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Ribulose-1,5-bisphosphate carboxylase-oxygenase, other Calvin-cycle enzymes, and chlorophyll decrease when glucose is supplied to mature spinach leaves via the transpiration stream

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Abstract

The inhibition of photosynthesis after supplying glucose to detached leaves of spinach (Spinacia oleracea L.) was used as a model system to search for mechanisms which potentially contribute to the “sink” regulation of photosynthesis. Detached leaves were supplied with 50 mM glucose or water for 7 d through the transpiration stream, holding the leaves in low irradiance (16 μmol photons · m−2 · s−1) and a cycle of 9 h light/15 h darkness to prevent any endogenous accumulation of carbohydrate. Leaves supplied with water only showed marginal changes of photosynthesis, respiration, enzyme levels or metabolites. When leaves were supplied with 50 mM glucose, photosynthesis was gradually inhibited over several days. The inhibition was most marked when photosynthesis was measured in saturating irradiance and ambient CO2, less marked in saturating irradiance and saturating CO2, and least marked in limiting irradiance. There was a gradual loss of ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco) protein, fructose-1,6-bisphosphatase, NADP-glyceraldehyde-3-phosphate dehydrogenase and chlorophyll. The inhibition of photosynthesis was accompanied by a large decrease of glycerate-3-phosphate, an increase of triose-phosphates and fructose-1,6-bisphospate, and a small decrease of ribulose-1,5-bisphosphate. The stromal NADPH/NADP ratio increased (as indicated by increased activation of NADP-malate dehydrogenase), and the ATP/ADP ratio increased. Chlorophyll-fluorescence analysis indicated that thylakoid energisation was increased, and that the acceptor side of photosystem II was more reduced. Similar results were obtained when glucose was supplied by floating leaf discs in low irradiance on glucose solution, and when detached spinach leaves were held in high light to produce an endogenous accumulation of carbohydrate. Feeding glucose also led to an increased rate of respiration. This was not accompanied by any changes of pyruvate kinase, phosphofructokinase, or pyrophosphate: fructose-6-phosphate phosphotransferase activity. There was a decrease of phosphoenolpyruvate, glycerate-3-phosphate and glycerate-2-phosphate, an increase of pyruvate and triose-phosphates, and an increased ATP/ADP ratio. These results show (i) that accumulation of carbohydrate can inhibit photosynthesis via a long-term mechanism involving a decrease of Rubisco and other Calvin-cycle enzymes and (ii) that respiration is stimulated due to an unknown mechanism, which increases the utilisation of phosphoenolpyruvate.

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Abbreviations

Ci :

CO2 concentration in the air space within the leaf

Fm :

fluorescence yield with a saturating pulse in dark-adapted material

Fo :

ground level of fluorescence using a weak non-actinic modulated beam in the dark

Fru1,6bisP:

fructose-1,6-bisphosphate

Fru1,6Pase:

fructose-1,6-bisphosphatase

Fru2,6bisP:

fructose-2,6-bisphosphate

IRGA:

infrared gas analyser

NAD-MDH:

NAD-dependent malate dehydrogenase

NADP-MDH:

NADP-dependent malate dehydrogenase

NADP-GAPDH:

NADP-dependent glyceraldehyde-3-phosphate dehydrogenase

PEP:

phosphoenolpyruvate

PFK:

phospho-fructokinase

PFP:

pyrophospate: fructose-6-phosphate-phosphotransferase

3-PGA:

glycerate-3-phospate

Pi :

inorganic phosphate

Ru1,5bisP:

ribulose 1,5-bisphosphate

Rubisco:

ribulose-1,5-bisphosphate carboxylase-oxygenase

triose-phosphates:

sum of glyceraldehyde-3-phosphate and dihydroxyacetone phosphate

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Author notes

  1. W. Paul Quick

    Present address: Department of Molecular Biology and Biotechnology, University of Sheffield, UK

  2. Mark Stitt

    Present address: Botanisches Institut, Im Neuenheimerfeld 360, W-6900, Heidelberg, Germany

Authors and Affiliations

  1. Lehrstuhl für Pflanzenphysiologie, Universität Bayreuth, W-8580, Bayreuth, Germany

    Anne Krapp, W. Paul Quick & Mark Stitt

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  1. Anne Krapp
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This research was supported by the Deutsche Forschungsgemeinschaft (SFB 137).

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Krapp, A., Quick, W.P. & Stitt, M. Ribulose-1,5-bisphosphate carboxylase-oxygenase, other Calvin-cycle enzymes, and chlorophyll decrease when glucose is supplied to mature spinach leaves via the transpiration stream. Planta 186, 58–69 (1991). https://doi.org/10.1007/BF00201498

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