TABLE S2

Interspecific Comparison of Constitutive Ash Phloem Phenolic Chemistry Reveals Compounds Unique to Manchurian Ash, a Species Resistant to Emerald Ash Borer

Justin G.A. Whitehill1,2*, Stephen O. Opiyo3, Jennifer L. Koch4, Daniel A. Herms5, Donald F. Cipollini6, and Pierluigi Bonello1

1Department of Plant Pathology, The Ohio State University, 2021 Coffey Road, Columbus, OH 43210

2Michael Smith Laboratories, University of British Columbia, 301-2185 East Mall, Vancouver, BC, Canada V6T 1Z4

3Molecular and Cellular Imaging Center-Columbus, Ohio Agricultural Research and Development Center, 2021 Coffey Road, Columbus, OH 43210

4Northern Research Station, USDA Forest Service, 359 Main Road, Delaware, OH 43015

5Department of Entomology, The Ohio State University, Ohio Agricultural Research and Development Center, 1680 Madison Ave., Wooster, Ohio 44691

6Department of Biological Sciences, Wright State University, 3640 Colonel Glenn Highway, Dayton, OH 45435

Justin G. A. Whitehill (*)

Michael Smith Laboratories

The University of British Columbia

301-2185 East Mall

Vancouver, BC V6T 1Z4

Canada

E-mail:

Table S2. Contents of individual phenolic compounds and lignin in susceptible white ash cv. ‘Autumn Purple’, green ash cv. ‘Patmore’, green ash seedling, blue ash seedling, European ash seedling, black ash cv. ‘Fallgold’, and the resistant Manchurian ash cv. ‘Mancana’ in samples collected on June 2nd, 2008. Compounds are separated into groups by phenolic compound class, while species are separated into the sections to which they belong within the genus Fraxinus (Wallander 2008). Contents are expressed in mg g-1 FW ± SEM (N = 8). Different letters within a row indicate significantly different means by the protected LSD test (α = 0.05). A solid black line separates black ash data from the other species because it was not part of the original experimental design, but can be visually compared to the other species.

/ Section Melioides / Section Dipetalae / Section Fraxinus /
Peak # / Compound Name / White / Green ‘Patmore’ / Green
Seedling / Blue / European / Manchurian / Black /
Phenolic Acids/Simple Phenolics
1 / Hydroxytyrosol hexoside / 0.2 ± 0.04 b / 0.2 ± 0.1 b / 0.4 ± 0.1 ab / 0.6 ± 0.2 a / 0.5 ± 0.1 ab / 0.6 ± 0.1 a / 0.6 ± 0.1
2 / Vanillic acid hexoside acetate adduct A / NDa / ND / ND / ND / ND / 0.4 ± 0.1 / 0.4 ± 0.1
3 / Tyrosol hexoside / 0.7 ± 0.1 b / 0.6 ± 0.1 b / 0.6 ± 0.1 b / ND / 1.4 ± 0.3 a / 0.4 ±0.1b / ND
4 / Unknown Phenolic Acid 1 / 0.1 ± 0.02 / ND / ND / ND / ND / ND / ND
5 / Vanillic acid hexoside acetate adduct B / ND / ND / 0.3 ± 0.1 / ND / ND / ND / ND
8 / Caffeoyl-quinic acid A / ND / ND / ND / ND / 3.5 ± 0.5 / ND / ND
11 / Tyrosol hexoside pentoside / 0.4 ± 0.1 a / 0.5 ± 0.04 a / 0.4 ± 0.1 a / ND / ND / ND / ND
23 / 3-Caffeoyl-quinic acid / ND / ND / ND / ND / 0.5 ± 0.1 a / 0.3 ± 0.04 a / 0.4 ± 0.02
Coumarins
6 / Fraxetin related compound / ND / ND / ND / ND / ND / ND / 0.9 ± 0.1
7 / Esculetin A / ND / ND / ND / 0.7 ± 0.2 / ND / ND / 0.7 ± 0.02
10 / Esculetin B / ND / ND / ND / 0.6 ± 0.2 / ND / ND / ND
12 / Esculin / ND / ND / ND / 32.9 ± 1.7 a / 0.7 ± 0.1 b / 0.8 ± 0.1 b / 16.3±0.4
13 / Esculetin C / ND / ND / ND / 1.1 ± 0.1 a / 0.1 ± 0.01 b / 0.1 ± 0.01 b / 0.7 ± 0.03
16 / Fraxidin A / ND / ND / ND / ND / 0.6 ± 0.1 a / 0.6 ± 0.1 a / 1.8 ± 0.2
18 / Fraxin / ND / ND / ND / 8.5 ± 1.9 a / 8.1 ± 0.8 a / 11.0 ± 0.6 a / 13.1 ± 0.7
19 / Fraxidin B / ND / ND / ND / 0.3 ± 0.1 b / 0.5 ± 0.1 ab / 0.6 ± 0.04 a / 0.03 ± 0.1
21 / Mandshurin / ND / ND / ND / ND / 0.8 ± 0.04 b / 3.0 ± 0.3 a / 5.7 ± 0.4
22 / Fraxetin / ND / ND / ND / ND / ND / 2.3 ± 0.2 / 4.3 ± 0.13
Monolignols
15 / Syringin / 1.9 ± 0.6 b / 4.4 ± 0.6 ab / 5.6 ± 1.3 a / ND / ND / ND / ND
17 / Monolignol 1 / 0.2 ± 0.1 a / 0.7 ± 0.1 a / 0.9 ± 0.2 a / ND / ND / ND / ND
Lignans
24 / Unknown Lignoid 1 / ND / ND / ND / ND / ND / ND / 1.9 ± 0.2
25 / Pinoresinol dihexoside + 2 H2O / ND / ND / ND / ND / ND / 1.0 ± 0.3 / ND
28 / (+)-1-Hydroxypinoresinol-4’-O-glucoside + 2H2O / ND / ND / ND / 0.6 ± 0.1 / ND / ND / ND
30 / Unknown Lignan 2 / ND / ND / ND / ND / 1.4 ± 0.3 / ND / ND
37 / Pinoresinol A / ND / ND / ND / 2.5 ± 0.5 / ND / ND / ND
39 / Pinoresinol / ND / 1.9 ± 0.2 b / 3.0 ± 0.9 b / ND / 8.3 ± 1.5 a / 8.4 ± 0.4 a / ND
Phenylethanoids
26 / Lugrandoside A / ND / ND / ND / ND / ND / ND / 0.7 ± 0.1
32 / Calceolarioside C / ND / ND / ND / ND / ND / 0.7±0.1 / 1.3 ± 0.3
34 / Forsythoside A O-glucoside - A / 2.2±0.4 a / 2.2±0.2 a / 2.4±0.5 a / ND / ND / ND / ND
40 / β-Methoxylferruginoside / ND / ND / ND / 1.8 ± 0.3 / ND / ND / 3.5 ± 0.5
41 / Forsythoside A O-glucoside - B / 0.8 ± 0.2 / ND / ND / ND / ND / ND / ND
42 / Calceolarioside A / ND / ND / ND / 3.9 ± 0.8 b / 3.1 ± 0.8 b / 11.7 ± 2.3 a / 33.0 ± 2.1
43 / Lugrandoside B / ND / ND / ND / ND / ND / 1.8 ± 0.4 / 5.3 ± 0.4
45 / Verbascoside / 4.5 ± 0.4 a / 3.6 ± 0.8 ab / 1.9 ± 1.1 ab / ND / 2.6 ± 1.1 ab / 1.5 ± 0.3 b / ND
50 / Calceolarioside B / ND / ND / ND / ND / ND / 5.0 ± 0.8 / 12.9 ± 0.7
64 / Verbascoside C / 0.1 ± 0.1 / ND / ND / ND / ND / ND / ND
Secoiridoids
29 / Oleuropeain Related Compound 1 / 2.7 ± 0.2 / ND / ND / ND / ND / ND / ND
27 / Ligustroside A / ND / 1.8 ± 0.2 a / 1.6 ± 0.4 a / ND / ND / ND / ND
31 / Unknown Secoiridoid 1 / ND / ND / ND / 1.2 ± 0.2 / ND / ND / ND
33 / Ligustroside B / ND / ND / ND / 0.7 ± 0.5 / ND / ND / ND
36 / 10-Hydroxyoleuropein / 2.2 ± 0.3 b / 4.6 ± 0.3 a / 4.4 ± 0.9 a / ND / ND / 5.1 ± 0.2 a / 3.9 ± 0.4
38 / Dimethyloleuropein / ND / ND / ND / ND / ND / ND / 4.8 ± 0.7
44 / Oleuropein A / ND / 3.8 ± 0.4 a / 2.8 ± 0.8 a / ND / ND / ND / ND
48 / Oleuropein B / ND / ND / ND / ND / ND / ND / 1.4 ± 0.2
49 / Demethylligstroside / ND / ND / ND / ND / ND / ND / 2.3 ± 0.5
51 / Oleuropein / 2.2 ± 0.3 c / 5.3 ± 1.8 bc / 10.9 ± 3.1 a / ND / 2.8 ± 0.5 c / 8.3 ± 0.9 ab / 8.8 ± 1.4
57 / Ligstroside / 9.0 ± 1.1 a / 8.9 ± 1.1 a / 7.7 ± 1.4 ab / ND / 6.3 ± 0.6 ab / 5.0 ± 0.5 b / ND
Flavonoids
55 / Quercetin diglycoside (rutin)/ Quercetin glucoside (isoquercetin) / ND / ND / 0.7 ± 0.2 / ND / ND / ND / ND
58 / Apigenin glucoside / ND / 0.1 ± 0.01 a / 0.1 ± 0.03 a / ND / ND / ND / ND
61 / Kaempherol galactoside / ND / ND / ND / ND / 0.3 ± 0.1 / ND / ND
65 / Luteolin / ND / ND / 0.1 ± 0.03 / ND / ND / ND / ND
66 / Apigenin / 0.01 ± 0.01 b / 0.1 ± 0.01 b / 0.1 ± 0.03 a / ND / ND / ND / ND
Phenolic Polymer
Lignin / 10.0 ± 1.9 ab / 7.2 ± 0.8 b / 7.6 ± 1.0 b / 11.7 ± 1.9 a / 3.1 ± 0.2 c / 9.0 ± 0.9 ab / 15.1 ± 1.7

aND = Not detected