Fertilization in Arabidopsis thaliana wild type: Developmental stages and time course
Article first published online: 22 MAY 2002
The Plant Journal
Volume 30, Issue 4, pages 481–488, May 2002
How to Cite
Faure, J.-E., Rotman, N., Fortuné, P. and Dumas, C. (2002), Fertilization in Arabidopsis thaliana wild type: Developmental stages and time course. The Plant Journal, 30: 481–488. doi: 10.1046/j.1365-313X.2002.01305.x
- Issue published online: 22 MAY 2002
- Article first published online: 22 MAY 2002
- Received 17 October 2001; revised 13 February 2002; accepted 25 February 2002.
- egg cell;
- pollen tube.
We describe some previously uncharacterised stages of fertilization in Arabidopsis thaliana and provide for the first time a precise time course of the fertilization process. We hand-pollinated wild type pistils with wild type pollen (Columbia ecotype), fixed them at various times after pollination, and analysed 600 embryo sacs using Confocal Laser Scanning Microscopy. Degeneration of one of the synergid cells starts at 5 Hours After Pollination (HAP). Polarity of the egg changes rapidly after this synergid degeneration. Karyogamy is then detected by the presence of two nucleoli of different diameters in both the egg and central cell nuclei, 7–8 HAP. Within the next hour, first nuclear division takes place in the fertilized central cell and two nucleoli can then be seen transiently in each nucleus produced. In a second set of experiments, we hand-pollinated wild type pistils with pollen from a transgenic promLAT52::EGFP line that expresses EGFP in its pollen vegetative cell. Release of the pollen tube contents into the synergid cell could be detected in living material. We show that the timing of synergid degeneration and pollen tube release correlate well, suggesting that either the synergid cell degenerates at the time of pollen tube discharge or very shortly before it. These observations and protocols constitute an important basis for the further phenotypic analysis of mutants affected in fertilization.