GENETIC TRANSFORMATION OF Beauveria bassiana USING Agrobacterium tumefaciens-MEDIATED TRANSFORMATION AND MUTANTS SCREENING WITH SPECIAL TRAITS

In this study,Beauveria bassiana YB-06 was successfully transformed using Agrobacterium tumefaciens-mediated transformation approach,and T-DNA insertion mutants were further obtained.The effects of pH and cultivation media on genetic transformation were studied under the condition of 28℃,1×106 spores/ml,A.tumefaciens(AGL1) OD600=0.8,200μmol/L acetosyringone and 48h co-cultivation in the presence of induction medium.The results showed that the transformants could be obtained in specific pH value ranging from 5.1 to 5.8 and the transformantion efficiency reached the highest level at pH5.3 with about 586 transformants per 106 spores.The results also revealed that transformation efficiency was higher on cellophane and NC millipore filter compared to filter paper.HPH+ resistance and PCR assay of the transformants showed that the T-DNA had been successfully integrated into the genome of the fungus.A few mutants were identified by colony growth,colony color,and conidium output of the 3,000 transformants.Altogether,this transformation system provided a basis for the study of development,pathogenicity mechanism and functional genetics study of the fungus.