Abstract
A low-starch mutant accumulating less than 5% of wild-type amounts was isolated after X-ray mutagenesis of Chlamydomonas reinhardtii cells. The recessive st-1-1 defect segregated as a single mendelian mutation through meiosis, and led to a severe decrease in starch accumulation under all culture conditions tested, whether in the light or in darkness. Adenosine 5′-diphosphoglucose pyrophosphorylase (in the absence of 3-phosphoglycerate), starch synthase, phosphoglucomutase, phosphorylase and starch-branching enzyme were all characterized and shown to be unaffected by the mutation. However, ADP-glucose pyrophosphorylase in the mutant had its sensitivity to activation by 3-phosphoglycerate lowered dramatically and became less responsive to orthophosphate. Our results are consistent both with a mutation in a structural gene of a multisubunit enzyme or in a regulatory gene responsible for switching ADP-glucose pyrophosphorylase from a 3-phosphoglycerate-insensitive to a 3-phosphoglycerate-sensitive form. These results provide definite proof of the in-vivo requirement for 3-phosphoglycerate activation to obtain substantial starch synthesis in plants. The conclusions hold both for synthesis from CO2 in the light or from exogenous organic carbon sources in darkness. A model is presented in which the existence of a 3-phosphoglycerate gradient explains localized starch synthesis around the pyrenoid of lower plants.
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Abbreviations
- glucose-1-P:
-
glucose-1-phosphate
- glucose-6-P:
-
glucose-6-phosphate
- Pi:
-
orthophosphate
- 3PGA:
-
3-phospho-glycerate
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This work was supported by the Université des Sciences et Techniques de Lille Flandres-Artois, the Ministère de l'Education Nationale and by the Centre National de la Recherche Scientifique (Unité Mixte de Recherche du CNRS n∘ 111, Director: Professor J. Montreuil). We also thank Brigitte Macquart and Michelle Autexier for typing the manuscript, A. Dhainaut (UFR de Biologie, Université des Sciences et Techniques de Lille, France) for helping us with the electron microscopy and R. Matagne (Département de Botanique, Université de Liège, Liège, Belgium) for helpful discussions.
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Ball, S., Marianne, T., Dirick, L. et al. A Chlamydomonas reinhardtii low-starch mutant is defective for 3-phosphoglycerate activation and orthophosphate inhibition of ADP-glucose pyrophosphorylase. Planta 185, 17–26 (1991). https://doi.org/10.1007/BF00194509
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DOI: https://doi.org/10.1007/BF00194509