Entomopathogenic fungi from Moroccan apple orchards: Molecular and enzymatic characterization of new biocontrol candidates against Cydia pomonella

The codling moth ( Cydia pomonella L.) is a major pome fruit pest urging sustainable control alternatives with increasing insecticide resistance. Entomopathogenic fungi (EPF) are emerging as effective biocontrol agents for pest management. This study evaluated the pathogenicity of EPF isolated from insect cadavers against two Moroccan C. pomonella populations and characterized their virulence-associated enzymatic activities (lipase, protease, phospholipase, chitinase) and virulence gene profiles. Seven fungal isolates were identified: Aspergillus tubingensis (2), Aspergillus niger (2), Penicillium olsonii , Penicillium glabrum , and Penicillium thomii . Neonate larvae were exposed to three conidial suspensions (1 × 10⁴, 1 × 10⁶, and 1 × 10⁸ conidia/mL), with two commercial Beauveria bassiana strains serving as reference controls. A. tubingensis (75.4 %) and P. olsonii (76.6 %) achieved highest mortality rates at 1 × 10⁸ conidia/mL for both populations, exceeding control strains. P. thomii , P. glabrum , and A. niger also achieved mortality rates above 60 % mortality. A. tubingensis and A. niger demonstrated superior enzymatic activity and metabolic flexibility compared to Penicillium isolates. The beauvericin gene was detected in all Penicillium isolates and two Aspergillus species. GGPP synthase was exclusive to Penicillium , while polyketide synthase occurred in P. olsonii , P. glabrum , and both Aspergillus isolates. Our results demonstrate high pathogenic potential of insect-derived fungi against C. pomonella , with genetic and enzymatic characteristics supporting their development as biocontrol agents.