Growing grape ferns from spore in axenic culture.

Dr. Whittier used Botrychium Jenmannii but this method or with slight variations may work for other Botrychium species. Dr. Whittier's method is one of the best developed to date.

Suface sterilize the spores with 20% Clorox (Whittier 1973) and sow on 15 ml of nutrient medium in culture tubes screw caps that can be tightened to reduce moisture loss. The cultures should be maintained at 22 degrees Celsius in darkness until after embryos are formed.

A liter of nutrient medium contained 25 mg arginine; 100 mg MgSO₄; 40 mg CaCl₂; and 100mg K₂HPO₄.

In addition, 8.5 ml of a FeEDTA solution and 0.5 ml of a minor element solution were added per liter. The medium was completed with 0.2% glucose and 0.8% agar and was adjusted to a pH of 6.1.

Spore germination should occur after 5 wks. in the dark producing cultures with numerous gametophytes that are white. If overcrowding occurs either weed some of them out or transfer them to a different container with the same solution. From sowing spore to mature gametophytes could take as long as 12 months. In any case flood actively growing gametophytes (3-4 months old in Whittiers case) with distilled water for 4 days removing excess water. After 2-3 more months roots will appear. If the young sporephytes are given light at this stage a green leaf will appear. Then you are on your own, no attempt was made to move the plants into more natural soil.