Figure 2
Inoculation of bumble bee pupae and larvae with RG-DWV. (a) qPCR analysis of DWV accumulation in bumble bee pupae injected with VVV, VDD and VVD DWV. Pupae were injected at white-eyed (P0-P1)—“we”—and brown-eyed (P7-P8)—“be”—stages and analyzed 48 h post injection. Each value corresponds to an individual sample analyzed, black lines show mean ± SD, “GE” —genome equivalents. (b) Detection of DWV RNA in bumble bee larvae from two different age groups fed with 108 GE VVD DWV: qPCR analysis of DWV levels in individual larvae samples, black-circled values correspond to samples which produced a positive result in (−)RNA assay; each value corresponds to an individual sample analyzed, error bars show mean ± SD, “GE” —genome equivalents. DWV (-)RNA assay—PCR products obtained after strand-specific reverse transcription and run in 1% agarose gel, “ + ”—positive PCR control for DWV, “M”—molecular weight DNA marker, “V”—PCR from the DWV inoculum used for larvae feeding.
