Plating Media: R - medium (contains only 10 grams of glucose per liter rather than 20 grams) + 2 uM metatopolin + 4, 1 cm3 cubes Russet Potato. Bleached 30 mins. in 0.525% chlorox bleach
Replate Media: R medium + 1, 1cm3 cube Russet Potato (200 ml replate medium in pint jars)
Number of Days between Plating and Replating: 41 days
Number of Days between Replating and Vernalization: 172 days
Number of Days between the start and end of Vernalization: 120 days
Date to Initiate Plating: April 30
Date to Replate: June 11
Date to Start to Vernalizae: Dec. 1
Seeds were plated Jan. 19, 2010, replated March 3, 2010, and started to vernalize on 9-19-2010.
Seedlings grew very well developing shoots 1/2 to 1.5 inches long with several roots 3-4 inches long. Adjust plating time to March 31 to better synchronize with the beginning of vernalization on Dec. 1. This protcol is very good for Epipactis gigantea. Some seedings treated with fung-onil to reduce fungus during vernalization. Some seedlings were fungused.
The second batch of seedlings taken from the replate jars on 10-2-2010 had shoots from 3-3.5 inches long and somewhat etiolated. I suggest taking the seedlings out of the jars 1 month earlier. I have adjusted the times above to reflect this change.