Arbuscular mycorrhizal fungus-promoted accumulation of two new triterpenoids in cucumber roots

Cucumber (Cucumis sativus L.) roots were analyzed by HPLC and TLC for their levels of secondary metabolites upon inoculation with the arbuscular mycorrhizal fungus, Glomus caledonium. Three compounds in EtOAc extracts from the mycorrhizal roots showed significant increases six weeks after inoculation. These compounds were isolated by column chromatography and determined to be two novel triterpenes, 2beta-hydroxybryonolic acid (2beta,3beta-dihydroxy-D:C-friedoolean-8-en-29-oic acid) and 3beta-bryoferulic acid [3beta-O-trans-ferulyl-D:C-friedooleana-7,9(11)-diene-29-oic acid], and the known triterpene, bryonolic acid, by spectroscopic methods. Time-course experiments showed that the levels of the three terpenoids in cucumber roots were significantly increased by the application of a 53-mum sieving from a soil inoculum of the arbuscular mycorrhizal fungus containing soil microbes but no mycorrhizal fungi, and that mycorrhizal colonization further promoted the terpenoid accumulation. Inoculation with Glomus mosseae also enhanced the accumulation of the triterpenes, whereas no accumulation was observed by inoculating with the fungal pathogen, Fusarium oxysporum f. sp. cucumerinum. 2beta-Hydroxybryonolic acid was also isolated from the roots of melon and watermelon.