Electronic Supplementary Material (ESI) for Natural Product Reports. This journal is © The Royal Society of Chemistry 2020 SUPPLEMENTARY INFORMATION Figure S1 Figure S2 Figure S3 Figure S4 Table S1 Table S2 Legends of the figures Figure S1: Biosynthetic pathways to aromatic amino acids and malonyl-CoA from glucose. Abbreviations used: DAHP, 2-keto-3-deoxy-D-arabino-heptulosonate-7-phosphate; ACC, acetyl-CoA carboxylase complex, CM1, chorismate mutase 1; PPA-AT, prephenate aminotransferase; ADT, arogenate dehydratase; ADH, arogenate dehydrogenase Figure S2: Putative relationships between 8-8’ dimeric naturally-occurring stilbenes (according to Stephenson’s group29) Figure S3: Examples of stilbene tetramers resulting from the 3-8’ condensation of e-viniferin. The two tetramers, vitisin A and vitisin B isolated from grapevine root extracts result from the 3-8’ coupling of two oxidized molecules of (-)--viniferin (according to Stephenson’s group29) Figure S4: Hypothetical mechanism for the formation of a stilbene heptamer. The stilbene heptamer pauciflorol D is formed by condensation between the resveratrol 8-10’ tetramer, vaticaphenol A, and an 8-8’ quinone trimer formed by the coupling of three resveratrol hydroxyphenyl radicals C (according to Stephenson’s group29) Pentose Phosphate GLUCOSE Glycolysis Pathway Pathway Erythrose-4-phosphate Phosphoenolpyruvate DAHP synthase Shikimate Pathway Pyruvate DAHP Pyruvate Dehydrogenase Complex Acetyl-CoA COOH ACC CO2 O O HO OH HO SCoA OH Malonyl CoA Shikimate COOH O COOH OH Chorismate Chorismate mutase 1 CM1 COOH COOH O HO Prephenate Arogenate Pathway Prephenate aminotransferase PPA-AT COOH COOH NH2 HO Arogenate Arogenate dehydratase Arogenate dehydrogenase ADT ADH COOH COOH NH2 NH2 HO Tyrosine Phenylalanine Figure S1 HO OH HO Phenanthrenic ring OH HO H OH OH HO H HO H h -2H+ H OH OH H H OH OH OH OH OH (-)-Quadrangularin A OH OH (-)-Parthenocissin A (+)-Laetevirenol A HO HO H H O H O H H HO -2H+ HO H H OH -H2O H OH OH OH O HO OH Vitisinol A Restrytisol A Figure S2 HO HO H H 3-8' COUPLING HO O O HO HO H OH H OH OH HRP / H2O2 A OH OH OH OH OH OH H B HO H OH HO H OH O OH O A H H HO H H H H O O 2 x -Viniferin HO HO O HO Hypothetical Intermediate Vitisin B (Tetramer) B HO H O HO H OH OH OH HO H H OH HO H O OH H OH Vitisin A (Tetramer) Figure S3 HEPTAMER FORMATION OH HO HO OH B1 C2 OH HO H H HO HO [O] B2 OH HO D HO H 2 OH O H OH H H H HO C2 H H HO H OH A1 A2 O D1 H H O OH H OH O HO H Vaticaphenol A HO H -Viniferin HO H OH HO H O OH OH HO O HO H OH E1 OH OH G2 HO H G1 H OH OH [O] H HO H H H E 2 H O OH H HO F2 H OH H F1 OH H HO OH O 102 OH 3 x Resveratrol HO OH Hypothetical Intermediate Pauciflorol D (Stilbene heptamer) Figure S4 Table S1 Antifungal activity of stilbene monomers and oligomers towards various microorganisms Tested stilbenes Microorganisms/structures Inhibiting activity References Cladosporium cuccumerinum (spores) Botrytis cinerea (spores) Resveratrol ED > 870 M 17 Pyricularia oryzae (spores) 50 Plasmopara viticola (zoospore motility) B. cinerea (mycelium) ED50 : 438 M Resveratrol P. oryzae (mycelium) ED50 : 276 M 17 Fusarium oxysporum (mycelium) ED50 : 88 M Resveratrol B. cinerea (spores) ED50 : 390 M 117 Fungal growth Stereum hirsutum inhibition at 490 M Fomitiporia punctata Resveratrol Ineffective or 283 Libertella blepharis stimulator on fungal Phaeomoniella chlamydospora growth at 490 M Phaeoacremonium aleophilum 57.2% growth decrease at 48 M Yeast 16.1% growth Resveratrol Penicillium expansum decrease at 48 M 284 Aspergillus niger 36.4% growth decrease at 48 M Table S1 Antifungal activity of stilbene monomers and oligomers towards various microorganisms (continued) P. viticola ED50 : 192 M (zoospore motility) Resveratrol 282 P. viticola ED50 : 145 M (disease development) Resveratrol B. cinerea (spores) ED50 : 438 M 280 P. viticola Resveratrol ED : 500 M 286 (zoospore motility) 100 P. viticola Resveratrol ED : 484 M 287 (sporulation) 50 Resveratrol B. cinerea (spores) ED50 : 1153 M 281 B. cinerea (spores) ED50 : 70 M Pterostilbene P. oryzae (spores) ED50 : 35 M 112 P. viticola (sporangia release) ED50 : 17.6 M Pterostilbene P. viticola ED50 : 9 M 288 (zoospore motility) Pterostilbene B. cinerea (spores) ED50 : 70 M 289 Pterostilbene B. cinerea (spores) ED50 : 78 M 117 Pterostilbene Esca disease-associated fungi Inhibition of fungal 283 growth (39 to 390 M) Table S1 Antifungal activity of stilbene monomers and oligomers towards various microorganisms (continued) P. viticola Pterostilbene ED : 14.6 M 282 (zoospore motility) 50 Piceatannol ED50 : 254 M Isorhapontigenin ED : 116 M Development of downy mildew 50 290 Astringin ED50 : 705 M Isorhapontin ED50 : 272 M -viniferin B. cinerea (spores) ED : 220 M 50 288 -viniferin B. cinerea (mycelium growth) ED50 : 230 M -viniferin P. viticola (sporulation) ED : 12.7 M 50 282,287 -viniferin P. viticola (sporulation) ED50 : 70 M Glucosylateddimers Piceasides I-L ED50 : 96-147 M Development of downy mildew 290 Ampelopsin A ED50 : 934 M Stilbene tetramers Hopeaphenol ED50 : 18 M Miyabenol C ED : 40 M Development of downy mildew 50 287 Vitisin A ED50 : 20 M Vitisin B ED50 : 12 M Table S2 Metabolic engineering of stilbene pathways in yeast Strain used for Incorporated genes Substrate Stilbenes produced References transformation 4CL from Populus Saccharomyces trichocarpa X Populus p-coumarate Resveratrol (1.45 μg/L) 394 cerevisiae FY23 deltoides, STS from Vitis vinifera 4CL from Nicotiana S.cerevisiae tabacum, STS from V. p-coumarate Resveratrol (5.8 mg/L) 395 CEN.PK113-3b vinifera TAL from Rhodobacter sphaeroides, 4CL::STS- S.cerevisiae 4CL from Arabidopsis p-coumarate Resveratrol (5.25 mg/L) 398 WAT11 thaliana and STS from V. vinifera TAL from Rhodotorula Yarrowia glutinis, 4CL from Tyrosine Resveratrol (1.46 mg/L) 408 lipolytica Streptomyces coelicolor, STS from V. vinifera PAL and CPR from P. S. cerevisiae trichocarpia x P. YPH499 deltoides, C4H and 4CL Phenylalanine Resveratrol (0.31 mg/L) 403 from Glycine max, STS from V. vinifera S. cerevisiae 4CL from A. thaliana, STS p-coumarate Resveratrol (391 mg/L) CEN.PK2–1 from V. vinifera 397 4CL from A. thaliana, STS S. cerevisiae from A. hypogea, ∆PAD1 p-coumarate Resveratrol (3.1 mg/L) 396 W303-1A in S. cerevisiae TAL from R. sphaeroides, S. cerevisiae 4CL::STS, 4CL from A. Tyrosine Resveratrol (3.1 mg/L) 406 WAT11 thaliana and STS from V. vinifera, araE from E. coli Table S2 Metabolic engineering of stilbene pathways in yeast (continued) 4CL::STS, 4CL from A. S. cerevisiae thaliana and STS from V. Tyrosine Resveratrol (14.4 mg/L) 402 WAT11 vinifera PAL from Rhodosporidium S. cerevisiae toruloides, C4H and 4CL Tyrosine Resveratrol (5.8 mg/L) 407 W303-1A from A. thaliana, STS from A. hypogea, ACC1 from S. cerevisiae TAL from Herpetosiphon aurianticus, 4CL from S. cerevisiae Arabidopsis thaliana and Glucose/ Resveratrol (415.65 with 386 CEN.PK102-5B STS from V. vinifera, Ethanol glucose/531.41 mg/L ScARO4K229L, ScARO7G141S, with ethanol) ACCS659A, S1157A PAL, C4H, ATR2 and 4CL from A. thaliana, STS from V. vinifera, ACS S. cerevisiae from Salmonella enterica, CEN.PK102-5B Resveratrol (800 mg/L) 387 AroL from E. coli, CYB5 Glucose Pinostilbene (5.5 mg/L) from S. cerevisiae Pterostilbene SbROMT from Sorghum (34.93 mg/L) bicolor/VvROMT from V. vinifera, ∆ARO10 in S. cerevisiae Abbreviations used: PAL, phenylalanine ammonia lyase, TAL, tyrosine ammonia lyase, C4H, cinnamate-4-hydroxylase; 4CL, p-coumaroyl-CoA ligase; STS, stilbene synthase; ACC, acetyl-CoA carboxylase complex; ROMT, resveratrol-O- methyltransferase; ARO10, gene encoding the phenylpyruvate decarboxylase; ScARO4K229L and ScARO7G141S genes encoding feedback-inhibition resistant versions of DAHP synthase and chorismate mutase, respectively; araE, arabinose- H+-transporter; PAD1 gene encoding the phenyl acrylic acid decarboxylase; CPR, cytochrome P450 reductases; aroL, shikimate kinase 2..