Bioinformatics Analysis of Soybean NAC Transcription Factor and Cloning and Subcellular Localization of GmNAC-1
To explore the soybean cyst nematode-resistant NAC gene,soybean Dongnong L10(SCN3-resistant)and Heinong 37(SCN3-sensitive)were subjected to SCN3 adversity stress treatment,and the differentially expressed GmNAC genes were screened by RNA-seq,and the physicochemical properties,phosphorylation,hydrophilicity,and protein structure of the soybean NAC proteins were analyzed by bioinformatic methods,and the subcellular localization,the subcellular localization,gene covariance analysis and promoter region element analysis were also performed.GmNAC-1 was cloned,pCAMBIA3300 plasmid overexpressing GmNAC-1 was constructed,and transformed into tobacco by agrobacterium injection method for subcellular localization.qRT-PCR was performed on roots,stems and leaves of resistant and susceptible soybean after SCN3 stress.Results showed that NAC family proteins were electrically neutral in general.The proportion of irregularly curled secondary and tertiary structures was the highest.Subcellular localization suggested that NAC family proteins were present in the nucleus.The proteins were hydrophilic in general,and there was no chromosomal preference,serine played a major role in maintaining the function of the proteins,the proteins were soluble proteins in general.Collinearity analysis showed that there were some homology of NAC genes in different species.qRT-PCR results showed that the GmNAC-1 gene was up-regulated in both resistant and susceptible soybeans after SCN3 stress,but the up-regulation of GmNAC-1 was more obvious in resistant than in susceptible soybean roots.The results of subcellular localization indicated that GmNAC-1 was localized in the nucleus,which was consistent with the bioinformatics prediction.In this study,we screened out the differentially expressed gene GmNAC-1,which was involved in SCN adversity stress by RNA-seq,and the results provided a basis for further research on cyst nematode resistance genes in resistant soybean.
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