Correlation Analysis of New Soybean [Glycine max (L.) Merr] Gene Gm15G117700 with Oleic Acid

Abstract

The fatty acid dehydrogenase gene plays an important role in regulating the oleic acid content in soybean. Genome-wide association study screened out soybean oleic acid related gene Gm15G117700. A fragment size of 693bp was obtained by PCR amplification of the gene and, it was connected by seamless cloning technology to the pMD18T cloning vector. Based on the gene sequence cloned, bioinformatic analysis of gene protein was performed. The overexpression vector of Gm15G117700 and the CRISPR/Cas9 gene editing vector were constructed. The positive plants were obtained by Agrobacterium-mediated transformation of soybean cotyledon nodes and T₂ plants were identified by conventional PCR, QT-PCR and Southern blot hybridization. 10 copies of high and low oleic acid seeds were selected for QT-PCR to identify the expression content of Gm15G117700 gene in different soybeans, and finally near-infrared spectroscopy analyzer was used to identify the oleic acid quality of soybeans. T₂ RT-PCR identification showed that overexpression was reduced by 3.94%, and gene editing was increased by 3.49%. It is determined that the Gm15G117700 gene may belong to a regulatory gene, a minor gene that can promote the conversion to linoleic acid content in soybean oleic acid synthesis. The gene cloning and its functional verification was not reported yet. This is the first report by PCR amplification of soybean Gm15G117700 genes and gene expression vector. Improving the content of oleic acid in soybean lay a foundation for researchers. Therefore;this study clearly identified the function of soybean Gm15G117700 gene and its role played in oleic acid synthesis and metabolism.