Skip to main content
Log in

Over-expression of chloroplast-targeted Mn superoxide dismutase in cotton (Gossypium hirsutum L., cv. Coker 312) does not alter the reduction of photosynthesis after short exposures to low temperature and high light intensity

  • Published:
Photosynthesis Research Aims and scope Submit manuscript

Abstract

Transgenic cotton plants from several independently-transformed lines expressing a chimeric gene encoding a chloroplast-targeted Mn superoxide dismutase (SOD) from tobacco exhibit a three-fold increase in the total leaf SOD activity, strong Mn SOD activity associated with isolated chloroplasts, and a 30% and 20% increase in ascorbate peroxidase and glutathione reductase activities, respectively. The Mn SOD plants did exhibit a slightly enhanced protection against light-mediated, paraquat-induced cellular damage but only at 0.3 µM paraquat. In addition, photosynthetic rates at 10°C and 15°C were similar to those of controls, and the immediate recovery of photosynthesis after a 35-min exposure to 5°C and full sun was only slightly better than that for wild-type plants. The recovery for longer exposure times was comparable for both genotypes as was the deactivation of the H2O2-sensitive, Calvin-cycle enzyme, stromal fructose 1,6-bisphosphatase (FBPase). Compared to the controls, Mn SOD plant leaves in full sun prior to chilling stress had a lower activation of FBPase, a higher ratio of oxidized to reduced forms of ascorbate, and a higher total glutathione content. After 35 min at 5°C in full sunlight, total glutathione had risen in control leaves to 88% of the Mn SOD plant values, and oxidized to reduced ascorbate ratios were higher for both genotypes. However, an 80% increase in the ratio of oxidized to reduced glutathione occurred for Mn SOD plant leaves with no change for controls. This increased demand on the ascorbate-glutathione cycle is circumstantial evidence that high Mn SOD activity in the chloroplast leads to increased H2O2 pools that could, in some manner, affect photosynthetic recovery after a stress period. We postulate that the pool sizes of reduced ascorbate and glutathione may restrict the ability of the ascorbate-glutathione cycle to compensate for the increased activity of SOD in cotton over-producing mitochondrial Mn SOD in chloroplasts during short-term chilling/high light stress.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Allen RD (1995a) Dissection of oxidative stress tolerance using transgenic plants. Plant Physiol 107: 1049–1054

    PubMed  Google Scholar 

  • Allen RD (1995b) Overexpression of chloroplastic Cu/Zn superoxide dismutase in plants. In: Gartland KMA and Davey MR (eds) Methods in Molecular Biology, Vol 44, pp 309–323. Humana Press, Totowa, NJ

    Google Scholar 

  • Allen RD, Sen Gupta A, Webb RP and Holaday AS (1995) Protection of plants from oxidative stress using SOD transgenes: Interactions with endogenous enzymes. In: Asada K and Yoshikawa T (eds) Frontiers of Reactive Oxygen Species in Biology and Medicine, pp 321–322. Excerpta Medica, Amsterdam

    Google Scholar 

  • Arnon DI (1949) Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol 24: 1–5

    Google Scholar 

  • Bayley C, Trolinder N, Ray C, Morgan M, Quisenberry JE and Ow DW (1992) Engineering 2,4-D resistance into cotton. Theor Appl Genet 83: 645–649

    Article  Google Scholar 

  • Beauchamp C and Fridovich I (1971) Superoxide dismutase: Improved assays and an assay applicable to acrylamide gels. Anal Biochem 44: 276–287

    PubMed  Google Scholar 

  • Berkowitz GA and Gibbs M (1982) Effect of osmotic stress on photosynthesis studied with the isolated spinach chloroplast. Plant Physiol 70: 1143–1148

    Google Scholar 

  • Bowler C, Slooten L, Vandenbranden S, De Rycke R, Botterman J, Sybesma C, Van Montagu M and Inzé D (1991) Manganese superoxide dismutase can reduce cellular damage mediated by oxygen radicals in transgenic plants. EMBO J 10: 1723–1732

    PubMed  Google Scholar 

  • Bowler C, Van Montagu M and Inzé D (1992) Superoxide dismutase and stress tolerance. Annu Rev Plant Physiol Plant Mol Biol 43: 83–116

    Article  Google Scholar 

  • 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: 248–254

    Article  PubMed  Google Scholar 

  • Brugnoli E and Björkman O (1992) Growth of cotton under continuous salinity stress: Influence on allocation pattern, stomatal and non-stomatal components of photosynthesis and dissipation of excess light energy. Planta 187: 335–347

    Article  Google Scholar 

  • Charles SA and Halliwell B (1981) Light activation of fructose bisphosphatase in isolated spinach chloroplasts and deactivation by hydrogen peroxide. A physiological role for the thioredoxin system. Planta 151: 242–246

    Google Scholar 

  • Foyer CH and Halliwell B (1976) The presence of glutathione and glutathione reductase in chloroplasts: A proposed role for ascorbic acid metabolism. Planta 133: 21–25

    Google Scholar 

  • Foyer CH, Rowell J and Walker DA (1983) Measurement of the ascorbate content of spinach leaf protoplasts and chloroplasts during illumination. Planta 157: 239–247

    Google Scholar 

  • Giersch C and Robinson SP (1987) Effect of photoinhibition on photosynthetic carbon metabolism in intact isolated spinach chloroplasts. Aust J Plant Physiol 14: 439–449

    Google Scholar 

  • Griffith OW (1980) Determination of glutathione and glutathione disulfide using glutathione reductase and 2-vinlypyridine. Anal Biochem 106: 207–212

    PubMed  Google Scholar 

  • Holaday AS, Martindale W, Alred R, Brooks AL and Leegood RC (1992) Changes in activities of enzymes of carbon metabolism in leaves during exposure of plants to low temperature. Plant Physiol 98: 1105–1114

    Google Scholar 

  • Horsch RB, Fry JE, Hoffman NJ, Eicholtz D, Rogers SG and Fraley RT (1985) A simple and general model for transferring genes into plants. Science 227: 119–123

    Google Scholar 

  • Kaiser WM (1979) Reversible inhibition of the Calvin Cycle in isolated chloroplasts by H2O2. Planta 145: 377–382

    Google Scholar 

  • Königer M and Winter K (1993) Reduction of photosynthesis in sun leaves of Gossypium hirsutum L. under conditions of high light intensities and suboptimal leaf temperatures. Agronomie 13: 659–669

    Google Scholar 

  • Lichtenthaler HK (1987) Chlorophylls and carotenoids: pigments of photosynthetic membranes. Methods Enzymol 148: 350–382

    Google Scholar 

  • McCord JM and Fridovich I (1969) Superoxide dismutase, an enzymatic function for erythrocuprein (hemocuprein). J Biol Chem 244: 6049–6055

    PubMed  Google Scholar 

  • McKersie BD, Chen Y, De Beus M, Bowley SR and Bowler C (1993) Superoxide dismutase enhances tolerance of freezing stess in transgenic alfalfa (Medicago sativa L.). Plant Physiol 103: 1155–1163

    Article  PubMed  Google Scholar 

  • Miyake C and Asada K (1992) Thylakoid-bound ascorbate peroxidase in spinach chloroplasts and photoreduction of its primary oxidation product monodehydroascorbate radicals in thylakoids. Plant Cell Physiol 33: 541–553

    Google Scholar 

  • Perera NH, Hartmann E and Holaday AS (1995) Regulation of cotton photosynthesis during moderate chilling. Plant Science 111: 133–143

    Article  Google Scholar 

  • Sassenrath GF, Ort DR and Portis AR (1990) Impaired reductive activation of stromal bisphosphatases in tomato leaves following low-temperature exposure at high light. Arch Biochem Biophys 282: 302–308

    PubMed  Google Scholar 

  • Schake SA (1995) Analysis of pea chloroplastic manganese superoxide dismutase overexpressed in tobacco. MS Thesis, Texas Tech University

  • Sen Gupta A, Heinen JL, Holaday AS and Allen RD (1993a) Increased resistance to oxidative stress in transgenic plants that overexpress chloroplastic Cu/Zn superoxide dismutase. Proc Natl Acad Sci USA 90: 1629–1633

    PubMed  Google Scholar 

  • Sen Gupta A, Webb RP, Holaday AS, and Allen RD (1993b) Overexpression of superoxide dismutase protects plants from oxidative stress. Plant Physiol 103: 1067–1073

    PubMed  Google Scholar 

  • Slooten L, Capiau K, Van Camp W, Van Montagu M, Sybesma C and Inzé D (1995) Factors affecting the enhancement of oxidative stress tolerance in transgenic tobacco overexpressing manganese superoxide dismutase in the chloroplasts. Plant Physiol 107: 737–750

    PubMed  Google Scholar 

  • Smirnoff N (1995) Antioxidant systems and plant response to the environment. In: Smirnoff N (ed) Environment and Plant Metabolism, pp 217–242. BIOS Scientific Publishers, Oxford, UK

    Google Scholar 

  • Smith IK, Vierheller TL, and Thorne CA (1989) Properties and functions of glutathione reductase in plants. Physiol Plant 77: 449–456

    Google Scholar 

  • Van Camp W, Capiau K, Van Montagu M, Inzé D and Slooten L (1996) Enhancement of oxidative stress tolerance in transgenic tobacco plants overproducing Fe-superoxide dismutase in chloroplasts. Plant Physiol 112: 1703–1714

    Article  PubMed  Google Scholar 

  • Vernon IP (1960) Spectrophotometric determination of chlorophyll and pheophytins in plant extracts. Anal Chem 32: 1144–1150

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Payton, P., Allen, R.D., Trolinder, N. et al. Over-expression of chloroplast-targeted Mn superoxide dismutase in cotton (Gossypium hirsutum L., cv. Coker 312) does not alter the reduction of photosynthesis after short exposures to low temperature and high light intensity. Photosynthesis Research 52, 233–244 (1997). https://doi.org/10.1023/A:1005873105596

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1005873105596

Navigation