Preliminary Construction Of Standard LMW-GS Profile In Common Wheat Using HPCE

Glutenin, as a kind of gluten in wheat endosperm, is heterogeneous polymeric protein, which formed by HMW-GS(as the main chain) and LMW-GS(as the side chain) that bonded by intermolecular disulfide and hydrogen. Wheat gluten plays an important role on the processing quality, which, LMW-GS, mainly affact gluten strength and extensibility. Rapid and accurate identification of LMW-GS composition is significant for system study influence of LMW-GS on wheat processing quality and quality prediction.In this study, high-performance capillary electrophoresis(HPCE) profile was preliminarily constructed to detect LMW-GS quickly and accurately. To construct the standard profile of HPCE, locations of LMW-GS were confirmed by comparing with same subunits. We implemented these following steps: First, SDS-PAGE and molecular markers were used to identify the compositions of LMW-GS in 14 common wheat varieties from Huang-Huai wheat region. Then, improved HPCE system was used to separate LMW-GS, controlling variable method was used to identify the corresponding peaks of LMW-GS in profiles by comparing same subunits from different varieties. The results are as follows:1. LMW-GS in 14 common wheat varieties were identified accuratly by combination of SDS-PAGE and molecular markers, Chinese Spring is Glu-A3 a, Glu-B3 a, Glu-D3a(the standard cultivar of SDS-PAGE); xinong538 is Glu-A3 a, Glu-B3 d, Glu-D3c; xinong 558 is Glu-A3 a, Glu-B3 j, Glu-D3c; xinong 528 and xinong 556 is Glu-A3 d, Glu-B3 c, Glu-D3b; shan 627 is Glu-A3 d, Glu-B3 j, Glu-D3c; shan 715 is Glu-A3 c, Glu-B3 h, Glu-D3c; xinong 979 is Glu-A3 c, Glu-B3 d, Glu-D3a; xiaoyan 22 is Glu-A3 a, Glu-B3 j, Glu-D3c; shanyou 225 is Glu-A3 d, Glu-B3 d, Glu-D3b; zhoumai 18 is Glu-A3 c, Glu-B3 j, Glu-D3a; shannong 138 is Glu-A3 c, Glu-B3 j, Glu-D3c; zhengmai 9023 is Glu-A3 c, Glu-B3 b, Glu-D3c; because of biology hybrid, zhengmai 366 is Glu-A3 a and Glu-A3 d, Glu-B3 d, Glu-D3 c.2. All varieties are seperated continuously, the electrophoresis results are highly repetitive. Among them, standard deviation of peaks migration time from 5 repetitions of Chinese Spring is 0.0149-0.0234, the relative standard deviation(RSD) is 0.13%-0.20%, the average RSD is 0.16%. RSD of migration time of other varieties are 0.30% or less; the HPCE profile can initially identify Glu-A3 a, Glu-A3 c, Glu-A3 d, Glu-B3 a, Glu-B3 d, Glu-B3 h rapidly by controlling variable method, the peak order and number of SDS-PAGE and HPCE are different due to the different separation principles.This study has made valuable exploration for constructing the HPCE profile of LMW-GS, but only partial LMW-GS can be identified due to the limitation of materials, thus, further research is still needed.