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Isolation and metabolism of Vigna unguiculata root nodule protoplasts

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Abstract

Axenic cultures of bacteroid-containing protoplasts were isolated from root nodules of Vigna unguiculata L. Walp. Dimensions of the protoplasts were 35 to 135 μm long x 35 to 95 μm wide. Yields were about 30 to 50 mg dry weight per gram fresh weight of nodules. About 5x108 protoplasts packed into 1 ml of basal medium under the influence of gravity. When incubated in hypertonic, nitrogen-free media, freshly isolated protoplasts began to reduce acetylene to ethylene after a lag period of 24 to 48 h. Various additions to the basal medium showed that the system possessed functional glycolytic and tricarboxylic acid pathways. Endogenous application of various intermediary metabolites stimulated both acetylene reduction and respiration, though not often equally. As acetylene reduction, but not respiration, was inhibitable by both asparagine and glutamine, the system appears suitable for the study of mechanisms controlling symbiotic nitrogen fixation.

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Abbreviations

BSA:

bovine serum albumine

HEPES:

N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid

PEP:

phospho(enol)pyruvate

UMKL 76:

University of Malaga, Kuala Lumpur, Rhizobium, No. 76

TCAC:

tricarboxylic acid cycle

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Author notes

  1. K. C. Wooi

    Present address: Clonal Oil Palm Research Unit, Unipamol Malaysia Sdn. Bhd., Layang-Layang, Johor, Malaysia

  2. W. J. Broughton

    Present address: Max-Planck-Institut für Züchtungsforschung, D-5000, Köln 30

Authors and Affiliations

  1. Department of Genetics and Cellular Biology, University of Malaya, Kuala Lumpur, Malaysia

    K. C. Wooi & W. J. Broughton

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  1. K. C. Wooi
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Wooi, K.C., Broughton, W.J. Isolation and metabolism of Vigna unguiculata root nodule protoplasts. Planta 145, 487–495 (1979). https://doi.org/10.1007/BF00380104

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  • DOI: https://doi.org/10.1007/BF00380104

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