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高分子量谷蛋白亚基(HMW-GS)是决定小麦食品加工品质的重要因素。为了快速而全面地鉴定小麦品种或杂交后代的高分子量谷蛋白亚基(HMW-GS),本研究以宁春4号、扬麦19号和宁春4号×扬麦19号F2分离群体种子为研究对象,采用半粒法进行综合鉴定。即先从小麦胚端1/3~1/2处切除种子胚乳提取DNA,用分子标记快速筛选出含5+10亚基的籽粒,再从这些籽粒提取的DNA残留物中提取蛋白质,采用SDS-PAGE电泳法从总体上确定其HMW-GS的组合,从中筛选出含1,17+18,5+10亚基的籽粒。结果表明,在96粒F2分离群体种子中,有31粒种子被PCR鉴定为5+10纯合亚基,再经SDS-PAGE电泳鉴定,确定有14粒含1,17+18,5+10优质亚基,其中4粒种子的亚基组合为1,17+18,5+10。与之前的研究相比,半粒法能充分利用实验样品,快速筛选到含有优质亚基组合的籽粒,有助于优质小麦育种。
High-molecular-weight glutenin subunit (HMW-GS) is an important factor in determining the quality of wheat food processing. In order to rapidly and comprehensively identify the high molecular weight glutenin subunits (HMW-GS) of wheat or hybrid progenies, seedlings were isolated from Ningchun 4, Yangmai 19 and Ningchun 4 × Yangmai 19 F2 As the research object, using half-grain method for comprehensive identification. Firstly, the endosperm was excised from 1/3 ~ 1/2 of wheat embryo and the 5 + 10 subunits were quickly screened out by molecular markers. Then, the protein was extracted from the DNA residues extracted from these seeds. SDS -PAGE electrophoresis to determine its HMW-GS combination, from which the seeds containing 1, 17 + 18, 5 + 10 subunits were screened out. The results showed that among the 96 seeds of F2 population, 31 seeds were identified as 5 + 10 homozygous by PCR, and then identified by SDS-PAGE electrophoresis, 14 seeds with 1, 17 + 18, 5 + 10 High-quality subunits, of which 4 seeds subunit combinations of 1,17 +18,5 +10. Compared with the previous study, the half-pellet method can make full use of the experimental samples and rapidly screen the kernels containing the high-quality subunit combinations for breeding high-quality wheat.