基于高通量筛选的蜜蜂生殖不相容性分子机制研究

项目名称： 基于高通量筛选的蜜蜂生殖不相容性分子机制研究

项目编号： No.31470123

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 畜牧学与草地科学

项目作者： 李兴安

作者单位： 吉林省养蜂科学研究所

项目金额： 78万元

中文摘要： 蜜蜂生殖隔离，即，蜜蜂种间完全生殖隔离和蜜蜂种内部分生殖隔离，在分子水平涉及交配行为后-受精卵前和受精卵后生殖不相容性分子机制。为了揭示蜜蜂生殖隔离分子机制，本文首先进行长白山东方蜜蜂、东北黑蜂雄蜂精液介质（即，异型附腺分泌物）的高通量蛋白质组学分析；其次，进行欧洲黑蜂、卡尼鄂拉蜜蜂雄蜂生殖细胞（即，异型精子和卵子）的高通量蛋白质亲和作用分析，以及它们体细胞（即，异型头、胸部组织）的高通量基因组微卫星长度多态性分析。上述分析可以给出反映蜜蜂交配行为后-受精卵前和受精卵后生殖不相容性分子机制的基础数据。特色与创新：本文在分子水平将蜜蜂生殖隔离机制归纳为交配行为后-受精卵前和受精卵后生殖不相容性分子机制的基本问题；为此，分别进行了基于双向电泳技术的蛋白质组学分析，基于生物膜层干涉技术的蛋白质亲和作用分析，以及基于集成芯片毛细管电泳技术的基因组微卫星长度多态性分析。

中文关键词： 高通量筛选；“交配行为后-受精卵前”和“受精卵后”生殖不相容性；分子机制；东方蜜蜂；西方蜜蜂

英文摘要： Complete and partial reproductive isolation in honeybees (Ginus Apis) is at molecular level involved in postcopulatory-prezygotic and postzygotic incompatibilities. To understand molecular characteristics with reproductive isolation, two independent analyses are conducted via high-throughput approaches: first, proteomics approach is applied to detect postcopulatory -prezygotic incompatibilities of heterotypic seminal fluid proteins (accessory gland proteins) between two types of honeybees (Apis cerana population in Changbai Mountain area and Apis mellifera population in Northeast China); furthermore, a assay for determining protein affinity and a assay for assessing genome-wide microsattlite polymorphism are respectively applied to detect postzygotic incompatibilities of heterotypic reproductive cells (sperms and drone eggs) between two types of honeybees (Apis mellifera mellifera and Apis mellifera carnica) and postzygotic incompatibilities of heterotypic somatic cells (head and thorax tissues) between two types of honeybees (Apis mellifera mellifera and Apis mellifera carnica). Above research produce molecular replicate datasets for mechanisms underlying postcopulatory-prezygotic and postzygotic incompatibilities. Original and novel: Complete and partial reproductive isolation in honeybee (Ginus Apis) is involved at molecular level in postcopulatory -prezygotic and postzygotic incompatibilities. For this purpose, three high-throughput tools, two dimensional gel electrophoresis, bio-layer interferometry and integrated capillary electrophoresis chip are incorporated respectively in proteomic anslysis, assay for determining protein affinity and assay for assessing genome-wide microsattlite polymorphism.

英文关键词： high-throughput screening;postcopulatory-prezygotic and postzygotic incompatibilities;molecular mechanisms;Apis cerana;Apis mellifera