Study On Identification And Chromosome Location Of Dasypyrum Villosum Storage Protein

Wheat wild relatives are an important gene bank for genetic improvement of common wheat.Dasypyrum villosum has strong resistance to biotic stress and abiotic stress,has high grain protein content,and its gliadin is similar to wheat gliadin in terms of classification,properties and protein primary structure characteristics.Providing excellent foreign genes for wheat quality improvement.In this study,the new Chinese Spring-Dasypyrum villosum germplasm was used as a material to establish a high-performance liquid chromatography separation system for gluten and gliadin subunits.Using proteomics sequencing to mine the excellent storage protein of Dasypyrum villosum,laying a foundation for the research on the intentional quality formation of Dasypyrum villosum.Below are key research findings:1?Optimal conditions for separation of high-molecular-weight glutenin subunits by HPLC waswere: 20?l injection volume,40? column temperature,0.5ml/min flow rate,and with The gradient of mobile phase B was increased from 5% to 50% within 30 minutes.Optimal conditions for separation of gliadin subunits by HPLC were: 20?l injection volume and,60? column temperature with 0.5ml/min flow rate,and the mobile phase B was increased from 5% to 39% in a gradient of 19 min.Add an isocratic gradient for 2 min,then 21 to 25 min of mobile phase B was raised to 44%,and then add an isocratic gradient of 2 min,27 to 35 min for mobile phase B was raised to 50%.Six subunits of a gliadin subunits were identified,and one of them was located on the 1V chromosome.2?Using grains of Dasypyrum villosum(TA10231)and common wheat Chinese spring(CS)as materials,i TRAQ proteomics analysis:1)A total of 883 differential proteins were identified,of which 357 were up-regulated proteins and 526 were down-regulated proteins.It was found that the differentially expressed proteins were mainly localized in chloroplast(32.05%),cytoplasm(25.82%),extracellular matrix(12.91%)and nucleus(12.34%).The differential proteins mainly involve the following biological pathways: Protein metabolism,hydrogen peroxide catabolism process,proteolysis regulation,monosaccharide decomposition,cell wall monosaccharide metabolism and polysaccharide decomposition process,etc.Annotated to 102 differentially expressed proteins using GO,COG,KEGG,involving 7 metabolic pathways,including phenylpropanol biosynthesis,starch and sucrose metabolism,degradation of other polysaccharides,glycolysis and glycogenogenesis,linolenic acid metabolism,phosphoinositide metabolism,and plant MAPK signaling pathways.2)Identified 11 differential proteins related to excellent quality formation,and 6 proteins that play a key role in the metabolic pathways that contribute to the formation of superior quality.