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The compartmentation of carboxylating and decarboxylating enzymes in guard cell protoplasts

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Abstract

Guard cell and mesophyll cell protoplasts of Vicia faba L. were purified and separated into cytoplasmic and plastid fractions by a selective silicone-oil filtration. Before fractionation, the protoplasts were ruptured by a low speed centrifugation through a narrow-aperture nylon net placed in a plastic vial. This protoplast homogenation and subsequently the silicone-oil fractionation offer the possibility of investigating the comparatmentation of the enzymatic carboxylating (ribulose bisphosphate carboxylase EC 4.1.1.39, phosphoenolpyruvate carboxylase EC 4.1.1.31, NAD+ and NADP+ linked malate dehydrogenase EC 1.1.1.37) and decarboxylating pathways of malic (malic enzyme EC 1.1.1.40, phosphoenolpyruvate carboxykinase EC 4.1.1.32, pyruvate orthophosphate dikinase EC 2.7.9.1) which occur during the swelling and shrinking of the guard cell protoplasts. A model is proposed which describes the transport processes of malic acid during the starch-malate balance as correlated to the volume changes of the protoplasts. As the enzymes and their compartmentation in the guard cell protoplasts seem to be consistent with those of crassulacean acid metabolism (CAM) plants, the metabolism of stomata and of CAM cells is compared.

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Abbreviations

AQ:

anthraquinone-2-sulfonic acid

CAM:

Crassulacean acid metabolism

DCPIPred :

2,6-dichlorophenol-indophenol

DTT:

dithiothreitol

EDTA:

ethylendiamine tetraacetic acid

GAPDH:

glyceraldehyde-3-phosphate dehydrogenase

HEPES:

N-2-hydroxyethyl-piperazine-N-2-ethane sulphonic acid

MDH:

malante dehydrogenase

MES:

2(N-morpholino) ethane sulphonic acid

OAA:

oxaloacetic acid

PEP:

phosphoenolpyruvate

PSI:

photosystem I

KuP2 :

ribulose bisphosphate

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  1. Institut für Botanik und Mikrobiologie, Technische Universität München, Arcisstraße 21, D-8000, München 2, Federal Republic of Germany

    H. Schnabl

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Schnabl, H. The compartmentation of carboxylating and decarboxylating enzymes in guard cell protoplasts. Planta 152, 307–313 (1981). https://doi.org/10.1007/BF00388254

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  • DOI: https://doi.org/10.1007/BF00388254

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