Gene expression analysis of mite-infested pupae and adult bees.

Relative expression of the immune-related genes Apidaecin (a), Defensin-1 (b), Dorsal-1A (c) and of the RNAi related genes Argonaute-2 (d) and Dicer-like (e) of pupae and adult bees challenged with V. destructor. (Two-way ANOVA test. Apidaecin: mite: df = 1, F = 0.192, p = 0.66; age: df = 1, F = 1.179, p = 0.29; mite*age: df = 1, F = 1.237, p = 0.28; Defensin-1: mite: df = 1, F = 9.680, p df = 1, F = 294.82, p df = 1; F = 0.185, p = 0.67; Dorsal-1A: mite: df = 1, F = 2.156, p = 0.15; age: df = 1, F = 37.624, p 0.01; df = 1, F = 3.058, p = 0.09).; Ago-2: mite: df = 1, F = 2.99, p = 0.09; age: df = 1, F = 0.09, p = 0.75; mite*age: df = 1, F = 1.07, p = 0.31,; Dicer-like: mite: df = 1, F = 1.90, p = 0.18; age: df = 1, F = 5.23, p = 0.03; mite*age: df = 1, F = 0.93, p = 0.34. Uninfested pupae: n = 6, mite-infested pupae: n = 6, uninfested adults: n = 6, mite-infested adults: n = 5).</p

Effects of the infestation’s time on bees’ survival.

Survival curves of the adult bees uninfested and artificially infested with one Varroa mite at the L5 stage or at the emergence and maintained under lab condition (Wilcoxon test. Infestation at larval stage vs control: chi square = 52.3, df = 1, p df = 1, p df = 1, p = 0.05, uninfested bees: n = 164, bees mite-infested at the larval stage: n = 140, bees mite-infested at the adult stage: n = 121).</p

Survival of the honey bees DWV-infected during the pupal stage.

Survival curves of adult honey bees infected or not with 10^3 DWV copies at the larval stage and maintained under lab conditions (Wilcoxon test. Chi square = 6.42, df = 1, p = 0.01, uninfected bees: n = 52, DWV-infected bees: n = 50).</p

DWV genotypes.

Percentage of viral reads mapping on the two genomes, type A and B DWV.</p

DWV infection in honey bees mite-infested during the pupal stage.

DWV relative quantification obtained by real time PCR in newly emerged honey bees infested or not with one mite at the L5 stage (Mann-Whitney U test. U = 22, df = 1, p = 0.02, uninfested bees: n = 10, mite-infested bees: n = 10).</p

Real time PCR primers.

Sequences of the primers used for real time PCR analysis.</p

DWV infection in newly emerged bees DWV-infected during the pupal stage.

DWV relative quantification obtained by real time PCR in newly emerged honey bees infected or not with 10^3 DWV copies at the larval stage (Mann-Whitney U test. U = 6, df = 1, p = 0.09, uninfected bees: n = 5, DWV-infected bees: n = 5).</p

Gene expression analysis of DWV-infected pupae and adult bees.

Relative expression of the immune-related genes Apidaecin (a), Defensin-1 (b), Dorsal-1A (c) and RNA interference related genes Argonaute-2 (d) and dicer-like (e) of pupae and adult bees artificially infected with 10^3 DWV copies. (Two-way ANOVA test. Apidaecin: DWV: df = 1, F = 2.36, p = 0.14; age: df = 1, F = 8.43, p df = 1, F = 1.45, p = 0.24; Defensin-1: DWV: df = 1, F = 4.39, p = 0.04; age: df = 1, F = 146.7, p df = 1; F = 1.00, p = 0.32; Dorsal-1A: DWV: df = 1, F = 1.18, p = 0.30; age: df = 1, F = 4.47, p df = 1, F = 1.01, p Ago-2: DWV: df = 1, F = 3.77, p = 0.06; age: df = 1, f = 3.90, p = 0.06; DWV*age: df = 1, F = 7.43, p = 0.01; Dicer-like: DWV: df = 1, F = 1.63, p = 0.21; age: df = 1, F = 2.11, p = 0.16; DWV*age: df = 1, F = 1.88, p = 0.18. Uninfected pupae: n = 6, DWV-infected pupae: n = 6, uninfected adults: n = 6, DWV-infected adults: n = 6).</p

DEGs bees infested at the adult stage.

List of the genes differentially expressed between adult bees infested after the emergence and uninfested. (DOCX)</p

Survival of honey bees mite-infested during the pupal stage.

Survival curves of the adult bees artificially infested or not with one Varroa mite at the L5 stage and maintained under lab conditions (Wilcoxon test. Chi square = 6.4; df = 1; p < 0.01, uninfested bees: n = 103, mite-infested bees: n = 76).</p