Preliminary Study On The Transfer Of Wheat High Molecular Weight Gluten Subunits 1Dx5+1Dy10 Into Maize

Corn(Zea mays L.)is the largest grain crop in China,and an important feed and industrial raw material,among which feed consumption accounts for 70%.However,as a ration and feed,there are two main problems.Firstly,the protein in corn seeds is mainly soluble,which cannot be absorbed and used by animals.In producing feed,soybean protein is added to increase the protein content,which leads to a high dependency on soybean.Second,corn grains do not contain glutenin,and their flour is hard to make dough when used for food processing,limiting the use of corn in the food processing industry.Wheat grains contain not only high protein content but also high molecular weight glutenin,which makes wheat flour mix.High molecular weight gluten is a special protein of wheat,which is the main component of wheat gluten protein.It determines the elasticity of dough and plays an important role in the rheological properties and baking quality of gluten.In this study,high molecular weight glutenin(HMW-GS)genes 1Dx5 and 1Dy10 were cloned from Zhengmai 366,and overexpression vectors were constructed.The 1Dx5 and1Dy10 genes were simultaneously transferred into B73 maize embryos by Agrobacterium-mediated transgenic method.It was cultured into plants by the technique of maize embryo tissue culture.The obtained plants were identified positively at the DNA level.The main research results are as follows:This study intends to overexpress wheat 1Dx5 and 1Dy10 genes in corn grains by transgenic method,and the main research results are as follows:1.The 1Dx5 and 1Dy10 genes of high molecular weight gluten were cloned from Zhengmai 366.The 1Dx5 gene was 2547 bp and the 1Dy10 gene was 1947 bp;2.The specific endosperm promoter Leg1 A of maize was cloned,and the original promoter on the vector was replaced with it.Plant overexpression vectors p HB-Leg1A-x5 and p HB-Leg1A-y10 were constructed respectively;3.The variety of maize B73 as transgenic receptor materials,young embryos of1.5-2mm 13 days after pollination were selected,and wheat high molecular weight gluten genes 1Dx5 and 1Dy10 were simultaneously transferred into young embryos of maize by Agrobacterium-mediated transgenic technology.Fifteen 1Dx5 T0 generation transgenic positive plants were obtained from maize embryo culture,and the transformation rate was 1%(15/1500=1%).The transformation rate of 13 1Dy10 T0 transgenic positive plants was 0.87%(13/1500=0.87%).The transformation rate of 13 T0 positive transgenic plants co-existing1Dx5 and 1Dy10 genes was 0.87%(15/1500=0.87%).4.Four T1 transgenic ears of 1Dx5 and 1Dy10 were harvested after field culture.By transferring the high molecular weight gluten genes 1Dx5 and 1Dy10 of wheat into corn,we hope to increase the protein content in corn grains,reduce the amount of protein added to the feed,and improve the corn quality,therefore,expanding its edible range.