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Laboratory Exercise 16
FUNGI: MOLDS

To identify fungi microscopically, it is important to observe certain structures (spores) and the
arrangement of the structures which produce the spores. When you begin culturing fungi and
observing them microscopically on microscopic slides, you will wonder how anything can be seen
and identified in such a tangled, broken up mess. You think that it would be good to see the fungi
microscopically exactly as they are growing in your medium. There is such a method called
Microculturing. Briefly, you will grow a fungus on a microscopic slide so when you are ready to
observe it, you disturb it very little.

Materials: PDA (Potato dextrose agar) plates, sterile toothpicks, sterile Petri plates, sterile distilled
water, glass slides, cover slips, sterile scalpel blade, bent wire needle, forceps

Procedure:
Part 1. Examination of Fungi (Molds)
1. Open and expose a potato dextrose agar plate to designated areas for 30 minutes. Close the
plate and seal it with a parafilm. Incubate the plate at room temperature for 7 days.
2. Take note of the colony morphology (colony diameter, color, color changes and the texture of the
colony surface whether loose, compact, wrinkled or buckled) from the agar surface every 24
hours.
3. Draw your observation (1st to 7th day culture).

Part 2. Fungal Culture: Slide Culture Technique


1. Place a bent glass rod or 2 sterile toothpicks in a petri plate. Sterilize.
2. After sterilization, carefully pour enough sterile water into the petri plate to just cover the bottom
of the petri dish.
3. Flame-sterilize a microscopic slide and
place it on top of the glass rod/
toothpicks (the rod keeps the slide out of
the water).
4. Using a sterile blade cut out an agar
block (7 x 7 mm) small enough to fit
under a cover slip.
5. Place the agar block on the center of the microscope slide which
is inside the sterile petri plate.
6. Inoculate the four sides of the agar block with spores or mycelial
fragments of the fungus
to be grown.
7. Hold a cover slip in a pair
of forceps and flame
sterilize it (this only takes
2-3 seconds). Let it cool
for a few seconds and
then place it on top of the
square piece of PDA agar that you just inoculated with the fungus.
8. Incubate the plate at 26°C until growth and sporulation have occurred.
9. The amount of incubation time required for this to happen varies with each fungus but usually 5-
10 days of incubation are required for contaminants and 1-3 weeks for pathogens.
10. Remove the cover slip from the agar block.
11. Optional: Apply a drop of 95% alcohol as a wetting agent.
12. Gently lower the cover slip onto a small drop of Lactophenol cotton blue on a clean glass slide.
13. Examine under the microscope. Draw and label all parts.
14. If you want to preserve the slide, carefully seal the cover slip to the glass slide with nail polish.
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STUDY GUIDE QUESTIONS:


1. Define:
a. Yeast
b. Molds
c. Budding
d. Thallus
e. Mycelium
f. Hyphae
g. Spore
2. What is the difference between vegetative and aerial mycelia?
3. Can bacteriological media be used for the cultivation of molds? Explain your answer.
4. What is the difference between vegetative and rhizoidal hyphae?

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