Shin Dongjin, Cha Jin-Kyung, Lee So-Myeong, Kabange Nkulu Rolly, Lee Jong-Hee
Department of Southern Area Crop Science, National Institute of Crop Science, RDA, Miryang 50424, Korea.
Plants (Basel). 2020 Nov 9;9(11):1517. doi: 10.3390/plants9111517.
Lab-on-a-chip technology is an emerging and convenient system to easily and quickly separate proteins of high molecular weight. The current study established a high-molecular-weight glutenin subunit (HMW-GS) identification system using Lab-on-a-chip for three, six, and three of the allelic variations at the , , and loci, respectively, which are commonly used in wheat breeding programs. The molecular weight of 1Ax1 and 1Ax2* encoded by locus were of 200 kDa and 192 kDa and positioned below 1Dx subunits. The HMW-GS encoded by locus were electrophoresed in the following order below 1Ax1 and 1Ax2*: 1Bx13 ≥ 1Bx7 = 1Bx7 > 1Bx17 > 1By16 > 1By8 = 1By18 > 1By9. 1Dx2 and Dx5 showed around 4-kDa difference in their molecular weights, with 1Dy10 and 1Dy12 having 11-kDa difference, and were clearly differentiated on Lab-on-a-chip. Additionally, some of the HMW-GS, including 1By8, 1By18, and 1Dy10, having different theoretical molecular weights showed similar electrophoretic mobility patterns on Lab-on-a-chip. The relative protein amount of 1Bx7 was two-fold higher than that of 1Bx7 or 1Dx5 and, therefore, translated a significant increase in the protein amount in 1Bx7. Similarly, the relative protein amounts of 8 & 10 and 10 & 18 were higher than each subunit taken alone. Therefore, this study suggests the established HMW-GS identification system using Lab-on-a-chip as a reliable approach for evaluating HMW-GS for wheat breeding programs.
芯片实验室技术是一种新兴的便捷系统，可轻松快速地分离高分子量蛋白质。当前研究建立了一种利用芯片实验室的高分子量谷蛋白亚基（HMW-GS）鉴定系统，分别针对小麦育种计划中常用的位于Glu-A1、Glu-B1和Glu-D1位点的三个、六个和三个等位基因变异。由Glu-A1位点编码的1Ax1和1Ax2的分子量分别为200 kDa和192 kDa，位于1Dx亚基下方。由Glu-B1位点编码的HMW-GS在1Ax1和1Ax2下方按以下顺序电泳：1Bx13≥1Bx7 = 1Bx7>1Bx17>1By16>1By8 = 1By18>1By9。1Dx2和Dx5的分子量相差约4 kDa，1Dy10和1Dy12相差11 kDa，在芯片实验室上能清晰区分。此外，一些具有不同理论分子量的HMW-GS，包括1By8、1By18和1Dy10，在芯片实验室上显示出相似的电泳迁移模式。1Bx7的相对蛋白量比1Bx7或1Dx5高两倍，因此1Bx7中的蛋白量显著增加。同样，8和10以及l0和18的相对蛋白量高于单独的每个亚基。因此，本研究表明所建立的利用芯片实验室的HMW-GS鉴定系统是评估小麦育种计划中HMW-GS的可靠方法。
