Cloning of the Soybean sHSP26 Gene and Analysis of Its Drought Resistance

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.

Cloning and Drought Resistance Analysis of Soybean GmHsps_p23-like Gene

Soybean(Glycine max(L.)Merr.)is an important cultivated crop,which requires much water during its growth,and drought seriously affects soybean yields.Studies have shown that the expression of small heat shock proteins can enhance drought resistance,cold resistance and salt resistance of plants.In this experiment,soybean GmHsps_p23-like gene was successfully cloned by RT-PCR,the protein encoded by the GmHsps_p23-like gene was subjected to bioinformatics analysis,and the pCAMBIA3301-GmHsps_p23-like overexpression vector and pCBSG015-GmHsps_p23-like gene editing vector were constructed.Agrobacterium-mediated method was used to transform soybeans to obtain positive plants.RT-PCR detection,rehydration experiment and drought resistance physiological and biochemical index detection were performed on the T2 generation positive transgenic soybean plants identified by PCR and Southern hybridization.The results showed that the overexpression vector plant GmHsps_p23-like gene expression increased.After rehydration,the transgenic overexpression plants returned to normal growth,and the damage to the plants was low.After drought 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.The drought resistance of the overexpressed soybeans under drought stress was higher than that of the control group,and had a stronger drought resistance.It showed that the expression of soybean GmHsps_p23-like gene can improve the drought resistance of soybean.The cloning and functional verification of soybean GmHsps_p23-like gene had not been reported yet.This is the first time that PCR technology has been used to amplify the soybean GmHsps_p23-like gene and construct an expression vector for this gene.This research has laid the foundation for transgenic technology to improve plant drought resistance and cultivate new drought-resistant transgenic soybean varieties.