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Isolation and collection of two populations of viable sperm cells from the pollen of Plumbago zeylanica

Published online by Cambridge University Press:  01 November 1998

Zhaojie Zhang ,
Huiling Xu ,
Mohan B. Singh  and
Scott D. Russell
Zhaojie Zhang
Affiliation:
Department of Botany and Microbiology, University of Oklahoma, Norman, Oklahoma, USA
Huiling Xu
Affiliation:
School of Botany, University of Melbourne, Parkville, Victoria 3052, Australia
Mohan B. Singh
Affiliation:
School of Botany, University of Melbourne, Parkville, Victoria 3052, Australia
Scott D. Russell
Affiliation:
Department of Botany and Microbiology, University of Oklahoma, Norman, Oklahoma, USA
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Abstract

A protocol is described for individually collecting two populations of sperm cells, Svn and Sua, from pollen of Plumbago zeylanica. Pollen grains were burst in 10 mM MOPS buffer containing 0.8 M mannitol (pH 4.6). Paired sperm cells released from pollen were separated using a microinjector. Svn and Sua were then collected individually with a microinjector, based upon known size differences. Collected sperm cells were washed with isolation medium and transferred to liquid nitrogen until use. Fluorochromatic reaction (FCR) test of isolated sperm cells showed a positive reaction, indicating that the isolated sperm cells are viable; most of the sperm cells retain viability for at least 2 h.

Keywords

Plumbago zeylanicaPollenPreferential fertilisationSperm cell isolation
Type
Research Article
Information
Zygote , Volume 6 , Issue 4 , November 1998 , pp. 295 - 298
Copyright
1998 Cambridge University Press

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