中华蜜蜂与意大利蜜蜂雄蜂胚胎发育差异蛋白质组与磷酸化蛋白质组分析  

Comparative Analysis of Proteome and Phosphoproteome of Drone Embryos Between Apis cerana cerana and A. mellifera ligustica

作　　者：周天娥[1,2] 房宇[2] 冯毛[2] 韩宾[2] 张兰[2] 李荣丽[2] 李建科[2] 

机构地区：[1]郑州大学生物工程系,郑州450001 [2]中国农业科学院蜜蜂研究所,北京100093

出　　处：《中国农业科学》2013年第2期394-408,共15页Scientia Agricultura Sinica

基　　金：国家现代农业产业技术体系(蜜蜂)(CARS-45-KXJ13);"十二五"国家科技支撑计划(2011BAD33B04);国家自然科学基金(30972148);中国农业科学院科技经费(201207)

摘　　要：【目的】通过对中华蜜蜂(中蜂)和意大利蜜蜂(意蜂)雄蜂胚胎发育期3个日龄差异蛋白质组和磷酸化蛋白质组进行分析,以探明中蜂、意蜂雄蜂胚胎发育特征并了解雄蜂胚胎发育生物学,为蜜蜂遗传改良提供理论依据。【方法】采用双向电泳方法建立中蜂、意蜂雄蜂胚胎发育3个日龄的蛋白质组及磷酸化蛋白质组表达谱,获得图谱中蛋白质的分子量、等电点和表达量等信息,并对部分差异蛋白质进行质谱分析、功能鉴定和生物信息学分析。【结果】在雄蜂胚胎发育的第1、2、3日龄,中蜂表达332、362、340个蛋白质,其中111、117、112个蛋白质发生了磷酸化修饰;意蜂表达302、331、311个蛋白质,其中有107、110、106个发生了磷酸化修饰。各日龄中蜂、意蜂雄蜂胚胎共同表达的蛋白质分别为214、239和220个。对30个差异表达蛋白质鉴定结果表明这些蛋白主要与碳水化合物代谢和能量生产、蛋白折叠、发育、调控以及骨架蛋白相关。【结论】中蜂雄蜂胚胎3个日龄所表达的全蛋白质、磷酸化蛋白质均多于意蜂,说明中蜂雄蜂胚胎发育较意蜂需要更多的蛋白质参与调控。中蜂、意蜂雄蜂胚胎各日龄约1/3的蛋白质发生了磷酸化修饰,这可能与胚胎发育过程中信号传导、细胞增殖分化等活动有关。意蜂较中蜂胚胎表达较多比例的管家蛋白质,说明意蜂较中蜂悠久的进化史使其胚胎形成了更为保守的发育特点。对差异蛋白功能分析表明,中蜂、意蜂雄蜂在长期进化过程中不仅形成了具有各自生物学特征的胚胎发育方式,而且各自不同生物学特征如中蜂善于利用零星蜜粉源、抗胡蜂和意蜂的高采蜜量、产浆量等性状已在各自蛋白质组中得到体现。[Objective] The objective of this study is to compare the proteome and phosphoproteomic during the three days development of Apis cerana cerana and .4. mellifera ligustica, investigate the characteristics of embryonic development and gain an in-depth understanding of the developmental biology of the drone embryos, and to lay a foundation for the future genetic improvement of honeybees. [Method] Two-dimensional gel electrophoresis （2-DE） was employed to compare the proteomic and phosphoproteomic differences between the drone embryogenesis of the two bee species at three time-points of 24, 48 and 72 h, respectively. The level of protein expression, pI and molecular weight were determined. Furthermore, some of the differentially expressed proteins were analyzed by mass spectrometry and bioinformatics. [Result] In total proteome, 332, 362 and 340 protein spots were detected in drone embryos of A. cerana cerana from day 1 to day 3. Of these, 111, 117 and 112 proteins were phosphorylated, respectively. In the drone embryos of A. mellifera ligustica, 302, 331 and 311 proteins were detected, in which 107, 110 and 106 proteins were phosphorylated. Meanwhile, 214, 239 and 220 proteins were commonly expressed during the first three days of drone embryonic development. Thirty differentially expressed proteins were involved in carbohydrate metabolism and energy production, protein folding, development, regulatory proteins and cytoskeletal proteins.[ Conclusion ]A larger number of proteins and phosphoproteins were expressed in A. cerana cerana than those in A. mellifera ligustica during the drone embryogenesis, indicating A. cerana cerana needs more protein to regulate its drone embryonic development. About 1/3 protein phosphorylated in both honeybee embryos might be associated with signal transduction, cell proliferation and differentiation during drone embryogenesis. Embryos of A. mellifera ligustica expressed higher proportion of housekeeping proteins than in A. cerana cerana suggest the embryogenesis of A. mellifera li

关 键 词：中华蜜蜂 意大利蜜蜂 雄蜂胚胎 双向电泳 蛋白质组 磷酸化蛋白质组 

分 类 号：S891[农业科学—特种经济动物饲养]