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ARS Home » Southeast Area » Baton Rouge, Louisiana » Honey Bee Lab » Research » Publications at this Location » Publication #300977
Title: A multiplex PCR assay for determination of mating type in isolates of the honey bee fungal pathogen, Ascosphaera apis

Author
item Aronstein, Katherine
item Colby, Deanna

Submitted to: Journal of Apicultural Research
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/15/2015
Publication Date: 1/1/2016
Citation: Aronstein, K.A., Colby, D.M. 2016. A multiplex PCR assay for determination of mating type in isolates of the honey bee fungal pathogen, Ascosphaera apis. Journal of Apicultural Research. doi: 10.1080.

Interpretive Summary: The ability to identify mating type idiomorphs in a spore-forming fungus, one of the major pathogens of the honey bee, is a critical step in research and management of bee diseases. A new PCR-based test developed in this study provides the first detection of Ascospahaera apis mating type idiomorphs in a single reaction. It can also be used for routine surveillance of chalkbrood and detection of early stages of the disease in honey bee colonies.

Technical Abstract: In this study we developed a multiplex PCR for identification of mating type idiomorphs in the filamentous fungus, Ascosphaera apis, the causative agent of chalkbrood disease in the honey bee (Apis melliffera). A combination of gene-specific primers was designed to amplify Mat1-1 and Mat1-2 gene fragments simultaneously in one reaction. Amplification of a single idiomorph DNA (Mat1-1 or Mat1-2) produces single PCR products of different sizes, 478 bp for Mat1-1and 214 bp for Mat1-2. Amplification of DNA from a mixed mating type culture results in two PCR bands of the corresponding sizes. Reaction conditions were tested using a variety of A. apis isolates collected in different states of the US and cultured in our laboratory. This new test can be used in research laboratories for studying fungal biology, reproduction and host virulence as well as for routine surveillance of chalkbrood disease in honey bee colonies.