Screening Of Specific Expression Protein Of Different Morphological Spores On Beauveria Bassiana D₁-5 Strain

As an entomopathogenic fungal insecticide, Beauveria bassiana was widely used and studied in pest control of agriculture and forestry。It exited at least three kinds of infectious propagules in their life cycles, which were known as aerial conidia, in vitro blastospore and submerged conidia, respectively, and they showed different morphology and pathogenicity. In the study, the optimal culture conditions of three kinds of conidias from Beauveria bassiana D1-5 strain, were studied. For aerial conidias, the optimal culture conditions was, 26℃, 168 h in dark, at the condition of pH 6.85, and on the solid culture medium PDA. For submerged conidia in the culture medium SDY and in vitro blastspores in culture medium TKI broth, was pH 6.85, 26℃, shaker speed of 150rpm, and 50mL liquid culture medium in 250mL conical cylinders.Surface SEM observation showed that it was greatly different for the external configuration of the three kinds of spores. Aerial conidia was hydrophobic, while in vitro blastospore was hydrophilic, and submerged conidia owned both the two characters. Crystals Preparation-Section SEM observation indicated that compared to the in vitro blastospore, there was a obvious hydrophobic cycle around the Aerial conidia, it was possible that hydrophobic protein existed on ektexine of the later, however, the in vitro blastospore was produced in liquid culture medium, it had no hydrophobic protein, which caused its good hydrophilicity.Two-dimensional gel electrophoresis method was employed to analyze the proteins expression of three kinds of spores, from Beauveria bassiana D1-5 strain. According to the analysis of two-way film electrophoresis by PDQuest software, 1241 protein spots were obtained from submerged conidia and in vitro blastospores, the expression level of 20 proteins were more than 2-fold change, in which 8 was up-regulated, and 12 were down-regulated. In 1304 protein spots of aerial conidia and in vitro blastospores, 43 were up-regulated, and 14 were down-regulated, came from 57 protein spots occurred more than 2-fold change. For submerged conidia and aerial conidia compared protein expression, 66 proteins occurred more than 2-fold change, in which 8 were up-regulated and 58 were down-regulated.It indicated that complicated changes took place on proteins expression of three different kinds of spores, from Beauveria bassiana D1-5 strain, some gene expression were shuffled, and some were induced. It was significant to study the peptide mass fingerprinting (PMF) of specific expression proteins, via above information, furthermore, it could be regarded as theoretical basis for the research of spores morphological development, insecticidal mechanism, and genetic engineered breeding.