摘要
小麦加工品质取决于籽粒面筋蛋白组分和含量,双向电泳技术是分离蛋白组分的一种有效技术手段。本研究以优质强筋材料郑麦366成熟籽粒为材料,从蛋白质提取方法、IPG预制胶条选择、蛋白质上样量3个方面进行双向电泳系统的优化,建立一套适用于小麦成熟籽粒的双向电泳技术。结果表明,利用直接裂解法提取蛋白质,使用17 cm、pH 5~8 IPG胶条,上样量为500μg的双向电泳体系,更为适合小麦成熟籽粒蛋白质的分离。利用该技术分离3种不同小麦品种成熟籽粒的蛋白质,经软件分析统计均可在电泳图谱上获得660个以上清晰的蛋白点,表明其在普通小麦中适用性较好。该体系的创建为小麦籽粒蛋白质学研究和品质相关基因的分离提供了技术支撑。
The processing quality of wh eat mainly depends on the composition and content of grain gluten,two-dimensional electrophoresis is an effective technical method for separating protein.In this study,the mature seed of Zhengmai 366,a high-quality and strong-gluten variety,was used to establish parameters of the twodimensional electrophoresis for wheat mature seed protein by comparing different protein extraction methods,IPG prefabricated strip and protein loading quantity.The results showed the optimized system of the mature wheat grain was as follows:total protein extracted by direct lyses method,separating the protein with 17 cm pH 5~8 IPG prefabricated strip,and the protein loading of 500μg.The mature seed proteins of three wheat varieties were separated using this technology,all of which obtained more than 660 clear protein spots,indicating that this method can be wildly used to separate protein of mature seed in common wheat.This method would provide technical support for wheat gain proteomic study and cloning quality-related genes in bread wheat.
作者
周正富
江雪
杨攀
秦毛毛
刘聪聪
常阳
王亚欢
李文旭
何盛莲
王美芳
晁岳恩
雷振生
田保明
吴政卿
Zhou Zhengfu;Jiang Xue;Yang Pan;Qin Maomao;Liu Congcong;Chang Yang;Wang Yahuan;Li Wenxu;He Shenglian;Wang Meifang;Chao Yueen;Lei Zhensheng;Tian Baoming;Wu Zhengqing(Henan Provincial Key Laboratory of Wheat Biology,Key Laboratory of Wheat Biology and Genetic Breeding in Central Huanghuai Region,Ministry of Agriculture,National Engineering Laboratory of Wheat,Wheat Research Institute,Henan Academy of Agricultural Sciences Zhengzhou,450002;School of Life Sciences,Zhengzhou University,Zhengzhou,450001)
出处
《分子植物育种》
CAS
CSCD
北大核心
2019年第22期7453-7458,共6页
Molecular Plant Breeding
基金
国家重点研发计划(2017YFD0100704)
河南省重大科技专项(161100110500
181100110200)
河南省科技攻关中原学者项目(162101510002)
河南省现代农业产业技术体系建设项目(S2010-01)共同资助
