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当前位置: 首页 > 文章 > 小麦种子高分子量谷蛋白亚基的高通量检测方法 分子植物育种 2015,13 (11) 2593-2598
Position: Home > Articles > High-throughput Method for Screening HMW-GS of Common Wheat Molecular Plant Breeding 2015,13 (11) 2593-2598

小麦种子高分子量谷蛋白亚基的高通量检测方法

作  者：
张平平;周淼平;马鸿翔
单  位：
江苏省农业科学院江苏省农业生物学重点实验室江苏省现代作物生产协同创新中心
关键词：
小麦;高分子量谷蛋白亚基;高通量检测
摘  要：
为建立小麦种子高分子量麦谷蛋白亚基(HMW-GS)高通量检测方法,本研究改进了传统聚丙烯酰氨凝胶电泳(SDS-PAGE)中的样品制备和电泳分析方法,提高了检测效率。具体方法为:在小麦种子无胚端切取其体积的1/4~1/5,置于单孔容积为200μL的96孔PCR扩增板中,并在每个微孔中放入1粒直径2 mm的钢珠,并加入浸泡缓冲液静置1~2 h,再加入提取缓冲液,经高通量研磨器(Retsch Tissue Lyser)研磨后,在60℃的烘箱静置30 min。对微孔板离心和上清液预处理后在DYCZ-30C高通量垂直版电泳槽进行电泳分离,获得清晰的HMW-GS分离图谱。该方法简易、快速、成本低,适用于小麦标记辅助选择和资源评价中的大规模HMW-GS组成的检测。
译  名：
High-throughput Method for Screening HMW-GS of Common Wheat
作  者：
Zhang Pingping;Zhou Miaoping;Ma Hongxiang;Provincial Key Laboratory for Agro-biology, Jiangsu Academy of Agricultural Sciences, Jiangsu Collaborative Innovation Center for Modern Crop Production;
关键词：
Common wheat,High molecular weight glutenin subunit,High-throughput screening
摘  要：
In ord er to establish a high-throughput method for screening HMW-GS of common wheat, this study modified and improved loading sample preparing and electrophoresis protocol. One fourth to one fifth in volume was separated at the end of single kernel, which was put into 96 well PCR plate. One steel ball with a diameter of 2mm was also put into each well of the plate. Soaking buffer was injected into each well, and let sit for 1~2 hours.Followed with injecting extracting buffer in each well, the plate was fixed on Retsch Tissue Lyser for grinding. Then the plate was put into the oven with 60℃ for 30 minutes. The supernatant was transferred and pretreated after the plate was centrifuged. HMW-GS was well separated by DYCZ-30 C electrophoresis bath. This methodology was characterized with rapid, simple and low-cost, which should be useful in wheat MAS breeding and germplasm evaluation.

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