SlNAP7基因调控番茄果实番茄红素合成的分子机理研究

Accumulation of lycopene is an important feature of ripening fruit, study on its synthesis and regulation has important significance for improving the quality of fruit. Our previous study found that SlNAP7 gene silencing has effect on the accumulation of lycopene in tomato fruit, but the mechanism is not clear. This project aims at screening the target genes and proteins of SlNAP7 regulation by iTRAQ, RNA-seq and tomato fruit yeast cDNA library screening, and using VIGS to do their functional analysis. By analyzing the metabolic intermediates of lycopene, the expression of key genes of the synthesis pathway of lycopene in SlNAP7 transgenic tomato fruit, it can be to determinate the key point of SlNAP7 regulating lycopene synthesis. Transmission electron microscope was used to observe the development of chromoplasts of SlNAP7 transgenic fruit. A comprehensive analysis of above results can determine the molecular mechanism of SlNAP7 gene regulating lycopene synthesis, which riches pigment synthesis theory of mature fruit; SlNAP7 also can provide a theoretical basis to increase fruit lycopene content through gene engineering technology in the future.

番茄红素积累是番茄果实成熟的重要特征，研究番茄红素的合成与调控对于改善果实品质具有重要意义。我们前期研究发现SlNAP7基因沉默影响了番茄果实番茄红素的合成与积累，但具体机制不清。本项目旨在通过蛋白组学（iTRAQ）、转录组学（RNA-seq）和番茄果实酵母cDNA文库筛选等技术筛选SlNAP7调控番茄红素合成与调控相关靶基因和蛋白，并通过VIGS对其进行功能验证；测定SlNAP7转基因番茄果实番茄红素合成代谢中间产物含量和关键酶基因的表达水平，确定SlNAP7基因参与调控的上述途径的关键位点；通过透射电镜观察SlNAP7转基因果实发育过程中有色体的变化趋势。综合分析确定SlNAP7基因调控番茄果实番茄红素合成的分子机理，丰富果实成熟色素合成理论，为今后通过基因工程技术提高番茄红素含量提供理论依据。

番茄红素在番茄果实成熟过程中大量积累，是一种重要的功能保健成分，也是番茄果实成熟的标志之一，番茄红素的合成与调控是重要的关键科学问题。我们的研究发现SlNAP7沉默导致了番茄叶片的叶绿素降解和果实番茄红素积累受限，但机制不清楚。本研究利用VIGS技术沉默SlNAP7基因，获得叶片和果实材料，开展叶绿素和番茄红素的测定，番茄红素和叶绿素代谢途径关键酶基因的检测，质体和叶绿体的电镜观察，RNA-seq测序，发现SlNAP的互作蛋白和参与调控叶绿素和番茄红素的相关基因，同时还分析了SlNAP7沉默对番茄材料代谢的影响。总之，SlNAP7对植物叶绿体的发育和果实质体的发育至关重要，会影响叶绿素的合成和番茄红素的积累。研究结果为果实的质体的调控提供了理论依据。