Function Analysis Of AbaA Gene Of Sporothrix Schenckii

Sporotrichosis is a zoonotic disease caused by Sporothrix.In recent decades,the incidence of Sporotrichosis has been increasing year by year,and it is widely distributed throughout the world.In China,it is mainly prevalent in three provinces of Northeast China and the middle and lower reaches of the Yangtze River,and it is endemic or occupational.Sporothrix schenckii is a biphasic fungus,it has two phases,hyphae phase and yeast phase,and the yeast phase is the pathogenic phase of the bacterium.The biphasic transformation of S.schenckii is a complex process involving multiple factors,and the key genes regulating this process still need to be further explored.We performed transcriptome analysis on the hyphae phase and yeast phase of S.schenckii and found that AbaA gene was differentially expressed in the two phases.Previous studies of the AbaA gene showed that it encodes aATTS/TEA DNA-binding domain transcriptional regulator and regulates biphasic transformation in the biphasic fungi Penicillium marneffei.However,the AbaA gene had never been studied in S.schenckii.Therefore,we constructed a knockout strain of the AbaA gene,to preliminarily study the function of AbaA gene in S.schenckii,the results are as follows:1.Comparison of the hyphae and yeast transcriptome of S.schenckii revealed that12,217 genes were differentially expressed,including 7271 genes up-regulated and4946 genes down-regulated.Our S.schenckii fungus to transcriptome and double phase transformation in differentially expressed genes related to signal channel for further statistical analysis to find AbaA genes differentially expressed in the dual phase,its|log₂FC|=-1.705;2.After analyzing the transcriptome data and consulting relevant papers,it was concluded that the AbaA gene may play an important role in the biphasic transformation and pathogenicity of S.schenckii.The knockout fragment was synthesized by overlapping extension PCR,and it was linked to the PCB309 vector to construct the knockout plasmid successfully.The knockout plasmid was transformed into S.schenckii by Agrobacterium tumefaciens,and the knockout strain was screened.3.Compare the differences of wild strain with AbaA knockout strain in many respects,the result showed that melanin generation in knockout strains decreased significantly,the growth rate of mycelial phase and yeast phase have both delayed,can't be transformed from mycelial phase to yeast phase completely.They can only be converted into a cigar transition state,reduced susceptibility to Congo red,but the sensitivity to SDS and yeast lyase increased,while the osmotic stress lowered.In conclusion,the AbaA gene plays an important role in the two-phase transformation of S.schenckii.It is also suggested that AbaA gene can further affect the pathogenicity of the strain by influencing the production of melanin,the growth of mycelium and yeast cells,the tolerance of osmotic stress and the composition of cell wall.This will lay a foundation for further study of gene function,pathogenesis and drug targets of S.schenckii.