‘鸭梨’SCF蛋白复合体组份功能解析

项目名称： ‘鸭梨’SCF蛋白复合体组份功能解析

项目编号： No.31201606

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 植物保护学、园艺学与植物营养学

项目作者： 李茂福

作者单位： 北京市农林科学院

项目金额： 21万元

中文摘要： 自交不亲和反应受控于花柱与花粉S基因。茄科和车前科自交不亲和反应中，花粉S基因产物除与S-RNase有物理互作外，还与SSK1及CUL1共同形成SCF蛋白复合体,但在与这两个科具较远亲缘关系且同属于S核酸酶类自交不亲和的蔷薇科中是怎样的至今无人研究。本项目拟以重要果树品种鸭梨为材料，克隆其候选花粉S基因，并通过转基因来确定其是否为花粉S基因；通过同源克隆法克隆SCF复合体组份的SSK1与CUL1，利用Southern杂交及RT-PCR来分析SCF复合体组份的表达模式，利用酵母双杂交、蛋白下拽来研究SCF复合体组份之间的相互作用关系，利用显性负效应的策略并通过转基因等方法研究SSK1基因的功能，最终探讨蔷薇科类自交不亲和反应中SCF复合体的作用机制及其与茄科和车前科的异同。预期结果对阐明蔷薇科自交不亲和性机理研究提供新的实验证明，也将为蔷薇科果树品种的选育奠定更充实的理论基础。

中文关键词： 梨；SCF复合体；功能分析；；

英文摘要： Generally speaking, Self-incompatibility(SI) is genetically controlled by a single polymorphic S locus, which harbors pistil S and pollen S determinant genes. The pollen S gene not only physically interact with pistil S gene, but also combine with SSK1 and CUL1 to form an SCF protein complex in Solanaceae and Plantaginaceae, and target at S-RNase destruction. However, it is unclear whether similar mechanism exist in Rosaceae (includes vast marjority of friut plants), which has a distant phylogenetic relationship with Solanaceae and Plantaginaceae, but they are blong to the same type of SI. The present study aims to: (1) clone the candidate pollen S gene by using YaLi as material, and confirm the existence of pollen S gene by transgene in Petunia; (2) isolate the orthologous genes of AhSSK1/PhSSK1 and CUL1 in Rosaceae by homologous cloning method; (3) identify the function of SSK1 by Southern blot, yeast two hybrid, protein pull-down and transgene; (4) verify the relationship among SSK1, SLF, CUL1, S-RNase in vitro by pull-down and yeast two hybrid. Finally, the mechanism of SCF complex in Rosaceae and its difference with Solanaceae and Plantaginaceae could be decerned. The present study will offer as new evidence in understanding the interspecific self-incompatibility in Rosaceae, as well as provide important th

英文关键词： Pear (Pyrus)；SCF complex；functional analysis；；