高效毛细管电泳(HPCE)对优质小麦HMW-GS的分离鉴定

Separation and Identification of HMW-GS in High Quality Wheat by HPCE

为了解应用高效毛细管电泳(HPCE)技术定量分析优质小麦高分子量谷蛋白亚基(HMW-GS)的效果,利用HPCE技术对13个小麦品种的HMW-GS进行亚基识别和定量分析,并与传统的SDS-PAGE电泳图进行了比较。结果表明,在HPCE电泳图中HMW-GS的亚基出现顺序与SDS-PAGE略有不同,亚基1Ax1早于1Bx7出峰,并且在主亚基之后还有小的亚基出现,尤其是1Dx2主亚基后面有3个小亚基。定量分析表明,优质小麦品种陕627的HMW-GS,不论单个亚基、亚基组合还是亚基总和的含量都高于其他小麦品种,且均达极显著水平。优质小麦品种陕715的HMW-GS含量除亚基1Ax1与陕627接近外,其余亚基和亚基组合都显著低于陕627。西农5211的HMW-GS因为不含有亚基1Ax1,尽管其他亚基、亚基组合含量接近于陕627,但HMW-GS的总含量显著低于陕627。优质小麦的优质亚基含量较高。HPCE具有高效、灵敏、再现性高等特点,能够快速识别和定量分析小麦HMW-GS,可以为小麦品质育种或组合选择提供参考。

Wheat high molecular weight glutenin subunits （HMW-GS） are important factors to deter- mine the quality of wheat,mainly to dough elasticity. In order to study the influence of the composi- tion and content of HMW-GS on wheat quality of high quality wheat, SDS-PAGE and high-perform- ance capillary electrophoresis （HPCE） analysis were used to identify and quantify the subunits of HMW. Comparison on the results of SDS-PAGE and HPCE found that the subunit of HMW-GS ap- peared in different order,in HPCE electrophoresis, the subunit of 1Axl appeared earlier than 1Bx7, a- long with one major and three minor isoforms were observed for the individual HMW-GS. Quantita- tive analysis showed that single subunit, subunit combination or sum of high quality wheat variety Shaan 627 were all higher significantly than other wheat varieties. The subunit lAx1 of high quality wheat variety Shaan 715 was close to Shaan 627, while the other subunits or subunit combinations were significantly lower than that of Shaan 627. As Xinong 5211 did not contain subunit 1Ax1, although other subunits or subunit combination content were close to Shaan 627,its content of subunit sum was significantly lower than that of Shaan 627. All together, subunit 1Ax1 has an important effect on the quality of wheat. The results of SDS-PAGE and HPCE analysis were similar,but the HPCE analysis is more efficient,sensitive and reproducable,and more quickly to identify and quantify the HMW-GS of wheat. HPCE can be used to discriminate,fingerprint,or monitor the purity of wheat cultivars,and to monitor protein synthesis during kernel development.