Facilitation and Inhibition: Changes in Plant Nitrogen and Secondary Metabolites Mediate

1 Supplementary tables (4) and figures (1)

3Table S1 Two-way MANOVA and ANOVAs showing the effects of adults (aboveground)

4and/or larvae (belowground) of a specialist flea beetle Bikasha collaris on tannins in leaves

5and roots of Triadica sebifera. We used Wilks’ lamda to assess significance of predictors in

6 the MANOVA. Significant results are shown in bold.

Adults [AG] Larvae [BG] Adults × Larvae Tissue Compoun df F P F P F P

d MANOVA 8, 48.88 <0.000 58.89 <0.0001 48.33 <0.0001

13 1 Leaves Gallic acid 1, 31.57 <0.000 0.23 0.6372 0.06 0.8034

20 1 Catechin 1, 40.92 <0.000 7.84 0.0111 0.24 0.6264

20 1 Tannic 1, 51.55 <0.000 7.70 0.0117 0.001 0.9689

acid 20 1 Ellagic 1, 30.06 <0.000 11.80 0.0026 0.68 0.4180

acid 20 1 Roots Gallic acid 1, 27.59 <0.000 12.25 0.0023 19.96 0.0002

20 1 Catechin 1, 132.5 <0.000 105.6 <0.0001 132.9 <0.0001

20 6 1 2 9 Tannic 1, 193.7 <0.000 114.6 <0.0001 135.7 <0.0001

acid 20 6 1 3 0 Ellagic 1, 9.69 0.0055 0.33 0.5739 2.67 0.1179

1 1 2 acid 20

3 2 4 8Table S2 Two-way MANOVA and ANOVAs showing the effects of adults (aboveground)

9 and/or larvae (belowground) of a specialist flea beetle Bikasha collaris on flavonoids in

10leaves and roots of Triadica sebifera. We used Wilks’ lamda to assess significance of

11predictors in the MANOVA. Significant results are shown in bold.

Adults [AG] Larvae [BG] Adults ×

Larvae Tissue Compound df F P F P F P MANOVA 10,1 6.2 0.00 5.40 0.0051 6.14 0.003

1 2 28 0 Leaves Quercetin 1,20 6.0 0.02 5.19 0.0339 1.98 0.174

Glycoside 9 27 9 Iso-Quercitrin 1,20 0.55 0.46 17.41 0.0005 4.32 0.050

69 7 Quercetin 1,20 0.00 0.96 3.19 0.0893 1.34 0.261

1 14 0 Kaempferitrin 1,20 19. 0.00 3.15 0.0913 3.23 0.087

56 03 5 Kaempferol 1,20 7.3 0.01 50.00 <0.000 0.00 0.968

0 37 1 1 2 Roots Quercetin 1,20 4.6 0.04 0.09 0.7612 11.0 0.003

Glycoside 3 37 1 4 Iso-Quercitrin 1,20 3.07 0.09 5.75 0.0263 1.42 0.247

53 6 Quercetin 1,20 8.4 0.00 0.52 0.4806 0.26 0.614

6 87 5 Kaempferitrin 1,20 2.34 0.14 0.04 0.8404 0.31 0.580

5 3 6 15 9 Kaempferol 1,20 4.26 0.05 0.41 0.5276 8.59 0.008

7 4 8 13Table S3 Two-way ANOVAs showing the effects of adults (aboveground) and/or larvae

14 (belowground) of a specialist flea beetle Bikasha collaris on tannins, flavonoids, carbon

15 and nitrogen in stems of Triadica sebifera. Significant results are shown in bold.

Adults [AG] Larvae [BG] Adults ×

Larvae Compound df F P F P F P Tannins Gallic acid 1,2 9.72 0.005 3.4 0.0765 5.97 0.024

0 4 9 0 Catechin 1,2 0.65 0.429 1.2 0.2819 5.28 0.032

0 2 2 5 Tannic acid 1,2 10.5 0.004 17. 0.0005 16.9 0.000

0 4 0 45 8 5 Ellagic acid 1,2 0.00 0.953 0.4 0.5189 1.40 0.250

0 3 3 2 Total tannins 1,2 5.03 0.036 9.0 0.0068 13.4 0.001

0 3 9 3 5 Flavonoids Quercetin 1,2 0.05 0.820 0.1 0.6694 7.65 0.011

Glycoside 0 0 9 9 Iso-Quercitrin 1,2 1.32 0.264 1.9 0.1808 0.23 0.638

0 5 2 3 Quercetin 1,2 0.26 0.619 1.3 0.2606 0.41 0.531

0 0 4 6 Kaempferitrin 1,2 0.00 0.952 0.0 0.8751 0.57 0.458

0 1 3 8 Kaempferol 1,2 3.15 0.091 1.6 0.2201 0.04 0.846

0 4 0 3 Total flavonoids 1,2 1.98 0.174 1.4 0.2449 1.52 0.231

9 5 10 0 3 4 9 Carbon 1,2 40.1 <0.00 28. <0.000 31.9 <0.00

0 3 01 17 1 3 01 Nitrogen 1,2 0.08 0.785 0.8 0.3559 3.88 0.062

0 7 9 9

11 6 12 17Table S4 Three-way repeated ANOVAs showing the effects of adults (aboveground) and/or

18 larvae (belowground) of a specialist flea beetle Bikasha collaris on tannins and flavonoids

19 in leaves versus roots (organ) of Triadica sebifera. Significant results are shown in bold.

Tannins Flavonoids Factor df F P F P Larvae [BG] 1,10 47.33 <0.0001 21.53 0.0009 Adults [AG] 1,10 14.03 0.0038 12.30 0.0057 Organ 1,10 30.37 0.0003 79.24 <0.0001 Larvae × Adults 1,10 17.04 0.0021 0.24 0.6334 Larvae × Organ 1,10 0.72 0.4160 19.47 0.0013 Adult × Organ 1,10 157.81 <0.0001 16.36 0.0023 Larvae × Adults × Organ 1,10 19.62 0.0013 1.95 0.1931 21

13 7 14 22Fig. S1 The effects of adults (aboveground) and/or larvae (belowground) of a

23specialist flea beetle Bikasha collaris on secondary chemistry (A-D, tannins; E-I,

24flavonoids) in leaves, stems, and roots of Triadica sebifera. (A) gallic acid, (B)

25catechin, (C) tannic acid, (D) ellagic acid, (E) quercetin-glycoside, (F) iso-quercitrin,

26(G) quercetin, (H) kaempferitrin, and (I) kaempferol. Values are means + 1 SE.

27Herbivory treatment: N = None (no herbivores); A = Adults [AG]; L = Larvae [BG]; B =

28Both [AG&BG].

0.035 A 0.7 B 4 C ) ) ) W

W 0.030 0.6 W D D

g 3 0.025 g 0.5 m m m 0 0 0 0 0 0 1 1 0.020 0.4 / 1 / / g g

g 2 m m ( m ( 0.015

d d i i n i c c h a a 0.010 0.2 c

c 1 c e i i t l n l a n a 0.005 0.1 C a

G T 0.000 0.0 0 N A L B N A L B N A L B N A L B N A L B N A L B N A L B N A L B N A L B Leaf Stem Root Leaf Stem Root Leaf Stem Root

0.16 ) 0.7 0.7

D E ) F W ) D W

W 0.14 D 0.6 g 0.6

g m g 0

0.12 m 0

0.5 0 1 0 0 / 0 1 0.10 g / 1 /

m 0.4 g (

0.08 m ( m e

n i 0.3 0.3 i d s r i

0.06 t i o c c c a r

y 0.2 0.2 l e c

i 0.04 u G g - Q a n l - 0.1 i 0.1 l

0.02 t o E e s

I c 0.00 r 0.0 0.0 e

N A L B N A L B N A L B u N A L B N A L B N A L B N A L B N A L B N A L B Q Leaf Stem Root Leaf Stem Root Leaf Stem Root

0.16 G 0.5 H 0.10 I ) ) ) W

0.14 W D W

D g 0.4 0.08

0.12 g g m m 0 m 0 0 0 0

0.10 1 0 /

0.3 1 0.06 / 1 g / g g 0.08 m ( m

( n l i 0.2 0.04 r 0.06 o n t r i i t r e f e e f c 0.04 p r p m e 0.1 0.02 m e u e

0.02 a Q a K K 0.00 0.0 0.00 N A L B N A L B N A L B N A L B N A L B N A L B N A L B N A L B N A L B 30 Leaf Stem Root Leaf Stem Root Leaf Stem Root

15 8 16