The characterization of entomopathogenic microorganisms is important for the selection of more effective strains for use in integrated pest-control programs. Five Nomuraea rileyi strains (SA86101, GU87401, SR86151, CG128 and VA9101) were characterized using random amplified polymorphic DNA (RAPD) analysis, virulence studies and assessment of chitinolytic and proteolytic activity. RAPD analysis divided the strains into two groups with a similarity coefficient of 0,76%, group 1 consisting of strains SA86101, GU87401 and SR86151 and group 2 of strains CG128 and VA9101. The LT50 varied from 165h with strain VA9101 to 246h with strain GU87401. Chitinolytic and proteolytic activity of the fungi after 144h growth in minimal medium were tested using colloidal chitin as substrate. All strains exhibited enzyme activity, with strain VA9101 having the highest chitinase activity (0,0040 mumol/mL/min the 40ºC) and strain SA86101 the highest proteolytic activity. No relationship was found between RAPD analysis, virulence and chitinase or protease activity. Caracterizaram-se cinco linhagens de N. rileyi por análise de RAPD, avaliação da virulência e atividade enzimática (quitinolítica e proteolítica). Pela técnica de RAPD foi possível distinguir, entre as cinco linhagens avaliadas, dois grupos com similaridade 0,76: grupo I (linhagens SA86101, GU87401, SR86151) e grupo II (linhagens VA9101 e CG128). O TL50 variou de 165 horas com a utilização da linhagem VA9101 a 246 horas com GU87401. Todas as linhagens apresentaram atividade quitinolítica e proteolítica, destacando-se quanto à produção de quitinases a linhagem VA9101 (com 0,0040 mimol/mL/min a 40ºC) e em relação à protease, a linhagem SA86101. Não foi possível evidenciar relações entre virulência, análise genética e atividade enzimática
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