Rapid characterization of wheat low molecular weight glutenin subunits by ultraperformance liquid chromatography (UPLC).
Rapid characterization of wheat low molecular weight glutenin subunits by ultraperformance liquid chromatography (UPLC).
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DOI：
10.1021/jf400472s
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发表时间：
2013-04
影响因子：
6.1
通讯作者：
Zitong Yu;Caixia Han;Xing Yan;Xiaohui Li;G. Jiang;Yueming Yan
中科院分区：
文献类型：
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作者：
Zitong Yu;Caixia Han;Xing Yan;Xiaohui Li;G. Jiang;Yueming Yan
低分子量麦谷蛋白亚基（LMW-GS）作为重要的种子贮藏蛋白，与高分子量麦谷蛋白亚基共同决定了面团独特的粘弹性能。本研究优化并建立了小麦LMW-GS的超高效液相色谱快速分离和表征方法。在流速为0.55 mL/min和分离温度为60 °C的条件下，通过在30 min内将洗脱梯度从21%逐渐增加到47%，可以获得快速、重现性和高分辨率的LMW-GS UPLC分离。通过该方法，一个样品的分析可以在<20 min内完成，显著少于传统的反相高效液相色谱（RP-HPLC）方法的时间。在优化条件下，成功分析了小麦LMW-GS的遗传特征及基因型×环境互作，快速鉴定出17个与不同品质性状相关的主要LMW-GS等位基因。结果表明，超高效液相色谱技术可作为小麦籽粒蛋白质遗传和蛋白质组学研究的有力工具，并可用于小麦品质改良中LMW-GS等位基因的快速鉴定和筛选。
Low molecular weight glutenin subunits (LMW-GS), as important seed storage proteins, together with HMW-GS significantly define unique dough viscoelastic properties. In this study, a rapid ultraperformance liquid chromatography (UPLC) method for the separation and characterization of LMW-GS in wheat was optimized and established. The fast, reproducible, and high-resolution UPLC separation of LMW-GS could be obtained by gradually increasing eluting gradient from 21 to 47% in 30 min at flow rate of 0.55 mL/min and 60 °C for separation temperature. By this method, analysis of one sample could be completed in <20 min, significantly less time than the traditional reversed-phase high-performance liquid chromatography (RP-HPLC) method. Under the optimized conditions, the genetic features of LMW-GS and genotype × environmental interaction were successfully analyzed, leading to a fast identification of 17 main LMW-GS alleles that were related to different quality properties in wheat. The results demonstrated that UPLC could be a powerful and alternative tool for genetic and proteomic studies of wheat grain proteins and fast identification or screening of desirable LMW-GS alleles in wheat quality improvement.