Fig. 4: Effect of Y-sat on alate formation.
From: A plant virus satellite RNA directly accelerates wing formation in its insect vector for spread
a Mean percentages of winged aphids in N. benthamiana transiently expressing Y-sat dsRNA (dsY-sat), plus-sense (Y-sat+) and minus-sense RNA (Y-sat−) relative to respective controls (one-way ANOVA, Dunnett’s test, n = 4, P = 0.0026). Agrobacterium carrying pBE2113-GUS was used as a control. b Mean percentages of winged aphids on transgenic plants expressing either dsY-sat, Y-sat+ or Y-sat− (one-way ANOVA, Tukey’s test, n = 4, P = 0.0030). c Proportion of alate morphs fed on diet containing dsY-sat (one-way ANOVA, Tukey’s test, n = 5, P = 0.0085). Controls: leek yellow stripe virus dsRNA; H2O. d Relative expression of Apns1-like in aphids fed on CMV-infected or [CMV + Y-sat]-infected plants. Third instars (N3) and early adult [A(1)] are as explained in Fig. 2a (one-way ANOVA, Tukey’s test, n = 4, P < 0.0001). e Relative expression of ABCG4 in N3 (two-sided t test, n = 4, P = 0.0122), A(1) (two-sided t test, n = 4, P = 0.0150) and late adults [A(4)] (two-sided t test, n = 4, P = 0.8923) fed on CMV-infected or [CMV + Y-sat]-infected tobacco plants. Each data point in Fig. 4e represents the level of mRNA isolated from aphids [N3, ten individuals; A(1) and A(4), one individual]. f Complementarity between miR9b and Y-sat or ABCG4. g RNA dot-blot hybridization. RNA oligonucleotides of Y-sat and ABCG4 complementary to miR9b shown in Fig. 4g were synthesized, blotted onto a nylon membrane (0.4–10 pmol) and probed by Dig-labelled miR9b. h Competitive hybridization assay. When ABCG4 RNA was probed with Dig-miR9b, Y-sat RNA was added at molar ratios indicated on right. Results were the same in two repeated experiments. Values on membrane are relative values determined using ImageJ, without Y-sat was set to 1.0. i RNA sequence alignment between Y-sat and T73 satellite (T73-sat). The complementary region to miR9b in Y-sat is indicated. j Mean percentages of alate morphs infesting CMV-infected or [CMV + T73]-infected tobacco. (GLM test in R, n = 4, z = −0.473, P = 0.636). Each data point in (a–c) and (j) represents the percentage of winged aphids. Values followed by the same letters in (b) and (d) are not significantly different. **, P < 0.01; *, P < 0.05; ns, not significant. Source data are provided as a Source Data file.
