番茄MYB72调控果实叶绿素和类胡萝卜素代谢的分子机理研究

果实是人类重要的食物和营养来源。阐明果实成熟过程中营养和色泽变化的分子机理对增强果实品质具有重要的科学意义。我们前期研究发现采用RNAi干扰方法降低MYB72基因表达的番茄果实绿果期呈现大量绿色斑点，红果期呈现黄色斑点，发现果实叶绿素含量升高，番茄红素含量降低，β-类胡萝卜素含量升高，表明MYB72基因调控了果实叶绿素和类胡萝卜素的代谢过程。本课题将进一步通过超表达和CRISPR/Cas9敲除方法研究MYB72基因在番茄果实叶绿素和类胡萝卜素代谢过程中的重要作用，同时通过染色质免疫共沉淀、凝胶阻滞实验、酵母双杂、双分子荧光互补等技术筛选MYB72下游靶标基因和与MYB72互作的蛋白，阐明MYB72基因在番茄果实叶绿素和类胡萝卜素代谢过程中的分子调控机理。本研究将扩展人们对番茄果实叶绿素和类胡萝素代谢过程中分子调控机理的认识，为基因工程手段调控类胡萝卜素代谢，改良果实品质提供新的基因资源。

The fruit is an important resource for food and nutrition of human. It is important scientific significance for enhancement of fruit quality to elucidate the molecular mechanism of fruit nutrition and color. Our previous studies found that there were a lot of green spots at green fruit and yellow spots at red fruit of MYB72 RNAi tomato. The MYB72 RNAi tomato showed increased content of chlorophyll, decreased content of lycopene, and increased content of β-carotenoids, which proved that MYB72 gene regulated the metabolism of chlorophyll and carotenoids in fruit. Based on the results, this project will further study the important roles of the SlMYB72 gene in chlorophyll and carotenoid metabolism regulation by overexpression and CRISPR/Cas9 knock out methods in tomato fruits. At the same time, chromation immunoprecipitation, electrophoretic mobility shift assay, bimolecular fluorescence complementation and yeast two hybrid methods will be used to select the SlMYB72 target genes and binding proteins and confirm the regulatory network of MYB72 in chlorophyll and carotenoid metabolism in tomato fruit. This study will expand the understanding of the molecular mechanism of chlorophyll and carotenoid mebabolism regulation in fruits and provide new genes for improving fruit quality by using genetic engineering method