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Fig. S1 Leaf area (mean ± SE) lost to attack from (A) different herbivore taxa over the growing season and (B) Lepidoptera at their peak abundance for accessions of Nicotiana attenuata from Arizona (AZ) and Utah (UT) planted into a natural Utah population in 2005. AZ and UT lines were arranged in pairs which were matched to produce trypsin protease inhibitors (TPI) and nicotine (+/+), only nicotine (+/−), or neither nicotine nor TPI (−/−) (via stable transformation; N = 20). (A) Plants were caged from days 3–15 post-planting. (B) Leaf area lost (bars and primary Y-axis) and percentage of AZ and UT plants attacked (line charts and secondary Y-axis) by Lepidoptera are shown for the period of the greatest amount of lepidopteran herbivory (day 37 post planting). Most damage from Lepidoptera was caused by Manduca sexta larvae applied to measure predation rates.

Table S1 Secondary metabolite production in VIGS plants. a,bDifferent letters within a column indicate significant differences at P < 0.05 as determined in the case of TPIs by t-tests between VIGS vectors (ev or lox) within treatments (before or after feeding), and in the case of nicotine, chlorogenic acid, rutin, and the most prominent diterpene glycoside (DTG) by two-way ANOVAs across all lines within a treatment (factors VIGS vector and line) followed by Fisher’s PLSD tests. *Marginal effects of VIGS vector on TPI levels, P = 0.06; on nicotine levels, P = 0.15; on chlorogenic acid levels, P = 0.06 with a significant interaction between line and VIGS vector, P < 0.01.

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