Cloning and Expression Pattern Analysis of GmSBPC Associated with Soybean Cyst Nematodes
This study used RNA-seq data from physiological race 3 of soybean cyst nematode,Heinong 37(susceptible)and Dongnong L10(resistant),to screen for the differentially expressed gene GmSBPC.Bioinformatics analysis was conducted on the spatial structure,protein physicochemical properties,hydrophilicity,and hydrophobicity of the protein encoded by this gene.Cloning GmSBPC using cDNA from the root system of disease resistant Dongnong L10.Transform the recombinant vector containing pCAMBIA1302-GmSBPC into Escherichia coli DH5a,Agrobacterium GV3101(psoup-p19)for subcellular localization analysis.Recombinant pCAMBIA3300-GmSBPC was transferred to Agrobacterium tumefaciens K599 for soybean hairy root infection.Planting Dongnong L10(resistant)and Dongnong 50(susceptible)in nematode soil,soybean cyst nematode stress treatment was performed on roots,stems,and leaves at0,3,6,9,12,and 15 days for qRT-PCR analysis of gene expression patterns.The results indicate that the GmSBPC protein encodes 146 amino acids and is an insoluble protein,a spiral zone accounts for 28.08％,extended structure accounts for 15.75％,and irregular curl accounts for 56.16％.The subcellular localization results indicate that the gene is located in the nucleus.Overexpression of hairy roots reduces the number of nematodes per unit area compared to wild-type soybeans.The gene expression pattern of the roots of Dongnong 50 and Dongnong L10 under soybean cyst nematode stress was initially increased and then decreased,with the overall expression level being higher in Dongnong L10 roots than in Dongnong 50 roots.The highest expression level was observed in Dongnong L10 roots after 12 days,which is the second instar larval stage of soybean infection by nematodes.Therefore,it is determined that this gene has a response to nematode stress,and it is speculated that this gene is involved in the stress response of soybean cyst nematodes.These research results contribute to further exploring the physiological function of SBPC gene in soybean resistance to cyst nematodes.
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