Temperature treatment / Water stress treatment / Population / Time (weeks) / N / SE / C:N / SE / Fiber / SE / Lignin / SE
25:18 / Water stress / BFS / 4 / 3.4 / 0.1 / 13.7 / 0.5 / 24.7 / 1.5 / 9.8 / 1.4
25:18 / No water stress / BFS / 4 / 2.9 / 0.1 / 15.6 / 0.5 / 25.6 / 1.4 / 10.6 / 1.4
25:18 / Water stress / CED / 4 / 3.6 / 0.1 / 12.6 / 0.4 / 27.2 / 1.3 / 14.1 / 1.3
25:18 / No water stress / CED / 4 / 2.9 / 0.1 / 15.6 / 0.4 / 26.7 / 1.3 / 11.4 / 1.3
25:18 / Water stress / KMP / 4 / 4.1 / 0.1 / 11.2 / 0.4 / 34.0 / 1.3 / 20.0 / 1.3
25:18 / No water stress / KMP / 4 / 3.1 / 0.1 / 14.5 / 0.4 / 28.2 / 1.3 / 12.7 / 1.3
25:18 / Water stress / MWH / 4 / 3.9 / 0.1 / 11.9 / 0.6 / 30.4 / 1.5 / 15.7 / 1.5
25:18 / No water stress / MWH / 4 / 3.0 / 0.1 / 15.0 / 0.6 / 27.4 / 1.5 / 11.9 / 1.5
25:18 / Water stress / TRU / 4 / 3.9 / 0.1 / 11.9 / 0.4 / 31.7 / 1.3 / 16.8 / 1.3
25:18 / No water stress / TRU / 4 / 2.9 / 0.1 / 15.8 / 0.4 / 34.3 / 1.3 / 16.5 / 1.3
30:23 / Water stress / BFS / 4 / 3.6 / 0.1 / 12.7 / 0.5 / 28.7 / 1.4 / 13.4 / 1.3
30:23 / No water stress / BFS / 4 / 3.3 / 0.1 / 13.8 / 0.5 / 30.5 / 1.4 / 14.5 / 1.3
30:23 / Water stress / CED / 4 / 4.0 / 0.1 / 11.6 / 0.4 / 32.2 / 1.3 / 16.4 / 1.3
30:23 / No water stress / CED / 4 / 3.2 / 0.1 / 14.6 / 0.4 / 33.3 / 1.3 / 17.0 / 1.3
30:23 / Water stress / KMP / 4 / 3.9 / 0.1 / 11.9 / 0.4 / 30.5 / 1.3 / 14.2 / 1.3
30:23 / No water stress / KMP / 4 / 3.4 / 0.1 / 13.5 / 0.4 / 29.6 / 1.3 / 13.7 / 1.3
30:23 / Water stress / MWH / 4 / 3.7 / 0.1 / 12.3 / 0.6 / 32.8 / 1.6 / 17.6 / 1.5
30:23 / No water stress / MWH / 4 / 3.2 / 0.1 / 14.3 / 0.6 / 35.6 / 1.5 / 20.6 / 1.4
30:23 / Water stress / TRU / 4 / 3.8 / 0.1 / 12.1 / 0.4 / 32.6 / 1.3 / 16.8 / 1.3
30:23 / No water stress / TRU / 4 / 3.5 / 0.1 / 13.3 / 0.4 / 34.5 / 1.3 / 18.0 / 1.3
25:18 / Water stress / BFS / 8 / 2.9 / 0.2 / 16.3 / 0.6 / 31.8 / 1.6 / 15.7 / 1.6
25:18 / No water stress / BFS / 8 / 2.8 / 0.1 / 16.9 / 0.6 / 34.1 / 1.6 / 16.3 / 1.5
25:18 / Water stress / CED / 8 / 3.2 / 0.1 / 14.6 / 0.5 / 32.6 / 1.4 / 17.2 / 1.3
25:18 / No water stress / CED / 8 / 2.9 / 0.1 / 16.1 / 0.5 / 32.4 / 1.4 / 15.9 / 1.3
25:18 / Water stress / KMP / 8 / 3.3 / 0.1 / 14.1 / 0.5 / 30.7 / 1.5 / 15.8 / 1.4
25:18 / No water stress / KMP / 8 / 3.0 / 0.1 / 15.8 / 0.5 / 30.5 / 1.4 / 14.4 / 1.3
25:18 / Water stress / MWH / 8 / 3.5 / 0.2 / 13.3 / 0.7 / 32.1 / 1.8 / 16.8 / 1.7
25:18 / No water stress / MWH / 8 / 2.7 / 0.2 / 17.6 / 0.8 / 31.8 / 1.9 / 14.1 / 1.8
25:18 / Water stress / TRU / 8 / 4.0 / 0.1 / 11.9 / 0.5 / 36.1 / 1.4 / 20.2 / 1.4
25:18 / No water stress / TRU / 8 / 2.7 / 0.1 / 17.2 / 0.5 / 34.3 / 1.4 / 17.3 / 1.4
30:23 / Water stress / BFS / 8 / 4.1 / 0.1 / 11.6 / 0.6 / 32.4 / 1.5 / 15.0 / 1.5
30:23 / No water stress / BFS / 8 / 3.1 / 0.1 / 15.4 / 0.5 / 33.7 / 1.5 / 16.3 / 1.4
30:23 / Water stress / CED / 8 / 3.4 / 0.1 / 13.9 / 0.5 / 35.4 / 1.4 / 16.6 / 1.4
30:23 / No water stress / CED / 8 / 3.1 / 0.1 / 15.1 / 0.5 / 32.8 / 1.4 / 15.5 / 1.3
30:23 / Water stress / KMP / 8 / 3.7 / 0.1 / 12.8 / 0.5 / 36.4 / 1.4 / 18.8 / 1.3
30:23 / No water stress / KMP / 8 / 3.4 / 0.1 / 13.8 / 0.5 / 33.6 / 1.4 / 16.7 / 1.3
30:23 / Water stress / MWH / 8 / 3.8 / 0.2 / 12.8 / 0.7 / 35.6 / 1.7 / 16.0 / 1.6
30:23 / No water stress / MWH / 8 / 3.3 / 0.1 / 14.3 / 0.6 / 35.8 / 1.6 / 17.5 / 1.5
30:23 / Water stress / TRU / 8 / 3.8 / 0.1 / 12.7 / 0.5 / 36.5 / 1.5 / 18.2 / 1.4
30:23 / No water stress / TRU / 8 / 3.5 / 0.1 / 13.8 / 0.5 / 35.9 / 1.4 / 16.7 / 1.3

Supporting Information Table 1 Foliar nitrogen (N), fiber, lignin, and cardenolide (CG) concentrations (% dry mass), C:N ratios, leaf mass per area (mg cm-1)and water content (NDWI) from all possible combinations of temperature and water treatments and plant population. SE, standard error.

Supporting Information Table 1 (cont.)

Temperature treatment / Water stress treatment / Population / Time (weeks) / LMA / SE / CG / SE / NDWI / SE
25:18 / Water stress / BFS / 4 / 43.6 / 5.6 / 1.5 / 0.2 / 0.035 / 0.005
25:18 / No water stress / BFS / 4 / 51.1 / 5.4 / 1.3 / 0.2 / 0.040 / 0.004
25:18 / Water stress / CED / 4 / 53.5 / 4.9 / 2.0 / 0.1 / 0.042 / 0.004
25:18 / No water stress / CED / 4 / 51.4 / 4.9 / 1.5 / 0.1 / 0.033 / 0.004
25:18 / Water stress / KMP / 4 / 51.4 / 5.0 / 2.1 / 0.1 / 0.029 / 0.004
25:18 / No water stress / KMP / 4 / 56.6 / 4.9 / 1.1 / 0.1 / 0.035 / 0.004
25:18 / Water stress / MWH / 4 / 52.3 / 5.8 / 2.3 / 0.2 / 0.033 / 0.005
25:18 / No water stress / MWH / 4 / 60.0 / 5.8 / 1.2 / 0.2 / 0.028 / 0.005
25:18 / Water stress / TRU / 4 / 63.3 / 4.9 / 2.0 / 0.1 / 0.030 / 0.004
25:18 / No water stress / TRU / 4 / 74.3 / 4.9 / 1.5 / 0.1 / 0.032 / 0.004
30:23 / Water stress / BFS / 4 / 31.1 / 5.2 / 1.9 / 0.2 / 0.026 / 0.004
30:23 / No water stress / BFS / 4 / 46.1 / 5.2 / 2.0 / 0.2 / 0.037 / 0.004
30:23 / Water stress / CED / 4 / 37.8 / 4.9 / 2.0 / 0.1 / 0.024 / 0.004
30:23 / No water stress / CED / 4 / 47.6 / 5.0 / 2.3 / 0.1 / 0.047 / 0.004
30:23 / Water stress / KMP / 4 / 34.7 / 4.9 / 2.1 / 0.1 / 0.026 / 0.004
30:23 / No water stress / KMP / 4 / 31.5 / 4.9 / 2.3 / 0.1 / 0.041 / 0.004
30:23 / Water stress / MWH / 4 / 41.1 / 5.9 / 1.7 / 0.2 / 0.026 / 0.005
30:23 / No water stress / MWH / 4 / 60.5 / 5.7 / 2.3 / 0.2 / 0.052 / 0.005
30:23 / Water stress / TRU / 4 / 41.9 / 5.0 / 2.3 / 0.1 / 0.040 / 0.004
30:23 / No water stress / TRU / 4 / 46.3 / 4.9 / 2.1 / 0.1 / 0.051 / 0.004
25:18 / Water stress / BFS / 8 / 75.6 / 6.3 / 1.8 / 0.2 / 0.034 / 0.005
25:18 / No water stress / BFS / 8 / 77.2 / 5.9 / 1.9 / 0.2 / 0.044 / 0.005
25:18 / Water stress / CED / 8 / 81.0 / 5.3 / 1.9 / 0.2 / 0.040 / 0.004
25:18 / No water stress / CED / 8 / 69.1 / 5.2 / 1.8 / 0.2 / 0.042 / 0.004
25:18 / Water stress / KMP / 8 / 66.6 / 5.6 / 1.9 / 0.2 / 0.034 / 0.005
25:18 / No water stress / KMP / 8 / 63.2 / 5.2 / 1.9 / 0.2 / 0.035 / 0.004
25:18 / Water stress / MWH / 8 / 66.0 / 6.8 / 2.1 / 0.2 / 0.029 / 0.005
25:18 / No water stress / MWH / 8 / 83.3 / 7.3 / 1.6 / 0.2 / 0.034 / 0.006
25:18 / Water stress / TRU / 8 / 86.1 / 5.3 / 2.3 / 0.2 / 0.026 / 0.004
25:18 / No water stress / TRU / 8 / 88.6 / 5.3 / 2.0 / 0.2 / 0.044 / 0.004
30:23 / Water stress / BFS / 8 / 48.4 / 5.8 / 2.3 / 0.2 / 0.023 / 0.005
30:23 / No water stress / BFS / 8 / 39.9 / 5.6 / 1.9 / 0.2 / 0.028 / 0.005
30:23 / Water stress / CED / 8 / 78.1 / 5.3 / 1.8 / 0.2 / 0.025 / 0.004
30:23 / No water stress / CED / 8 / 45.2 / 5.2 / 2.0 / 0.2 / 0.031 / 0.004
30:23 / Water stress / KMP / 8 / 69.3 / 5.2 / 1.8 / 0.2 / 0.034 / 0.004
30:23 / No water stress / KMP / 8 / 44.6 / 5.2 / 1.9 / 0.2 / 0.035 / 0.004
30:23 / Water stress / MWH / 8 / 64.7 / 6.5 / 2.0 / 0.2 / 0.031 / 0.005
30:23 / No water stress / MWH / 8 / 58.3 / 6.0 / 1.7 / 0.2 / 0.025 / 0.005
30:23 / Water stress / TRU / 8 / 82.7 / 5.5 / 2.1 / 0.2 / 0.040 / 0.004
30:23 / No water stress / TRU / 8 / 57.9 / 5.2 / 1.8 / 0.2 / 0.040 / 0.004

Supporting Information Table 2 Summary of F and Pvalues for the effects of temperature, water stress, plant population source, time, and their interactions on leaf chemical and morphological traits and water content. P values <0.05 are bolded and P values 0.05 <P <0.10 are italicized. C:N, ratio of carbon to nitrogen; NDWI, normalized differential water index; LMA, leaf mass per area.

Nitrogen / C:N / NDWI / Fiber / Lignin / LMA / Cardenolides
Treatments and interactions / df / F / P / df / F / P / df / F / P / df / F / P / df / F / P / df / F / P / df / F / P
Temperature / 1,2 / 24.1 / 0.034 / 1,2 / 19.4 / 0.042 / 1,2 / 0.1 / 0.737 / 1,2 / 44.6 / 0.015 / 1,2 / 33.2 / 0.014 / 1,2 / 29.4 / 0.029 / 1,2 / 3.7 / 0.193
Water stress / 1,2 / 61.9 / 0.014 / 1,2 / 72.5 / 0.010 / 1,2 / 22.4 / 0.042 / 1,2 / 0.0 / 0.901 / 1,2 / 0.6 / 0.519 / 1,2 / 0.1 / 0.842 / 1,2 / 10.8 / 0.067
Temperature x water stress / 1,2 / 2.0 / 0.285 / 1,2 / 4.3 / 0.163 / 1,2 / 0.6 / 0.513 / 1,2 / 0.1 / 0.768 / 1,2 / 1.1 / 0.403 / 1,2 / 1.5 / 0.349 / 1,2 / 20.1 / 0.035
Population / 4,13 / 6.6 / 0.003 / 4,12 / 7.4 / 0.003 / 4,17 / 1.8 / 0.181 / 4,16 / 9.0 / <0.001 / 4,16 / 6.2 / 0.003 / 4,18 / 10.9 / <0.001 / 4,19 / 1.7 / 0.194
Temperature x population / 4,13 / 2.0 / 0.155 / 4,12 / 2.5 / 0.102 / 4,17 / 6.0 / 0.004 / 4,16 / 1.9 / 0.160 / 4,16 / 1.8 / 0.183 / 4,18 / 1.6 / 0.230 / 4,19 / 0.8 / 0.555
Water stress x population / 413 / 1.4 / 0.279 / 4,12 / 1.8 / 0.193 / 4,17 / 0.2 / 0.952 / 4,16 / 1.9 / 0.157 / 4,16 / 1.7 / 0.203 / 4,18 / 3.2 / 0.036 / 4,19 / 1.2 / 0.333
Temperature x water stress x population / 4,13 / 6.2 / 0.004 / 4,12 / 6.5 / 0.005 / 4,17 / 1.9 / 0.151 / 4,16 / 0.4 / 0.811 / 4,16 / 0.7 / 0.586 / 4,18 / 0.7 / 0.588 / 4,19 / 2.9 / 0.051
Time / 1,675 / 27.4 / <0.001 / 1,675 / 66.0 / <0.001 / 1,676 / 5.1 / 0.025 / 1,678 / 146.2 / <0.001 / 1,678 / 30.1 / <0.001 / 1,679 / 293.3 / <0.001 / 1,674 / 1.2 / 0.284
Temperature x time / 1,675 / 16.4 / <0.001 / 1,675 / 13.1 / <0.001 / 1,676 / 34.2 / <0.001 / 1,678 / 2.4 / 0.121 / 1,678 / 12.7 / <0.001 / 1,679 / 1.8 / 0.185 / 1,674 / 40.4 / <0.001
Water stress x time / 1,675 / 3.1 / 0.080 / 1,675 / 0.6 / 0.453 / 1,676 / 9.0 / 0.003 / 1,678 / 1.1 / 0.298 / 1,677 / 0.1 / 0.743 / 1,679 / 58.8 / <0.001 / 1,675 / 2.0 / 0.160
Temperature x water stress x time / 1,675 / 3.6 / 0.057 / 1,675 / 0.3 / 0.554 / 1,676 / 62.1 / <0.001 / 1,678 / 10.1 / 0.001 / 1,677 / 6.3 / 0.012 / 1,679 / 30.7 / <0.001 / 1,675 / 37.9 / <0.001
Population x time / 4,675 / 3.2 / 0.012 / 4,674 / 1.3 / 0.274 / 4,676 / 2.4 / 0.049 / 4,678 / 5.4 / <0.001 / 4,678 / 4.6 / 0.001 / 4,678 / 1.6 / 0.168 / 4,674 / 5.4 / <0.001
Temperature x population x time / 4,675 / 3.8 / 0.004 / 4,674 / 2.6 / 0.034 / 4,676 / 1.7 / 0.148 / 4,678 / 15.6 / <0.001 / 4,678 / 15.3 / <0.001 / 4,678 / 9.2 / <0.001 / 4,674 / 1.6 / 0.175
Water stress x population x time / 4,675 / 5.9 / <0.001 / 4,674 / 4.3 / 0.002 / 4,676 / 3.2 / 0.013 / 4,678 / 4.1 / 0.002 / 4,677 / 3.1 / 0.015 / 4,678 / 2.0 / 0.099 / 4,674 / 2.1 / 0.081
Temperature x water stress x population x time / 4,675 / 3.5 / 0.007 / 4,674 / 3.5 / 0.007 / 4,676 / 3.8 / 0.004 / 4,678 / 3.0 / 0.019 / 4,677 / 2.3 / 0.062 / 4,678 / 1.1 / 0.355 / 4,674 / 2.2 / 0.063

Numerator and denominator degrees of freedom (df: numerator, denominator) were calculated using the Satterthwaite approximation.

Supporting Information Table 3 Foliar latex (mg) and trichomes (mm2) from all possible combinations of temperature and water treatments and plant population.SE, standard error.

Temperature treatment / Water stress treatment / Population / Latex / SE / Trichomes / SE
25:18 / Water stress / BFS / 0.54 / 0.02 / 5.7 / 1.9
25:18 / No water stress / BFS / 0.93 / 0.02 / 13.6 / 1.7
25:18 / Water stress / CED / 1.50 / 0.02 / 9.6 / 1.7
25:18 / No water stress / CED / 1.54 / 0.02 / 9.4 / 1.6
25:18 / Water stress / KMP / 1.29 / 0.02 / 13.2 / 1.7
25:18 / No water stress / KMP / 0.63 / 0.02 / 9.7 / 2.1
25:18 / Water stress / MWH / 1.98 / 0.02 / 10.2 / 2.2
25:18 / No water stress / MWH / 1.91 / 0.02 / 20.7 / 1.8
25:18 / Water stress / TRU / 0.44 / 0.02 / 11.1 / 1.8
25:18 / No water stress / TRU / 1.04 / 0.02 / 15.8 / 1.7
30:23 / Water stress / BFS / 0.51 / 0.02 / 9.5 / 1.7
30:23 / No water stress / BFS / 1.52 / 0.02 / 16.5 / 1.8
30:23 / Water stress / CED / 1.98 / 0.02 / 13.8 / 1.6
30:23 / No water stress / CED / 1.88 / 0.02 / 14.8 / 1.6
30:23 / Water stress / KMP / 1.18 / 0.02 / 14.2 / 1.5
30:23 / No water stress / KMP / 1.33 / 0.02 / 13.1 / 1.6
30:23 / Water stress / MWH / 1.84 / 0.02 / 13.2 / 2.4
30:23 / No water stress / MWH / 2.17 / 0.02 / 17.3 / 1.8
30:23 / Water stress / TRU / 0.54 / 0.02 / 13.6 / 1.6
30:23 / No water stress / TRU / 1.07 / 0.02 / 16.7 / 1.6

Supporting Information Table 4 Summary of F and Pvalues for the effects of temperature, water stress, plant population source, and their interactions on leaf trichome densities and latex production. P values <0.05 are bolded and P values 0.05 <P <0.10 are italicized.

Treatments and interactions / Trichomes / Latex
df / F / P / df / F / P
Temperature / 1,2 / 3.1 / 0.213 / 1,2 / 2.2 / 0.272
Water stress / 1,2 / 9.7 / 0.079 / 1,2 / 21.6 / 0.043
Temperature x water stress / 1,2 / 0.3 / 0.646 / 1,2 / 9.4 / 0.092
Population / 4,178 / 4.2 / 0.002 / 4,179 / 44.3 / <0.001
Temperature x population / 4,178 / 1.5 / 0.206 / 4,179 / 1.2 / 0.295
Water stress x population / 4,178 / 8.5 / <0.001 / 4,178 / 7.0 / <0.001
Temperature x water stress x population / 4,178 / 1.0 / 0.396 / 4,178 / 1.9 / 0.108

Numerator and denominator degrees of freedom (df: numerator, denominator) were calculated using the Satterthwaite approximation

Supporting Information Figure 1 Model diagnostic statistics generated through the evaluation of 500 random subsets, split 70-30 for calibration and validation, respectively, of the data set from Couture et al. (2013) amended with an additional 63 samples from the low-end of the calibration range (< 1.0 µg mg-1) for cardenolide prediction. Modeling approach followed Couture et al. (2013). Total number of samples used to build the model was 143, wavelength range used was 400-2400, and model was built using partial least squares regression. Top left: Observed vs. predicted (± SD) cardenolide concentrations from the calibration coefficients generated from the 500 iterations of the data set. Solid black line is best fit; dashed gray line is 1:1 line. The remaining boxes depict the 95 percentile range for the R2(top right), root mean square error (RMSE, bottom left), and bias (bottom right) from the 500 permutations of the model for calibration (Cal.) and validation (Val.).

Supporting Information Figure 2 Absolute growth of five different populations of A. syriaca exposed to ambient or elevated temperatures experiencing no water stress (black bars) or periodic water stress (gray bars).

Supporting Information Figure 3 Absolute growth of monarch larvae feeding on five different plant populations exposed to ambient or elevated temperatures experiencing no water stress (black bars) or periodic water stress (gray bars).