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Formation of wheat (Triticum aestivum L.) embryogenic callus involves peroxide-generating germin-like oxalate oxidase

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Abstract

In wheat (Triticum aestivum L.), embryogenic callus formation comprises suppression of precocious germination by the zygotic embryo and the initiation of dedifferentiated cellular proliferation within it. Embryogenic calli are induced by treating immature embryos with 2,4-dichlorophenoxyacetic acid (2,4-D). Upon withdrawal from 2,4-D, somatic embryos develop from the periphery of the callus. Prior to visible callus formation, there is a striking induction of “germin-like” oxalate oxidase (“gl-OXO”: EC 1.2.3.4) gene expression. Accumulation of gl-OXO mRNA is rapidly stimulated upon auxin treatment, with a consequent development of apoplastic enzyme activity producing H2O2 within the cell wall. Within the dedifferentiated calli, gl-OXO enzyme activity becomes widespread over the surface of embryogenic calli. Differentiation of somatic embryos is initiated in regions of densely cytoplasmic, meristematic cells that are marked by highly localised expression of gl-OXO activity within these embryogenic cell masses. We suggest that this localised generation of H2O2 by gl-OXO promotes peroxidative cross-linking of cell wall components, thereby preventing cellular expansion and maintaining these cell masses in an embryogenically competent condition.

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Fig. 1a–d
Fig. 2a–e
Fig. 3a–f
Fig. 4a–e
Fig. 5a–f

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Notes

  1. When measured on a per unit protein basis, the decline in OXO specific activity is much more marked, as the quantity of protein accumulating in the growing callus increases.

Abbreviations

daf :

Days after fertilisation

2,4-D:

2,4-Dichlorophenoxyacetic acid

gl-OXO :

Germin-like oxalate oxidase

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Acknowledgements

M.T. gratefully acknowledges the support a studentship from the Scientific and Technical Research Council of Turkey, and M.C. is indebted to a scholarship from Mustafa Kemal University, Antakya, Turkey. We thank Adrian Hick for his skill in preparation of the scanning electron micrographs, and Dr. Paul Knox (Leeds University) for assistance and instruction in immunofluorescence microscopy. As ever, we are pleased to acknowledge the assistance of Prof. Byron Lane (University of Toronto) who supplied the anti-germin serum and the germin cDNA clone in addition to valuable criticism, encouragement and insight in equal measure.

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Author notes

  1. Mahmut Caliskan

    Present address: Department of Biology, Faculty of Arts and Sciences, Mustafa Kemal University, 31024, Hatay, Turkey

  2. Müge Turet

    Present address: Department of Molecular Biology and Genetics, Bogazici University, 80815, Bebek, Istanbul, Turkey

Authors and Affiliations

  1. Centre for Plant Sciences, Leeds University, Leeds, LS2 9JT, UK

    Mahmut Caliskan, Müge Turet & Andrew C. Cuming

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  1. Mahmut Caliskan
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  2. Müge Turet
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  3. Andrew C. Cuming
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Correspondence to Andrew C. Cuming.

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This paper is dedicated to Professor Byron Lane, on the occasion of his retirement, in celebration of a distinguished scientific career.

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Caliskan, M., Turet, M. & Cuming, A.C. Formation of wheat (Triticum aestivum L.) embryogenic callus involves peroxide-generating germin-like oxalate oxidase. Planta 219, 132–140 (2004). https://doi.org/10.1007/s00425-003-1199-9

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