollinated ovary at the seven-leaf stage of soybean Jinong 18(JN18)and Jinong 18 mutant(JB18)soybeans,using reverse transcription,one gene in the sHSP family was cloned using PCR(RT-PCR)and it was named sHSP26.In this experiment,the soybean sHSP26 gene was successfully cloned by RT-PCR,the protein encoded by the sHSP26 gene was analyzed by bioinformatics,and the sHSP26 gene overexpression vector and CRISPR/Cas9 gene-editing vector were constructed.The positive plants were derived from Agrobacterium-mediated transformation of soybean cotyledon nodes,and T2 plants were identified through conventional PCR,QT-PCR,and Southern blot hybridization.Finally,through the determination of drought-related physiological and biochemical indicators and the analysis of agronomic traits,further research on gene function was conducted.The results indicated that the overexpression vector plant GmsHSP26 gene expression increased.After stress,the SOD and POD activities,and the PRO content of the transgenic overexpression plants increased,while the MDA content decreased.The reverse was true for soybean plants with genetically modified editing vectors.A survey of agronomic traits indicated that the fourpod ratio and yield per plant of the transgenic overexpression plants were higher than those of the control and transgenic editing vector soybean plants.It indicates that the expression of the sHSP26 gene can enhance drought resistance and soybean yield.The soybean sHSP26 gene cloning and its functional verification have not yet been reported.This is the first report where PCR amplification of soybean sHSP26 genes and gene expression vector were applied.It lays the foundation for creating new drought-resistant transgenic soybean lines through genetic engineering technology and is essential for improving soybean yield and quality.展开更多
基金This project was supported by National Hi-Tech Research and Development Program of China ("863"Project , No2001AA624090 and No2003AA624090)the National Natural ScienceFoundation of China (No30171103)CPUpeople of talent recommend-ingfor 211 item(No211032)
文摘目的:从鲨鱼肝脏中分离纯化得到了一种鲨鱼肝刺激多肽(sHSP)。方法:将天然健康的条纹斑竹鲨鱼肝脏经过粗提,超滤,柱层析得到sHSP。通过采用MTT法,利用人肝癌SMMC-7721细胞和STZ损伤的小鼠胰岛β细胞瘤细胞来检测其生物活性。结果:通过MALDI-TOF-MS检测,sHSP的分子量为4 899 .715 Da ,其紫外特征吸收波长为273.1 nm。通过MTT法检测sHSP的刺激肝细胞再生的生物活性,结果显示,sHSP剂量为100μg.mL-1时,刺激指数为3.85 ,并且具有热稳定性和耐酸碱性。对STZ损伤的NIT-1β细胞,sHSP具有保护细胞完整性和修复细胞形态的作用。结论:我们从鲨鱼肝脏中分离纯化得到了一种能够显著刺激人肝癌细胞SMMC-7721再生,并且可以部分对抗STZ导致的NIT-1β细胞毒性损伤的活性肽。
文摘棉铃虫Helicoverpa armigera是一种重要的农业害虫,本文旨在研究小分子热激蛋白在其生长发育过程、抵御高温及Cry1Ac杀虫蛋白中的功能。利用PCR结合RACE技术克隆了棉铃虫sHSP22.0(small heat shock protein 22.0)基因,通过生物信息学软件分析了棉铃虫sHSP22.0基因序列,利用实时荧光定量qRT-PCR分析了该基因在棉铃虫不同生长发育阶段和组织中的表达模式;并分析了该基因受高温及Cry1Ac全长蛋白的诱导效应。获得了棉铃虫sHSP22.0(GenBank登录号:XP_021196802.1)761 bp cDNA片段,其开放阅读框576 bp,编码191个氨基酸,具有小分子热激蛋白典型的α-晶体结构域(α-crystallin domain,ACD)。sHSP22.0在棉铃虫4龄和5龄期特异性表达,尤其在5龄幼虫的表皮、中肠和后肠内特异性表达。该小分子热激蛋白受温度和Cry1Ac全长蛋白的诱导后显著高表达。sHSP22.0不仅在暴食期及肠道内特异性表达,而且响应Cry1Ac全长蛋白的诱导,表明它在棉铃虫消化吸收及抵御外源物的活动中可能起到重要的作用。
基金the Jilin Province Education Department Science and Technology Research Project[JJKH20210350KJ]the Jilin Province Science and Technology Guidance Program Project[20200402023NC]+1 种基金the Jilin Provincial Natural Science Foundation Project[20200201027JC]the Innovation and Entrepreneurship Training Program for College Students in Jilin Province[2021]。
文摘Exploring the molecular mechanism of soybean response to drought stress,providing a basis for genetic improvement and breeding of heat-resistant varieties,relying on the transcriptome sequencing data of unpollinated ovary at the seven-leaf stage of soybean Jinong 18(JN18)and Jinong 18 mutant(JB18)soybeans,using reverse transcription,one gene in the sHSP family was cloned using PCR(RT-PCR)and it was named sHSP26.In this experiment,the soybean sHSP26 gene was successfully cloned by RT-PCR,the protein encoded by the sHSP26 gene was analyzed by bioinformatics,and the sHSP26 gene overexpression vector and CRISPR/Cas9 gene-editing vector were constructed.The positive plants were derived from Agrobacterium-mediated transformation of soybean cotyledon nodes,and T2 plants were identified through conventional PCR,QT-PCR,and Southern blot hybridization.Finally,through the determination of drought-related physiological and biochemical indicators and the analysis of agronomic traits,further research on gene function was conducted.The results indicated that the overexpression vector plant GmsHSP26 gene expression increased.After stress,the SOD and POD activities,and the PRO content of the transgenic overexpression plants increased,while the MDA content decreased.The reverse was true for soybean plants with genetically modified editing vectors.A survey of agronomic traits indicated that the fourpod ratio and yield per plant of the transgenic overexpression plants were higher than those of the control and transgenic editing vector soybean plants.It indicates that the expression of the sHSP26 gene can enhance drought resistance and soybean yield.The soybean sHSP26 gene cloning and its functional verification have not yet been reported.This is the first report where PCR amplification of soybean sHSP26 genes and gene expression vector were applied.It lays the foundation for creating new drought-resistant transgenic soybean lines through genetic engineering technology and is essential for improving soybean yield and quality.
