Characterization of nitrogenous solutes in tissues and xylem sap of Leucaena leucocephala

Amino acid profiles of leaf, stem, and root tissues from nodulated and nonnodulated Leucaena leucocephala (Lam.) de Wit plants were determined by gas chromatography-mass spectrometry. High concentrations of mimosine and several other potentially toxic nonprotein amino acids, including pipecolic acid and two isomers of hydroxypipecolic acid, were identified in the tissues. Five metabolites remain unidentified. Of the foliar free amino acid nitrogen, 57-66% was associated with the potentially toxic amino acids. Major constituents in the leaf tissues of nonnodulated plants were mimosine and hydroxypipecolic acid (isomer 1). Mimosine was recovered in both the neutral plus basic and acidic amino acid fractions. Major differences between amino acid profiles of nodulated and nonnodulated roots were the low percentages of asparagine + aspartate (3.6% of the total pool compared to 33% in nodulated plants) and pipecolic acid in nonnodulated roots (1% of the total compared to 12.5% in nodulated plants). A novel plant betaine (dihydroxypipecolic acid betaine) was identified by fast-atom-bombardment mass spectrometry in leaf tissues, albeit at relatively low concentrations (< 1 micro mol per gram fresh weight). Analyses of the xylem sap collected from nodulated plants confirmed that Leucaena is an asparagine transporter, as suggested by the high concentrations of asparagine and the low concentrations of ureides in its root nodules. Amino acid profiles of xylem sap from nonnodulated plants showed extremely low concentrations of asparagine + aspartate (0.12 micro mol ml(-1)), whereas asparagine + aspartate was the major constituent (4.38 micro mol ml(-1)) in the xylem sap of nodulated plants. Two nonprotein amino acids, pipecolic acid and hydroxypipecolic acid, were major constituents of the xylem sap of nodulated and nonnodulated plants, respectively. Three unidentified compounds detected in xylem sap samples from both nodulated and nonnodulated plants did not correspond with any of the peaks characterized from tissue samples.