Study On The Mechanism Of Photomorphogenesis Regulatory Factor SmCOP1 Regulating Eggplant Fruit Development And Anthocyanin Synthesis

Eggplant(Solanum melongena L.)is regarded as an important vegetable crop by the United Nations.It has a wide cultivation area in China.Eggplant is rich in anthocyanins and is beneficial to human health.In the process of eggplant cultivation,anthocyanins in the fruit peel cannot be effectively synthesized and accumulated due to uneven illumination,which seriously affects the commercial value of eggplant.How to increase the accumulation of anthocyanins in the peel of eggplant under low light conditions is very important.Therefore,this study explored the mechanism of the photomorphogenesis factor Sm COP1 in regulating eggplant fruit development and anthocyanin synthesis.Under low light conditions,the synthesis of anthocyanins in eggplant fruit peel is improved.In this paper,the RNA interference technology was used to construct the Sm COP1-RNAi silencing vector and a stable genetically modified transgenic line was obtained through genetic transformation.It combines q RT-PCR,WB,yeast two hybrid,HPLC and p H differential method at the three levels of transcription,protein and metabolism The differential detection technique was used to analyze the specific mechanism of photomorphogenesis factor Sm COP1 in regulating eggplant fruit development and anthocyanin synthesis.The main results are as follows:(1)The synthesis of anthocyanins in purple eggplant is strictly regulated by light.With the increase of light time,the stronger the fruit coloration,the higher the anthocyanin content.q RT-PCR detection found that the key enzyme genes early,middle and later Sm PAL,Sm F3 H,Sm3GT in the regulation of anthocyanin synthesis were detected by q RT-PCR.is regulated by light induction,and the protein abundance of Sm MYB1 protein will increase with the increase of light time,which is positively correlated with the accumulation of anthocyanins.(2)Through the analysis of tissue expression pattern of anthocyanins,it was found that the synthesis of anthocyanins is tissue-specific.The expression of anthocyanins is highest in the peel,and is hardly expressed in the root and flesh.Sm COP1 protein is expressed in leaves,flowers,and peel without tissue specificity.The highest accumulation of Sm MYB1 protein in the peel is highly correlated with anthocyanin expression.The expression of photosensitin receptor is positively correlated with the accumulation of anthocyanins.The blue light receptor Sm CRY1 is The highest expression was in the stamen,and there was no difference in the expression of Sm CRY2 in each tissue.(3)The accumulation of anthocyanins in Sm COP1-RNAi seedlings and eggplant fruit peel was significantly up-regulated,and the increase of Sm MYB1 protein level was conducive to the synthesis and accumulation of anthocyanins.Sm MYB1 protein abundance was significantly up-regulated,which promoted the significant up-regulation of anthocyanin biosynthesis related genes Sm PAL,Sm CHS,Sm CHI,Sm DFR,Sm3 GT and Sm ANS and chlorogenic acid to synthesize the genes Sm C3 H,Sm4CL and Sm HQT,promoting anthocyanins and chlorogenic acid content was significantly increased.(4)Sm COP1-RNAi eggplant fruit peel Sm MYB1 protein changes significantly under low light conditions,Sm MYB1 protein promotes anthocyanin biosynthesis related genes Sm PAL,Sm CHS,Sm CHI,Sm DFR,Sm3 GT and Sm ANS and chlorogenic acid synthesis structural genes Sm C3 H,Sm4CL,Sm HQT was significantly up-regulated to promote the up-regulation of anthocyanins and chlorogenic acid on the fruit epidermis.(5)By cloning the Sm COP1 gene and predicting its protein structure,it was found that it has a conserved WD40 repeat sequence domain near the C-terminus.In vitro experiments with yeast two hybrid showed that the Sm MYB1 R3 domain interacted with the Sm COP1 WD40 domain.(6)In this study,we also found that Sm COP1-RNAi seedlings showed short hypocotyls in the dark,and their aerial parts also shortened significantly under normal light conditions,showing a photomorphogenesis phenotype.Silencing the Sm COP1 gene revealed that Sm CIP7 was significantly up-regulated,and the eggplant fruit became larger,the number of seeds increased,the seeds became larger,the grain weight increased,the lignin synthesis gene was significantly up-regulated,and the lignin content increased.This paper not only explored the regulation of anthocyanin biosynthesis at the protein level by Sm COP1,but also enabled us to achieve the goal of improving pigment yield and improving fruit quality,and provided a new theoretical model in the analysis of anthocyanin accumulation in other crops.