基于赤霉素初始响应的水稻糊粉层基因表达调控机制-扬州大学.doc

基于赤霉素初始响应的水稻糊粉层 基因表达调控机制 张文虎（M05305） 导师：王忠教授 扬州大学生物科学与技术学院植物生理教研室 225009 摘要：在谷物种子萌发过程中，糊粉层细胞通过赤霉素作用，合成并分泌水解酶水解贮存的营养物质，实验证实，赤霉素基因表达的早期产物，如转录因子GAMyb，在上述过程中具有关键作用。GAMyb在水解酶基因表达过程中的作用已被证实，但GAMyb自身的表达调控却不甚明了。为了搞清赤霉素基因媒介——GAMyb的真实表达机制，我们使用一个糊粉层瞬时表达的水稻体系（Oryza sativa L.），检测其糊粉层中GA基因早期响应的响应域。启动子上游不能赋予GA响应。大量分析表明，第一基因内区（内含子1）具有类似增强子的功能区域。现已确定：拟南芥内含子的增强子是MADS框中的同源异型基因，AGAMOUS（AG）和FLOWERING　LOCUSC（FLC）。这两种大型的内含子不但可以让每个基因得以表达，而且通过与组蛋白DNA共价结合而与基因沉默相联系。依据这些事实，我们可以推测，基本染色体调控在的GA表达初始行为方面相当重要。基于这种假定，我们已经证明在萌发的水稻种子中GAMyb的甲基化DNA定位。 关键词：糊粉层；DNA甲基化；GAMyb；赤霉素；水稻；种子萌发 Common mechanisms regulating expression of rice aleurone genes thatcontribute to the primary response for gibberellin Abstract：During germination of cereal seeds, aleurone cells respond to gibberellins (GA) by synthesizing and secreting hydrolytic enzymes that mobilizethe reserved nutrients. It has been shown that products of early GA response genes, like a transcription factor GAMyb, act as key moleculesleading to this regulation. Pivotal roles of GAMyb on expression of hydrolase genes have been well documented, whereas regulation of GAMybexpression itself remains obscure. In order to understand virtual mechanisms of the GA-mediated expression of genes, it is important to know howGA control expression of early GA response genes. Using an aleurone transient expression system of rice (Oryza sativa L.), we examined GAresponsive domains of early GA response genes in the aleurone, such as GAMyb and OsDof3. The upstream promoter could not confer GAresponse. Extensive analyses revealed the presence of enhancer-like activities in a large first intron. In Arabidopsis, intron enhancers have beenidentified in MADS-box homeotic genes, AGAMOUS (AG) and FLOWERING LOCUS C (FLC), in which large introns should not only conferproper gene expressions, but also associate with gene silencing by covalent modifications of both DNA and histone. These evidences prompt us toassign that chromatin-based control might be important for initial GA