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为了分析黑龙江省小麦主要栽培品种是否含有超量表达的优质亚基,利用SDS-PAGE和RP-HPLC在2009年和2011年分析了48份黑龙江省小麦主要栽培品种(系)。研究中共使用了同一厂家生产的同一型号不同批次的3个色谱柱,由于批次不同它们在分离8*亚基时所需柱温是不一致的。其中一个色谱柱在60℃时可以将各HMW-GS有效分离,另两个在60℃时8*亚基淹没在2或5亚基的峰中,只有在柱温提高到70℃时各HMW-GS才取得了较好的分离效果。因此,在应用RP-HPLC对小麦贮藏蛋白分析时,要充分考虑色谱柱性能指标的差异来确定最适柱温才能取得较好的分离效果。结果表明:黑龙江省小麦品种(系)中超量表达的亚基仅限于来自加拿大超强筋小麦Glenlea的Bx7oe亚基。在具有5个HMW-GS的不同品系中,该亚基相对含量(单个亚基RP-HPLC曲线峰面积占全部HMW-GS曲线峰面积的百分比)的平均值为46%(变化范围为43%～49%),其它Glu-B1x位点亚基相对含量的平均值为31%(变化范围为26%～35%)。因此在强筋小麦育种及生产中利用Bx7oe亚基可以有效地改善小麦品质。
In order to analyze whether the major cultivars of wheat in Heilongjiang province contain overexpressed high quality subunits, 48 major cultivars (lines) of wheat in Heilongjiang Province were analyzed by SDS-PAGE and RP-HPLC in 2009 and 2011. In the study, three columns of different batches of the same type manufactured by the same manufacturer were used, and the column temperatures required for the separation of 8 * subunits were different due to different batches. One of the columns effectively separated each HMW-GS at 60 ° C and the other two sub-sub-groups subunit 8 * at 60 ° C in the 2 or 5 subunit. Only when the column temperature was raised to 70 ° C, each HMW -GS only achieved a better separation effect. Therefore, in the application of RP-HPLC for wheat storage protein analysis, we must take full account of the differences in performance indicators to determine the optimum column temperature in order to achieve better separation. The results showed that the subunits overexpressed in wheat cultivars (lines) in Heilongjiang Province were restricted to the Bx7oe subunit from Glenlea, a Canadian super gluten wheat. The average relative content of this subunit (percentage of peak area of single subunit RP-HPLC curve to total peak area of HMW-GS curve) was 46% for different lines with 5 HMW-GS (range 43% ~ 49%). The average relative content of other Glu-B1x subunits was 31% (ranged from 26% to 35%). Therefore, the use of Bx7oe subunits in gluten-resistant wheat breeding and production can effectively improve wheat quality.