Online Resource 1 s2

Online Resource 1

TABLE 1 DETAILS OF STATISTICAL TESTS OF PLANT PARAMETERS FROM BRASSICA OLERACEA

PLANTS WITHOUT HERBIVORES (CONTROL), WITH APHIDS, NEMATODES (NEMA), OR BOTH.

This is supporting information for Table 2 in the main text. General test: lme(parameter ~ Aphids + Nema, random=~1|Block/Diversity, method = ”ML”). Number of observations = 84. For phloem data the number of observations was 83 since one sample was lost during processing. Harvest time was included as covariate for the phloem data. Glucosinolates in the phloem were measured in 4 of the 7 blocks.

Transformation t-value t-value Df Covariate

Aphids Nema Shoot biomass log -1.45 -4.71 61 Root biomass log 0.08 0.15 61 Water % shoot no 1.35 -4.09 61 N % shoot log 0.49 -5.07 61 N % root log 0.29 2.68 61 N shoot allocation no -0.71 -7.08 61 Carbon shoot log -2.54 0.16 61 Carbon root no -0.02 0.36 61 C:N shoot no -0.94 5.14 61 C:N root yes -0.95 -6.45 61

Sugars Leaf Total no -0.57 2.83 61 Glucose no -0.81 3.24 61 Fructose no -0.49 2.43 61 Sucrose no 0.66 -0.28 61 Sugars Phloem Total log 0.44 -4.60 59 Harvest time

Amino acids Leaf Total no -0.48 0.98 61 Arginine no -0.42 -4.10 61 Leucine log 0.08 -0.48 61 Threonine no -0.85 2.48 61 Tryptophan no 2.06 2.40 61 Valine no -0.54 1.57 61 Amino acids Phloem Total log 0.81 -4.30 59 Harvest time

Glucosinolates Leaf Total log 0.33 -1.28 61 IBE log 0.09 -1.89 61 PRO log + 1 -0.07 -1.47 61 RAPH log + 1 -0.26 -0.32 61 SIN log -0.04 -1.77 61 GNA log + 1 -0.74 -3.14 61 TROP log + 1 0.70 0.62 61 4OH log 0.96 0.56 61 GBC log -0.53 -0.35 61 4MeOH GBC log 2.67 -0.82 61 Glucosinolates Phloem Total log 1.75 -0.22 31 Harvest time Online Resource 2

FIG 1 Concentrations of aminoacids (a) and sugars (b) in the phloem changed in relation to harvest time and day. On harvest day 1 the measured concentrations were much lower than on harvest day 2. From morning till midday, the concentrations declined on both harvest days. Since pots were harvested per block, containing all treatments, detection of treatment effects was not affected by the temporal fluctuations. In the statistical analyses block was used as random factor. Online Resource 3

TABLE 2 P-VALUES AND F-VALUE OF ANCOVAS TO TEST WHETHER THE RELATIONSHIP

BETWEEN APHID DOUBLING TIME AND PLANT PARAMETERS DEPENDED ON NEMATODE

PRESENCE.

Plant parameter Aphid Nematod Adt * Np Adt * Np Pearson Correlation between Aphid doubling e Doubling times and plant parameters time presence (Adt) (Np) F1,29 - Nema + Nema P P P F-value slope R2 slope R2 Water % shoot 0.495 0.972 0.313 1.056 Nitrogen % shoot 0.020 0.609 0.036 4.817 Glucose 0.046 0.286 0.045 4.381 Fructose 0.060 0.355 0.083 3.230 Sucrose 0.437 0.836 0.564 0.341 Valine 0.004 0.073 0.009 7.867 -0.08 0.37 -0.04 0.01 Threonine 0.001 0.036 0.002 12.035 -0.08 0.37 0.01 0.00 Tryptophan 0.263 0.458 0.465 0.549 Arginine 0.000 0.000 0.000 21.530 -0.37 0.53 0.60 0.03 Leucine 0.183 0.438 0.269 1.273 Glucoiberin 0.010 0.004 0.006 8.836 -2.26 0.41 1.56 0.02 Progoitrin 0.009 0.003 0.008 8.229 -5.35 0.30 6.89 0.03 Glucoraphanin 0.006 0.010 0.007 8.319 -6.09 0.43 2.11 0.01 Sinigrin 0.005 0.006 0.008 8.038 -4.87 0.32 0.11 0.00 Gluconapin 0.001 0.001 0.002 11.562 -20.04 0.41 0.67 0.00 Glucotropaeolin 0.440 0.937 0.876 0.025 4-hydroxyglucobrassicin 0.467 0.962 0.545 0.375 4-methoxyglucobrassicin 0.768 0.819 0.567 0.336 Phloem total sugars 0.769 0.899 0.776 0.082 Phloem total amino acids 0.997 0.861 0.563 0.342

General model: plant parameter~aphid doubling time * nematode presence. N = 15 for plants without nematodes,

N = 18 for plants with nematodes. For significant interactions between aphid doubling time and nematode presence, we show the Pearson correlation coefficients between aphid doubling time and plant parameter in presence and absence of nematodes. Significant P-values after False Discovery Rate control (Benjamini and

Hochberg 2000) are indicated in bold.

REFERENCES

BENJAMINI, Y. and HOCHBERG, Y. 2000. On the adaptive control of the false discovery rate in multiple testing

with independent statistics. J. Educ. Behav. Stat. 25:60-83. Online Resource 4

For the complete dataset including aphid doubling times from plants with and without nematodes, the 1st latent structure explained 31% of the variation in aphid doubling time which was similar to models using permutations of the aphid doubling times. Hence we concluded that plant parameters did not explain any significant amount of variation in aphid doubling time using data from plants with and without nematodes. When separate PLSR models were made for the datasets in absence and presence of nematodes, no model was found for the aphid doubling times on plants with nematodes. On the other hand, in absence of nematodes the first component of the model explained 59% and the second component another 18% of the variation in aphid doubling times (2 PLS components (R2Xcum 52%, R2Ycum 77%, Figure below). The variables with the most influence were similar to those showing up in the ANCOVAs in Online Resource 1: the amino acids arginine, valine, threonine and the glucosinolates sinigrin, glucoiberin, progoiterin, glucoraphanin, gluconapin were all negatively correlated with aphid doubling time.

FIG 2 Multivariate data analysis of the relationship between plant metabolites and aphid doubling times by

PLSR (partial least squares regression). Loading plot of the first two components showing the contribution of primary (open diamonds) and secondary (closed squares) metabolites to explaining aphid doubling times. %C =

% carbon in shoot, %N = % nitrogen in shoot, glu = glucose, fru = fructose, suc = sucrose, sug_phl = total sugars in phloem, aa_phl = total amino acids in phloem, arg = arginine, leu = leucine, thr = threonine, try = tryptophan, val = valine, IBE = glucoiberin, PRO = progoitrin, RAPH = glucoraphanin, SIN = sinigrin, GNA = gluconapin,

TROP = glucotropaeolin, 4OH = 4-hydroxyglucobrassicin, GBC = glucobrassicin, 4-MeOH = 4- methoxyglucobrassicin, DoubTime = aphid doubling time.