Rapid HPLC screening of jasmonate-induced increases in tobacco alkaloids, phenolics, and diterpene glycosides in Nicotiana attenuata

A rapid, HPLC-based screening procedure for the main classes of secondary m etabolites in Nicotiana attenuata leaves (alkaloids, phenolics, and diterpe ne glycosides) is reported. In a single step, leaves are extracted in aqueo us acidified (0.5% acetic acid) methanol, and the extracted compounds are s eparated by reversed-phase HPLC with an acidic water/acetonitrile gradient in < 30 min. The utility of the method in quantifying changes in the second ary metabolites after methyl jasmonate treatment of the plants, a treatment known to elicit resistance to herbivores in nature, is illustrated. Methyl jasmonate treatment elicited dramatic increases in some secondary metaboli tes (caffeoylputrescine, nicotine, and diterpene glycosides increased 12.5- , 1.4-, and 1.9-fold, respectively) but left others, such as rutin, unchang ed. Such broad-based analytical screens will help characterize environmenta l and genetic changes in secondary metabolite profiles.