Expression Patterns And Characterization Of Transcription Factors VdCHTF1 And VdCHTF7 During Microsclerotia Formation In The Smoke Tree Wilt Fungus Verticillium Dahliae

Smoke-tree wilt is a destructive disease,which seriously affectes the beautiful red-leaf scenery,and its pathogen is Verticilliun dahliae Kleb.Since the 1990s,the disease has casused mortality of smoke trees in Xiangshan.The Verticilliun dahlia is a soil-borne pathogen which will forme the dormant structure:microsclerotia with strong resistance and adaptability.Microsclerotia are the main source of infection and show high adapatability in the soil.Microslerotial is most difficult reason to prevent it effectively.In order to study the molecular mechanism of microsclrotial formation in V.dahliae,this study is based on genome information of Verticilliun dahlias,using the bioinformatics and genetics technique to exploring expression modal and functional of C2H2-Homeobox transcription factor family,to exploring the biological significance of gene expansion of C2H2-Homeobox transcription factor family.(1) C2H2-Homeobox transcription factor family gene identification and sequence analysis:Through sequence alignment and phylogenetic tree analysis method to determine the location and size of the structure of the family-genes domain,concluded that VdCHTF4 is the strongest specifity gene, VdCHTFl is most conserved gene in the family.The study found that although in the same family,there are significant differences between genes;(2) C2H2-Homeobox transcription facor family gene expression modal:By constructing Pro::GFP vector with the resistance marke geneticin,seven genes GFP expression vector were successfully obtained by PEG-mediated protoplast transformation method.Observed VdCHTF1 and VdCHTF7 fluorescence expression patterns and the GFP transformation of VdCHTF2, VdCHTF3 and VdCHTF4 during microsclerotia development.(3) Exploring CHTF4 function of C2H2-Homeobox gene transcription facor family:using Double-joint method to build C2H2-Homeobox transcription factor family gene knockout vector,with the resistance marke genenticin,received a total of 173 knockout transcfonnation by PEG mediated genetic transformation method.after screening,to gain a VdCHTF4mutants.