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Planta

, Volume 185, Issue 3, pp 287–296 | Cite as

Carbon metabolism enzymes and photosynthesis in transgenic tobacco (Nicotiana tabacum L.) having excess phytochrome

  • Thomas D. Sharkey
  • Terry L. Vassey
  • Peter J. Vanderveer
  • Richard D. Vierstra
Article

Abstract

J.M. Keller et al. (1989, EMBO J. 8, 1005–1012) introduced a phytochrome gene controlled by a cauliflower mosaic virus 35S promoter into tobacco (Nicotiana tabacum L.) providing material to test whether several photosynthesis enzymes can be increased by one modification to the plant. We report here that this transgenic tobacco had greater amounts of all enzymes examined as well as greater amounts of total protein and chlorophyll per unit leaf area. Fructose bisphosphatase (E.C. 3.1.3.11), glyceraldehyde 3-phosphate dehydrogenase (E.C. 1.2.1.12), and sucrose-phosphate synthase (E.C. 2.4.1.14) were also higher when expressed per unit protein. However, ribulose-1,5-bisphosphate carboxylase (E.C. 4.1.1.39) amount per unit leaf protein was the same in transgenic and wild-type (WT) plants. Photosynthesis in the transgenic plants was lower than in WT at air levels of CO2, but higher than in WT above 1000 μbar CO2. The photosynthesis results indicated a high resistance to CO2 diffusion in the mesophyll of the transgenic plants. Examination of electron micrographs showed that chloroplasts in the transgenic plants were often cup-shaped, preventing close association between chloroplast and cell surface. Chloroplast cupping may have caused the increase in the mesophyll resistance to CO2 diffusion. We conclude that it is possible to affect more than one enzyme with a single modification, but unexpected physical modifications worsened the photosynthetic performance of this plant.

Key words

Mesophyll resistance Nicotiana (photosynthesis and phytochrome in transgenic -) Photosynthesis (transgenic plant) Phytochrome and photosynthesis Ribulose-1,5-bisphosphate carboxylase Sucrose phosphate synthase Transgenic plant 

Abbreviations

CABP

2-carboxyarabitinol 1,5-bisphosphate

FBP

fructose-1,6-bisphosphate

FBPase

fructose-1,6-bisphosphatase

GAP

glyceraldehyde 3-phosphate

Rubisco

ribulose-1,5-bisphosphate carboxylase

SPS

sucrose-phosphate synthase

WT

wild type

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References

  1. Barnett, L.K., Clugston, C.K., Jenkins, G.I. (1987) Two phytochrome-mediated effects of light on transcription of genes encoding the small subunit of ribulose-1,5-bisphosphate carboxylase-oxygenase in dark-grown pea (Pisum sativum) plants. FEBS Lett. 2, 287–290Google Scholar
  2. Björkman, O. (1981) Responses to different quantum flux densities. In: Encyclopedia of plant physiology, N.S. vol. 12A: Physiological plant ecology I-Responses to the physical environment, pp.57–107, Lange, O.L., Nobel, P.S., Osmond, C.B., Zeigler, H., eds. Springer, Berlin HeidelbergGoogle Scholar
  3. Bongi, G., Loreto, F. (1989) Gas-exchange properties of saltstressed olive (Oka europa L.) leaves. Plant Physiol. 90, 1408–1416Google Scholar
  4. Cardon, Z.G., Mott, K.A. (1989) Evidence that ribulose 1,5-bis-phosphate (RuBP) binds to inactive sites of RuBP carboxylase in vivo and an estimate of the rate constant for dissociation. Plant Physiol. 89, 1253–1257Google Scholar
  5. Chao, S., Raines, C.A., Longstaff, M., Sharp, P.J., Gale, M.D., Dyer, T.A. (1989) Chromosomal location and copy number in wheat and some of its close relatives of genes for enzymes involved in photosynthesis. Mol. Gen. Gen. 218, 423–430Google Scholar
  6. Cherry, J., Hershey, H.P., Vierstra, R.D. (1991) Characterization of tobacco expressing functional oat phytochrome: Domains responsible for rapid degradation of PFR are conserved between monocots and dicots. Plant Physiol. 96, 775–785Google Scholar
  7. Cowan, I.R. (1986) Economics of carbon fixation in higher plants. In: On the economy of plant form and function, pp. 133–170, Givnish, T.J., ed. Cambridge, CambridgeGoogle Scholar
  8. Dietz, K.-J. (1986) an evaluation of light and CO2 limitation of leaf photosynthesis by CO2 gas-exchange analysis. Planta 167, 260–263Google Scholar
  9. Downton, W.J.S., Loveys, B.R., Grant, W.J.R. (1988a) Stomatal closure fully accounts for the inhibition of photosynthesis by abscisic acid. New Phytol. 108, 263–266Google Scholar
  10. Downton, W.J.S., Loveys, B.R., Grant, W.J.R. (1988b) Nonuniform stomatal closure induced by water stress causes putative non-stomatal inhibition of photosynthesis. New Phytol. 110, 503–509Google Scholar
  11. Evans, J.R., Sharkey, T.D., Berry, J.A., Farquhar, G.D. (1986) Carbon isotope discrimination measured concurrently with gas exchange to investigate CO2 diffusion in leaves of higher plants. Aust. J. Plant Physiol. 13, 281–292Google Scholar
  12. Farquhar, G.D., von Caemmerer, S., Berry, J.A. (1980) A biochemical model of photosynthetic CO2 assimilation in leaves of C3 species. Planta 149, 78–90Google Scholar
  13. Farquhar, G.D., O'Leary, M.H., Berry, J.A. (1982) On the relationship between carbon isotope discrimination and the intercellular carbon dioxide concentration in leaves. Aust. J. Plant Physiol. 9, 121–137Google Scholar
  14. Fluhr, R., Chua, N.H. (1987) Developmental regulation of two genes encoding ribulose-bisphosphate carboxylase small subunit in pea and transgenic petunia plants: phytochrome response and blue-light induction. Proc. Natl. Acad. Sci. USA 83, 2358–2362Google Scholar
  15. Guang-yao, W., Xiao-bo, Z., Xiang-yu, W. (1987) Light regulation of the synthesis of ribulose 1,5-bisphosphate carboxylase and fructose 1,6-bisphosphatase. Acta Bot. Sin. 29, 388–396Google Scholar
  16. Heber, U., Neimanis, S., Dietz, K.-J. (1988) Fractional control of photosynthesis by the QB protein, the cytochrome f/b 6complex and other components of the photosynthetic apparatus. Planta 173, 267–274Google Scholar
  17. Hoagland, D.R., Arnon, D.I. (1938) The water culture method for growing plants without soil. Univ. of Cal. Exp. Sta. Circ. 347Google Scholar
  18. Kaufman, L.S., Thompson, W.F., Briggs, W.R. (1984) Different red light requirements for phytochrome-induced accumulation of cab RNA and rcbS RNA. Science 226, 1447–1449Google Scholar
  19. Keller, J.M., Shanklin, J., Vierstra, R.D., Hershey, H.P. (1989) Expression of a functional monocotyledonous phytochrome in transgenic tobacco. EMBO J. 8, 1005–1012Google Scholar
  20. Leegood, R.C. (1985) Regulation of photosynthetic CO2-pathway enzymes by light and other factors. Photosynth. Res. 6, 247–259Google Scholar
  21. Nagy, B.F., Fluhr, R., Kuhlemeier, C., Kay, S., Boutry, M., Green, P., Poulsen, C., Chua, N.-H. (1986) Cis-acting elements for selective expression of two photosynthetic genes in transgenic plants. Phil. Trans. R. Soc. London Ser. B 314, 493–500Google Scholar
  22. Neuhaus, H.E., Kruckeberg, A.L., Feil, R., Stitt, M. (1989) Reduced-activity mutants of phosphoglucose isomerase in the cytosol and chloroplast of Clarkia xantiana II. Study of the mechanisms which regulate photosynthate partitioning. Planta 178, 110–122Google Scholar
  23. Nobel, P.S. (1977) Internal leaf area and cellular CO2 resistance: Photosynthetic implications of variations with growth conditions and plant species. Physiol. Plant. 40, 137–144Google Scholar
  24. Sage, R.F. (1990) A model describing the regulation of ribulose-1,5- bisphosphate carboxylase, electron transport, and triose phosphate use in response to light intensity and CO2 in C3 plants. Plant Physiol. 94, 1728–1734Google Scholar
  25. Sasaki, T., Sakihama, T., Kamikubo, T., Shinozaki, K. (1983) Phytochrome-mediated regulation of two mRNAs, encoded by nuclei and chloroplasts of ribulose-1,5-bisphosphate carboxyl-ase/oxygenase. Eur. J. Biochem. 133, 617–620Google Scholar
  26. Schmidt, G.W., Mishkind, ML. (1983) Rapid degradation of unassembled ribulose 1,5-bisphosphate carboxylase small subunits in chloroplasts. Proc. Natl. Acad. Sci. USA 80, 2632–2636Google Scholar
  27. Schmidt, S., Drumm-Herrel, H., Oelmuller, R., Mohr, H. (1987) Time course of competence in phytochrome-controlled appearance of nuclear-encoded plastidic proteins and messenger RNAs. Planta 170, 400–407Google Scholar
  28. Sharkey, T.D. (1988) Estimating the rate of photorespiration in leaves. Physiol. Plant. 73, 147–152Google Scholar
  29. Sharkey, T.D. (1989) Evaluating the role of rubisco regulation in C3 photosynthesis. Phil. Trans. R. Soc. London Ser. B 323, 435–448Google Scholar
  30. Sharkey, T.D., Seemann, J.R. (1989) Mild water stress effects on carbon-reduction-cycle intermediates, RuBP carboxylase activity, and spatial homogeneity of photosynthesis in leaves. Plant Physiol. 89, 1060–1065Google Scholar
  31. Sharkey, T.D., Berry, J.A., Sage, R.F. (1988a) Regulation of photosynthetic electron-transport as determined by roomtemperature chlorophyll a fluorescence in Phaseolus vulgaris L. Planta 176, 415–424Google Scholar
  32. Sharkey, T.D., Kobza, J., Seemann, J.R., Brown, R.H. (1988b) Reduced cytosolic fructose-1,6-bisphosphatase activity leads to loss of O2 sensitivity in a Flaveria linearis mutant. Plant Physiol. 86, 667–671Google Scholar
  33. Sharkey, T.D., Loreto, F., Vassey, T.L. (1990) Effects of stress on photosynthesis. In: Current research in photosynthesis, pp. 549–556, Baltscheffsky, M., ed. Kluwer, Dordrecht (The Netherlands)Google Scholar
  34. Sharkey, T.D., Savitch, L.V., Butz, N.D. (1991) Photometric method for routine determination of kcat and carbamylation of rubisco. Photosynth. Res., in pressGoogle Scholar
  35. Silverthorne, J., Tobin, E.M. (1984) Demonstration of transcriptional regulation of specific genes by phytochrome action. Proc. Natl. Acad. Sci. USA 81, 1112–1116Google Scholar
  36. Stiekema, W.J., Wimpee, C.F., Silverthorne, J., Tobin, E.M. (1983) Phytochrome control of the expression of two nuclear genes encoding chloroplast proteins in Lemna gibba L. G-3. Plant Physiol. 72, 717–724Google Scholar
  37. Terashima, I., Wong, S.-C., Osmond, C.B., Farquhar, G.D. (1988) Characterisation of non-uniform photosynthesis induced by abscisic acid in leaves having different mesophyll anatomies. Plant Cell Environ. 29, 385–394Google Scholar
  38. Valentin, K., Zetsche, K. (1989) The genes of both subunits of ribulose-1,5-bisphosphate carboxylase constitute an operon on the plastome of a red alga. Curr. Genet. 16, 203–209Google Scholar
  39. Vassey, T.L. (1988) Phytochrome mediated regulation of sucrose phosphate synthase activity in maize. Plant Physiol. 88, 540–542Google Scholar
  40. Vassey, T.L. (1989) Light/dark profiles of sucrose phosphate synthase, sucrose synthase, and acid invertase in leaves of sugarbeet. Plant Physiol. 89, 347–351Google Scholar
  41. Vassey, T.L., Sharkey, T.D. (1989) Mild water stress of Phaseolus vulgaris plants leads to reduced starch synthesis and extractable sucrose phosphate synthase activity. Plant Physiol. 89, 1066–1070Google Scholar
  42. Wehmeyer, B., Cashmore, A.R., Schafer, E. (1990) Photocontrol of the expression of genes encoding chlorophyll a/b binding proteins and small subunit of ribulose-1,5-bisphosphate carboxylase in etiolated seedlings of Lycopersicon esculentum (L.) and Nicotiana tabacum (L.). Plant Physiol. 93, 990–997Google Scholar
  43. Winter, U., Feierabend, J. (1990) Multiple coordinate controls contribute to a balanced expression of ribulose-1,5-bisphosphate carboxylase/oxygenase subunits in rye leaves. Eur. J. Biochem. 187, 445–453Google Scholar
  44. Wintermans, J.G.F.M., DeMots, A. (1965) Spectrophotometric characteristics of chlorophylls a and b and their pheophytins in ethanol. Biochim. Biophys. Acta 109, 448–453Google Scholar
  45. Woodrow, I.E., Berry, J.A. (1988) Enzymatic regulation of photosynthetic CO2 fixation in C3 plants. Annu. Rev. Plant Physiol. Plant Mol. Biol. 39, 533–594Google Scholar

Copyright information

© Springer-Verlag 1991

Authors and Affiliations

  • Thomas D. Sharkey
    • 1
  • Terry L. Vassey
  • Peter J. Vanderveer
  • Richard D. Vierstra
    • 2
  1. 1.Department of BotanyUniversity of WisconsinMadisonUSA
  2. 2.Department of HorticultureUniversity of WisconsinMadisonUSA

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