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Metabolic response of Chlorella emersonii to the herbicide sulfometuron methyl

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Abstract

Although it is clear that acetohydroxy acid synthase (AHAS; EC 4.1.3.18) is the target for sulfonylurea herbicides such as sulfometuron methyl (SMM), there is considerable uncertainty as to the mechanism(s) by which inhibition of AHAS inhibits or kills cells. We have further studied the mode of action of SMM, and its effects on metabolism and physiology in the unicellular green alga Chlorella emersonii var. emersonii. Addition of SMM to cells synchronized to a cycle of 16 h light-8 h dark showed that they were very sensitive to SMM toxicity in the first 16 h of the cell cycle, during which cell mass, protein and DNA increased. The increase in protein, DNA and chlorophyll was halted rapidly after SMM addition. Sulfometuron methyl prevented cell division even if added late in the light stages, when most of the protein and DNA were already synthesized, but did not affect cell division and autospore release if added after protein and DNA synthesis were complete. This suggests that SMM interferes with processes involved in preparation for division, beyond what would be expected if the cells were starved of the branched-chain amino acids needed as precursors for synthesis of proteins in general. The accumulation of α-ketobutyrate (αKB) in the cells in response to addition of SMM, and its possible role in the growth inhibition, was also investigated (in continually illuminated cultures). Intracellular αKB accumulated rapidly within 30 min of SMM addition, but declined nearly to basal levels in several hours. This paralleled the decrease and subsequent recovery of extractable AHAS activity. Despite this, growth of the algal culture did not recover. We suggest that metabolites formed by misincorporation of αKB in place of α-ketoisovalerate (e.g., in the ketopantoate hydroxymethyl transferase reaction) might be responsible for the persistence of growth inhibition. We note that an important difference between the effect of SMM and that observed with externally added αKB is that the ratio between intracellular αKB and α-ketoisovalerate is expected to be high in the first case, but not necessarily in the second.

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Abbreviations

AHAS:

acetohydroxy acid synthase

BCAA:

branched-chain amino acids

IM:

imidazolinone

αKB:

α-keto-butyrate

SMM:

sulfometuron methyl

SU:

sulfonyl urea

References

  1. Arnon, D.I. (1949) Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol. 24, 1–15

  2. Chaleff, R.S., Mauvais, C.J. (1984) Acetolactate synthase is the site of action of two sulfonylurea herbicides in plants. Science 224, 1443–1445

  3. Chaleff, R.S., Ray, T.B. (1984) Herbicide resistant mutants from tobacco cell cultures. Science 223, 1148–1151

  4. Clayton, D.S., Reynolds, T.L. (1991) Chlorsulfuron effect on protein synthesis and accumulation in cultured root tips of Pisum sativum L. J. Plant Physiol. 137, 337–341

  5. Durner, J., Gailus, V., Boger, P. (1991) New aspects on inhibition of plant acetolactate synthase by chlorsulfuron and imazaquin. Plant Physiol. 95, 1144–1149

  6. Epelbaum, S., Landstein, D., (Malis) Arad, S., Barak, Z., Chipman, D.M., LaRossa, R.A., VanDyk, T.K. (1992) Is the inhibitory effect of the herbicide sulfometuron methyl indeed due to 2-ketobutyrate accumulation? In: Biosynthesis and molecular regulation of amino acids in plants, pp. 51–52, Flores, H., Shanon, J., Singh, B. eds. American Society of Plant Physiologists

  7. Giles, K.M., Mayers, A. (1965) An improved diphenylamine method for the estimation of deoxyribonucleic acid. Nature 206, 93

  8. Hartnett, M.E., Newcomb, J.R., Hodsen, R.C. (1987) Mutations in Chlamydomonas reinhardtii conferring resistance to the herbicide sulfometuron methyl. Plant Physiol. 85, 898–901

  9. Hawkes, T.R., Howard, J.L., Pontin, S.E. (1989) Herbicides that inhibit the biosynthesis of branched chain amino acids. In: Herbicides and plant metabolism (Seminar series), pp. 113–137, Dodge, A. ed, Cambridge University Press, Cambridge

  10. Kishore, G.M., Shah, D.M. (1988) Amino acid biosynthesis inhibitors as herbicides. Annu. Rev. Biochem. 57, 627–663

  11. Landstein, D., Chipman, D.M., (Malis) Arad, S., Barak, Z. (1990) Acetohydroxyacid synthase activity in Chlorella emersonii under auto-and heterotrophic growth conditions. Plant Physiol. bd94, pp614-620

  12. LaRossa, R.A., VanDyk, T.K. (1989) Leaky pantothenate and thiamin mutations of Salmonella typhimurium conferring sulphometuron methyl sensitivity. J. Gen. Microbiol. 135, 2209–2222

  13. LaRossa, R.A., VanDyk, T.K., Smulsky, D.R. (1987) Toxic accumulation of 2KB caused by inhibition of the branched chain amino acid biosynthetic enzyme acetolactate synthase in Salmonella typhimurium. J. Bacteriol. 169, 1372–1378

  14. Lorenzen, H., Hesse, M. (1974) Synchronous cultures. In: Algal physiology and biochemistry, pp. 894, Stewart W.D.P. ed, University of California Press, Berkeley and Los Angeles

  15. Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, H.J. (1951) Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265–275

  16. Miflin, B.J., Cave, P.R. (1972) The control of leucine isoleucine and valine biosynthesis in a range of higher plants. J. Exp. Bot. 23, 511–516

  17. Rhodes, D., Hogan, A.L., Deal, L., Jamieson, G.C., Haworth, P. (1987) Amino acid metabolism of Lemna minor L. II. Responses to chlorsulfuron. Plant Physiol. 84, 775–780

  18. Rost, T.L. (1984) The comparative cell cycle and metabolic effects of chemical treatments on root tip meristems. III. chlorsulfuron. J. Plant Growth Regul. 3, 51–63

  19. Rost, T.L., Reynolds, T. (1985) Reversal of chlorsulfuron-induced inhibition of mitotic entry by isoleucine and valine. Plant Physiol. 77, 481–482

  20. Rost, T.L., Gladish, D., Steffen, J., Robbins, J. (1990) Is there a relationship between branched chain amino acid pool size and cell cycle inhibition in roots treated with imidazolinone herbicides? J. Plant Growth Regul. 9, 227–232

  21. Shaner, D.L., Singh, B.K. (1993) Phytotoxicity of acetohydroxyacid synthase inhibitors is not due to accumulation of 2-ketobutyrate and/or 2-aminobutyrate. Plant Physiol. 103, 1221–1226

  22. Shaner, D.L., Reider, M.L. (1986) Physiological responses of corn (Zea mays) to AC 243,997 in combination with valine, leucine, and isoleucine. Pestic. Biochem. Physiol. 24, 248–257

  23. Shaner, D.L., Anderson, P.C., Stidham, M.A. (1984) Imidazolinones. Potent inhibitors of acetohydroxyacid synthase. Plant Physiol. 76, 545–546

  24. Soeder, C.J., Ried, A., Strotmann, H. (1964) Hemmwirkung von CO2 auf späte Stadien der Zellentwicklung von Chlorella. Beitr. Biol. Pflanz. 46, 150–171

  25. Subramanian, M., Hung, H., Dias, J.M., Miner, V.M., Butler, J.H., Jachettia, J.J. (1990) Properties of mutant acetolactate synthase resistant to triazolopyrimidine sulfonanilide. Plant Physiol. 94, 239–244

  26. VanDyk, T.K., LaRossa, R.A. (1990) Prevention of endogenous 2-ketobutyrate toxicity in Salmonella typhimurium. In: Biosynthesis of branched chain amino acids, pp. 123–130, Barak, Z., Chipman, D.M., Schloss, J.V. eds, VCH, Weinheim

  27. Wallsgrove, R.M. (1990) The biochemistry and genetics of branched chain amino acid biosynthesis in higher plants. In: Biosynthesis of branched chain amino acids, pp. 43–52, Barak, Z., Chipman, D.M., Schloss, J.V. eds. VCH, Weinheim.

  28. Wang, Z.-J., Zaitsu, K., Ohkura, Y. (1988) High performance liquid Chromatographic determination of 2-keto acids in human serum and urine using 1,2-diamino-4,5-methylenedioxybenzene as a precolumn fluorescence derivatization reagent. J. Chromatogr. 430, 223–231

  29. Wanka, F., Mulders, P.F.M. (1967) The effect of light on DNA synthesis and related processes in synchronous cultures of Chlorella. Arch. Mikrobiol. 58, 257–269

  30. Wittenbach, V., Aulabaugh, A., Schloss, J.V. (1991) Herbicidal activity of a ketol-acid reductoisomerase inhibitor. (Abstr) Plant Physiol. 96, Suppl. 951

  31. Yadav, N., McDevitt, R.E., Benard, S., Falco, S.C. (1986) Single amino acid substitutions in the enzyme acetolactate synthase confer resistance to the herbicide sulfometuron methyl. Proc. Natl. Acad. Sci. USA 83, 4418–4422

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Author information

Correspondence to David M. Chipman.

Additional information

This research was supported in part by grant 338/92 from the Israel Science Foundation. It was also supported by the Lily and Sidney Oelbaum Chair in Applied Biochemistry, of which D.M.C. is the incumbent.

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Landstein, D., Epelbaum, S., Arad, S.(. et al. Metabolic response of Chlorella emersonii to the herbicide sulfometuron methyl. Planta 197, 219–224 (1995). https://doi.org/10.1007/BF00202640

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Key words

  • Acetohydroxy acid synthase
  • Acetolactate synthase
  • Ketobutyrate
  • Chlorella
  • Sulfonylurea
  • Synchronized growth