Biotechnology Applications of Microinjection, Microscopic Imaging, and Fluorescence pp 11-17 | Cite as
How to Make Glass Microtools for the Injection of Isolated Plant Sperm Cells into Embryo SAC Cells Using a Microforge
Abstract
Preceding fertilization in higher plants, the pollen tube penetrates a synergid of the embryo sac in order to shed its contents, including the two sperm cells. In order to analyse additional functions of the pollen tube cytoplasm in the synergid (apart from sperm cell transfer), we directly injected isolated sperm cells into synergids of Torenia fournieri, using micromanipulation (Keijzer et al., 1988a; Keijzer, 1992). The rate of successful sperm cell transfers in these studies was, however, low (Keijzer, 1992) but serve to illustrate the potential usefulness of the technique to be described below.
Keywords
Pollen Tube Sperm Cell Sperm Cell Transfer Heated Filament Injection PipettePreview
Unable to display preview. Download preview PDF.
References
- De Fonbrune, P. (1949) Technique de Micromanipulation, Masson, Paris.Google Scholar
- Keijzer, CJ., Reinders, M.C., and Leferink-ten Klooster, H.B. (1988a) A micromanipulation method for artificial fertilization in Torenia, in: “Sexual Reproduction in Higher Plants”, M. Cresti, P. Gori and E. Pacini, eds., Springer, Berlin, 119–124.Google Scholar
- Keijzer, C.J., Wilms, H.J., and Mogensen, H.L. (1988b) Sperm cell research, the current status and applications for plant breeding, in: “Plant Sperm Cells as Tools for Biotechnology”, H.J. Wilms and CJ. Keijzer, eds., Pudoc, Wageningen, 3–8.Google Scholar
- Keijzer, C.J. (1992) The isolation of sperm cells, their microinjection into the egg apparatus and methods for structural analysis of the injected cells, in: “Promotion of Sexual Plant Reproduction”, M. Cresti and A. Tiezzi, eds., Springer, Berlin.Google Scholar
- Theunis, C.H., Pierson, E.S., and Cresti, M. (1991) Isolation of male and female gametes in higher plants. Sex. Plant Reprod. 4: 145–154.CrossRefGoogle Scholar