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. 2015 Oct 7;6:8525. doi: 10.1038/ncomms9525

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BG1 transcripts (relative to ubiquitin) in midguts of fourth-instar larvae feeding on (a) Lyc4-containing EV and Lyc4-depleted irGGPPS plants (F_1,10=833.5, P≤0.0001; significant difference (P≤0.05) between means (±s.e.) determined by Fisher's LSD test (one-way ANOVA); n=6) and (b) AD containing water (control), 6 mM Lyc4 or 6 mM RGHGL (F_2,15=5812.13, P≤0.0001; significant differences (P≤0.05) between means (±s.e.) determined by Games Howell test (Welch's ANOVA); n=6). (c) Plant-mediated RNAi: pSOL8 binary vector constructed to express 301 bp MsBG1 dsRNA in N. attenuata and the trophic transfer of dsRNA from plant to larvae. (d) BG1 transcripts (relative to ubiquitin) in various tissues of fourth-instar larvae feeding on EV, irBG1 and irGGPPS plants (midgut: F_2,15=9.53 P≤0.002; hindgut: F_2,15=248, P≤0.0001; significant differences (P≤0.05) between means (±s.e.) determined by Games Howell test (Welch's ANOVA, separately conducted for each tissue); n=6 in all bars). (e) RGHGL formed in vitro enzyme assays containing midgut extracts of fourth-instar EV- and irBG1-feeding larvae and either (1 mM) Lyc4 (F_3,8=104.9, P≤0.0001; significant differences (P≤0.05) between means (±s.e.) determined by Fisher's LSD test (one-way ANOVA); n=3) or 1 mM Lyc4+1 mM BG inhibitor, δ-gluconolactone (n=3). Excretion (% of total Lyc4 ingested) of (f) Lyc4 (F_{1, 25}=13.95, P≤0.001; significant difference (P≤0.05) between means (±s.e.) determined by Fisher's LSD test (one-way ANOVA); n=13 (EV) and 14 (irBG1)) and (g) RGHGL (F_{1, 25}=62.65, P≤0.0001; significant difference (P≤0.05) between means (±s.e.) determined by Fisher's LSD test (one-way ANOVA); n=13 (EV) and 14 (irBG1)) by fourth-instar EV- or irBG1-feeding larvae. (h) Phenotype (in irBG1-feeding larva) and (i) percentage of moulting impairment (significant difference (P≤0.05) determined by Fisher's exact test of frequencies; n=30) and (j) mortality (%) in larvae feeding (for 12 d) on EV or irBG1 plants (significant difference (P≤0.05) determined by Fisher's exact test of frequencies; n=30). ANOVA, analysis of variance; d, days; LSD, least significant difference.