裸仁美洲南瓜种皮发育PAL基因分子调控机理

项目名称： 裸仁美洲南瓜种皮发育PAL基因分子调控机理

项目编号： No.31460518

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 徐秉良

作者单位： 甘肃农业大学

项目金额： 53万元

中文摘要： 裸仁美洲南瓜（Hull-less Cucurbita pepo）是自然界中罕见的一种变异类型。通过前两个项目对种皮木质素含量、PAL活性和PAL基因表达的研究，证实了PAL基因在裸仁美洲南瓜种皮木质素合成过程中具有重要的作用。本研究将以有壳和裸仁美洲南瓜自交系为材料，对裸仁美洲南瓜种皮PAL基因家族成员进行克隆和分析；用TAIL-PCR克隆PAL基因启动子，预测和分析启动子功能区域、转录起始位点和作用元件；构建5'端缺失表达载体，转化烟草验证PAL基因启动子功能；构建PAL基因原核和真核表达载体，分别转化大肠杆菌和烟草，进一步验证PAL基因功能。研究结果对于阐明裸仁美洲南瓜种皮PAL基因的表达调控机理和种皮发育特性，拓宽南瓜裸粒种质资源和提升裸粒南瓜种质创新能力等方面均具有重要的意义。

中文关键词： 裸仁美洲南瓜；种皮发育；PAL基因家族；启动子；调控机理

英文摘要： Hull-less Cucurbita pepo is a rare type of variant in nature. The content of lignin, PAL activity and PAL gene expression were studied in the first two research projects, and the results demonstrated that PAL gene plays an important role in the seed coat of lignin synthesis. Inbred lines of hull-less Cucurbita pepo and hulled Cucurbita pepo were used as the materials, and the members of PAL gene family will be cloned and analyzed in this study; PAL gene promoter was cloned by TAIL-PCR, and the function area, transcription start site and gene cis-acting element will be predicted and analyzed in this study; 5 'end missing expression vectors were constructed, and transformed into tobacco to confirm the function of PAL gene promoters; The prokaryotic and eukaryotic expression vector of PAL gene were constructed, and transformed into Escherichia coli and tobacco to further identify the function of PAL gene. The results of this study have significant role in clarifying the mechanism of PAL gene expression and its regulation and seed development features, which is useful for broadening the naked pumpkin seed germplasm resources and improving the naked pumpkin seed germplasm innovation.

英文关键词： Hull-less Cucurbita pepo;Seed-coat development;PAL gene family;Promoters;Regulation mechanism