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Plant and Cell Physiology, 1992, Vol. 33, No. 5 569-575
© 1992

Artificial Insemination ‘Regulated by EDTA’ in the Monoecious Brown Alga Fucus evanescens

Isamu Wakana1,3 and Mamoru Abe2

1 Institute of Algological Research, Faculty of Science, Hokkaido University Muroran, 051 Japan
2 Department of Biology, Faculty of Science, Yamagata University Yamagata, 990 Japan

As part of an attempt to control the fertilization of a monoecious brown alga Fucusevanescens, the effects of EDTA on the release and fertilization of gametes were studied. When receptacles were treated for liberation of gametes by soaking in plain seawater, egg-packets and spermatophores were discharged from oogonia and antheridia, respectively, with the gametes retained within their packing envelopes. After a while, the envelopes were disrupted and the gametes were released. Thirty min after the soaking, over 99% of eggs were fertilized. However, when receptacles were soaked in seawater that contained 0.5 mg m1–1 Na2.EDTA at 4°C, the release and fertilization of gametes were prevented after egg-packets and spermatophores had been released from the receptacles. Release of gametes from such egg-packets and spermatophores occurred rapidly when the medium was diluted with an excess of plain seawater. The chelating agent affected the disruption of egg-packets and spermatophores, but it did not affect the subsequent fertilization and development of the fertilized eggs. On the basis of these results, normal unfertilized eggs and sperm were isolated separately by filtration and centrifugation of the released packing envelopes in the presence of 0.5 mg ml–1 Na2EDTA at 4°C. Artificial insemination using the isolated gametes was successful.

3 Present address: Akan Board of Education, Akan, 085-02 Japan.

(Received August 13, 1991; Accepted April 27, 1992)
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