Study On The Mechanism Of EIN3 Regulating Seed Dormancy Through DOG1 In Arabidopsis Thaliana

Seed germination is a critical stage in the plant life cycle that largely determines crop yield.Seed germination is regulated by many factors,including temperature,light,water,and plant hormones.Ethylene is an important plant hormone that plays an important regulatory role in plant growth and development,including seed dormancy.However,the regulation of ethylene on seed dormancy and germination is still at the physiological level,and the specific molecular mechanism remains to be studied.EIN3(Ethylene-Insensitive3)is an important transcription factor in the ethylene signaling pathway,which can transmit and respond to ethylene response.In our previous experimental studies,we found that EIN3 can bind to the promoter regions of DOG1(Delay of Germination 1)and asDOG1(antisense-DOG1),and EIN3 is involved in the regulation of seed germination.DOG1 is a quantitative trait locus that regulates seed dormancy and is a positive regulator of seed dormancy.We further verified and analyzed the molecular mechanism of EIN3 regulating seed dormancy by binding to the DOG1 promoter through molecular biology and genetic methods.The main results are as follows:1.It was found that EIN3 can bind to the promoter regions of DOG1 and asDOG1 through CUT&Tag experiments.This result was further verified by yeast single hybrid experiment and EMSA experiment.And through promoter analysis,it was found that the promoter regions bound by EIN3 all had EIN3 binding sites.2.Through the analysis of DOG1 transcription and protein levels in ein3-1 mutant dry seeds and imbibed seeds,it was found that EIN3 can inhibit the transcription of DOG1 and the protein expression of DOG1,thereby regulating seed dormancy.3.By analyzing the germination phenotypes of ein3-1,dog1-3,dog1-5,dog1-3ein3-1and dog1-5ein3-1,it was found that EIN3 could promote seed germination by inhibiting the expression of DOG1.4.By measuring the expression level of DOG1 in dog1-5 and dog1-5ein3-1 seeds treated with ethylene precursor ACC and testing the germination phenotype,it was found that ethylene negatively regulates the expression of DOG1 through EIN3 and then regulates seed dormancy.5.By analyzing the seed germination phenotype under salt stress,it was found that EIN3 could positively regulate seed germination under salt stress,while DOG1 played a negative role.In conclusion,ethylene can promote seed germination through negative regulation of DOG1 expression by EIN3,which gives us a new understanding of the molecular mechanism of ethylene regulation of seed germination.At the same time,EIN3 and DOG1 are involved in regulating seed germination under salt stress conditions,which enables us to have a deeper understanding of the regulation of seed dormancy by DOG1.