Ultra-high performance liquid chromatography method for identifying high molecular weight glutenin subunits in wheat and application thereof

Abstract

The invention discloses a method for the rapid identification of high molecular weight glutenin subunits in wheat by utilizing the ultra-high performance liquid chromatography technology. By adopting the method, the rapid and accurate identification of the high molecular weight glutenin subunits in wheat seed storage proteins can be realized; and mobile phase conditions for ultra-high performanceliquid chromatography, added ingredients, chromatographic condition parameters and a chromatographic column washing procedure are optimized. By adopting the method, the composition of the high molecular weight subunits in different wheat varieties can be fast identified, and the high molecular weight glutenin subunits which can not be separated and identified by the HPLC (high performance liquid chromatography) method can be accurately identified. By adopting the method, fast and accurate identification and screening can be performed on wheat quality and hybrid early generation high-quality storage protein subunits.

Description

technical field [0001] The invention relates to a method for identifying wheat high-molecular-weight glutenin subunits, in particular to a method for quickly identifying wheat high-molecular-weight glutenin subunits through ultra-high performance liquid chromatography (UPLC) technology. Background technique [0002] The quality of wheat is affected by many factors, such as growth environment, agronomic measures and internal genotype, etc. These factors determine the content and quality of wheat protein. Among them, protein content is a quantitative trait, which is mainly affected by environmental and agronomic measures; while protein quality is a qualitative trait, which is mainly controlled by genotype. The composition, content and ratio of different subunits of glutenin are closely related to the processing quality of wheat flour, especially the high molecular weight glutenin subunits, whose different coding sites, different subunit compositions and different expression le...

AI Technical Summary

Problems solved by technology

High-performance capillary electrophoresis (HPCE) separation is fast and reproducible, but some subunits with similar molecular weights cannot be distinguished, such as Glu-1Dy10 and Glu-1Dy12 (Gao et al., 2010) (Gao LY, Ma WJ, Chen J, Wang K, Li J, Wang SL, Bekes F, Appels R, Yan YM (2010) Characterization and Comparative Analysis of Wheat High Molecular Weight Glutenin Subunits by SDS-PAGE, RP-HPLC, HPCE, and MALDI-TOF-MS. J Agri Food Chem 58: 2777–2786.)

Matrix-assisted laser desorption ionization / time-of-flight mass spectrometry (MALD I-TOF-MS) has the characteristics of simple operation, sample saving, short measurement time, sensitivity, rapidity, accuracy and wide range of measurement mass. Polymer application has special advantages; however, the requirements for sample pre-treatment are very high, and mass spectrometers are extremely expensive, making it difficult to be widely used

Although reversed-phase high-performance liquid chromatography (RP-HPLC) can perform quantitative analysis of proteins and overcome some weaknesses of SDS-PAGE, it cannot separate subunits with similar hydrophobicity (Dong et al., 2009) (Dong K, Hu CY , Wang AL, Cai MH, Yan YM (2009) Characterization of HMW glutenin subunits in bread and tetraploid wheats by reversed-phase high-performance liquid chromatography. Cereal Res Commun 37: 65-72.)

Images