Preliminary Analysis Of GmWRI1 Gene Expression And Function In Soybean

As one of the most important oil crops in people's lives,soybeans affect agriculture at all times.Vegetable oils are getting more and more attention.Increasing the oil content of soybean seeds is also becoming more and more important.In this study,the full-length cDNA sequence of GmWRI1 was cloned from soybean kernels,and the constructed recombinant plasmid pBASTA-GmWRI1 was transformed into mutant Arabidopsis thaliana mutants with deletion wri gene by floral Dip method to obtain various transgenic Arabidopsis thaliana.Complement the plant and analyze the seed oil content of wild-type Arabidopsis thaliana,Arabidopsis thaliana overexpressing,Arabidopsis thaliana mutant and replenished Arabidopsis thaliana to verify the function of GmWRI1 gene;Infected by Agrobacterium inoculation Transgenic tobacco was used to study the cell localization function of GmWRI1;a knockout vector of GmWRI1 gene was constructed using CRISPR/cas9 system,and two vectors of CRISPR-GmWRI1 and pBASTA-GmWRI1 were transformed into soybean using soybean cotyledonary node transformation to obtain overexpressed soybean.And knockout soybeans.The findings are as follows:1.Plant expression vector pBASTA-GmWRI1 was transformed into Arabidopsis thaliana mutants lacking the wri gene by floral Dip method to obtain Arabidopsis thaliana plants.Soxhlet extraction and real-time fluorescence quantitative PCR were used to analyze and quantitatively analyze seed oil content in wild-type Arabidopsis thaliana,Arabidopsis thaliana,Arabidopsis thaliana mutant and replenished Arabidopsis.The results showed that the oil content in seeds of over-expressed Arabidopsis thaliana increased by 2.52%over the Arabidopsis thaliana.The Arabidopsis thaliana mutant had a 4.06%decrease in seed oil content compared to the wild Arabidopsis thaliana.Compared with the wild-type Arabidopsis seeds,the oil content of replenished Arabidopsis decreased by 1.74%.It was initially confirmed that GmWRI1 gene promoted the oil content of Arabidopsis seeds,and had a certain role in complementing the oil content of low oil seeds caused by gene deletion.Through comparison of the results of oil content and gene expression,Arabidopsis thaliana was found.The oil content in seeds and the expression level of GmWRI1 gene may be positively correlated.2.The transient expression vector pCAMBIA1302-GmWRI1 was successfully constructed,and the plasmid was transferred into Agrobacterium tumefaciens EHA105.The subcellular localization function of GmWRI1 was observed by injecting tobacco leaves by Agrobacterium injection.The results showed that GmWRI1 was localized in the nucleus and cell membrane.3.Recombinant DNA technology was used to construct plant expression vector CRISPR-GmWRI1 and transferred into Agrobacterium EHA105 with infective plant function.Overexpression vector and gene knockout vector were transformed into soybean by soybean cotyledon node transformation method,and get 5 T₁ generation overexpressed transgenic soybean and 6 T₀ generation knockout transgenic soybeans.