US 2002O132021A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2002/0132021 A1 Raskin et al. (43) Pub. Date: Sep. 19, 2002 (54) ELICITED PLANT PRODUCTS (60) Provisional application No. 60/045,220, filed on Apr. 30, 1997. Provisional application No. 60/050,441, (76) Inventors: Ilya Raskin, Manalapan, NJ (US); filed on Jun. 27, 1997. Alexander Poulev, Highland Park, NJ (US) Publication Classification Correspondence Address: (51) Int. Cl. ................................................. A61K 35/78 MARSHALL, GERSTEIN & BORUN (52) U.S. Cl. .............................................................. 424/773 6300 SEARS TOWER 233 SOUTHWACKER CHICAGO, IL 60606-6357 (US) (57) ABSTRACT (21) Appl. No.: 09/929,328 Intact living plants or plant parts are contacted with water to extract from the plant or plant part exuded chemical com (22) Filed: Aug. 13, 2001 pounds, with the extracted chemical compounds Subse quently being recovered from the water. The plant is treated Related U.S. Application Data with an elicitor or inducer to initiate or increase production of a chemical compound. The roots may be harvested for (63) Continuation-in-part of application No. 09/130,185, recovery of the chemical compounds. Valuable Substances filed on Aug. 6, 1998, now abandoned. Continuation exuded from or onto a plant Surface, Such as a plant cuticle in-part of application No. 09/203,772, filed on Jun. or the root of a plant, can be identified as biologically active. 23, 1998, now abandoned, which is a continuation Libraries of Substances exuded or Secreted from various in-part of application No. 09/067,836, filed on Apr. plant Species can be elicited or induced to produce one or 28, 1998, now abandoned. more of Such Substances. Patent Application Publication Sep.19, 2002. Sheet 1 of 27 US 2002/0132021 A1 3. 5 6 7 10 21 24 25 37 38 40 42 45 47 55 0.4 O0.23 O. 1. 3 5 6 7 10 21 24 25 37 38 40 42 45 47 55 Treatment Figure 1 Patent Application Publication Sep.19, 2002 Sheet 2 of 27 US 2002/0132021 A1 0.01 0. OO 7 5 0. O05 0.0025 3 4 5 6 7 10 21 24 25 37 42 45 55 0.3 0.25 O. 2 O 5 O. 3 4 5 6 7 10 21 24 25 37 42 45 55 Treatment Figure 2 Patent Application Publication Sep.19, 2002 Sheet 3 of 27 US 2002/0132021 A1 O. O. Phaseolus vulgaris 0.20 icotiana tabacun 0.60 20.40 0.2 O. O. 0.80 Phytolacca anericana 0.40 W COO - Lupinus luteus 0.00 O. 60 Lactuca satilva a 0.40 0.20 OOO ... 00 Lupinus polyphyllus 2 OSO 0.00 OOO 20 x Patent Application Publication Sep. 19, 2002 Sheet 4 of 27 US 2002/0132021 A1 O. O. C. SO control ro.ao 0.20 0.00 30.50 0.80 2 0.20 l G O. Co 0.00 2009 tautas Figure 4 Patent Application Publication Sep.19, 2002 Sheet 5 of 27 US 2002/0132021 A1 Control 0.00 00000 0.00 O. O.O. 2000 000 OOOO Arachidonic acid (7.5 mM) 0.00 OOO 20.00 3.00 and | \ M. AgNO3 (1.0 M) o VW-M Y---JuJ 0.00 0.00 cites OO2OO H2O2 (52) 2 OOOOO 0.00 OOO 20.00 3000 Mirtas Effect of elicitation on the chemical composition of root exudates of Brassica iuncea. HPLC-profiles with UV detection at 254 nm. Figure 5 Patent Application Publication Sep.19, 2002 Sheet 6 of 27 US 2002/0132021 A1 Control 0.10 O 0.00 OO 20.00 Kautes AgNO3 (1.0 mM) Bo. 10 GOO 0.00 20.00 aute Wy Arachidonic acid (7.5 mM) B 0.10 . - - re. ----- 0.00 0.00 2000 Jastaonic acid (1.66 nM) OOO to do 20.00 titute ofEffect Datura of aetel.elicitation on the chemical composition of root exudates HPLC-profiles with UW detection at 254 nm. Figure 6 Patent Application Publication Sep.19. 2002. Sheet 7 of 27 US 2002/0132021 A1 O. O.200 \ Control V 0.0000.00 0.00 Ys-i-TiY. 2000 so,00 2 0.0200 0000 0.00 Ox 2000 0.00 Kicutes 0.0200 AgNO3 (1.0 a.M) A. | - - - A. 3. ... -- ~~ - - - - - - 0.00 O. Oxo 2000 30.00 Carted ofEffect Lupinus of elicitationpolyphyllus on the chemical composition of root exudates HPLC-profiles with UV detection at 254 nm. Figure 7 Patent Application Publication Sep.19, 2002 Sheet 8 of 27 US 2002/0132021 A1 t Control 0,000 OOO s 20. o sited - Arachidonic acid (7.5 M) a 0.0500 OOO 0.00 0.00 23. so Cite AgNO3 (1.0 m) :- - - , ------------------ o ob 200 0.00 edita Effect of elicitation on the chemical composition of root exudates of Melilotus credicaginoides. HPLC-profiles with UV detection at 254 inn. Figure 8 Patent Application Publication Sep.19, 2002 Sheet 9 of 27 US 2002/0132021 A1 o,000 HO phase 0.0200 0.0000 O.00 o, too 2000 30.06 COO 0.00 2000 30.00 0.00 MeOH/MeCl phase 0.0200 Oxo O. Oro OOO 2000 3000 sta Chemical diversity in different extraction solvents. Root extracts from Solanum melongena (eggplant). HPLC-profiles with UV detection at 254 nm. Figure 9 Patent Application Publication Sep.19, 2002 Sheet 10 of 27 US 2002/0132021 A1 OC200 0000 HO phase 00000 0.00 0.00 20.00 3.00 heritas o EtOAC 0000 phase tood T-- - a VY -----" " 000 OOO 20.00 30.00 Cata OO200 MeOH/MeCl 2 0000 phase 0.0000 O00 OOO 20.00 30.00 cuta Chemical diversity in different extraction solvents. Root extracts from Solanum melongena (eggplant), elicited with 1 mM Salicylic acid. HPLC-profiles with UV detection at 254 nm. Figure 10 Patent Application Publication Sep.19, 2002 Sheet 11 of 27 US 2002/0132021 A1 0.200 HO phase s 0.100 O00 OOO 0.00 20.00 30.00 tae 200 EtOAC ..ice phase OOOO OO 0.00 29CO 30.00 Certas 0200 o.100 MeOH/MeCl phase OOOO OO 0.00 200 2000 autas Chemical diversity in different extraction solvents. Root extracts from Daucus Carota (carrot), elicited with 1 mM AgNO. HPLC-profiles with UV detection at 254 nm. Figure 11 Patent Application Publication Sep.19. 2002. Sheet 12 of 27 US 2002/0132021 A1 eco .azoo Control O.co. 0.00 0.00 O. O io.ozoo000 1 mM AgNO. 8000 COO i.e. M : st-w histas o.O.300 1 mM e Arachidonic io.ozoo acid Oood 0.00 0.00 2000 300 aths old 2 mM io.ozoo Methyl Ooloo jasmonate 0.00 lo) 2000 3.0 irrata Effect of elicitation on chemical diversity of root extracts. EtOAC phases of extracts from Glycyne max (soybean). eTotalion - Elicitor Current peak of chromatograms scanned from 70 m/z to 400 m/z. Figure 12 Patent Application Publication Sep.19, 2002. Sheet 13 of 27 US 2002/0132021 A1 0.0200 0.000 Control 0.0000 o 80 20,00 0.00 Crata 1 mM AgNO 3 OO 0. 0.00 0.00 20.00 300 0.020 0.3 M w ACetic 0.000 acid oxo 0.00 500 2. Oc trated O29 0.75 mM Arachidonic acid 000 O.0 2006 3000 heate Effect of elicitation on chemical diversity of root extracts. MeOH/MeClaphases of extracts from Daucus carota (carrot). HPLC-profiles with UV detection at 254 nm. Figure 13 Patent Application Publication Sep.19, 2002 Sheet 14 of 27 US 2002/0132021 A1 0.000 0.040 Control D.00 0.0000 0.00 O 29.00 3000 Cratas C. CO 1 mM AgNO, R O. d. doo f s -...- lated 0.3 M OOOC Aceticacid 2 OSOC - OOOOO O o hittas2000 3CO 2.2 mM Olood Arachidonicacid 20. Osoo 00000 COO 1000 20.00 30.00 brutad Effect of elicitation on chemical diversity of root extracts. EtOAC phases of extracts from Daucus Carota (carrot). HPLC-profiles with UV detection at 254 nm. Figure 14 Patent Application Publication Sep.19, 2002. Sheet 15 of 27 US 2002/0132021 A1 0.0500 control 0.000 0.00 OGO 2000 3000 lateau 1 mM AgNO a 0.0soo 00000 OOC OOO 2008 900 artites - ser Methyl R0.0500 jasmonate 00000 O.00 0.09 29.00 300 estab 2.2 mM Arachidonic o.Osoo acid 00000 0.00 0.00 2000 3.00 stad Effect of elicitation on chemical diversity of root extracts. EtOAC phases of extracts from Lycopersicon aesculentum (tomato). HPLC-profiles with UV detection at 254 nm. Figure 15 Patent Application Publication Sep.19, 2002. Sheet 16 of 27 US 2002/0132021 A1 does Control O. Cado 000 O) 0.00 20.80 soo beats irradiation of shoots for 6 hours 0.200 prers Mru-m-on-- r -n 0.00 O. O. 2.0 3000 birtas irradiation of roots O200 for 6 hours OOOOO 0.00 OOO 20. O 30 do birutae Effect of UV irradiation on chemical diversity of root extracts. EtOAC phases of extracts from Lupinus polyphylus (lupine). HPLC-profiles with UV detection at 254 nm. Figure 16 Patent Application Publication Sep.19, 2002 Sheet 17 of 27 US 2002/0132021 A1 1513 1515 1517 519 521 1523 Patent Application Publication Sep.19, 2002. Sheet 18 of 27 US 2002/0132021 A1 1781 1783 1785 1787 1789 179 Patent Application Publication Sep.19, 2002. Sheet 19 of 27 US 2002/0132021 A1 1883 1885 1887 escareer ran s -e o Patent Application Publication Sep.19, 2002.