Research On Soybean GY1Gene RNAi Expression Vectors Of Genetic

Soybean oil is the main cooking oil in our country. Soybean is Soybean oil production raw materials, and soybean in our country has broad planting area. But the80%of the domestic sources soybean rely on imports. The import of soybean yield efficiency higher than domestic soybean yield efficiency, and the price is low, this is the main cause of our country soybean rely on imports. Therefore, how to improve the competitiveness of our local soybean, is now we are faced with the primary issue.At present, there are a lot of research on improve soybean fat content. Some researches proof that soy protein and fat was negatively related, if the soybean protein content is reduced, the fat content will be increased. There are some researches have confirmed this phenomenon. But most of the researches’direction was how to restrain the enzyme activity of the way on soybean fat metabolism, few studies’ direction have reported from inhibition soybean protein expression to improve soybean fat content.This experiment through RNA interference, construct the soybean11S globulin GY1gene RNAi plants vector. And transform the RNAi plant vector into soy-cotyledonary node by genetic transformation system mediated with agrobacterium and pollen-tube pathway, inhibit the expression of soybean11S globulin GY1gene, make soybean storage protein content reduced, and increase the soybean fat content. Provides a new research direction of increase the soybean fat content.The main research results are as follows:To construct the RNAi plants vector p3301-GY1with the selection marker of Bar gene. Cut the function sequence(seed specific promoter+Gy1gene sense segment+functional interval sequence GFP+Gyl gene antisense segment) in plant expression vector p7aP-Gy1with the enzyme of EcoR I and BstE II, insert the function sequence into pcAMBIA3301which have the selection marker of Bar gene. After identification of PCR and enzyme cut, the results was corresponds with the expected. Successfully constructed plants express p3301-GY1vector.Test the soybean tolerance of Glufosinate. Make Sure the five soybean varieties selection pressure of Glufosinate:JiNong28critical concentration is3.5mg-L-1; Jilin30and JiNong27is3.0mg-L-1; DE2259is2.5mg-L-1; JiNong17is2.0mg-L-1.By genetic transformation system mediated with agrobacterium, Get two plants into JiNong28soybean To generation,12plants into T1generation,16paints into T2generation.By genetic transformation system mediated with pollen-tube pathway,got312grains of soybean seed, including "JiNong27"103grain,"JiXinDou1"111grain,"Tong nong13"98grains. Through the PCR and Southern blot test, the result indicated that the RNAi plant expression vector-p3301-Gyl already successful transformed into the soybean plant genomic DNA, and the RT-PCR detection analysis shows that the transcription of RNA interference levels have obvious interference effect on the soybean11S globulin Gy1gene in the level of mRNA expression. SDS-PAGE results show that the11S globulin expression levels are low. Tast the content of the transformation soybean protein and fat, the results show that the transformation plant protein content is36.07%on average, and low1.43%than the control group plants which the protein content is37.50%, the biggest difference is2%from36.07%to37.50%; And the transformation plant fat content was increased, and the fat average content is21.28%, higher than the control group which fat content was20.52%by0.76%, the biggest increase by1.45%from21.97%to20.52%.