Differential metabolic response of narrow leafed lupine (Lupinus angustifolius) leaves to infection with Colletotrichum lupini

Flavones and isoflavones are a major group of phenolic secondary metabolites which occur in leaves of narrow leafed lupine (Lupinus angustifolius) either as free aglycones or in a form of glycosides and malonyl-glycosides. Profiles of phenolic compounds in leaves of seedlings infected with anthracnose causing fungus Colletotrichum lupini were compared to those of healthy plants. A HPLC with diode array UV detector was used as the analytical method and identification of these secondary metabolites was confirmed with a HPLC/MSn instrument. Isomers of several target compounds differing in the glycosilation and/or malonylation pattern were detected in the studied samples. However, the application of standard HPLC with C18 columns resulted in the co-elution of several glyconjugates in single chromatographic peaks whereas for isoflavonoid aglycones complete resolution was achieved. Lupine plants grown in a greenhouse were either sprayed with the C. lupini spore suspension or the suspension was spotted on to wounded leaves. Profiles of the isoflavones were altered in result to infection with both methods. In particular, the concentration of isoflavone free aglycones detected in extracts from diseased plants was substantially increased in all of the studied samples. However, the pattern of these compounds depended on the age of lupine leaves as well as on the method of infection. Synthesis of luteone and 2'-hydroxygenistein was enhanced in the youngest leaves of plants sprayed with spores as well as in wound-infected leaves. Wighteone synthesis was induced mainly in older leaves of plants sprayed with the spore suspension.