3.1@ Observation of protonemata and gametophores
Freshly inoculated and cultured protonemata are suitable for
observation. When protonemata are cultured more than two weeks on a seat of
cellophane on BCDATG or BCDAT media under continuous white light at 25˚C,
protonemata start to change brownish and likely die. If you cultivate on the
cellophane, it is better to observe protonemata in a week after inoculation
especially on BCDATG.
Cell and
organelle sizes change in different media. Chloroplasts become larger on BCDATG
or BCDAT media than on BCD medium.
Chloronemata
are easily observed in BCDATG or BCDAT media. When you inoculate protonemata on
BCD medium, protonemata change to caulonemata earlier than on BCD.
For observation
of gametophores, protonemata are inoculated in BCD medium rather than BCDATG
and BCDAT media and the plate is sealed with surgical tape to prevent
evaporation of water. For observation of bud formation, BCD medium is
appropriate because addition of ammonium in the medium enhances growth of a
callus-like bud.
1. To observe a
gametophore with more than 10 leaves.
1) Place a
gametophore on solid medium.@
2) Drop water on
the gametophore and cover with cover slip
3) Observe with
stereomicroscope.@
2. To observe
protonemata
1) Drop water on
a glass slide and place a few protonemata in water. For living cells, do not
use glycerol solution.
2) Cover with a coverslip
and observe immediately.@
Material
· Microscope
· Glass slide
· Coverslip
· Forceps
· Water
·
Solid medium (ex. BCDATG plate)