miR828和miR858调控猕猴桃果实花青苷合成的机制研究

色泽是影响果实商品性的一项重要品质指标。相较于绿肉/黄肉猕猴桃而言，红肉猕猴桃更受消费者青睐，而花青苷是红肉猕猴桃区别于其他猕猴桃的特征色素。因此，提高果肉花青苷含量可提升果实品质。有研究表明，大多数猕猴桃的花青苷合成途径是健全的，因此项目重点关注猕猴桃花青苷代谢上游调控因子。依据国内外研究进展，本项目拟重点关注miRNA对猕猴桃果实花青苷代谢的调控机制，研究不同品种猕猴桃中花青苷积累模式；利用小RNA组，鉴定猕猴桃中miR828和miR858成员，并分析其与花青苷代谢的相关性；通过靶基因预测、双荧光素酶体系、可视荧光体系等体系，明确miR828/miR858对靶基因（MYB等）的调控机制；通过猕猴桃瞬时过表达体系、猕猴桃转基因体系，明确miR828/miR858对靶基因和果实花青苷代谢的调控效应，以期进一步完善果实花青苷代谢的调控机制。

Colour is an essential quality index affecting the commodity of fruit. Compared with green/yellow kiwifruit, red kiwifruit is more attractive for consumers. Moreover, anthocyanin is the characteristic pigment of red kiwifruit, which is different from other kiwifruit. Therefore, increasing the content of anthocyanin in the flesh can improve the quality of kiwifruit, and help to improve the competitiveness of domestic kiwifruit in the international market. Based on the previous findings, this project focuses on the upstream regulatory factors of anthocyanin metabolism in kiwifruit. Various cultivar will be used to study the accumulation pattern of anthocyanin; small RNA sequencing will be carried out to identify miR828 and miR858 members in kiwifruit, and analyse their correlation with anthocyanin metabolism; clarify the regulation mechanism of miR828/miR858 on the target gene (MYB etc) by target gene prediction, dual-luciferase assay, visible fluorescence system, etc; the regulatory effects of miR828/miR858 on targets, as well as anthocyanin accumulation, will be verified by kiwifruit transient assay and stable over-expression system. This study aims to improve the regulatory mechanism of miRNA for coloration in kiwifruit.