Cloning And Function Analysis Of ANovel Valsa Mali Transcription Factor VmSom1

The canker disease caused by Valsa mali is the most serious biological disaster to the apple tree in China.It is important for the effective prevention and control of apple valsa canker disease to define molecular pathogenesis mechanism of the pathogen.,This paper mainly screened the phenotype and got a mutant among G1-G50 isolates based on constructed ATMT mutant library before of Valsa mali,the mutant gene was cloned and sequenced,and the same time,the transcriptome analysis of mutant G23 and wild strains sdau11-175 were carried out and cleared out the mutant genes expression and regulation role at the transcriptional level.The results were as follows:G23 mutant was found significantly difference on the colony morphology,colony growth rate and the ability of fruiting body production and pathogenicity among 50 isolates.which showed slow colony growth rate,mycelium sparse,an easily wettable phenotype,and non-pathogenic compared with the wild type sdau11-175,A modified hiTAIL-PCR method was established based on TAIL-PCR,hiTAIL-PCR and FPNI-PCR.The nested special primers were selected from the same sequences of the T-DNA's both borders of binary vector pBHt2 and pKO1-HPH.Modified hiTAIL-PCR method has higher efficiency compared with the traditional TAIL-PCR.The T-DNA both flanking sequences of G23 were cloned and sequenced,the T-DNA insertion in G23 was positioned among 69537-69547,which was located on in the first exon of a hypothetical gene VMIG₀9677 of chromosome 12 and named after VmSom1,it had a LisH domain at the N-terminal portion and a nuclear location signal in the VmSom1 protein.the amino acid identity analysis and phylogenetic analysis showed that Som1 in different filamentous fungi genus was not conserved.V.mali wild type sdau11-175 and mutant G23 transcriptome were analyzed by using Illumina Hiseq2500 high-throughput sequencing platform.A total of 857 genes were identified and differentially expressed,295 gene were up-regulated and 562 were down-regulated.GO analysis,KEGG analysis and COG analysis were found that many different expression genes associated with material transport and metabolism,elevendown-regulated gene were Cytoskeleton protein including four myosin and two actin in G23 transcriptome.All the result showed that VmSom1 mainly directly or indirectly regulated the hypha polar growth and seven transcription factors?two of C2H2 Zinc finger transcription factor,one Zn2Cys6 type transcription factors?Myb-like transcription factors?HMG-box transcription factors,Bromodomain transcription factors and Velvet transcription factors?were maybe responsible for phenotype of G23 mutant.