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Procedure to Isolate Viable Sperm Cells from Corn (Zea mays L.) Pollen Grains
I Dupuis1, P Roeckel, E Matthys-Rochon
1Université Claude Bernard-Lyon 1, Laboratoire Reconnaissance Cellulaire et Amélioration des Plantes, Bät 741, 43 Boulevard du 11 Novembre 1918, F-69622 Villeurbanne Cedex, France.
Abstract:
Sperm cells were isolated from corn (Zea mays L.) tricellular pollen grains. They were released using a light osmotic chock, and separated from pollen contaminants (especially starch grains) by a Percoll gradient centrifugation. Isolated sperm cells (3 x 10(6) per milliliter) show a high viability score (90%) as demonstrated with the fluorochromatic reaction. They appeared as spherical cells which lack cell wall and plastids, and can be considered as haploid protoplasts.
