山葡萄雄株性别CKX基因家族分析与VaCKX的性别转换功能研究

项目名称： 山葡萄雄株性别CKX基因家族分析与VaCKX的性别转换功能研究

项目编号： No.31501747

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 农业科学

项目作者： 李晓艳

作者单位： 内蒙古农业大学

项目金额： 20万元

中文摘要： 在前期构建的山葡萄雄株花序性别转换关键期的转录组测序（RNA-Seq）数据库中，发现编码细胞分裂素氧化酶/脱氢酶（CKX）的6个差异表达基因（DEGs）在细胞分裂素CPPU处理后的雄株花序（已转变成两性花）中全部上调表达。根据测序结果我们初步分析了山葡萄雄株性别CKX基因家族与影响性别转换的关键基因。本项目以此为切入点，通过基因克隆、生物信息学分析、亚细胞定位、时空表达特异性分析、基因表达调控和转基因植株功能研究，对山葡萄雄株性别转换过程中CKX基因家族进行分析，从中选择1-2个与性别转换密切相关的关键基因（VaCKX）进行功能研究。揭示山葡萄VaCKX基因在雄株花序性别转换过程中的功能，为今后利用山葡萄雄株性别转化为雌花，获得可结实的山葡萄新种质提供基因依据。

中文关键词： 山葡萄雄株；性别转换；细胞分裂素氧化酶/脱氢酶；基因功能；生殖发育

英文摘要： In order to reveal the molecular mechanism of the sex reversal of Vitis. amurensis Rupr (The sex of the male Vitis.amurensis flower can be converted to a hermaphrodite form after treatment with cytokinin.) at the transcriptional level, RNA-Seq technique was used to collect the transcriptome data of V. amurensis male flower buds after treatment with cytokinin. Differentially expressed genes (DEGs) encoded cytokinin oxidase/dehydrogenase (CKX) were found in this database and all up-regulated in the hermaphrodite flower buds. We hypothesize CKX family members may play an important role during the sex of V. amurensis male flower buds converted to a hermaphrodite form. In this study, the CKX family genes related with sex conversion were analyses by using the methods of gene cloning, bioinformatic analysis, subcellular localization, expression patterns of specific temporal and spatial, gene expression and regulation and function of transgenic plants. One or two CKX genes closely related with sex conversion were chosen for functional analysis by transforming into Arabidopsis. And the expressional level and their response to exogenous hormones in different flower organs were studied. These data may provide additional germplasm information for evaluation of VaCKX gene in the male V. amurensis, and laid the foundation for further mechanistic studies on V. amurensis male convert female. Moreover, the identification of VaCKX functional will be useful for generating superior varieties of V.amurensis.

英文关键词： Male of Vitis.amurensis;Sex conversion;CKX;Gene function;Reproductive development