CROSSING ARABIDOPSIS PLANTS (FLOWER EMMANSCULATION AND FLOWER PREPARATION FOR FERTILIZATION
 
Last Update: December 2006
 
 
PROCEDURE
Prior to performing the experiments, maturing flowers must be present in the bolting Arabidopsis plants
1. Preparation of recepient flower (ovary). The objective is to remove all the flower parts except the ovary
2. Choose an inflourescence and remove all the flowers that are too young (too small) and the ones that already show white petals (opening flowers will tend to have started self-fertilization). Cut both too young and too old flowers from inflourescence, leaving 3-10 flowers in the middle to work with

3. Cut of all other plant parts in the immediate vicinity, specually siliques. The idea is to have as free a work environment as possible
4. While cutting parts of from a flowers, DO NOT tare parts off. Flowers are delicate and be easily damamged. Practice will give a good feel for how much they can take
NOTE: This procedure can be done using very fine forceps: INOX1
5. In between flowers, clean forceps by dipping them in 95% ethanol followed by distilled water
6. Use a kim-wipe as surface while viewing the flowers on a disecting scope. This helps in holding the flower parts to the paper and not the forceps

7. Obtain pollen by getting fully mature flowers and removing the stamens. Use these stamens to brush the prepared ovaries. Repeat this at least twice to make sure thre is plenty of pollen at the tip of the flower. This should be evident when looking at the ovaries through the diescting scope as the pollen looks like a grainy brownish surface on top of the green ovary.
8. Label the cross accordingly and wrap the ovaries with Raynolds 905 sahran-wrap to make sure to cross contamination takes place
9. Leave ovaries developing until they start yellowing before harvesting. If too dry, they may shed their seeds