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3 protocols using 2 carboxycinnamic acid

1

Identification of Plant Defense Compounds

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Genistein, genistin, daidzin, daidzein, jasmonic acid (JA), coronatine, benzo(1,2,3) thiadiazole-7-carbothioic acid S-methyl ester (acibenzolar S-methyl), and benzo(1,2,3) thiadiazole-7-carbothioic acid (acibenzolar acid) were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Coumestrol, 2′-hydroxyGenistein, dalbergioidin, phaseollin, phaseollidin, kievitone, and phaseollinisoflavone were obtained and identified according to Durango et al. (2002) . Methyl jasmonate (MeJA), cis-jasmone and 2-carboxy cinnamic acid were from Alfa-Aesar Co. (Ward Hill, MA, USA). Methanol, ethyl acetate, NaOH, silica gel and all other chemicals were purchased from Merck KGaA (Darmstadt, Germany). 1-oxoindanoyl-amino acid conjugates were prepared from 2-carboxy cinnamic acid according to Krumm et al. (1995) (link) and Botero et al. (2021) .
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2

Synthesis of Bioactive Compounds

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L-isoleucine methyl ester, L-leucine methyl ester, 3-(dimethylamino)propylamine, 3-(diethylamino)propylamine, benzylhydrazine, 3-bromobenzaldehyde, 3-bromo-1-propanol, MeJA, palladium on carbon, N,N′-dicyclohexylcarbodiimide (DCC), 1-hydroxybenzotriazole (HOBt), N,N-dimethylformamide (DMF), genistein, daidzein, and coumestrol were purchased from Sigma-Aldrich (St. Louis, MO, USA). 2-carboxycinnamic acid, L-valine methyl ester, glycine methyl ester, and anhydrous aluminum chloride (AlCl3) were obtained from Alfa Aesar Co. (Ward Hill, MA, USA). 2′-hidroxygenistein, dalbergioidin, kievitone, phaseollidin, phaseollinisoflavan, and phaseollin were isolated from a previous work described in [1 (link)]. Dichloromethane (DCM), ethyl acetate (EtOAc), and n-hexane (n-hex) were acquired from Protokimica (Medellín, Colombia) and doubly distilled. Solvents for HPLC were obtained from Merck (Darmstadt, Germany). Ultrapure water (type 1 and 2) was produced by Synergy® UV and Elix Technology (Millipore, Merck) water purification systems (Darmstadt, Germany). 1-oxo-indane-4-carboxylic acid was prepared according to processes described in [9 (link)] and [13 (link)].
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3

Isoflavones Extraction and Identification

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Isoflavones, genistein and daidzein, were purchased from Sigma-Aldrich (St Louis, MO, USA). Dalbergioidin, 2′-hydroxygenistein, coumestrol, phaseollidin, phaseollinisoflavan, kievitone and phaseollin were obtained and identified from a previous work described by Durango et al. [22 ]. Methyl jasmonate (MeJA), palladium on carbon and oxalyl chloride was acquired from Sigma-Aldrich Co. (St Louis, MO, USA), whereas jasmonic acid (JA), anhydrous aluminum chloride (AlCl3) and 2-carboxycinnamic acid were bought in Alfa Aesar Co. (Ward Hill, MA, USA). 1-oxo-4-indanoyl carboxylic acid was prepared by conventional organic reactions [23 (link)].
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