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© 1989 Oxford University Press

RESEARCH-ARTICLE

Inhibition of NaCI-Induced Proline Biosynthesis by Exogenous Proline in Halophilic Distichlis spicata Suspension Cultures

J. W. HEYSER1, D DE BRUIN, M L. KINCAID, R. Y. JOHNSON, M. M. RODRIGUEZ and N J ROBINSON 2

Genetics Group (LS-3), Los Alamos National Laboratory Los Alamos, New Mexico 87545, USA

1 To whom correspondence should be addressed.

Cell suspensions of the halophilic grass, Distichlis spicata L., accumulate up to 50 µmol g–1 fresh weight proline when transferred to medium containing 260 mol m–3 NaCl. The metabolism of L-[5-13C]-glutamic acid to L-[5-13C]-proline was followed by 13C-NMR analysis to detect 13C-enrichment of carbon-5 of proline. Chemical and enzymatic analysis of cell extracts and media were used to determine L-proline and L-glutamic acid concentrations, respectively. 13C-Glutamic acid metabolism to proline was greater in cells adapting to 260 mol m-3 NaCl compared with cells not adapted to NaCl, or grown long-term on NaCl medium. Metabolism of 13C-glutamic acid (5.0 mol m–3) to proline was inhibited when 5.0 mol m–3 proline was also included in the growth medium. Exogenous proline did not sufficiently reduce glutamic acid transport to account for this inhibition. The inhibition was thus either regulated by feedback-inhibition of existing enzymes, or by repression of transcription and/or translation of genes encoding enzymes involved in proline biosynthesis.

Key words: Halophyte, tissue culture, proline, NMR, Distichlis spicata


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