CmLEC1调控菊花胚胎发育的机理研究

目前国内菊花品种高达数千个，但多数品种抗性差，而许多菊花野生资源抗性强，运用栽培品种与野生资源进行杂交是改良菊花抗性的有效途径。研究表明菊花远缘杂交中胚胎大量败育经常发生，严重影响育种效果。在刚结题国家基金资助下，我们针对菊花远缘杂交胚胎败育现象进行了研究，阶段性结果已在Scientific Reports等期刊上发表，并发现CmLEC1与菊花胚胎发育密切相关，但其作用机制尚不清楚。为此，本项目在良好研究基础上，采用RACE、RT-PCR技术克隆该基因，利用酵母双杂技术筛选互作蛋白，运用蛋白体外表达、BiFC及体内瞬时表达系统研究其机制和功能；通过超表达、RNAi转基因植株筛选出受CmLEC1调控的下游关键功能基因，并进行体内外验证。结果将揭示CmLEC1调控菊花胚胎发育的作用机制，为菊花远缘杂交胚胎发育的分子设计育种提供理论基础和优异基因储备。因此，本研究具有重要的科学意义和应用前景。

There are thousands of chrysanthemum cultivars in China now, but most of them are poor in biotic and abiotic resistance. However, many chrysanthemum wild resources usually have strong biotic and abiotic resistance. Distant hybridization between chrysanthemum cultivars and the wild resources is an effective way to improve chrysanthemum resistance. Previous studies have shown that embryo abortion occurs frequently in chrysanthemum distant hybridization, and seriously affects its breeding effect. We invesitigated the mechanism of embryo abortion in chrysanthemum distant hybridization from 2012 to 2015 under the support of a project from National Natural Science Foundation of China, and some stage results have been published in Scientific Reports and some other journals. In particular, we found that one gene, CmLEC1, is closely related to chrysanthemum embryo development, while its function and mechanism is unclear. Based on our good foundation,we will clone CmLEC1, screen its interaction proteins, and study its mechanism and function by RACE, RT-PCR, yeast two hybrid, bimolecular fluorescence complementation, and in vivo transient expression system. In addition, we will screen downstream key functional genes regulated by CmLEC1 and carry out internal and external validation expreiments. Results will shed light on the mechanism of CmLEC1 regulating chrysanthemum embryo development, and provide theoretical foundation and excellent gene pool for molecular design breeding during chrysanthemum distant hybridization embryo development. Therefore, the study in this project is of great scientific significance and prospective application.