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.
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。