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Preparation and properties of pollen sporoplasts

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Summary

Methods for the removal of exine from mature, ungerminatedLilium longiflorum pollen and release of intact gametophytes (sporoplasts) have been developed. These methods rely on the low temperature solvolytic activity of 4-methylmorpholine N-oxide (MMNO), which allows partial or complete detachment of exine from intine during subsequent washing procedures. These methods are: aqueous MMNO combined with cyclohexylamine (method I), aqueous MMNO at alkaline pH (method II), and aqueous MMNO containing a high Ca2+ concentration with added cellulysin and macerase (method III). Sporoplasts produced by methods I and II are most frequently completely separated from exine and, as shown by histochemical tests, enveloped by the intine layer. Selected enzyme activities in method II sporoplasts are measurable but, as indicated by other tests, considerable damage to the plasma membrane accompanies this treatment. Sporoplasts produced by melhod III largely remain attached to their ruptured exine layer and retain substantial biological competence in terms of extractable enzyme activities, membrane integrity, and respiration.

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Abbreviations

MMNO:

4-methylmorpholine N-oxide

SEM:

scanning electron microscope

TEM:

transmission electron microscope

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Author notes

  1. B. G. Baldi

    Present address: Plant Hormone Laboratory, U.S. Department of Agriculture, Beltsville Agricultural Research Center, B050, 20705, Beltsville, MD, USA

Authors and Affiliations

  1. Graduate Program in Plant Physiology, Washington State University, USA

    B. G. Baldi

  2. Department of Botany, Washington State University, USA

    V. R. Franceschi

  3. Institute of Biological Chemistry, Washington State University, 99164-6340, Pullman, WA, USA

    B. G. Baldi & F. A. Loewus

Authors
  1. B. G. Baldi
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  2. V. R. Franceschi
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  3. F. A. Loewus
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Baldi, B.G., Franceschi, V.R. & Loewus, F.A. Preparation and properties of pollen sporoplasts. Protoplasma 141, 47–55 (1987). https://doi.org/10.1007/BF01276787

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  • DOI: https://doi.org/10.1007/BF01276787

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