低温诱导甜菜春化基因BvRAV的功能研究

Sugar beet (Beta vulgaris) is used as the materials in this research. The BvRAV sequence members are explored by BLAST against the database and EST database of Beta vulgaris, and then transgenic BvRAVRNAi lines will be constructed after structure identification. BvRAV, BvFT1 and BvFT2 proteins and their traits will be analyzed. The molecular mechanism of vernalization gene BvRAV interacted with BvFT1 and BvFT2 will be investigated by subcellular localization and yeast two-hybrid system. In particular, methylation, chromatin and histone modifications of BvRAV will be analyzed by Bisulphite Sequencing and Chromatin immunoprecipitation. Furthermore, long noncoding RNAs (lncRNAs) of sugar beet under vernalization and nonvernalization conditions will be explored. The interaction among lncRNAs, BvRAV, BvFT1 and BvFT2 will also be analyzed. The molecular mechanism of BvRAV interacted with BvFT1 and BvFT2 will be elucidated at the level of epigenetics. In addition, the mechanism of vernalization gene BvRAV interaction with endogenous and exogenous Gibberellin(GA) will also be investigated under vernalization and nonvernalization conditions. This research will be very important to understand the mechanism of sugar beet bolting and flowering.

以二年生糖甜菜（Beta vulgaris）为材料，通过BLAST搜索基因库数据，索引甜菜春化基因（BvRAV）和甜菜EST数据库鉴定BvRAV基因家族成员序列。分析BvRAV基因结构，建立BvRAVRNAi基因沉默体系验证BvRAV基因功能；GST pull-down筛选BvRAV, BvFT1和BvFT2基因的蛋白，分析其特点及结构；亚细胞定位和酵母双杂交探索BvRAV对开花调节基因BvFT1、BvFT2作用机制；重亚硫酸盐测序和染色质免疫沉淀研究BvRAV, BvFT1和BvFT2基因甲基化和染色质组蛋白修饰的变化；高通量测序发掘春化与非春化lncRNA与BvRAV，BvFT1和BvFT2基因作用机制，从表观遗传学水平探明BvRAV基因的功能。同时，解析BVRAV在春化和非春化条件下与内源和外源植物激素赤霉素（GA）调控机理。本项目对明确甜菜抽薹、开花分子机理具有重要意义。

项目以二年生糖甜菜（Beta vulgaris）为试验材料，通过长日照光周期、短日照光周期与春化、非春化相结合的方法处理甜菜母根。经过16W、19W、20W光周期处理，探索春化长日照、春化短日照、非春化长日照、非春化短日照甜菜植株的抽薹、开花表型表现、激素变化，利用转录组学技术、蛋白组学技术分析及qPCR技术其分子水平差异。利用BLAST搜索基因库数据，索引甜菜春化基因（BvRAV）和甜菜EST数据库鉴定BvRAV基因家族成员序列。分析了BvRAV基因结构。初步明确了BvRAV、BvFT1和BvFT2 基因的结构及功能；明确了不同春化和光照处理对甜菜抽薹、开花的影响及分子机制。进一步研究了植物激素赤霉素（GAs）、吲哚-3-乙酸（IAA） 和脱落酸（ABA）对甜菜春化、抽薹、开花的调控机制及时空表达。项目发表相关科研论文25篇。申请国家发明专利4项（未授权）。培养博士研究生2名，硕士研究生3名。