Inflorescence In Vitro Culture, and Polyploidization in Dikaryotype MT-A Hybrids of Spider Lily (Lycoris spp.)
Date Issued
2010
Date
2010
Author(s)
Wang, Mei-Chen
Abstract
Lycoris aurea Taiwanese native flower has been grown for cut flower trade. Lycoris species also have high economic value for their medicinal alkaloids. They require four to five cycles from seed to flower, and the propagation rate is restrained to nature. The purpose of this research was not only to develop a method for propagation of dikaryotype MT-A hybrids and testprogenies of Lycoris spp, but also to use different culture container for rapid mass propagation.
Experimental materials were dikaryotype MT-A hybrids and testprogenies of Lycoris spp. including of L.radiata × L.sprengeri, 2n = 22 (22A), L.aurea × L.radiata, 2n = 18 (4M+3T+11A) and L.aurea × L.sprengeri, 2n = 18 (4M+3T+11A). Immature inflorescenes segments were used for callus and adventitious buds initiation. The immature perianth produced the highest regeneration rate, and the adventitious bud inducing rates in the medium with 2 mg/L 2,4-D and 5 or 10 mg/L BA were more than other treatments. The medium with 2 mg/L NAA and 10 mg/L BA also showed good regeneration effect in some lines. After 8 weeks of subculture, the immature perianth-induced buds regenerated 40 to 80 buds, and the multiple rate was 1.
In vitro grown bulblets with 5 mm diameters were cut into four segments for adventitious buds initiation. After 8 weeks of solid culture, the A specific karyotype and trispecific origin dikaryotype hybrid inoculated in medium with 0.5 mg/L NAA and 5 mg/L BA regenerated 10.36 and 13.88 buds, and the MT-A dikaryotype hybrids inoculated in medium with 0.5 mg/L 2,4-D and 5 mg/L BA regenerated 9.08 buds. By using TIS as culture container for segments adventitious buds initiation, the A specific karyotype (LRM8709 × LS) and MT-A dikaryotype hybrids (LA × LRK2) had much more adventitious buds regeneration than solid culture in all treatments. And the greatest result showed in the medium with 0.5 mg/L 2,4-D and 5 mg/L BA which induced 108 buds, and the multiple rate was 21.6 of A specific karyotype. MT-A dikaryotype hybrids had the greatest adventitious buds regeneration in the medium with 0.5 mg/L 2,4-D and 5 mg/L 2-ip which induced 76 buds, and the multiple rate was 16.89.
The different karyotype buds with1 to 2 mm diameters in the medium with 90g/L sucrose after 8 weeks, the buds of A specific karyotype and MT-A dikaryotype grew 10.2 and 10.75 mm individually. After culturing in medium with activated charcoal , the bulblets formed root system. The survival rate of plantlets were more than 90%, and the leaf number grew to more than 3.
For restoring the fertility of MT-A dikaryotype hybrids, the scale segments were used as explants for artificial polyploidy induction. The chromosome of root tips were tested after the adventitious buds formed plantlet. The regenerated leaves of pre-tetrapolyploidies were tested DNA content by flow cytometry. Besides the plantlet treated with 100 μM Oryzalin 72hr at pH 5.7 was chimera, the other were reverted to diploidy.
Subjects
Lycoris
in vitro propagation
inflorescene culture
proliferation
temporary immersion system
polyploidization
Type
thesis
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