Planta

, Volume 211, Issue 1, pp 112-119

First online:

Decrease of phosphoribulokinase activity by antisense RNA in transgenic tobacco: definition of the light environment under which phosphoribulokinase is not in large excess

  • Matthew J. PaulAffiliated withBiochemistry and Physiology Department, IACR-Rothamsted, Harpenden, Hertfordshire, AL5 2JQ, UK
  • , Simon P. DriscollAffiliated withBiochemistry and Physiology Department, IACR-Rothamsted, Harpenden, Hertfordshire, AL5 2JQ, UK
  • , P. John AndralojcAffiliated withBiochemistry and Physiology Department, IACR-Rothamsted, Harpenden, Hertfordshire, AL5 2JQ, UK
  • , Jacqui S. KnightAffiliated withDepartment of Plant Sciences, University of Cambridge, Downing Street, Cambridge, CB2 3EA, UK
  • , John C. GrayAffiliated withDepartment of Plant Sciences, University of Cambridge, Downing Street, Cambridge, CB2 3EA, UK
  • , David W. LawlorAffiliated withBiochemistry and Physiology Department, IACR-Rothamsted, Harpenden, Hertfordshire, AL5 2JQ, UK

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Abstract.

 To test the hypothesis that the contribution of phosphoribulokinase (PRK) to the control of photosynthesis changes depending on the light environment of the plant, the response of transgenic tobacco (Nicotiana tabacum L.) transformed with antisense PRK constructs to irradiance was determined. In plants grown under low irradiance (330 μmol m−2 s−1) steady-state photosynthesis was limited in plants with decreased PRK activity upon exposure to higher irradiance, with a control coefficient of PRK for CO2 assimilation of 0.25 at and above 800 μmol m−2 s−1. The flux control coefficient of PRK for steady-state CO2 assimilation was zero, however, at all irradiances in plant material grown at 800 μmol m−2 s−1 and in plants grown in a glasshouse during mid-summer (alternating shade and sun 300–1600 μmol m−2 s−1). To explain these differences between plants grown under low and high irradiances, Calvin cycle enzyme activities and metabolite content were determined. Activities of PRK and other non-equilibrium Calvin cycle enzymes fructose-1,6-bisphosphatase, sedoheptulose-1,7-bisphosphatase and ribulose-1,5-bisphosphate carboxylase-oxygenase were twofold higher in plants grown at 800 μmol m−2 s−1 or in the glasshouse than in plants grown at 330 μmol m−2 s−1. Activities of equilibrium enzymes transketolase, aldolase, ribulose-5-phosphate epimerase and isomerase were very similar under all growth irradiances. The flux control coefficient of 0.25 in plants grown at 330 μmol m−2 s−1 can be explained because low ribulose-5-phosphate content in combination with low PRK activity limits the synthesis of ribulose-1,5-bisphosphate. This limitation is overcome in high-light-grown plants because of the large relative increase in activities of sedoheptulose-1,7-bisphosphatase and fructose-1,6-bisphosphatase under these conditions, which facilitates the synthesis of larger amounts of ribulose-5-phosphate. This potential limitation will have maintained evolutionary selection pressure for high concentrations of PRK within the chloroplast.

Key words: Calvin cycle – Light –Nicotiana (Calvin cycle) – Phosphoribulokinase – Photosynthesis – Transgenic tobacco