Laboratory Topic I Cells Of Crystal And Color
EXERCISE A: PLANT CELLS WITHOUT WALLS
- Buffer solution is approximately 13% mannitol or 0.625 M mannitol (FW 182.2). Prepare by dissolving 56.94 g of mannitol and add distilled water to bring to tinal volume of 500 ml. Adjust the pH to 6-7 by adding drops of 0.1 N NaOH. Distribute 10 ml of buffer solution to each student team.
- For each student group, measure out 0.1 g of pecti-nase and 0.2 g of ccllulysin. These should be added to 10 ml of buffer solution immediately' before use to make an enzyme concentration of 1% pcctinase and 2% ccllulysin (cellulase). Purchase pccrinasc and eel-lulysin from Sigma.
- Protoplasts can be viewed directly through the petri dish or purchase microslidcs from Ward's to view at higher magnifications.
- Ward's sells a Protoplast Isolation Kit (86W 8009) and Protoplast Fusion Kit <86W 8008 to use with geranium petals. The kits work well, and all you need to supply are the geranium plants with contrasting flower colors. Ward's will also sell the plants for additional cost.
EXERCISE B: COMPONENTS OF THE PLANT CELL
To prepare Sudan 111 solution for staining elaioplasts in avocado fruit, mix 70 ml of 95% ethanol in 30 ml of distilled water in a beaker. Heat gently. Add Sudan III crystals slowly with stirring until no more will dissolve. Decant. Place in dropper bottles.
EXERCISE C: PLANT ULTRASTRUCTURE
If you have an Electron Microscope facility, collcct SFM and TEM micrographs for student folders. Add a qucs tion or worksheet for cach micrograph lo ensure that students carefully examine the collection. If not, bring in SF.M and TEM adascs for students to examine.
EXERCISE D: CRYSTAL PERSUASION
If you wish, Triarch and Ward's sells prepared slides of
Fiats clastica leaf cystoliths.
EXERCISE E: PLANTS TO DYE FOR
- Note that betacvanins, found in beets Chcnopo-diaccae) and species within the Aizoaccac, Amaran-thaceae, Cactaccac. Nyctaginaccae, Phytolaccaccae, Portulacaccac and a few other families within the Caryophllales, are alkaloids and not natural pH indicators. Thcv will not show die color change as the andiocyanins in red cabbage juicc will.
- Ii is best to pre-mordant the wool before dyeing. Alum and cream of tartar can be purchased from the spice section in the supermarket. Use natural (undyed) wool or wool diat has been dyed natural or wrhitc. Using a 1,000-ml beaker, dissolve 105 g of alum and 1 g of cream to tartar in 500 ml of distilled water that has been heated to approximately 60 C. Add a length of wool (about 15 g), and simmer for an hour. Let cool. Dye direcdy or store the wool in a sealed plastic bag to keep moist until ready to dye.
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