Patterns of occurrence and activity of Entomopathogenic Fungi in the Algarve (Portugal) using different isolation methods

Entomopathogenic fungi (EPF) are distributed in natural and agricultural soils worldwide. To investigate EPF occurrence in different botanical habitats and soil-ecoregions, we surveyed 50 georeferenced localities in the spring of 2016 across the Algarve region (South Portugal). Additionally, we compared three EPF isolation methods: insect baiting in untreated or pre-dried-soil and soil dilution plating on a selective medium. We hypothesized that forest habitats (oak and pine semi-natural areas) and the acidic soil ecoregion may favor EPF occurrence. Overall, EPF species were present in 68% of sites, widely distributed throughout the Algarve. The use of selective media resulted in higher recovery of EPF than did either soil-baiting method. Contrary to our hypothesis, neither vegetation type nor ecoregion appeared to influence EPF occurrence. Traditional and molecular methods confirmed the presence of five EPF species. Beauveria bassiana (34% of sites), was the most frequently detected EPF, using pre-dried soil baiting and soil dilution methods. However, baiting untreated soil recovered Fusarium solani more frequently (26% of sites), demonstrating the utility of using multiple isolation methods. We also found Fusarium oxysporum, Purpureocillium lilacinum and Metarhizium anisopliae in 14%, 8% and 2% of the sites, respectively. Three abiotic variables (pH, soil organic matter and Mg) explained 96% of the variability of the entomopathogen community (EPF and entomopathogenic nematodes) in a canonical correspondence analysis, confirming the congruence of the soil properties that drive the assemblage of both entomopathogens. This study expands the knowledge of EPF distribution in natural and cultivated Mediterranean habitats.The Government of Portugal, thanks to the “Starting Grant” associate funds (Grant Number IF/00552/2014/CP1234/CT0007), supported this work. R.C.-H. was awarded an Investigator Program contract (Grant Number IF/00552/2014) and currently is supported by the Ramón y Cajal Program (Spanish Government, RYC-2016-19939). R.B.-P. and F.A.B.-P. were financed by the scientific assistantship fellowships associated with the grant IF/00552/2014/CP1234/CT0007 (BI calls UAlg-2016/004 and UAlg-2016/003, respectively). R.B.-P. is currently supported by the Department of Economic Development and Innovation of the Government of La Rioja (Spain) with a pre-doctoral contract (CAR-2018). I.V.-D. is currently supported by an ADER postgraduate grant (Government of La Rioja, Spain)Peer reviewe