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Induction of a ricinosomal-protease and programmed cell death in tomato endosperm by gibberellic acid

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Abstract

Several examples of programmed cell death (PCD) in plants utilize ricinosomes, organelles that appear prior to cell death and store inactive KDEL-tailed cysteine proteinases. Upon cell death, the contents of ricinosomes are released into the cell corpse where the proteinases are activated and proceed to degrade any remaining protein for use in adjacent cells or, in the case of nutritive seed tissues, by the growing seedling. Ricinosomes containing pro-SlCysEP have been observed in anther tissues prior to PCD and ricinosome-like structures have been observed in imbibed seeds within endosperm cells of tomato. The present study confirms that the structures in tomato endosperm cells contain pro-SlCysEP making them bona fide ricinosomes. The relative abundance of pro- versus mature SlCysEP is suggested to be a useful indicator of the degree of PCD that has occurred in tomato endosperm, and is supported by biochemical and structural data. This diagnostic tool is used to demonstrate that a sub-region of the micropylar endosperm surrounding the emerged radical is relatively long-lived and may serve to prevent loss of mobilized reserves from the lateral endosperm. We also demonstrate that GA-induced reserve mobilization, SlCysEP accumulation and processing, and PCD in tomato endosperm are antagonized by ABA.

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Abbreviations

PCD:

Programmed cell death

ABA:

Abscisic acid

GA:

Gibberellic acid

CysEP:

Cysteine endopeptidase

NG:

Not germinated

SS:

Short seedling

LS:

Long seedling

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Acknowledgments

We thank Usher Posluszny for the use of his epi-illumination microscope, and Bob Harris for his assistance with the TEM. This work was funded through the award of a Natural Sciences and Engineering Research Council of Canada Discovery Grant to J.S.G.

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Authors and Affiliations

  1. Department of Plant Agriculture, University of Guelph, Guelph, ON, N1G 2W1, Canada

    Christopher P. Trobacher

  2. Department of Biology, University of Waterloo, Waterloo, ON, N2L 3G1, Canada

    Adriano Senatore

  3. Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON, N1G 2W1, Canada

    Christine Holley & John S. Greenwood

Authors
  1. Christopher P. Trobacher
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  2. Adriano Senatore
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  3. Christine Holley
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  4. John S. Greenwood
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Corresponding author

Correspondence to Christopher P. Trobacher.

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425_2012_1780_MOESM1_ESM.jpg

Supplementary material 1 Transcript analysis of cysteine proteinase genes in germinating and post-germinative seeds. Gene-specific primers were used to amplify regions of several cysteine proteinase transcripts via RT-PCR performed on total RNA extracted from LS embryos/seedlings and from NG, SS and LS seeds, all at 144 HAI. A representative ethidium bromide stained agarose gel, selected from three biological replicates, is shown; actin is included as a loading control (JPEG 80 kb)

Supplementary material 2 (DOCX 14 kb)

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Trobacher, C.P., Senatore, A., Holley, C. et al. Induction of a ricinosomal-protease and programmed cell death in tomato endosperm by gibberellic acid. Planta 237, 665–679 (2013). https://doi.org/10.1007/s00425-012-1780-1

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  • DOI: https://doi.org/10.1007/s00425-012-1780-1

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