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.
