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Storage Protein Analysis of High Grain-protein-content Wheat Varieties

Author: YeCongYun
Tutor: QinGuangYong
School: Zhengzhou University
Course: Biophysical
Keywords: Wheat High molecular weight glutenin subunits Gliadin Storage protein SDS - sedimentation value High protein
CLC: S512.1
Type: Master's thesis
Year: 2007
Downloads: 70
Quote: 1
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Abstract


Our laboratory using ion beam radiation mutagenesis mediated transgenic wheat genetic improvement, has received a number of protein content compared with the control (CK) three to five percentage points higher, and the highest protein content of more than 20 % of fine materials. These mutations material has been stably inherited to the fifth generation is still unclear, but the reasons for the increase of the protein content and quality traits. Storage protein is a major component of the wheat protein, including about 40% of the gluten and gliadin, from each of wheat protein. Gluten can be divided into high molecular weight glutenin subunits (HMW-GS) and low molecular weight glutenin subunits (LMW-GS) according to their mobility in SDS-PAGE of HMW-GS is a key factor in determining wheat quality . Therefore, this study were to SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and acid polyacrylamide gel electrophoresis (A-PAGE) analysis of high protein wheat Variant Strains of HMW-GS and gliadin variation . First, from the gel concentration, catalyst, plus sample volume, voltage, and sample buffer five aspects to optimize the experimental conditions of the SDS-PAGE gel electrophoresis, to lay the foundation for the analysis of the high protein wheat HMW-GS variation. Determined with the SDS-PAGE gel electrophoresis after optimization Yumai 52 HMW-GS subunit composition of 1,7 9 and 512. The analysis showed that the variation on the high-protein wheat strains HMW-GS: 74 different monoclonal HMW-GS and CK did not increase or missing bands, but significant increase in subunit expression. In addition, the use of A-PAGE gel electrophoresis analysis of the variability of the gliadin, first, but also from the gel concentration, the amount of the added sample, the voltage, the sample buffer and the catalyst in five aspects optimize experimental conditions in order to achieve a satisfactory result of the electrophoresis. Variants of high-protein wheat gliadin A-PAGE electrophoresis conditions optimized analysis variation in the study of 74 high-protein wheat variation monoclonal bands α, β, γ-gliadin area frequency and increase or missing, only the expression levels of different, but in ω-gliadin 40 per plant compared with CK increase two bands, the overall difference frequency is 54.05%, and the difference bands appear The protein content was negatively correlated. Protein content and SDS-sedimentation value measure is an important indicator of the quality of wheat, this study measured the SDS-sedimentation value of high-protein variant strains, and analysis of the relationship between SDS-sedimentation value and the average protein content of its HMW-GS subunits on SDS-sedimentation value contribution. The results showed that the SDS-sedimentation value of high protein wheat mutant strains was significantly higher compared with the control, and its lines of average protein content was positively correlated.

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CLC: > Agricultural Sciences > Crop > Cereal crops > Wheat > Wheat
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