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Regulatory effects of exogenous branched-chain amino acids in Nicotiana plumbaginifolia cell suspension cultures

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Abstract

Nicotiana plumbaginifolia suspension cultured cells were grown on medium supplemented with valine, leucine and isoleucine, singly or in combination. The effects of the three branched-chain amino acids on cell growth rate and on the activity of acetohydroxyacid synthase (AHAS), the first enzyme (and the main regulative site) of their biosynthetic pathway, were studied. Results showed that valine and leucine, at concentrations ranging from 10−4 to 10−3 M, inhibit growth, and at higher doses (from 10−2 to 10−1 M) AHAS activity. Growth, but not AHAS activity, was affected also by isoleucine. The addition of ammonium succinate to the culture medium, in order to counteract a possible general inhibitory effect of these compounds on nitrogen metabolism, relieved only partially their cytotoxicity. Feeding cells with equimolar mixtures of the three amino acids resulted in a minor but reproducible decrease in AHAS level, which was proportional to the dose. A similar result was obtained also on N. plumbaginifolia seedlings, suggesting that in this species a modulation of enzyme level could play a role in controlling the flow of metabolites through the pathway.

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Abbreviations

AHAS:

acetohydroxyacid synthase

BCAA:

branched-chain amino acids

FAD:

flavin adenine dinucleotide

GS:

glutamine synthetase

TPP:

thiamine pyrophosphate

References

  1. Barak Z, Kogan N and Gollop N and Chipman DM (1990) Importance of AHAS isozymes in branched chain amino acid biosynthesis. In: Barak Z, Chipman DM, Schloss JV (eds) Biosynthesis of branched chain amino acids, pp91–107, VCH, Weinheim

    Google Scholar 

  2. Behrend J, Mateles R (1975) Nitrogen metabolism in plant cell suspension culture. I. Effect of amino acids on growth. Plant Physiol 56: 584–589

    Google Scholar 

  3. Bonner CA, Rodrigues AM, Miller JA, Jensen RA (1992) Amino acids are generally growth inhibitors of Nicotiana silvestris in tissue culture. Physiol Plant 84: 319–328

    Google Scholar 

  4. Borstlap AC (1981) Interaction between the branched-chain amino acids in the growth of Spirodela polyrhiza. Planta 151: 314–319

    Google Scholar 

  5. Bourgin JP (1976) Valine-induced inhibition of growth of haploid tobacco protoplasts and its reversal by isoleucine. Z Naturforsch 31: 337–338

    Google Scholar 

  6. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254

    Google Scholar 

  7. Bryan JK (1980) Synthesis of the aspartate family and branched-chain amino acids. In: Miflin BJ (ed) The biochemistry of plants: a comprehensive treatise, Vol. V, Amino acids and derivatives, pp403–452. Academic Press, New York

    Google Scholar 

  8. De Felice M, Lago CT, Squires CH, Calvo JM (1982) Acetohydroxy acid synthase isoenzymes of Escherichia coli K12 and Salmonella typhimurium. Ann Microbiol (Paris) 133A: 251–256

    Google Scholar 

  9. Dougall K (1970) Threonine deaminase from Paul's scarlet rose tissue cultures. Phytochemistry 9: 959–964

    Google Scholar 

  10. Durner J, Böger P (1990) Oligomeric forms of plant acetolactate synthase depend on flavin adenine dinucleotide. Plant Physiol 93: 1027–1031

    Google Scholar 

  11. Eoyang L, Silverman PM (1984) Purification and subunit composition of acetohydroxyacid synthase I from Escherichia coli K12. J Bacteriol 157: 184–189

    Google Scholar 

  12. Forlani G, Riccardi G, De Rossi E, De Felice M (1991) Biochemical evidence for multiple forms of acetohydroxyacid synthase in Spirulina platensis. Arch Microbiol 155: 298–302

    Google Scholar 

  13. Gamborg OL, Millar RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50: 151–158

    Google Scholar 

  14. Hayzer DJ, Moses V (1978) The enzymes of proline biosynthesis in Escherichia coli. Biochem J 173: 219–228

    Google Scholar 

  15. Heimer YM, Filner P (1970) Regulation of the nitrate assimilation pathway of cultured tobacco cells. Biochim Biophys Acta 215: 152–165

    Google Scholar 

  16. Holmberg S, Petersen JGL (1988) Regulation of isoleucinevaline biosynthesis in Saccharomyces cerevisiae. Curr Genet 13: 207–218

    Google Scholar 

  17. Miflin BJ (1969) A technique for the sterile culture of germinating barley embryos. J Exp Bot 20: 805–809

    Google Scholar 

  18. Miflin BJ (1969) The inhibitory effects of various amino acids on the growth of barley seedlings. J Exp Bot 20: 810–819

    Google Scholar 

  19. Miflin BJ (1971) Cooperative feedback control of barley acetohydroxyacid synthetase by leucine, isoleucine and valine. Arch Biochem Biophys 146: 542–550

    Google Scholar 

  20. Miflin BJ, Cave PR (1972) The control of leucine, isoleucine and valine biosynthesis in a range of higher plants. J Exp Bot 23: 511–516

    Google Scholar 

  21. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio-assay with tobacco tissue cultures. Physiol Plant 15: 473–497

    Google Scholar 

  22. Nielsen E, Forlani G, Cella R, Parisi B (1986) Biochemical characterization of the natural resistance of rice to the proline analogue azetidin-2-carboxylic acid. Plant Sci 44: 147–154

    Google Scholar 

  23. Nielsen E, Rollo F, Parisi B, Cella R, Sala F (1979) Genetic markers in cultured plant cells: differential sensitivities to amethopterin, azetidin-2-carboxylic acid and hydroxyurea. Plant Sci Lett 15: 113–125

    Google Scholar 

  24. Pinto JEBP, Dyer WE, Weller SC, Herrmann KM (1988) Glyphosate induces 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase in potato (Solanum tuberosum L.) cells grown in suspension culture. Plant Physiol 87: 891–893

    Google Scholar 

  25. Rognes SE, Wallsgrove RM, Kueh JSH, Bright SWJ (1986) Effects of exogenous amino acids on growth and activity of four aspartate pathway enzymes in barley. Plant Sci 43: 45–50

    Google Scholar 

  26. Sakano K (1979) Derepression and repression of lysinesensitive aspartokinase during in vitro culture of carrot root tissue. Plant Physiol 63: 583–585

    Google Scholar 

  27. Squires CT, Levinthal M, De Felice M (1981) A role for threonine deaminase in the regulation of α-acetolactate biosynthesis in Escherichia coli K12. J Gen Microbiol 127: 19–26

    Google Scholar 

  28. Thompson GA, Datko AH, Mudd SH, Giovanelli J (1982) Methionine biosynthesis in Lemna. Plant Physiol 69: 1077–1083

    Google Scholar 

  29. Umbarger HE (1983) The biosynthesis of isoleucine and valine and its regulation. In: Herrmann KM and Somerville RL (eds) Amino acids: biosynthesis and genetic regulation, pp245–266. Addison-Wesley, Reading, MA

    Google Scholar 

  30. Umbarger HE (1987) Biosynthesis of the branched-chain amino acids. In: Neidhardt FC, Ingraham JL, Low BL, Magasanick B, Schaechter M and Umbarger HE (eds) Escherichia coli and Salmonella typhimurium. Cellular and molecular biology, Vol. I, pp352–367. American Society for Microbiology, Washington

    Google Scholar 

  31. Wallsgrove RM (1990) The biochemistry and genetics of branched chain amino acid biosynthesis in higher plants. In: Barak Z, Chipman DM and Schloss JV (eds) Biosynthesis of branched chain amino acids, pp43–51. VCH, Weinheim

    Google Scholar 

  32. Wallsgrove RM, Risiott R, King J, Bright SJW (1986) Biochemical characterisation of Nicotiana plumbaginifolia auxotrophs that require branched-chain amino acids. Plant Cell Rep 3: 223–226

    Google Scholar 

  33. Westerfeld WW (1945) A colorimetric determination of blood acetoin. J Biol Chem 161: 495–502

    Google Scholar 

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Authors and Affiliations

  1. Department of Genetics and Microbiology, University of Pavia, I-27100, Pavia, Italy

    G. Forlani, M. C. Suardi, B. Parisi & E. Nielsen

Authors
  1. G. Forlani
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  2. M. C. Suardi
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  3. B. Parisi
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  4. E. Nielsen
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Forlani, G., Suardi, M.C., Parisi, B. et al. Regulatory effects of exogenous branched-chain amino acids in Nicotiana plumbaginifolia cell suspension cultures. Plant Growth Regul 14, 203–209 (1994). https://doi.org/10.1007/BF00024794

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  • DOI: https://doi.org/10.1007/BF00024794

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