Gametophyte culture, Sporophyte culture – Carolina C-FERN Express Extended Culture User Manual
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Gametophyte Culture
Follow the basic procedure described in the culture instructions or online at
www.c-fern.org. If cultured at 82ºF, mature gametophytes can be observed at
10 days from sowing spores. While standard C-FERN gametophytes exhibit
clear distinctions between the small males and the larger hermaphrodites, the
distinguishing features may be a little less conspicuous with C-FERN Express
gametophytes. The basic distinction still exists between the males (smaller,
without meristem, and with many antheridia) and the hermaphrodites (larger,
with meristem, and
with both archegonia
and a few antheridia).
However, some
smaller C-FERN
Express gametophytes
with several
antheridia may look
like males initially but
then develop
a meristem
and become
hermaphrodites with
several archegonia.
Water added to
gametophyte cultures
at 10 days or later will allow fertilization by swimming sperm.
One week later, young sporophytes will be visible at the first leaf stage.
Sporophyte Culture
Sporophytes can be grown to maturity within the same size petri dishes
(60 mm) used for gametophyte culture. Dishes should be two-thirds full with
Basic C-FERN Medium. When sporophytes are from 1 to 2 weeks old, use a
sterile toothpick to transfer them to fresh dishes (four per dish). Maintain the
cultures in a Growth Pod or similar container under the same conditions as for
gametophytes. Be aware that cooler temperatures (as low as 75°F) may
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C-FERN Express
male (left) and
C-FERN 10-day-old
hermaphrodite
gametophyte
(right).
Note: These two images approximate actual size
differences between males and hermaphrodites.