转录因子MADS-MC对番茄果实成熟及品质形成的调控机制研究

Tomato RIN and MC are adjacent genes in the genome, which encode important MADS-box factors MADS-RIN and MADS-MC, namely RIN and MC in brief. MC functioned mainly in flower and sepal development, and RIN was a key regulator in fruit ripening, whose research was based on tomato rin mutant. Latest research about gain of function in rin mutant proved RIN-MC inhibited ripening, which indicated MC transcription factor might function as an inhibitor in the ripening process. The program is proposed to ensure the function of MC in fruit ripening and quality formation by using CRISPR/Cas9 knocking out and over-expressing MC expression; Then using RNA-seq, proteomics and metabolomics, its target genes related to ripening and fruit quality formation could be identified; Further characterizing the coordinative or competitive regulation between MC and other MADS-box factor such as RIN on target genes, the regulating mechanism of MC could be cleared, which could clarify the regulation on expression of functional genes in ripening and fruit quality, and further helping improve storage and quality of tomato fruit.

番茄RIN和MC是基因组上相邻的两个基因，分别编码番茄中重要的MADS-box转录因子MADS-RIN和MADS-MC（分别简称RIN和MC）。其中MC作用于花和萼片的发育，而RIN是调控果实成熟的重要转录因子。RIN的功能研究主要基于rin突变体，关于rin的最新研究发现，该突变体是功能获得型突变，其形成的RIN-MC融合蛋白对果实成熟过程具有很强的抑制作用，因此推测MC可能是番茄果实成熟的重要抑制子。本项目拟通过CRISPR/Cas9敲除和过表达MC基因，明确其对番茄果实成熟的作用方式；采用转录组、蛋白组和代谢组分析，确定MC调控果实成熟和品质形成的相关靶基因，进一步通过MC与MADS-box因子（RIN等）互作对靶基因的协同或竞争作用机制解析，阐述MC的精细调控机制，为全面、系统阐明番茄果实成熟和品质形成过程中功能基因表达调控机制奠定基础，并为培育耐贮藏、高品质的番茄提供新思路。

rin突变体是功能获得型突变，其形成的RIN-MC融合蛋白对果实成熟过程具有很强的抑制作用，RIN对于番茄果实成熟的抑制作用已经研究的较为透彻，但MC在番茄果实成熟过程中的功能仍然需要进一步的探究。本研究CRISPR/Cas9敲除和过表达技术获得MC基因敲除和过表达的番茄材料，采用转录组、蛋白组和代谢组分析确定了MC调控果实成熟和品质形成的相关靶基因如RIN、TAGL1、ACS2等，进一步通过烟草分子荧光互补、竞争性转录激活分析等阐述MC的精细调控机制。为进一步阐明番茄果实成熟和品质形成过程中功能基因表达调控机制奠定基础。