Sporophyte Culture – Transplantation

Transplant young sporophytes from agar culture to a soil mix in the greenhouse

Materials

  • Greenhouse Containers, Cubes or Pots
  • Commercial Potting Soil or Mixture: ProMix® Potting Soil (used as is) OR Potting mixture (2 parts peat moss, 1 part vermiculite, 1 part perlite, adjusted to ca. pH 6.5 with 100 g agricultural lime per 10 gallons of the potting mixture)
  • C-Fern sporophytes, 2 weeks old grown from agar-based cultures

Methods

Multispore culture with young sporophytes

SPROPHYTE SELECTION Two-week old sporophytes grown from agar culture can be transplanted to the greenhouse. Sporophytes should have leaves of at least 2-3 cm. If sporophytes are generated in multispore cultures, subculturing very young sporophytes into fresh nutrient medium may be necessary to generate sporophytes that are large enough for successful transfer to the greenhouse. The C-Fern Nutrient Medium used for gametophytes will not support long term sporophytic growth.

CONTAINERS AND POTTING MIXTURES For initial transplanting, transfer sporophytes to small potting containers such as 2″x2″ cell well trays or equivalent that contain moist ProMix® Potting Soil or a potting mixture. Pro-Mix gives excellent results when used as-is. Other potting mixtures should be adjusted to pH 6.5 as indicated above.

TRANSPLANT Carefully remove the sporophyte from the agar-based medium. Gently and quickly remove as much excess agar as feasible, being careful to not damage roots. Place sporophyte in potting mix depression and carefully fill in and completely bury roots in potting mix. Lightly pack potting mix around roots.

GREENHOUSE CULTURE Place potting containers in a greenhouse tray containing 1-2 cm of water. Newly transplanted sporophytes should be covered with a clear plastic dome (humidity dome) for 3 days. As sporophytes grow they should be transplanted when roots begin to emerge from the bottom of the potting containers. Transplant sporophytes to a 4″ diameter pot and subsequently to a 6″ diameter pot.

Soil should be kept moist to wet at all times – an automated watering system helps. At all stages of growth, Ceratopteris sporophytes require warm temperatures and high humidity; remember that it’s a tropical fern.

Thomas Warne