Biology and Fertility of Soils

, Volume 44, Issue 7, pp 909–916 | Cite as

Low levels of ferredoxin, ATP and leghemoglobin contribute to limited N2 fixation of peas (Pisum sativum L.) and alfalfa (Medicago sativa L.) under S deficiency conditions

  • Heinrich W. Scherer
  • Svea Pacyna
  • Katrin R. Spoth
  • Margot Schulz
Original Paper


Sulphur (S) has become a major limiting factor for plant production in industrial as well as in remote industrial rural areas. Limitation of S can reduce legume N2 fixation by affecting nodule development and function. In pot experiments with pea (Pisum sativum L.) and alfalfa (Medicago sativa L.), we investigated the influence of S on growth, ferredoxin, ATP and leghemoglobin concentrations. Addition of 200 mg S pot−1 increased yield of shoots, roots and nodules of both plant species significantly. However, the influence of S on nodule yield formation was most pronounced. Pea and alfalfa roots were found to have higher S concentrations than shoots and being up to 2.9 times the S concentration in the shoots of peas under S-sufficient conditions. Sulphur addition also increased N2 fixation significantly. The ferredoxin concentration in bacteroids of root nodules of pea was increased significantly by S only 10 weeks after planting and in bacteroids of root nodules of alfalfa 10 and 17 weeks after planting, while on per pot base the amounts of ferredoxin were higher throughout the experimental period of time. The ATP concentration of bacteroids of root nodules of both plant species as well as of mitochondria of root nodules of pea were significantly higher with optimum S supply. The effects of S deficiency on N2 fixation are likely to be caused by the shortage of ferredoxin and ATP. The amount of leghemoglobin was reduced in comparison to nodules of the S-sufficient plants.


ATP Ferredoxin Legumes N2-fixation Sulphur 



The authors are much indebted to the Deutsche Forschungsgemeinschaft (DFG) for financial support and Fa. Jost for providing Rhizobium bacteria.


  1. Anonymous (2006) SAS-Makro Box-cox-transformation. Universität Hohenheim, Institut für Bioinformatik.
  2. Bergersen FJ (1991) Physiological control of nitrogenase and uptake hydrogenase. In: Dilworth MJ, Glenn AR (eds) Biology and biochemistry of nitrogen fixation. Elsevier, Amsterdam, NL, pp 76–102Google Scholar
  3. Carter KR, Rawlings J, Orme-Johnson WH, Becker RR, Evans HJ (1980) Purification and characterization of ferredoxin from Rhizobium japonicum bacteroids. J Biol Chem 255:4213–4223PubMedGoogle Scholar
  4. Ching TM, Hedtke S, Russell SA, Evans HJ (1975) Energy state and dinitrogen fixation in soybean nodules of dark-grown plants. Plant Physiol 55:796–798PubMedGoogle Scholar
  5. Dalton DA, Joyner SL, Becana M, Iturbe-Ormaetxe I, Chatfield JM (1998) Antioxidant defenses in the peripheral cell layers of legume root nodules. Plant Physiol 116:37–43PubMedCrossRefGoogle Scholar
  6. Dietz KJ (1989) Recovery of spinach leaves from sulphate and phosphate deficiency. J Plant Physiol 134:551–557Google Scholar
  7. Duke SH, Reisenauer H (1986) Roles and requirements of sulphur in plant nutrition. In: Tabatabai MA (ed) Sulfur in agriculture. Agronomy Monograph no. 27. American Society of Agronomy, Madison, Wisconsin, USA, pp 123–168Google Scholar
  8. Fox RL, Olson RA, Rhoades HF (1964) Evaluating the sulfur status of soils by plants and soil tests. Soil Sci Soc Am Proc 28:243–246CrossRefGoogle Scholar
  9. Fukuyama K (2004) Structure and function of plant-type ferredoxin. Photosynth Res 81:289–301PubMedCrossRefGoogle Scholar
  10. Gonzalez EM, Galves L, Royuela M, Aparicio-Tejo PM, Arrese-Igor C (2001) Insights into the regulation of nitrogen fixation in pea nodules: lessons from drought, abscisic acid and increased photoassimilate availability. Agronomy 21:607–613CrossRefGoogle Scholar
  11. Gordon AJ, Minchin FR, Skot L, James CL (1997) Stress-induced declines in soybean N2 fixation are related to nodule sucrose synthase activity. Plant Physiol 114:937–946PubMedGoogle Scholar
  12. Hrivna L, Richter R, Losak T (2001) The effect of the content of water-soluble sulphur in the soil on the utilisation of nitrogen and on the yields and quality of winter rape. Rostl Vyroba 47:18–22Google Scholar
  13. Hrivna L, Richter R, Losak T, Hlusek J (2002) Effect of increasing doses of nitrogen and sulphur on chemical composition of plants, yields and seed quality in winter rape. Rostl Vyroba 48:1–6Google Scholar
  14. Johnson MD, Wang X (1996) Differentially expressed forms of 1-L-myo-inositol-1-phosphate synthase (EC) in Phaseolus vulgaris. J Biol Chem 271:17215–17218PubMedCrossRefGoogle Scholar
  15. Kawashima K, Suganuma N, Tamaoki M, Kouchi H (2001) Two types of pea leghemoglobin genes showing different O2-binding affinities and distinct patterns of spatial expression in nodules. Plant Physiol 125:641–651PubMedCrossRefGoogle Scholar
  16. Lange A (1998) Einfluß der Schwefel-Versorgung auf die biologische Stickstoff-Fixierung von Leguminosen. Ph.D. thesis, University of Bonn, GermanyGoogle Scholar
  17. Lodwig EM, Hosie AHF, Bourdès A, Findlay K, Allaway D, Karunakaran R, Downie JA, Poole PS (2003) Amino-acid cycling drives nitrogen fixation in the legume-Rhizobium symbiosis. Nature 422:722–726PubMedCrossRefGoogle Scholar
  18. Marschner H (1995) Mineral nutrition of higher plants, 2nd edn. Academic Press, LondonGoogle Scholar
  19. Matamoros MA, Baird LM, Escuredo PR, Dalton D, Minchin FR, Iturbe-Ormaetxe I, Rubio MC, Moran JF, Gordon AJ, Becana M (1999) Stress-induced legume root nodule senescence. Physiological, biochemical and structural alterations. Plant Physiol 121:97–111PubMedCrossRefGoogle Scholar
  20. Mengel K, Kirkby EA (2001) Principles of plant nutrition, 5th edn. Kluwer Academic PublishersGoogle Scholar
  21. Mengel K, Haghparast M, Koch K (1974) The effect of potassium on the fixation of molecular nitrogen by root nodules of V. faba. Plant Physiol 54:535–538PubMedCrossRefGoogle Scholar
  22. Mortensen LE, Thornley RNF (1979) Structure and function of nitrogenase. Ann Rev Biochem 48:387–418CrossRefGoogle Scholar
  23. Murphy MD, Brogan JC, Noonan DG (1983) Sulphur fertilization of pasture improves cattle performance. Sulphur Agric 7:2–6Google Scholar
  24. Neuburger M, Journet E-P, Bligny R, Carde J-P, Douce R (1982) Purification of plant mitochondria by isopycnic centrifugation in density gradients of Percoll. Arch Biochem Biophys 217:312–323PubMedCrossRefGoogle Scholar
  25. Pacyna S, Schulz M, Scherer HW (2006) Influence of sulphur supply on glucose and ATP concentrations of inoculated broad beans (V. faba minor L.). Biol Fertil Soils 42:324–329CrossRefGoogle Scholar
  26. Price GD, Day DA, Gresshoff PM (1987) Rapid isolation of intact peribacteroid envelopes from soybean nodules and demonstration of selective permeability to metabolites. J Plant Physiol 130:157–164Google Scholar
  27. Ryden JC (1984) Fertilizers for grassland. Chem Ind 18:652–657Google Scholar
  28. Scherer HW, Lange A (1996) N2-fixation and growth of legumes as affected by sulphur fertilization. Biol Fertil Soils 23:449–453CrossRefGoogle Scholar
  29. Scherer HW, Pacyna S, Manthey N, Schulz M (2006) Sulphur supply to peas (Pisum sativum L.) influences symbiotic N2 fixation. Plant Soil Environ 52:72–77Google Scholar
  30. Schwinghamer EA (1980) A method for improved lysis of some gram negative bacteria. FEMS Microbiol Lett 7:157–162CrossRefGoogle Scholar
  31. Scott NM, Watson ME, Caldwell KS, Inkson RHE (1983) Response of grassland to the application of S at two sites in north-east Scotland. J Sci Food Agric 3:357–361CrossRefGoogle Scholar
  32. Simonsen ACW, Rosendahl L (1999) Origin of de novo synthesized proteins in the different compartments of pea-Rhizobium sp. symbiosomes. Mol. Plant-Microbe Interact 12:319–327CrossRefGoogle Scholar
  33. Singh P, Raj B (1988) Sulphur fertilization in relation to yield and trend of production of leghemoglobin in the nodules of pea (Pisum sativum var. Arvense). Ann Agric Res 9:13–19Google Scholar
  34. Sunarpi S, Anderson JW (1997) Effect of nitrogen nutrition on remobilization of protein sulfur in the leaves of vegetative soybean and associated changes in soluble sulfur metabolites. Plant Physiol 115:1671–1680PubMedGoogle Scholar
  35. Vance CP, Gantt JS (1992) Control of nitrogen and carbon metabolism in root nodules. Physiol Plant 85:266–274CrossRefGoogle Scholar
  36. Vance CP, Heichel GH (1991) Carbon in N2 fixation: limitation or exquisite adaption. Ann Rev Plant Physiol Plant Mol Biol 42:373–392CrossRefGoogle Scholar
  37. Vikman PA, Vessey JK (1993) Gas-exchange activity, carbohydrate status and protein turnover in root nodule subpopulations of field pea (Pisum sativum Century). Plant Siol 151:31–38CrossRefGoogle Scholar
  38. Wilson DO, Reisenauer HM (1963) Determination of leghemoglobin in legume nodules. Anal Biochem 6:27–30CrossRefGoogle Scholar
  39. Wooding FJ, Paulsen GM, Murphy LS (1970) Response of nodulated and nonnodulated soybean seedlings to sulfur nutrition. Agron J 62:277–280CrossRefGoogle Scholar
  40. Yoch D (1973) Purification and characterization of ferredoxin-nicotinamide adenine dinucleotide phosphate reductase from a nitrogen-fixing bacterium. J Bacteriol 118:384–391Google Scholar
  41. Zhao FJ, Wood AP, McGrath SP (1999) Effects of sulphur nutrition on growth and nitrogen fixation of pea (P. sativum L.). Plant Soil 212:209–219CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Heinrich W. Scherer
    • 1
  • Svea Pacyna
    • 1
  • Katrin R. Spoth
    • 1
  • Margot Schulz
    • 2
  1. 1.INRES—Institute of Plant NutritionBonnGermany
  2. 2.Institute of Molecular Biotechnology and Physiology of PlantsBonnGermany

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