Cloning And Identification Of FAD3 Gene In Soybean

Soybean(Glycine max)is one of the main sources of nutrients in our lives.Under the current social conditions that promote healthy eating,soybeans account for an increasing proportion of our lives.As China's soybean production is generally low,the acquisition of soybean raw materials is mainly dependent on imports.Since 2017,the United States imposed trade restrictions on China,which has led to serious restrictions on China's soybean imports.Therefore,it is imperative to improve China's soybean production and quality.The factors affecting soybean yield are nothing more than varieties and environment,and environmental factors are more variable.It is especially important to cultivate excellent soybean varieties.At present,the cultivation of varieties includes traditional breeding and genetic engineering breeding.In order to obtain corresponding high yield and high resistance to soybean varieties,Firstly,it is essential for the expression and analysis of soybean itself.At present,more and more studies have been conducted on the FAD7 gene family of FAD in plant resistance.Excessive expression of FAD7 gene in arabidopsis thaliana and FAD3 gene in brassica napus increased the tolerance of tobacco plants to drought and osmotic stress,and studies have proved that FAD8 gene plays an important role in plant response to low temperature.Therefore,this experiment transformed the Arabidopsis thaliana by cloning the soybean FAD3 gene,and changed the cold tolerance of Arabidopsis thaliana.To verify that soybean gene can improve the cold tolerance of plants,and provide basic research for the development and utilization of soybean excellent genes.The specific research results are as follows:1.According to the analysis of soybean expression profile determined by our laboratory,the gene related to unsaturated fatty acid GmFAD3 was selected and cloned.The length of the fragment was 1143 bp.The analysis indicated that GmFAD3 encodes 380 amino acids,and GmFAD3 protein has higher Conservative.2.The pCHF-3300-GmFAD3 overexpression vector and pEGC5941-GmFAD3 interference vector were successfully constructed,and the T3 generation Arabidopsis thaliana plants were obtained by simultaneously transforming the overexpressionvector and the interference vector into Arabidopsis by Agrobacterium-mediated method.3.The germination rate of T3 GmFAD3 overexpressing Arabidopsis thaliana plants was higher than that of wild type under 4 °C treatment and salt stress,and the germination rate of the interfering lines was lower than that of wild type.It showed a certain salt sensitivity,indicating that the expression of GmFAD3 gene can increase the tolerance of plants to cold and salt.The overexpressing lines showed higher survival rates in the antifreeze experiment,while the interfering lines showed lower survival rates than the wild type,indicating that the expression of the GmFAD3 gene can promote plants to have some resistance to low temperature stress.4.The determination of physiological and biochemical indexes of transgenic Arabidopsis thaliana,SOD,POD activity and PRO content showed that the Arabidopsis thaliana plants overexpressing GmFAD3 gene had high SOD,POD activity and PRO content.In wild-type and interfering plants,the SOD,POD activity and PRO content of the interfering plants were lower than those of the wild type.This indicates that the expression of the GmFAD3 gene can increase the cold tolerance of Arabidopsis plants.