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Characterisation of a glutathione reductase gene and its genetic locus from pea (Pisum sativum L.)

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Abstract

A cDNA encoding the chloroplast/mitochondrial form of glutathione reductase (GR; EC 1.6.4.2) from pea (Pisum sativum L.) was used to map a single GR locus, named GOR1. In two domesticated genotypes of pea (cv. Birte and JI 399) it is likely that the GOR1 locus contains a single gene. However, in a semi-domesticated land race of pea (JI 281) two distinct but closely related sets of GR gene sequences were detected at the GOR1 locus. The extra GR sequences in JI 281 represent either a second intact gene or a partial or pseudogene copy. A GR gene was cloned from cv. Birte, sequenced and its structure analysed. No feature of the transcription or structure of the gene suggested a mechanism for generating any more than one form of GR. From these data plus previously published biochemical evidence we suggest that a second, distinct gene encoding for the cytosolic form of GR should be present in peas. The GOR1-encoded GR mRNA can be detected in all main organs of the plant and no alternative spliced species was present which could perhaps account for the generation of multiple isoforms of GR. The mismatch between the number of charge-separable isoforms in pea and the proposed number of genes suggests that different GR isoforms arise by some form of post-translational modification.

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Abbreviations

GR:

glutathione reductase

PCR:

polymerase chain reaction

RACE:

rapid amplification of cDNA ends

RFLP:

restriction fragment length polymorphism

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Correspondence to Philip Mullineaux.

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The pea glutathione reductase gene sequences will appear in the EMBL/Genbank databases with the accession number X90996

The authors are grateful to Dr. S. Karpinski for critical reading of the manuscript. This work was supported by a grant-in-aid from the Biotechnology and Biological Sciences Research Council (BBSRC) to the John Innes Centre. C.E. is grateful for a fellowship from the European Molecular Biology Organisation. We acknowledge the support of the BBSRC Global Environmental Change Programme and the support of the Commission of the European Union Framework 3 Agro-Industrial Programme (contract no. AIR-CT92-0205).

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Mullineaux, P., Enard, C., Hellens, R. et al. Characterisation of a glutathione reductase gene and its genetic locus from pea (Pisum sativum L.). Planta 200, 186–194 (1996). https://doi.org/10.1007/BF00208308

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