Use of RAPD Analysis forin SituIdentification ofAscosphaera aggregataandAscosphaera larvisin Larval Cadavers of the Alfalfa Leafcutting Bee,Megachile rotundata

Autor: Lu, Rui, Goerzen, D.Wayne, Rank, Gerald H.
Zdroj: Journal of Invertebrate Pathology; July 1996, Vol. 68 Issue: 1 p78-83, 6p
Abstrakt: Chalkbrood of the alfalfa leafcutting bee,Megachile rotundata,is caused by the fungusAscosphaera aggregata.We used random amplified polymorphic DNA (RAPD) analysis for thein situidentification ofA. aggregataand a related species,Ascosphaera larvis,in larval cadavers ofM. rotundata.A simple DNA extraction method was developed to preferentially isolate DNA from fungal spores on the cadaver surface, or from ascocysts beneath the cuticle. Similar banding patterns were obtained inA. aggregata-infected larval cadavers from different sources and geographic areas. The RAPD banding pattern of cadavers infected withA. aggregatadiffered from that of healthy leafcutting bee prepupae. RAPD analyses of cadavers infected withA. aggregataandA. larvisresulted in similar banding profiles as those obtained from corresponding pure fungal cultures of the two species. This suggests that the RAPD bands of infected cadavers were amplified from fungal DNA, rather than from other DNA associated with the leafcutting bee cadaver. The banding patterns of “sporulating” and “non-sporu-lating” chalkbrood cadavers exhibited no differences; this provides the first definitive evidence that both forms of the disease result from infection withA. aggregata.
Databáze: Supplemental Index