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In this article we will discuss about the principle, requirements and procedure for physical dissection of basidiomycotina.
Principle:
Mycelium present in soil particles are picked up and cultured after washing soil several times with water.
Requirements:
1. Soil crump.
2. Petri plates, beakers.
3. Agar medium.
4. Forceps.
5. Binocular.
Procedure:
1. Place 1-1.5 g of soil crump in water in a beaker and leave for 4-5 minutes. Remove supernatant.
2. With a rapid jet break the crump, allow heavier soil particles to settle, remove supernatant. Repeat the process till the supernatant becomes colourless.
3. Distribute soil particles in Petri dish in a little quantity of water.
4. Observe under a binocular and remove hyphal portions with a sterile forceps from among soil particles and place in a drop of sterile distilled water in a sterile Petri dish.
5. Tease hyphal masses since they may contain hyphae of more than one fungus.
6. Add 10-12 ml of cooled (45°C) malt agar medium to the plate rotate and allow the agar medium to solidify and incubate at room temperature (28°-30°C). for three to four days. (72 to 96 hours).
7. Examine the plate under microscope (120x) for fungal growth.
8. Cut out tips of growing hyphae and transfer to fresh media and observe for growth.
Czapek-Dox agar plus 0.5% Yeast extract (pH 5.6-5.8) diluted to 1/6 normal strength is recommended. Malt agar medium also is good.
Malt extract Agar:
