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Embryo cell wall properties in relation to development and desiccation in the recalcitrant-seeded Encephalartos natalensis (Zamiaceae) Dyer and Verdoorn

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Abstract

Plant cell walls are dynamic entities that may change with development, differ between plant species and tissue type and play an important role in responses to various stresses. In this regard, the present investigation employed immunocytochemistry to determine wall composition and possible changes during development of immature and mature embryos of the recalcitrant-seeded cycad Encephalartos natalensis. Fluorescent and gold markers, together with cryo-scanning and transmission electron microscopy (TEM) were also used to analyse potential changes in the cell walls of mature embryos upon desiccation. Immature cell walls were characterised by low- and high methyl-esterified epitopes of pectin, rhamnogalacturonan-associated arabinan, and the hemicellulose xyloglucan. Arabinogalactan protein recognised by the LM2 antibody, along with rhamnogalacturonan-associated galactan and the hemicellulose xylan, were not positively localised using immunological probes, suggesting that the cell walls of the embryo of E. natalensis do not possess these epitopes. Interestingly, mature embryos appeared to be identical to immature ones with respect to the cell wall components investigated, implying that these may not change during the protracted post-shedding embryogenesis of this species. Drying appeared to induce some degree of cell wall folding in mature embryos, although this was limited by the abundant amyloplasts, which filled the cytomatrical space. Folding, however, was correlated with relatively high levels of wall plasticisers typified by arabinose polymers. From the results of this study, it is proposed that the embryo cell walls of E. natalensis are constitutively prepared for the flexibility required during cell growth and expansion, which may also facilitate the moderate cell wall folding observed in mature embryos upon drying. This, together with the abundant occurrence of amyloplasts in the cytomatrix, may provide sufficient mechanical stabilisation if water is lost, even though the seeds of this species are highly desiccation-sensitive.

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Acknowledgments

We wish to thank Vishal Bharuth, Phillip Christopher and Nelisha Murugan (the microscopy and Microanalysis Unit, University of KwaZulu-Natal, Westville Campus) for technical assistance with the microscopy. This work was supported by the National Research Foundation (NRF) of South Africa.

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Authors and Affiliations

  1. School of Life Sciences, University of KwaZulu-Natal (Westville Campus), Durban, 4001, South Africa

    Wynston Ray Woodenberg, N. W. Pammenter, Jill M. Farrant & Patricia Berjak

  2. Department of Molecular and Cell Biology, University of Cape Town, Rondebosch, 7701, South Africa

    Jill M. Farrant

  3. Laboratoire ‘Glycobiologie et Matrice Extracellulaire Végétale’, Glyco-MEV, IFRMP23-PRIMACEN IBiSA, Université de Rouen, 76821, Mont-Saint-Aignan, France

    Azeddine Driouich

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  1. Wynston Ray Woodenberg
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  2. N. W. Pammenter
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Correspondence to Wynston Ray Woodenberg.

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Fig. 7

Fig. 7
figure 7

The figure demonstrates fluorescence microscopy images of negative and positive controls used to verify results. a Some autofluorescence appeared to be present in the cells with faint fluorescence evident in the nuclei and certain areas of the cytomatrix; bar 50 μm. b A similar picture can be seen in a primary antibody only control; bar 50 μm. c Fluorescence was somewhat more intense in the nuclei and cytomatrix when the protocol was performed with the secondary antibody only; bar 50 μm. d Relatively intense fluorescence was evident in the root tip cell walls [especially in the xylem walls (arrowed)] of Daucus spp. when the LM2 antibody was employed; bar 50 μm. e Epidermal and subepidermal cell walls display fluorescence when the LM5 antibody was used on the pericarp tissue of Lycopersicon spp.; bar 50 μm. f Intense fluorescence is apparent when the LM11 antibody was used on Z. mays leaf tissue; bar 100 μm

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Woodenberg, W.R., Pammenter, N.W., Farrant, J.M. et al. Embryo cell wall properties in relation to development and desiccation in the recalcitrant-seeded Encephalartos natalensis (Zamiaceae) Dyer and Verdoorn. Protoplasma 252, 245–258 (2015). https://doi.org/10.1007/s00709-014-0672-6

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