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Optimization of isolation and storage of sperm cells from pollen of perennial ryegrass (Lolium perenne L.)

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Summary

A procedure for isolating sperm cells of perennial ryegrass (Lolium perenne L.) was developed. The sperm cells were released from the pollen grains by osmotic shock, with the right combination of pH and osmolality being important for optimal release. Various combinations of vitamins E, C and fetal calf serum were tested with the aim of improving yield and long-term viability, and their possible mode of action as important components for improvement of these two parameters is discussed. Under optimized conditions, a yield of 12% was established, and the storage time after which 50% of the sperm cells were still viable was improved to 60 h. Cytological observations demonstrated that sperm cells of perennial ryegrass are true protoplasts, which may allow future fusion experiments to be carried out.

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Authors and Affiliations

  1. Department of Cell Biology, Centre for Plant Breeding and Reproduction Research, CPRO-DLO, PO Box 16, 6700 AA, Wageningen, The Netherlands

    H. M. van der Maas, M. A. C. M. Zaal, E. R. de Jong & F. A. Krens

  2. Department of Plant Cytology and Morphology, Wageningen Agricultural University, Arboretumlaan 4, 6703 BD, Wageningen, The Netherlands

    J. L. van Went

Authors
  1. H. M. van der Maas
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  2. M. A. C. M. Zaal
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  3. E. R. de Jong
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  4. J. L. van Went
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  5. F. A. Krens
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van der Maas, H.M., Zaal, M.A.C.M., de Jong, E.R. et al. Optimization of isolation and storage of sperm cells from pollen of perennial ryegrass (Lolium perenne L.). Sexual Plant Reprod 6, 64–70 (1993). https://doi.org/10.1007/BF00227585

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