Narcissus and Daffodil

Production of Narcissus bulbs 105
leaf tissue at alternate transfers (Chow et al., 1992b). Staikidou et al. (1994) compared
bulbil formation by single leaf explants and by shoot clump cultures. Bulbil
formation on the former was slow and inefficient compared with using shoot
clump cultures, but was responsive to growth regulators and so might be a useful,
simple system for investigating the regulation of narcissus bulbil initiation and
development.
Other reports on the micropropagation of narcissus include those of Popov and
Cherkasov (1984), Paek et al. (1987), Kozak (1991), Langens-Gerrits and Nashimoto
(1997) and Sochacki et al. (1997), who used various explants including those
consisting of scale bases with attached base plate tissue. Infection of cultures with
Fusarium is a problem in the tissue culture of narcissus, and Hol and van der Linde
(1989) demonstrated that infection could be controlled by storing bulbs at 30 °C
from lifting and giving bulb HWT at 54 °C for 1 hour before setting up cultures.
Riera et al. (1993) reported the beneficial effect of the polyamine diaminopropane
on regeneration from twin-scales in culture. Langens-Gerrits and Nashimoto
(1997) demonstrated that the presence of roots on bulblets was needed to obtain
good subsequent growth when they were planted out. Joung et al. (1997) reported
regeneration from different parts of the flower stem. Santos et al. (1998) reported
tissue culture starting with twin-scale explants of N. bulbocodium, in which 90% of
the bulbs obtained flowered during the first growing season. There are some
advantages in using a shaken liquid culture over using solid agar media. Bergoñón
et al. (1992) described shake cultures of N. papyraceus (see also Chapter 7, this
volume). In other tissue culture investigations with narcissus, ovule culture produced
viable seeds (Lu et al., 1988), as did excised placentae pollinated in vitro
(Balatková et al., 1977).
For Tazetta cultivars, successful induction of plantlets has also been reported
using young stem explants (Hosoki and Asahira, 1980), stem and scale explants
(Nagai, 1999a,b) and cultured twin-scale bases (Dachs et al., 1979; Steinitz and
Yahel, 1982; Gu and Gao, 1987), and Li and Tang (1982) reported callus induction
on twin-scale segments. In the case of cultivar ‘Grand Soleil d’Or’, under the
most appropriate conditions 80–100 explants were obtained from each parent
bulb, giving 200–300 bulblets in 6 months, and 80–90% of bulblets weighing over
0.25 g with two leaves and abundant roots were transplanted successfully (Dachs
et al., 1979; Steinitz and Yahel, 1982). Gu and Zhang (1991) compared production
from micro-propagated Tazetta narcissus derived from callus cultures with naturally
produced bulblets. Micropropagated plants flowered after 4 to 5 years, and
there were no differences between them or naturally derived plants in the number
of flowers or in flower morphology.
Sage et al. (2000) produced somatic embryos from narcissus cultivars. Somatic
embryos were obtained from leaf lamina, leaf base, bulb scale and, especially, stem
explants. Leaf explants from shoot cultures produced somatic embryos and
converted to plantlets efficiently on an appropriate medium following a cold treatment,
the plantlets subsequently transferring to ex vitro conditions readily.
Chipping, twin-scaling and related propagation techniques
Several simple propagation methods suitable for on-farm use have been investigated
for the multiplication of choice cultivars and VT stocks of narcissus. Scooping