• 作物遗传育种·种质资源·分子遗传学 •

### 高分子量麦谷蛋白亚基HPCE高效分离及图谱鉴定

1. 西北农林科技大学农学院, 陕西杨凌 712100
• 收稿日期:2017-11-24 接受日期:2018-03-26 出版日期:2018-07-10 网络出版日期:2018-07-19
• 通讯作者: 高翔
• 基金资助:
本研究由国家现代农业产业技术体系建设专项(CARS-3-2-47)和国家高技术研究发展计划(863计划)项目(2011AA100501)资助

### Efficient Separation and Identification of High Molecular Weight Glutenin Subunits by HPCE

Wei-Dong WANG,Xiang GAO(),Dan-Yang ZHAO

1. College of Agronomy, Northwest A&F University, Yangling 712100, Shaanxi, China
• Received:2017-11-24 Accepted:2018-03-26 Published:2018-07-10 Published online:2018-07-19
• Contact: Xiang GAO
• Supported by:
This study was supported by the China Agriculture Research System (CARS-3-2-47) and the National High Technology Research and Development Program of China (2011AA100501).

Abstract:

High molecular weight glutenin subunit (HMW-GS) is important for processing quality of wheat. High performance capillary electrophoresis (HPCE) is increasingly used in the work of separation and identification due to its advantages of small sample, rapidness and high precision. However, this technique has been seldom used in wheat HMW-GS study and its separation system needs improvement in analytical speed and discernibility. On the basis of HMW-GS identification with SDS-PAGE and molecular markers, an high-efficiency separation system of HPCE was set up using Chinese Spring as the standard. The system components (pH 2.5) were 75 mmol L -1 IDA, 0.05% HPMC, and 15% ACN. The electrophoresis parameters were 25 μm of inner diameter of the capillary, 200 nm of detection wavelength, 20 kV of separation voltage, and 30°C of operating temperature. Using mixed injection method, the standard spectrums were obtained for 18 subunits. Their migration order was 1Dy12→ 1Dy10→ 1By9→ 1By8→ 1By18→ 1By16→ 1By20→ 1Bx17→ 1Bx20→ 1Bx13→ 1Bx6→ 1Bx7→ 1Ax2*→ 1Ax1→ 1Dx5→ 1Dx4→ 1Dx3→ 1Dx2, and standard peak time of these subunits was 9.39, 9.69, 10.30, 11.70, 11.89, 12.09, 12.22, 12.36, 12.62, 12.83, 13.08, 13.18, 13.50, 13.73, 14.04, 14.24, 14.46, and 14.73 min, respectively. The relative standard deviation was smaller than 0.2%. The y-type and x-type subunits appeared in the phases of 9.39-12.36 min and 12.36-14.76 min, respectively, between which there was 1Bx17 as the boundary. These results indicate that the HPCE separation system is applicable in rapid identification of HMW-GS in wheat germplasm resources when we simultaneously consider migration order, standard peak time, and HPCE spectrum.