Cloning And Functional Analysis Of FvHY5 Gene In Fragaria Vesca

Transcription factor HY5(ELONGATED HYPOCOTYL5)is a positive regulator of photomorphogenesis,a molecular switch for light regulation of plant development,and regulates the expression of thousands of genes in the plant genome in light-induced gene expression.In addition to its role in photomorphogenesis,HY5 protein also plays an important role in the signaling process of plant hormones.It integrates light signaling and signaling of plant hormones.In this study,woodland strawberry was used as a test material.Fv HY5 gene was isolated and cloned from woodland strawberry by PCR technology and its expression characteristics were analyzed.The function of Fv HY5 gene was analyzed by overexpression technology.The main results are as follows:1.Fv HY5 gene was cloned from woodland strawberry.The length of CDS region of Fv HY5 gene is 501 bp and it encodes 166 amino acids.The evolutionary tree analysis of the Fv HY5 protein showed that Fv HY5 has close homology with the HY5 protein of sweet cherry,peach,pear and apple,and far homology with the HY5 protein of date,grape,grapefruit,and Arabidopsis thaliana.2.The promoter region of Fv HY5 gene was cloned.Sequence analysis shows that there are many cis-acting elements in the promoter of Fv HY5 gene,such as CAAT,TATA,light response element,hormone response element and stress response element.3.The expression characteristics of Fv HY5 gene were confirmed by q RT-PCR analysis.Fv HY5 was highly expressed in flowers and fruits,but low in young leaves,old leaves and roots.4.An overexpression vector of Fv HY5 gene was constructed,and 'Rugen' was genetically transformed by an Agrobacterium-mediated method.Three Fv HY5 gene overexpression transgenic lines were obtained.These three genes were identified by q RT-PCR analysis.The expression of Fv HY5 gene in each transgenic line was 21-23 times that of the control.The chlorophyll content and anthocyanin content of the leaves of transgenic strawberry plants overexpressed with Fv HY5 gene were found to be different,but the differences were not the same at different times.The chlorophyll content in mature and old leaves was 1.4-1.5 in the control.Times.The content of anthocyanins in petioles was found to be 7-8 times higher in transgenic plants than in transgenic plants.And the determination of phosphorus content found that in tissue culture seedlings,the phosphorus content of the transgenic plant leaves was lower than the control.During the trait investigation,it was found that the height of the transgenic plants became shorter,the crown diameter became smaller,and the content of soluble solids decreased.