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Figure S2: Levels (mean ? SE) of different secondary metabolites in wild-type (WT) Nicotiana attenuata and 2 independently transformed lines (1 & 2) silenced for putrescine N-methyl transferase (pmt), protease inhibitor (pi), or both.(A) nicotine (B) anatabine, (C) anabasine,(D) caffeoylputrescine,(E) chlorogenic acid, and (F) rutin.

Figure S3: Constitutive (white bars) and induced (black bars) levels (mean ? SE) of different secondary metabolites in wild-type (WT) Nicotiana attenuata and 2 independently transformed lines (1 & 2) silenced for putrescine N-methyl transferase (pmt), protease inhibitor (pi), or both. Levels of (A) jasmonic acid conjugates to isoleucine and leucine JA-Ile + JA-Leu and (B) SA (N = 3) were determined in leaves of untreated control plants and plants elicited with M. sexta oral secretion 30 min prior to being harvested for analysis.(C) Levels of an abundant diterpene glycoside in unelicited and methyl jasmonate (MeJA)-elicited plants. Plants were hydroponically grown in the climate chamber. T-tests did not detect significant differences between each transformed line and WT (P < 0.05).

Table S1: Comparisons (unpaired t-tests; a adjusted according to sequential Bonferroni procedure) of levels of secondary metabolites between wild type (WT) Nicotiana attenuata and each of 2 independently transformed lines (1 & 2) silenced for either putrescine N-methyl transferase (pmt), protease inhibitor (pi), or both. Plants were hydroponically grown in the climate chamber and phytohormones (jasmonic acid ? JA, salicylic acid ? SA, and JA conjugates) were analyzed in unelicited plants and plants that were elicited with Manduca sexta oral secretions 30 min prior to analysis. All other metabolites were analyzed in control plants and plants that were treated with methyl jasmonate 4 days prior to analysis.

Table S2: Comparisons (unpaired t-tests; a adjusted according to sequential Bonferroni procedure) of levels of secondary metabolites between wild type (WT) N. attenuata and each of 2 independently transformed lines (1 & 2) silenced for either putrescine N-methyl transferase (pmt), protease inhibitor (pi), or both. Plants were grown in the glasshouse under same conditions as the Spodoptera exigua assay and secondary metabolites were analyzed in control plants and plants that were treated with jasmonic acid conjugated to isoleucine 4 days before.

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ELE+1045Figures+S1.doc249KSupporting info item
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ELE+1045TableS1.doc53KSupporting info item
ELE+1045TableS2.doc45KSupporting info item
ELE1045+Figure+S2.doc45KSupporting info item

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