A -GLS Arabidopsis Cuscuta gronovii

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1 -GLS Arabidopsis Cuscuta gronovii internal standard 4MS Wildtype Arabidopsis Cuscuta gronovii base Cuscuta gronovii apex 3MSP internal standard MSP 4MT 8MSO 4MO 1MO Figure S1. Representative HPLC-DAD chromatograms from analysis of glucosinolates in dodder (Cuscuta gronovii) and Arabidopsis thaliana Col- hosts. A. Glucosinolates are not detectable in -GLS mutant Arabidopsis nor in parasites grown on this host, indicating that C. gronovii does not produce glucosinolates endogenously.. In contrast to A, wildtype hosts and their parasites contain eight glucosinolates that we detected in this assay. Glucosinolate levels are generally greater in the parasites than in their hosts. A decline in glucosinolate content is evident between the base and the apices of the parasite vines. Glucosinolate Key: 3MSP = 3-methylsulfinylpropyl-; 4MS = 4-methylsulfinylbutyl-; MSP = -methylsulfinylpentyl-; 4MT = 4-methylthiobutyl-; 8MSO = 8-methylsulfinyloctyl-; = indol-3-ylmethyl-; 4MO = 4-methoxyindol-3-ylmethyl-; 1MO = 1-methoxyindol-3- ylmethyl-

2 Camalexin ng g -1 FW ± s.e N.D. Cuscuta Arabidopsis (infested) Arabidopsis (healthy) Figure S2. Camalexin content in dodder and wildtype Arabidopsis. Parasites were grown from seedlings on flowering wildtype Arabidopsis hosts for three weeks. Samples consisting of entire vines (excluding tissue at point of connection to hosts) and Arabidopsis flower stems (excluding leaves and siliques) were flash-frozen in liquid nitrogen and stored at -8ºC. Then, frozen samples were ground to powder, extracted with 3!l mg -1 cold 7% methanol with 1!M chlorpropamide as internal standard, thermomixed at 7ºC and 13 rpm for 2 min, and centrifuged at 2,8 x g. Supernatants were transferred to glass sample bottles and analyzed by LC-MS/MS with positive ionization. Major ion peaks were used for normalization (chlorpropamide: m/z 139.; camalexin: m/z 142.1). Identification and quantification was based upon serial dilutions of a camalexin standard (Sigma Aldrich). Camalexin in infested and healthy Arabidopsis was not significantly different. Statistics: Mann-Whitney test: W = 33, P =.24, N = 7 replicates per treatment.

3 Leaf Glucosinolates 1 3MSP 4MS MSP 4MT 8MSO aliphat. total Leaf Glucosinolates C Stem Glucosinolates 1 1 3MSP 4MS MSP 4MT 8MSO aliphat. total D Stem Glucosinolates 1.. N.D. N.D. Figure S3. Systemic (non)effects of dodder on glucosinolate levels in Arabidopsis. Dodder infestation of one stem did not induce systemic glucosinolate content changes in non-infested leaves (A and ) or stems (C and D) of the same wildtype Arabidopsis plants. In contrast, dodder infestation induced localized increases of aliphatic and indole glucosinolates (Figure 7). Statistics: Pairs were compared with T-tests, Wilcoxon Sign Rank tests or Exact Permutation T- test according to the properties of the data. No pairs were significant different (all had P >! =.). Glucosinolate Key: 3MSP = 3-methylsulfinylpropyl-; 4MS = 4-methylsulfinylbutyl-; MSP = -methylsulfinylpentyl-; 4MT = 4-methylthiobutyl-; 8MSO = 8-methylsulfinyloctyl-; = indol-3-ylmethyl-

4 Aliphatic glucosinolates a b c a b c wildtype atr1d Indole glucosinolates 1 a a b wildtype b b c atr1d Figure S4. Glucosinolates in dodder and Arabidopsis plants in Myzus persicae population assay. Wildtype and atr1d hosts and parasites are charted together in A and since levels are comparable, whereas -GLS plant data is plotted on a much smaller scale (C). Samples comprised about 2 cm of dodder or Arabidopsis stem tissue immediately distal to aphid cages in the Figure 4 aphid assay. Aphids were caged to either healthy Arabidopsis or dodder. Statistics: N = 13-1 replicates per treatment. A. Factorial ANOVA: Tissue F (2,84) =166, P < 2E- 16; Genotype F (1,84) =.796, P =.37. Tukey tests: v. Cuscuta with aphids P <.1; v. P =.117; v. P <.1.. Ranked factorial ANOVA: Tissue F (2,84) = 6.3, P =.282; Genotype F (1,84) = 63.3, P = 7.E- 12. Tukey tests: Letters above columns indicate statistical grouping at the! =. significance level. C. Aliphatics ANOVA: Tissue F (2,43) = 36.3, P =.76E-1; Tukey tests: v. P <.1; v. P =.337; v. P <.1. Indole Ranked ANOVA: Tissue F (2,43) =.136, P =.873. C Glucosinolates 1. a a b aliphatic -GLS indole

5 Table SI. Tukey tests for amino acids with significant plant*genotype interactions. Arabidopsis tissue Cuscuta tissue -GLS v atr1d -GLS v WT WT v atr1d -GLS v atr1d -GLS v WT WT v atr1d alanine ~ ns ** ns ns ns glutamic acid ns ns ns * ns ns leucine ns ns ns *** * ns Significance test categories: ns P!.1 ~ P <.1 * P <. ** P <.1 *** P <.1