Identification And Expression Characteristics Analysis Of Strawberry FvMIF2 Gene

MIFs(mini zinc finger proteins)contain zinc finger domains,which are involved in hormone regulation,affect cell differentiation,affect flower development.So far,only MIF in Arabidopsis,tomato,and gerbera have been reported,and the function of MIF in strawberry is not clear.In this study,the coding sequence region of the FvMIF2 gene is cloned from the diploid woodland strawberry(Fragaria vesca).The functions and expression characteristics of the FvMIF2 gene are revealed through subcellular localization,transcriptional activation,and phenotypic analysis of overexpressing plants.The main results are as follows:(1)The FvMIF2 gene was cloned from the diploid woodland strawberry resource 'Ruegen' using PCR technology.CDS contains 267 bp and encods 88 amino acids.The sequence identity between FvMIF2 gene and Fa MIF2 gene of cultivated strawberry(F.× ananassa)is 99.25%.Using NCBI analysis,it is found that the domain of FvMIF2 is ‘ZH-HD?dimer'.(2)The pRI101-FvMIF2-eGFP fusion vector was constructed and introduced into the protoplasts of Arabidopsis thaliana for subcellular localization.The result showed that FvMIF2 was localized in the nucleus.(3)The qRT-PCR method was used to analyze the expression levels of FvMIF2 gene in different organs of strawberry.It is found that the expression level in red fruit is the highest,and its relative expression in leaves was the lowest,especially in new leaves.(4)Tissue culture plantlets of cultivated strawberry genotype 'D-10' tissue culture seedlings were treated with 3 kinds of hormones including methyl jasmonate,salicylic acid,abscisic acid and 4 kinds of stresses inclduing 4°C low temperature,37°C high temperature,salt,osmotic stresses,respectively.Samples are taken at 0 h,12 h,24 h,48 h,72 h,and 96 h,and the relative expression levels of MIF2 gene under different treatments and different treatment times were identified by qRT-PCR.It was found that strawberry MIF2 responded to hormone treatment and stress treatment(5)The pGBT9-FvMIF2 fusion vector was constructed and introduced into yeast to perform a transcription activation test.The result showed that FvMIF2 didn't have transcriptional self-activating activity.(6)The pRI101-AN-FvMIF2 overexpression fusion vector was constructed,and two 'D-10' FvMIF2 gene overexpression lines were obtained by Agrobacterium-mediated genetic transformation.Two transgenic lines and non-transgenic control plants have been domesticated and transplanted to survive and grew in a solar greenhouse.So far,the transgenic plants have begun to show differential phenotypes.Their leaf color is darkened and the degree of leaf unfolding is reduced.