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      Screening of Metarhizium anisopliae UV-induced mutants for faster growth yields a hyper-virulent isolate with greater UV and thermal tolerances.

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          Abstract

          The insect pathogenic fungus Metarhizium anisopliae is an important insect biological control agent commercialized for use worldwide. Fungal infection is percutaneous, and rapid germination and growth has been linked to virulence. Using a simple in vitro growth screen to isolate mutants with increased virulence, M. anisopliae SM04 conidia were exposed to UV radiation for 20, 40, and 60 min, and mutants were subsequently screened for more rapid growth on standard potato dextrose agar. From a screen of >6,000 colonies, mutants were selected based on larger colony diameters as compared to the wild-type parent. Insect bioassays using the diamondback moth, Plutella xylostella, revealed one mutant, designated as MaUV-40.1 as displaying both more rapid growth and increased virulence. The mean lethal time to kill (LT50 using 106 conidia/ml) was 57.6 and 115.4 h for the MaUV-40.1 mutant and wild-type strains, respectively. Total conidial production, UV and thermal tolerances of the MaUV-40.1 strain were increased, but a reduced secretome was seen for the mutant compared to wild type. Analyses of culture supernatants indicated significant shifts in secondary metabolite production in the mutant. The insecticidal activity of EthOAc extracts derived from MaUV-40.1 mutant cell-free culture supernatants were ~20 times more potent that wild-type extracts. These data indicate that mutagenesis coupled to a growth screen can be a simple approach to isolate strains with greater stress resistance and virulence and that cell-free extracts may hold promise as an alternative to the living organism for insect control.

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          Author and article information

          Journal
          Appl. Microbiol. Biotechnol.
          Applied microbiology and biotechnology
          Springer Nature America, Inc
          1432-0614
          0175-7598
          Nov 2016
          : 100
          : 21
          Affiliations
          [1 ] Key Laboratory of Bio-Pesticide Innovation and Application of Guangdong Province, College of Agriculture, South China Agricultural University, Guangzhou, 510642, China.
          [2 ] Department of Microbiology and Cell Science, Institute of Food and Agricultural Sciences, University of Florida, Bldg. 981, Museum Rd, Gainesville, FL, 98195-2700, USA. keyhani@ufl.edu.
          [3 ] Key Laboratory of Bio-Pesticide Innovation and Application of Guangdong Province, College of Agriculture, South China Agricultural University, Guangzhou, 510642, China. hzscau@126.com.
          [4 ] Department of Microbiology and Cell Science, Institute of Food and Agricultural Sciences, University of Florida, Bldg. 981, Museum Rd, Gainesville, FL, 98195-2700, USA. hzscau@126.com.
          Article
          10.1007/s00253-016-7746-7
          10.1007/s00253-016-7746-7
          27521024
          1d0d6590-167d-424c-8ab3-4922e08f855e
          History

          UV-induced mutation,Destruxin,Insecticidal activity,Metarhizium anisopliae,Plutella xylostella,Secretome

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