Micromanipulation of male and female gametes ofNicotiana tabacum: I. Isolation of gametes
- 1 May 1997
- journal article
- Published by Springer Nature in Plant Cell Reports
- Vol. 16 (8), 555-560
- https://doi.org/10.1007/bf01142323
Abstract
The isolation of male and female gametes is a precondition for the micromanipulation of flowering plant gametes. To reflect their condition at fertilization, isolated gametes need to be physiologically mature and vigorous. Sperm cells are isolated from pollen tubes grown on cut styles using the “in vivo/in vitro” technique. Embryo sacs are isolated 2 days after anthesis using brief treatments of minimal concentrations of cell-wall-digesting enzymes on ovules of emasculated flowers. Egg cells are then mechanically separated from the embryo sac, allowing unambiguous identification of cells. Two days is usually the minimum required for the pollen tube to penetrate the ovule and effect fertilization in vivo.Keywords
This publication has 14 references indexed in Scilit:
- Electro-fused isolated wheat (Triticum aestivum L.) gametes develop into multicellular structuresPlant Cell Reports, 1995
- Deleterious effect of minimal enzymatic treatments on the development of isolated maize embryo sacs in cultureSexual Plant Reproduction, 1995
- Early cytological events after induction of cell division in egg cells and zygote development following in vitro fertilization with angiosperm gametesThe Plant Journal, 1995
- In vitro fertilisation of maize by single egg and sperm cell protoplast fusion mediated by high calcium and high pHZygote, 1994
- Regeneration of Fertile Barley Plants from Mechanically Isolated Protoplasts of the Fertilized Egg Cell.Plant Cell, 1994
- An in Vitro System for Adhesion and Fusion of Maize GametesScience, 1994
- The dynamics of calcium distribution in the synergid cells of wheat after pollinationSexual Plant Reproduction, 1992
- Electrofusion-mediated transmission of cytoplasmic organelles through the in vitro fertilization process, fusion of sperm cells with synergids and central cells, and cell reconstitution in maizeSexual Plant Reproduction, 1991
- In vitro fertilization of single, isolated gametes of maize mediated by electrofusionSexual Plant Reproduction, 1991
- Evidence for the role of Ca2+ ions in tip extension in pollen tubesProtoplasma, 1983