Study On Molecular Detection Technology And Its Application Of HMW-GS In Wheat

Wheat is one of the world’s three major food crops,and it is also the most widely distributed cereal with the largest sown area and yield.With the rapid increase in the demand for functional foods,the focus of wheat breeding in China gradually shifts from yield breeding to quality breeding.HMW-GS is a major component of glutenin and plays a decisive role in the processing quality of wheat.In recent years,in the breeding of wheat quality in Shanxi Province and even in the whole country,the variation types of the various high-quality HMW-GS and their respective proportions have increased,but the overall improvement of the quality of wheat has not changed substantially.The main reason is the lack of high-quality breeding materials.This study improved the traditional method of wheat genomic DNA extraction,improved the molecular detection technology of high-quality subunits of wheat high-molecular-weight glutenin,and analyzed,detected and compared the related wheat material HMW-GS.The results provided the basis for wheat quality breeding.The results of this study are as follows:1.In this study,we sought to find out a new high-throughput DNA extraction method to detect HMW-GS in wheat.Using wheat seeds as DNA extraction material,based on traditional CTAB method and magnetic bead method,the study created a new method of high-throughput and low-cost DNA extraction method of wheat seeds.The yield and quality of DNA were verified by ultraviolet spectrop Hotometer,agarose gel electrop Horesis and AS-PCR.The results showed that the quality and yield of the genomic DNA extracted by the new method were stable and need the experimental requirements of wheat HMW-GS molecular detection.2.The HMW-GS components of 40 wheat cultivars were detected by SDS-PAGE method and AS-PCR method respectively,and the results were compared and analyzed.Based on this,a rapid and accurate molecular detection method of high-quality high molecular weight glutenin in wheat was established.3.113 international wheat materials were analyzed by using the developed AS-PCR technique and SDS-PAGE gel electrop Horesis:(1)A total of 13 subunit types were detected.There were three kinds of subnits in the GluA1 site and the dominant subunit is 2*,accounting for 49.56%;there were 7 species in the Glu-B1 site and the dominant subunits were 7+8 and 7+9,accounting for The proportions were 29.20%.There were three kinds in the Glu-D1 site and the dominant subunit was 5+10,accounting for 55.75%.(2)Two high quality subunits,1 and 2 *,were detected at A locus,accounting for 84.96%.Four high quality subunits of 7+8,17+18,13+16 and 14+15 were detected in B site,accounting for 59.29%;5+10 subunits were detected in D site,accounting for 55.75%.(3)A total of 32 subunit combinations were detected,of which 85 had a score of 8 points or more,accounting for 75.22%.