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Development of cycad ovules and seeds. 1. Implication of the ER in primary cellularisation of the megagametophyte in Encephalartos natalensis Dyer and Verdoorn

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Abstract

Very little is known about the pre- and post-shedding megagametophyte development and utilisation of accumulated reserves, respectively, in cycads (Zamiaceae). In the present study on developing ovules of the recalcitrant-seeded species, Encephalartos natalensis, cells of the megagametophyte were found to become progressively packed with starch and protein as the two main storage reserves, a limited number of discrete lipid bodies, and occasional mitochondria all of which appeared to be embedded in a homogeneous matrix. ER-derived vesicles (and not Golgi-derived vesicles) appeared to be the principal contributors of the primary cell wall components, pectin and xylan, during megagametophyte cellularisation. This was confirmed by the use of enzyme-gold localisation. High-pressure freezing (HPF) and freeze substitution (FS) of samples the following season showed that while the apparently featureless cytomatrix of the megaspore was an artefact of conventional fixation, there was still an insignificant occurrence of Golgi bodies during primary wall formation. When enzyme-gold localisation was employed on the HPF-FS material, label for pectin and xylan was found only in the regions of ER and vesicles and not in any of the few Golgi bodies or their associated vesicles. Immunocytochemistry revealed that pectin and xylan were restricted to the ER and not to any vesicles or to the occasional Golgi body that was found. This suggests that the ER exclusively, is involved in the deposition of these primary cell wall components during the cellularisation of the E. natalensis megagametophyte. While cellularisation took place over approximately 1–2 weeks, subsequent development of the megagametophyte cells involved the accumulation of storage reserves, this phase lasting approximately 8 months—after which the seeds were shed whether pollination/fertilisation had recently occurred, or not.

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Notes

  1. In the context of the present work, however, this is described as the central compartment, as it is not a vacuole in terms of the accepted definition (e.g. Lamb and Berjak 1981).

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Acknowledgments

We would like to thank Ms P. Maartens and Dr J. Wesley-Smith (both of the Electron Microscope Unit, University of KwaZulu-Natal, Westville Campus) for assistance with the microscopy, and Mr C. van der Merwe (Laboratory for Microscopy and Microanalysis, University of Pretoria) for facilitating the HPF and FS work. This work was supported by the National Research Foundation (NRF) of South Africa.

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  1. Biological and Conservation Sciences Building, University of KwaZulu-Natal, South Ring Road, Westville Campus, Durban, 4001, South Africa

    Wynston Ray Woodenberg, Patricia Berjak & Norman William Pammenter

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  1. Wynston Ray Woodenberg
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Correspondence to Wynston Ray Woodenberg.

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Woodenberg, W.R., Berjak, P. & Pammenter, N.W. Development of cycad ovules and seeds. 1. Implication of the ER in primary cellularisation of the megagametophyte in Encephalartos natalensis Dyer and Verdoorn. Plant Growth Regul 62, 265–278 (2010). https://doi.org/10.1007/s10725-010-9469-6

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