Espression And Phyligenetic Analysis Of Glycyrrhiza DNA Methyltransferase Gene Under High Voltage Electrostatic

DNA methylation is one of the important ways of epigenetic modification,participating in and regulating the processes of plant growth,development and evolution.The establishment and maintenance of DNA methylation in the methylation process relies on the DNA methyltransferase.In this paper,licorice was used as the material to isolate the partial sequences of three-type of methyltransferase genes by PCR technique.The bioinformatics analysis was used to confirm the preliminary sequence.Using phylogenetic analysis to construct a phylogenetic tree of genes related to 20 species such as Medicago and Arabidopsis.Ther real-time quantitative technique(RT-qPCR)was used to study the expression of three methyltransferase genes in licorice seedlings treated by high voltage electrostatic field.The main results show that:1.Using the method of PCR amplification of degenerate primers,one MET1,DRM and CMT methyltransferase gene fragments were isolated from licorice genome.The target fragment of MET1-typemethyltransferase is 951 bp,and the nucleic acid sequence homology with pea MET1 gene is 92.63%;the target fragment of DRM-type methyltransferase is 357 bp,and the nucleic acid sequence homology with Medicago DRM gene is 92.37%;CMT-like methyltransferase target fragment length is 692 bp,and the nucleic acid sequence homology with Medicago CMT gene is 83.97%.2.The phylogenetic analysis method was used to construct the phylogenetic tree of MET1,DRM-like and CMT-like methyltransferase genes and other 20 species.The distance between MET1 methyltransferase of licorice and the chickpeas was very close;The closest to the DRM-like methyltransferase is the branch of the Medicago;The branches of Medicago and chickpeas are closest to CMT-like methyltransferases.3.High voltage electrostatic field treatment of licorice in the methyltransferase gene expression in different growth periods have different change,three kinds of methyltransferase to the response of different electric field intensity.(1)When the electric field was E3(18kV/cm,25min),the expression of the three methyltransferases was up-regulated.(2)Under the treatment of high voltage electrostatic field,the expressions of three kinds of methyltransferase genes were up-regulated and down-regulated,indicating that the electrostatic field promoted and inhibited the methylation regulation of licorice.