An Isaria fumosorosea SCAR marker for evaluation of soil, insect, and airborne samples

Abstract

Molecular detection markers are needed for ecological studies of entomopathogenic fungi. In this study, a sequence-characterized amplified region (SCAR) marker of Isaria fumosorosea was used to detect the fungus in soil, insects, and airborne samples. These were artificially added with different fungal conidial concentrations. Specificity and sensitivity were tested with semi-nested PCR using oligonucleotides E-AA/M-CTA124 F and E-AA/M-CTA124 R for the first amplification and E-AA/M-CTA124 F and E-AA/M-CTA103 R for the second amplification. Specificity assays showed a specific band of 103 bp for DNA samples from 10 I. fumosorosea strains used. Negative results were observed for DNA samples from other species of Isaria, including I. amoene-rosea, I. farinosa, and Paecilomyces carneus as well as with other entomopathogens such as Metarhizium acridum, M. anisopliae, M. majus, M. flavoviride Type E, and Lecanicillium lecanii. Sensitivity assays showed that the specific SCAR marker detected 104 conidia from I. fumosorosea EH-511/3 that were artificially mixed with soil, from 1 to 104 conidia artificially mixed with Galleria mellonella (Lepidoptera: Pyralidae), and from 10 to 105 conidia in Melinex tape for airborne samples. The marker was also able to detect conidia in airborne samples from cotton wicks in a mini wind tunnel. These SCAR markers for I. fumosorosea had excellent specificity and sensitivity and are relevant tools for ecological studies of this fungus.