Random amplified polymorphic DNA (RAPD) markers were utilized for screening the clonal fidelity of in vitro-raised bulblets of Lilium sp. (Asiatic hybrids) produced through adventitious mode of propagation. The first set of 14 bulblets was randomly chosen from about 175 micropropagated bulblets regenerated from 1 Â 1 cm 2 bulbscale segments (explants) of cultivar`Gran Paradiso' after four subcultures (after 6 mo.). The second set of 15 bulblets was selected again randomly after 12 subcultures (after one and a half years) when the bulblets were to be taken out for transplantation. Of the 20 primers used to screen the samples, only 14 primers gave clear reproducible bands. The 14 primers produced a total of 163 (an average of 11.6 bands per primer) scorable bands. Analysis of individual primers revealed the RAPD patterns produced were all shared by both the in vitro-raised bulblets (randomly selected after four and 12 subcultures) and the mother bulb. There was no variation observed within the tissue culture-raised progenies. Thus, our results show that adventitiously propagated in vitro bulblets of Asiatic hybrids of lilies are clonally uniform and stable.
A simple, rapid and cost-effective in vitro scheme has been proposed for mass propagating two cultivars of Asiatic lily hybrids. An average of seven bulblets was formed after 17 d when 1 Â 1 cm 2 bulb scale segments (explants) were cultured on Murashige and Skoog (MS) medium with 3% sucrose and 0.5 mM a-naphthaleneacetic acid (NAA). On MS medium containing 0.5 mM NAA and 6 or 9% sucrose, depending on the cultivar, large numbers of bulblets of increased size (3.5± 5.0 cm in circumference) were formed under a 16/8 h photoperiod. A continuous system of mass propagation of bulblets was achieved through in vitro scale formation (secondary explants) on MS medium supplemented with 23 mM kinetin and 0.5 mM NAA, as well as scale proliferation on MS basal liquid stationary medium. Upon transplantation all bulblets sprouted, of which 40% flowered in the first season. Under ideal conditions, ca. 9X68 Â 10 5 bulblets can be produced from a single scale segment in 1 yr by following the systematic propagation steps proposed here.
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