Core Practicals :

1) Describe the stages of mitosis and how to prepare and stain a root tip squash in order to
observe them practically.

2) Describe how totipotency can be demonstrated practically using plant tissue culture
techniques

3) Describe how to determine the tensile strength of plant fibres practically.

4) Describe how to investigate plant mineral deficiencies practically

5) Describe how to investigate the antimicrobial properties of plants

How to prepare and stain a root tip squash in order to observe them practically.

Method:

1) Cut off end 1 cm of a root and fix in methylated spirit and glacial ethanoic acid for 2
hours
2) Treat the root tip then in HCl at 60c for 6 to 7 minutes. (If treated for longer the
staining reaction becomes weaker)
3) Remove the acid and rinse in distilled water
4) Add acetic orcein reagent and leave for 1 hour
5) Rinse again in distilled water
6) Place the root tip on microscope slide, cut off ends, add few frops of glycerol , using
mounted needle, break up the root tip.
7) Apply a coverslip , blot dry, squash gently ( to spread the cells ) and view under the
microscope.

Describe how totipotency can be demonstrated practically using plant tissue culture
techniques

In this practical you will take the tops of plant seedlings and grow them into complete plants
on an agar medium. The explants (the parts of the removed
seedling) should grow new leaves, stems, roots and more leaves.

METHOD
1. Dampen some cotton wool with water and place in a plastic tray.
2. Sprinkle some seeds onto the cotton wool and cover with transparent cling
film.
3. Place in a warm light place to germinate. Use the light bank as a light
source. Leave for 3-4 days.
4. Measure 2.5g of agar powder into a beaker and add 250ml of distilled water.
Boil and stir gently with a glass rod until all the agar has dissolved.
!
5. Pour the molten agar into several test tubes. Allow to cool and solidify.
6. Cut off the top of the seedlings just below the shoot apex with scissors.
These are the explants.
7. Carefully push the cut end of each seedling into the agar (make sure the
cotyledons do not touch the agar).
8. Cover the tubes with cling film and place under a light rack.
9. Observe and note any developments in the seedlings over the next 12 days.