Inducing Apothecia
- Cover bottom of glass petri dishes (10 cm ∅ ) thinly with vermiculite and saturate that with water. Autoclave. Fifty ml sclerotia fill about nine petri plates. Also autoclave 2 L deionized water, paper towels and cheesecloth.
- Surface sterilize sclerotia: Add 46 mL household bleach (new 6%) to total of 500ml water. For smaller amount, add 20 mL bleach to 180 mL of water. Treat sclerotia for 5 min. Cover beaker with cheese cloth, secure with rubber band and pour out the bleach. Rinse 3x with sterile water, 5 min each. Last, cover sclerotia with 70 % alcohol, 5 min. Dry sclerotia in the hood on sterile paper towels for a several hours to overnight.
- Check moisture in the vermiculite dishes and place sclerotia on surface, close together.
- Place plates into –20°C for 24 hours and then room temperature for 24 h. Repeat two more times. After third time in –20°C , place in a +15°C incubator with constant lights, fluorescent, cool white.
- First initials are observed after three weeks in +15°C. Apothecia will be mature about 2 weeks later.
Updated 28 May 2007 by Jeffrey Rollins.
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