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C O N T E N T S
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Contact the C-Fern Project
cfern@utk.edu
 Supported, in part, by the National Science Foundation - Division of Undergraduate Education
Development of C-Fern is supported
by the National Science Foundation (NSF-DUE)
Copyright © 1997-2000
Thomas R. Warne and
Leslie G. Hickok.
All rights reserved.
 Culturing Sporophytes

  C-Fern sporophyte in a mini-terrarium
Culturing C-Fern sporophytes in a low-maintenance mini-terrarium
C-Fern sporophytes can be easily grown in plastic drink bottles; ranging in size from small 8 oz. types to the more common 2L variety. Sporophytes will effectively "size themselves" to fit whatever size bottle they are planted in. For convenience and economy of space, we like to use the small 8 oz. size bottles. To start, all one needs is a young sporophyte, a small amount of a commercial potting soil such as ProMix®, a time-release fertilizer such as Osmocote® 14-14-14, and a clean clear plastic bottle with the label removed. Since most users employ agar cultures, young sporophytes can be generated in multi-spore cultures and then isolated to new agar plates and cultured until they develop several leaves and at least a few roots.

With small bottles it is possible to use a pair of large forceps or the eraser end of a pencil to place/push the sporophyte into the bottle without cutting the top off the bottle. Prior to inserting the sporophyte, add premoistened soil mix to a depth of 1–2 cm and add 1-2 pellets of the time-release fertilizer. Water well and gently seat the sporophyte into the soil with the end of the forceps or the pencil. Be sure not to damage the central meristem area! Leave the screw top off of the bottle and keep the mini-terrarium under lights and water as needed. Continual 24 hour light at the same or, better, a higher intensity than that used for gametophyte cultures works very well. Although it is hard to overwater C-Fern (it’s semi-aquatic!), it is best to just keep the soil moist. This usually only requires watering every 1-2 weeks. Use a small amount of distilled or de-ionized water, if available.

Under these conditions sporophyte growth is quite rapid and within a few weeks one should be able to see a progression from relatively simple leaves to more highly divided fern-like leaves. The fern-like leaves are fertile and contain sporangia within which meiosis occurs to produce spores. If good growth is evident but this progression in leaf form does not take place, the terrarium may be too moist. Cut back on watering and/or cut the top of the bottle off to dry it out some. C-Fern progresses to a fertile leaf stage in response to drier conditions.