A Systematic Study On The Gene Function Of Dof Transcription Tactor Family In Rice

Dof transcription factor is a kind of protein which is involved in growth and development processes of plants.At present,the studies of Dof genes were mainly concentrated in Arabidopsis,their function can be divided into tissue differentiation,seed development and metabolic regulation and so on.Rice Dof transcription factor family contains a total of 30 genes.Through evolutionary analysis,we divided them into seven subfamilies,and each family has a large number of conservative motifs.Dof genes had no significant difference between cultivated and wild rice.Dof genes had obvious purification selection effect in cultivated and wild rice,and the differentiation degree of Dof genes in cultivated rice was small.Analysis of Dof transcription factor expression patterns found that different subfamily expression patterns are different.Therefore,according to the above systematic analysis,we constructed different subfamily gene lose-function mutants in order to analyze the relationship between phylogenetic branch and gene functional differentiation.We constructed a total of 30 CRISPR/Cas9 vectors.After transformation of Agrobacterium tumefaciens,a total of 12 gene lose-function mutations were obtained.10 genes in the 12 mutants showed phenotypic phenotypes,which involved plant height,growth period,leaf shape,spike shape,fertility and so on.Among them,the phenotype of plant height was spread in subfamily A,C,D and F,while fertility-related phenotypes were mainly concentrated in subfamily C and subfamily E,and leaf-related phenotypeswere concentrated in subfamilies A and F.What's more,the subfamily A and subfamily F affect many kinds of phenotypes.The results showed that different branches were responsible for regulating the different phenotypes of rice during the evolution of Dof transcription factor family,and there was redundancy of gene function of repeat gene.In this study,the Dof gene was successfully knocked out by CRISPR/Cas9 technique in rice the first time and the function of rice was analyzed systematically by mutant phenotype and evolutionary relationship.This work laid the foundation of the Dof gene mechanism in plants.