Plant Growth Regulation

, Volume 53, Issue 2, pp 107–115

Oxidative stress and antioxidant activity as the basis of senescence in chrysanthemum florets

Original Paper


Stems of chrysanthemum (Chrysanthemum morifolium Ramat.) cv. Maghi were harvested when half of the buds showed colour and were put in distilled water at 21°C. Flowers showed visible senescence symptoms after 12–15 d. Reactive oxygen species (ROS) concentration and lipid peroxidation increased from young floret stage to the senescent stage. Activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), peroxidase (POD) and catalase (CAT) showed uniform increases from young floret through to the mature stage and thereafter, declined. Among the SOD isoforms, Fe-SOD and Cu/Zn-SOD were induced during the onset of senescence. Similarly different isoforms of APX and glutathione reductase (GR) also appeared during the senescence process. The capacity of the antioxidative defence system increased during the onset of senescence but the imbalance between ROS production and antioxidant defences ultimately led to oxidative damage. It is proposed that a decrease in the activity of a number of antioxidant enzymes that normally prevent the build up of free radicals can at least partially account for the observed senescence of chrysanthemum florets.


Antioxidant enzymes Chrysanthemum Florets Senescence Reactive oxygen species 



Ascorbate peroxidase




Glutathione reductase






Guaiacol peroxidase


Reactive oxygen species


Relative water content


Superoxide dismutase


  1. Able AJ, Guest DI, Sutherland MW (1988) Use of a new tetrazolium-based assay to study the production of superoxide radicals by tobacco cell cultures challenged with avirulent zoospores of Phytophthora parasitica var nicotianae. Plant Physiol 117:491–499CrossRefGoogle Scholar
  2. Aebi H (1974) Catalases. In: Bergmeyer HU (ed) Methods of enzymatic analysis, vol 2. Academic Press, New York, pp 673–684Google Scholar
  3. Alexieva V, Sergiev I, Mapelli S, Karanov E (2001) The effect of drought and UV on growth and stress markers in pea and wheat. Plant Cell Environ 24:1337–1344 CrossRefGoogle Scholar
  4. Alscher RG (1989) Biosynthesis and antioxidant function of glutathione in plants. Plant Physiol 77:457–464CrossRefGoogle Scholar
  5. Anderson MD, Prasad TK, Stewart CR (1995) Changes in the isozyme profiles of catalase, peroxidase and glutathione reductase during acclimation to chilling in mesocotyls of maize seedlings. Plant Physiol 109:1247–1257PubMedGoogle Scholar
  6. Asada K (1992) Ascorbate peroxidase—a hydrogen peroxide scavenging enzyme in plants. Physiol Plant 85:235–241CrossRefGoogle Scholar
  7. Axerold B, Chesbrough TM, Laakso S (1981) Lipoxygenase from soybean. In: Lowenstein JM (ed) Methods enzymology. Academic press, New York, pp 441–451Google Scholar
  8. Bailly C, Benamar A, Corbineau F, Dôme D (1996) Changes in malondialdehyde content and in superoxide dismutase, catalase and glutathione reductase activities in sunflower seed as related to deterioration during accelerated aging. Physiol Plant 97:104–110CrossRefGoogle Scholar
  9. Bailly C, Corbineau F, Doorn WG (2001) Free radical scavenging and senescence in Iris tepals. Plant Physiol Biochem 39:649–656CrossRefGoogle Scholar
  10. Bartoli CG, Simontacchi M, Guiamet JJ, Montaldi E, Puntarulo S (1995) Antioxidant enzymes and lipid peroxidation during aging of Chrysanthemum morifolium RAM petals. Plant Sci 104:161–168CrossRefGoogle Scholar
  11. Bayer WF, Fridovich I (1987) Assaying for superoxide dismutase activity: some large consequences of minor changes in condition. Annals Biochem 161:559–566CrossRefGoogle Scholar
  12. Beauchamp C, Fridovich I (1971) Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Annals Biochem 44:276–287CrossRefGoogle Scholar
  13. Bowler CM, van Montagu, Inze D (1992) Superoxide dismutase and stress tolerance. Ann Rev Plant Physiol Plant Mol Biol 43:83–116CrossRefGoogle Scholar
  14. Bowler C, Van Camp W, Van Montagu M, Inze D (1994) Superoxide dismutase in plants. CRC Crit Rev Plant Sci 13:199–218CrossRefGoogle Scholar
  15. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Annals Biochem 72:248–254CrossRefGoogle Scholar
  16. Broadbent P, Creissen GP, Kular B, Wellburn AR, Mullineaux P (1995) Oxidative stress responses in transgenic tobacco containing altered levels of glutathione reductase activity. Plant J 8:247–255CrossRefGoogle Scholar
  17. Calbert I, Mannervik B (1985) Glutathione reductase. Meth Enzymol 113:484–490Google Scholar
  18. Chen GX, Asada K (1989) Ascorbate peroxidase in tea leaves: occurrence of two isozymes and the differences in their enzymatic and molecular properties. Plant Cell Physiol 30:987–998Google Scholar
  19. Corpas FJ, Barroso JB, del Rio LA (2001) Peroxisomes as a source of reactive oxygen species and nitric oxide signal molecules in plant cells. Trends in Plant Sci 6:145–150CrossRefGoogle Scholar
  20. del Rio LA, Pastori GM, Palma JM, Sandalio LM, Sevilla F, Corpas FJ, Jimenez A, Lopez-Huertas E, Hernandez JA (1998) The activated oxygen role of peroxisomes in senescence. Plant Physiol 116:1195–1200PubMedCrossRefGoogle Scholar
  21. del Rio LA, Sandalio LM, Altomare DA, Zilinskas BA (2003) Mitochondrial and peroxisomal manganese superoxide dismutase: differential expression during leaf senescence. J Exp Bot 54:923–933PubMedCrossRefGoogle Scholar
  22. Dhindsa RA, Plumb-Dhindsa P, Thorpe PA (1981) Leaf senescence: correlated with increased permeability and lipid peroxidation, and decreases levels of superoxide dismutase and catalase. J Exp Bot 126:93–101CrossRefGoogle Scholar
  23. Edwards EA, Enard C, Creissen GP, Mullineaux PM (1994) Synthesis and properties of glutathione reductase in stressed peas. Planta 192:137–143Google Scholar
  24. Fielding JL, Hall JL (1978) A biochemical and cytological study of peroxidase activity in roots of Pisum sativum. J Exp Bot 29:969–981CrossRefGoogle Scholar
  25. Halliwell B (1987) Oxidative damage, lipid peroxidation and antioxidant protection in chloroplasts. Chem Phys Lipids 44:327–340CrossRefGoogle Scholar
  26. Halliwell B, Gutteridge JMC (1989) Free radicals in biology and medicine. Clarendon Press, OxfordGoogle Scholar
  27. Heath RL, Packer L (1968) Photoperoxidation in isolated chloroplasts. I. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys 125:189–198PubMedCrossRefGoogle Scholar
  28. Hossain Z, Mandal AKA, Datta SK, Biswas AK (2006) Decline in ascorbate peroxidase activity- a prerequisite factor for tepal senescence in gladiolus. J Plant Physiol 167:186–194CrossRefGoogle Scholar
  29. Hung SH, Yu CW, Lin CH (2005) Hydrogen peroxide functions as a stress signal in plants. Bot Bull Acad Sin 46:1–10Google Scholar
  30. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685PubMedCrossRefGoogle Scholar
  31. Larson RA (1988) The antioxidants of higher plants. Phytochemistry 27:969–978CrossRefGoogle Scholar
  32. Lesham YY (1992) Membrane-associated phospholytic and lipolytic enzymes. In: Lesham YY (ed) Plant membranes: a biophysical approach to structure, development and senescence. Kluwer Academic Publishers, Dordrecht, pp 174–191Google Scholar
  33. Leverentz MK, Rogers CW, Stead JH, Usawadee ADC, Silkowski H, Thomas B, Weichert H, Feussner I, Griffiths G (2002) Characterization of a novel lipoxygenase-independent senescence mechanism in Alstroemeria peruviana floral tissue. Plant Physiol 130:273–283PubMedCrossRefGoogle Scholar
  34. Levine A, Tenhaken R, Dixon R, Lamb C (1994) H2O2 from the oxidative burst orchestrates the plant hypersensitive disease resistance response. Cell 79:583–593PubMedCrossRefGoogle Scholar
  35. McCarthy I, Romero-Puertas MC, Palma JM, Sandalio LM, Corpas FJ, Gomez M, del Rio LA (2001) Cadmium induces senescence symptoms in leaf peroxisomes of pea plants. Plant Cell Environ 24:1065–1073CrossRefGoogle Scholar
  36. Meister A (1981) Metabolism and functions of glutathione. Trends Biochem Sci 6:231–234CrossRefGoogle Scholar
  37. Mittler R, Zilinskas BA (1993) Detection of ascorbate peroxidase activity in native gels by inhibition of the ascorbate dependent reduction of nitroblue tetrazolium. Annals Biochem 212:540–546CrossRefGoogle Scholar
  38. Ogawa K, Tasaka Y, Mino M, Tanaka Y, Iwabuchi M (2001) Association of glutathione with flowering in Arabidopsis thaliana. Plant Cell Physiol 42:524–530PubMedCrossRefGoogle Scholar
  39. Panavas T, Rubinstein B (1998) Oxidative events during programmed cell death of daylily (Hemerocallis hybrid) petals. Plant Sci 133:25–138CrossRefGoogle Scholar
  40. Paulin A, Droillard M, Bureau JM (1986) Effect of a free radical scavenger, 3,4,5-trichlorophenol, on ethylene production and on changes in lipids and membrane integrity during senescence of petals of cut carnations (Dianthus carvophyllus). Physiol Plant 67:465–471CrossRefGoogle Scholar
  41. Peary JS, Prince TA (1990) Floral lipoxygenase: activity during senescence and inhibition by phenidone. J Am Soc Hortic Sci 115:455–457Google Scholar
  42. Prochazkova D, Sairam RK, Srivastava GC, Singh DV (2001) Oxidative stress and antioxidant activity as the basis of senescence in maize leaves. Plant Sci 161:765–771CrossRefGoogle Scholar
  43. Pütter J (1974) Peroxidases. In: Bergmeyer HU (ed) Methods of enzymatic analysis, vol 2. Academic Press, New York, pp 685–690Google Scholar
  44. Rao MV, Paliyath G, Ormrod DP (1996) Ultraviolet-B- and ozone induced biochemical changes in antioxidant enzymes of Arabidopsis thaliana. Plant Physiol 110:125–136PubMedCrossRefGoogle Scholar
  45. Reed DJ (1990) Glutathione: toxicological implications. Annu Rev Pharmacol Toxicol 30:603–631PubMedCrossRefGoogle Scholar
  46. Sairam RK, Deshmukh PS, Shukla DS (1997) Tolerance to drought and temperature stress in relation to increased antioxidant enzyme activity in wheat. J Agron Crop Sci 178:171–177CrossRefGoogle Scholar
  47. Schöner S, Krause GH (1990) Protective systems against active oxygen species in spinach: response to cold acclimation in excess light. Planta 180:383–389CrossRefGoogle Scholar
  48. Smith IK, Vierheller TL, Thorne CA (1988) Assay of glutathione reductase in crude tissue homogenates using 5, 5′-dithiobis (2-nitrobenzoic acid). Annals Biochem 175:408–413CrossRefGoogle Scholar
  49. Thompson JE, Froese CD, Madey E, Smith MD, Hong YW (1998) Lipid metabolism during plant senescence. Progr Lipid Res 37:119–141CrossRefGoogle Scholar
  50. Weatherley PE (1950) Studies in the water relations of the cotton plant. 1. The field measurement of water deficit in leaves. New Phytology 49:8CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Debasis Chakrabarty
    • 1
  • J. Chatterjee
    • 1
  • S. K. Datta
    • 1
  1. 1.Floriculture SectionNational Botanical Research InstituteLucknowIndia

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