叶色变异是体现墨兰观赏和经济价值的重要性状，对其分子机制的研究将对墨兰叶色变异形成机理的解析及叶色变异品种的选育具有十分重要的意义。项目前期研究结果发现所有乙烯响应因子CsERFs基因表达量及乙烯前体ACC含量在叶色变异叶片中显著高表达，其中CsERF2差异表达量最高。基于以上研究基础，本项目将对CsERF2进行分离克隆，通过竹叶兰遗传转化体系对CsERF2基因在叶色变异形成中的功能进行研究，并通过对转基因植株进行转录组测序，分析CsERF2基因可能调控的下游基因及参与的信号通路，同时利用交联质谱技术对其互作蛋白进行分离，并对互作蛋白进行墨兰同源基因克隆及互作验证，用以解析CsERF2基因调控墨兰叶色变异形成的分子机制，为墨兰叶色变异机理的研究及叶色变异品种的选育提供重要依据。

Leaf variegation is an important horticultural characteristic for Cymbidium Sinense presenting ornamental and economic value. Molecular mechanism study will be of great significance for analyzing the formation mechanism of leaf variegation and the breeding of leaf variegation varieties. Our previous study revealed that the expression of all identified ethylene response factor CsERFs and the content of ethylene synthesis precursor ACC (1-aminocyclopropane-1- carboxylic acid) were significantly higher in variegated leaves than control leaves, and the expression level of CsERF2 was the highest one in all CsERFs. According to these results, we will clone the CsERF2 gene in this study, and transform its over-expression and RNA interference vectors into Arundina graminifolia by genetic transformation system to research the gene function in leaf variegation of Cymbidium sinense. Moreover, in order to understand the molecular mechanism regulated by CsERF2 gene in leaf variegation, we will carry out transcriptome sequencing for transgenic plants to analyze downstream genes and signal pathway that maybe regulated by the CsERF2, and isolate its interacting proteins by cross-linking mass spectrometry. The homologous genes of the isolated proteins from Cymbidium sinense will be cloned and demonstrated by yeast two-hybrid and bimolecular fluorescence complementation system. The results of this study will provide a basis for further research in molecular mechanism of leaf variegation and the breeding of leaf variegation varieties of Cymbidium sinense.