, Volume 179, Issue 2, pp 228–234 | Cite as

Localization of enzymes of assimilatory sulfate reduction in pea roots

  • C. Brunold
  • M. Suter


The localization of enzymes of assimilatory sulfate reduction was examined in roots of 5-d-old pea (Pisum sativum L.) seedlings. During an 8-h period, roots of intact plants incorporated more label from 35SO42-in the nutrient solution into the amino-acid and protein fractions than shoots. Excised roots and roots of intact plants assimilated comparable amounts of radioactivity from 35SO42-into the amino-acid and protein fractions during a 1-h period, demonstrating that roots of pea seedlings at this stage of development were not completely dependent on the shoots for reduced sulfur compounds. Indeed, these roots contained activities of ATP-sulfurylase (EC, adenosine 5′-phosphosulfate sulfotransferase, sulfite reductase (EC and O-acetyl-l-serine sulfhydrylase (EC at levels of 50, 30, 120 and 100%, respectively, of that in shoots. Most of the extractable activity of adenosine 5′-phosphosulfate sulfotransferase was detected in the first centimeter of the root tip. Using sucrose density gradients for organelle separation from this part of the root showed that almost 40% of the activity of ATP-sulfurylase, adenosine 5′-phosphosulfate sulfotransferase and sulfite reductase banded with the marker enzyme for proplastids, whereas only approximately 7% of O-acetyl-l-serine sulfhydrylase activity was detected in these fractions. Because their distributions on the gradients were very similar to that of nitrite reductase, a proplastid enzyme, it is concluded that ATP-sulfurylase, adenosine 5′-phosphosulfate sulfotransferase and sulfite reductase are also exclusively or almost exclusively localized in the proplastids of pea roots. O-Acetyl-l-serine sulfhydrylase is predominantly present in the cytoplasm.

Key words

ATP-sulfurylase-adenosine 5′-phosphosulfate sulfotransferase Pisum (sulfate reduction) Proplastid Sulfite reductase 



adenosine 5′-phosphosulfate sulfotransferase


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Anderson, L.E., Advani, V.R. (1970) Chloroplast and cytoplasmic enzymes. Plant Physiol. 45, 583–585Google Scholar
  2. Becker, M.A., Kredich, N.M., Tomkins, G.M. (1969) The purification and characterization of O-acetlyserine sulfhydrylase-A from Salmonella typhimurium. J. Biol. Chem. 244, 2418–2427Google Scholar
  3. Bonas, U., Schmitz, K., Rennenberg, H., Bergmann, L. (1982) Phloem transport of sulfur in Ricinus. Planta 155, 82–88Google Scholar
  4. Bradford, M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254Google Scholar
  5. Brunold, C., Schiff, J.A. (1976) Studies of sulfate utilization by algae. Enzymes of assimilatory sulfate reduction in Euglena and their cellular localization. Plant Physiol. 57, 430–436Google Scholar
  6. Brunold, C., Suter, M. (1983) Aktivitätsmessung der Adenosin 5′-phosphosulfat-sulfotransferase und ihre Anwendung bei der Untersuchung der de novo-Synthese des Enzyms. Bot. Helv. 93, 105–114Google Scholar
  7. Brunold, C., Suter, M. (1984) Regulation of sulfate assimilation by nitrogen nutrition in the duckweed Lemna minor L. Plant Physiol. 76, 579–583Google Scholar
  8. Clarkson, D.T., Smith, F.W., Van den Berg, P.J. (1983) Regulation of sulphate transport in a tropical legume, Macroptilium atropurpureum, cv. Siratro. J. Exp. Bot. 34, 1463–1483Google Scholar
  9. Ellis, R.J. (1963) Cysteine biosynthesis in beet discs. Phytochemistry 2, 129–136Google Scholar
  10. Ellis, R.J. (1969) Sulphate activation in higher plants. Planta 88, 34–42Google Scholar
  11. Emes, M.J., Fowler, M.W. (1979) The intracellular location of the enzymes of nitrate assimilation in the apices of seedling pea roots. Planta 144, 249–253Google Scholar
  12. Fankhauser, H., Brunold, C. (1978) Localization of adenosine 5′-phosphosulfate sulfotransferase in spinach leaves. Planta 143, 285–289Google Scholar
  13. Fankhauser, H., Brunold, C. (1979) Localization of O-acetyl-l-serine sulfhydrylase in Spinacia oleracea L.. Plant Sci. Lett. 14, 185–192Google Scholar
  14. Fankhauser, H., Brunold, C., Erismann, K.H. (1976) Subcellular localization of O-acetylserine sulfhydrylase in spinach leaves. Experientia 32, 1494–1496Google Scholar
  15. Fujita, M., Kawanishi, T. (1987) Cd-binding complexes from the root tissues of various higher plants cultivated in Cd2+-containing medium. Plant Cell Physiol. 28, 379–382Google Scholar
  16. Grill, E., Winnacker, E.-L., Zenk, M.H. (1985) Phytochelatins: the principal heavy-metal complexing peptides of higher plants. Science 230, 674–676Google Scholar
  17. Klapheck, S., Latus, C., Bergmann, L. (1987) Localization and distribution of glutathione in leaf cells of Pisum sativum L.. J. Plant Physiol. 131, 123–131Google Scholar
  18. Krueger, R.J., Siegel, L.M. (1982) Spinach siroheme enzymes: isolation and characterization of ferredoxin-sulfite reductase in comparison of properties with ferredoxin-nitrite reductase. Biochemistry 21, 2892–2904Google Scholar
  19. Mayer, A.M. (1967) Subcellular location of sulphite reductase in plant tissues. Plant Physiol. 42, 324–326Google Scholar
  20. Miflin, B.J. (1974) The location of nitrite reductase and other enzymes related to amino acid biosynthesis in the plastids of roots and leaves. Plant Physiol. 54, 550–555Google Scholar
  21. Nussbaum, S., Schmutz, D., Brunold, C. (1988) Regulation of assimilatory sulfate reduction by cadmium in Zea mays L.. Plant Physiol. 88, 1407–1410Google Scholar
  22. Pate, J.S. (1965) roots as organs of assimilation of sulfate. Science 149, 547–548Google Scholar
  23. Pieniacek, N.J., Stephien, P.P., Pazewsky, A. (1973) An Aspergillus nidulans mutant lacking cystathionin-synthase. Biochim. Biophys. Acta. 297, 37–47Google Scholar
  24. Saidha, T., Stern, A.I., Lee, D.-H., Schiff, J.A. (1985) Localization of a sulphate-activating system within Euglena mitochondria. Biochem J. 232, 357–365Google Scholar
  25. Schiff, J.A. (1983) Reduction and other metabolic reactions of sulfate. In: Encyclopedia of plant physiology, NS, vol. 15A:, pp. 401–421, Läuchli, A., Bieleski, R.L., eds. Springer, Berlin Heidelberg New York TokyoGoogle Scholar
  26. Schmidt, A. (1976) Development of the adenosine 5′-phosphosulfate-sulfotransferase in sunflower Helianthus annuus L. Z. Pflanzenphysiol. 78, 164–168Google Scholar
  27. Schmidt, A. (1986) Regulation of sulfur metabolism in plants. Progr. Bot. 48, 133–150Google Scholar
  28. Schmutz, D., Brunold, C. (1982) Rapid and simple measurement of ATP-sulfurylase activity in crude plant extracts using an ATP meter for bioluminescence determination. Anal. Biochem. 121, 151–155Google Scholar
  29. Schwenn, J.D., Trebst, A. (1976) Photosynthetic sulfate reduction by chloroplasts. In: The intact chloroplast, pp. 315–334, Barber, J., ed. Elsevier, AmsterdamGoogle Scholar
  30. Suzuki, A., Oaks, A., Jacquot, J.-P., Vidal, J., Gadal, P. (1985) An electron transport system in maize roots for reactions of glutamate synthase and nitrite reductase. Plant Physiol. 78, 374–378Google Scholar
  31. Tamura, G., Hosoi, T. (1979) The occurence of ferredoxin-sulfite reductase in barley roots. Agric. Biol. Chem. 43, 1601–1602Google Scholar
  32. Tamura, G., Iwasawa, T., Masada, M., Fukushima, K. (1976) Some properties of cysteine synthase from radish roots. Agric. Biol. Chem. 40, 637–638Google Scholar
  33. Trebst, A., Schmidt, A. (1969) Photosynthetic sulfate and sulfite reduction by chloroplasts. Prog. Photosynth. Res. 3, 1510–1516Google Scholar
  34. Tsang, M.L., Lemieux, J., Schiff, J.A., Bojarksi, T.B. (1976) Preparation of adenosine 5′-phosphosulfate (APS) from 3′-phosphate 5′-phosphosulfate (PAPS) prepared by an improved method. Anal. Biochem. 74, 623–626Google Scholar
  35. von Arb, C., Brunold, C. (1983) Measurement of ferredoxin-dependent sulfite reductase activity in crude extracts from leaves using O-acetyl-l-serine sulfhydrylase in a coupled assay system to measure the sulfide formed. Anal. Biochem. 131, 198–204Google Scholar
  36. Wallace, W. (1986) Distribution of nitrate assimilation between the root and shoot of legumes and a comparison with wheat. Physiol. Plant. 66, 630–636Google Scholar
  37. Wyss, H.-R., Brunold, C. (1979) Regulation of adenosine 5′-phosphosulfate sulfotransferase activity by H2S and cyst(e)ine in primary leaves of Phaseolus vulgaris L. Planta 147, 37–42Google Scholar

Copyright information

© Springer-Verlag 1989

Authors and Affiliations

  • C. Brunold
    • 1
  • M. Suter
    • 1
  1. 1.Pflanzenphysiologisches Institut der Universität BernBernSwitzerland

Personalised recommendations