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The role of malate in ammonia assimilation in cotyledons of radish (Raphanus sativus L.)

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Abstract

The relationships between the metabolism of malate, nitrogen assimilation and biosynthesis of amino acids in response to different nitrogen sources (nitrate and ammonium) have been examined in cotyledons of radish (Raphanus sativus L.). Measurements of the activities of some key enzymes and pulse-chase experiments with [14C]malate indicate the operation of an anaplerotic pathway for malate, which is involved in the synthesis of glutamine during increased ammonia assimilation. It is most likely that the tricarboxylicacid cycle is supplied with carbon through entry of malate, formed via the phosphoenolpyruvate (PEP)-carboxylation pathway, when 2-oxoglutarate leaves the cycle to serve as precursor for an increased synthesis of glutamine via glutamate. This might occur predominantly in the cytosol via the activity of the glutamine synthetase/glutamate synthase (GS/GOGAT) cycle, the NADH-dependent GOGAT being the rate-limiting activity.

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Abbreviations

DTT:

dithiothreitol

EDTA:

ethylenediamine-tetraacetic acid

GDH:

glutamate dehydrogenase

GOGAT:

glutamate synthase (glutamine: 2-oxoglutarate aminotransferase)

GOT:

aspartate aminotransferase (glutamate: oxaloacetate transaminase)

GS:

glutamine synthetase

HPLC:

high-performance liquid chromatography

MCF:

extraction medium of methanol: chloroform: 7M formic acid, 12:5:3, by vol.

MDH:

malate dehydrogenase

MSO:

L-methionine, sulfoximine

PEPCase:

phosphoenolpyruvate carboxylase

TLC:

thin-layer chromatography

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  1. Botanisches Institut der Universität zu Köln, Gyrhofstrasse 15, D-5000, Köln 41, Federal Republic of Germany

    B. Dahlbender & D. Strack

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  1. B. Dahlbender
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  2. D. Strack
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Dahlbender, B., Strack, D. The role of malate in ammonia assimilation in cotyledons of radish (Raphanus sativus L.). Planta 169, 382–392 (1986). https://doi.org/10.1007/BF00392135

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