Construction method of colored Zantedeschia hybrida tissue culture in-vitro rapid propagation system
A technology of tissue culture and construction method, which is applied in the field of construction of an in vitro rapid propagation system for tissue culture of color calla lilies, can solve the problems of insufficient systematicness and poor repeatability, and achieves the effect of high application value.
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Embodiment 1
[0042] Explant Selection, Disinfection and Sterilization Method Screening
[0043] ① Bulb shoots: Select bulbs with germinated buds, first wash off the soil with tap water, then remove the outermost brown skin, add neutral detergent to soak for 15 minutes, and then rinse with running water for 1 hour (sterilization method A); or, Select bulbs with germinated buds, first wash off the soil with tap water, soak with 3g / L 50% carbendazim wettable powder for 24-36 hours, then remove the outermost brown skin, add neutral detergent and soak for 15 minutes , rinse with running water for 1h (sterilization method B);
[0044] Then select the young buds on the tubers and cut them into small pieces with a length of about 1 cm. There should be a small amount of tubers at the bottom of the buds to ensure the integrity of the buds;
[0045] Surface disinfection was carried out on the ultra-clean workbench, soaked in 0.1% Hgcl2 solution for 7 minutes, then soaked in 2% NaClO solution for 10 ...
Embodiment 2
[0057] Effects of Different Concentrations of 6-BA on Induction of Cluster Buds
[0058] Bulb bud explants (sterilized by method B) can be induced to form clustered buds after 45 days of primary culture in medium supplemented with different concentrations of 6-BA (1.0-4.0mg / L), and the clustered bud induction rate is uniform. Significant differences exist, see figure 1 ,table 3:
[0059] Table 3: Effects of different 6-BA concentrations on the induction of clustered buds in the first generation
[0060] 6-BA concentration (mg / l) Cluster bud induction rate (%) 1.0 14.1±1.1c 2.0 28.9±0.8a 3.0 20.2±0.6b 4.0 15.6±1.8c
[0061] The induction rate of clustered buds was the highest in the medium supplemented with 2.0mg / L 6-BA, which was 28.9%, and the growth of clustered plantlets was the best. Therefore, the optimal induction medium was MS+6-BA2.0mg / L+NAA 0.2 mg / L; when the concentration of 6-BA was low, the volume of the explant incision was e...
Embodiment 3
[0063] Optimization of Subculture Propagation of Adventitious Buds
[0064] The adventitious buds were subcultured for 30 days, and the influence of different 6-BA concentrations (1.0-4.0mg / L) on the subculture of adventitious buds was investigated with the adventitious bud proliferation area, the adventitious bud multiplication multiple and the number of plants greater than 1cm. see results figure 2 ,Table 4:
[0065] Table 4: Effects of different 6-BA concentrations on proliferation of adventitious buds
[0066]
[0067] The proliferation area of adventitious buds was the best at 2.0 mg / L 6-BA, and the worst at 1.0 mg / L; The multiple can reach more than 7 times; followed by 3.0mg / L, there is no significant difference between the proliferation multiple and 4.0 mg / L 6-BA concentration, and the most suitable medium is 1 / 2MS+6-BA 2.0 mg / L+ NAA0.2mg / L.
[0068] There are significant differences in the number of plants larger than 1cm, and they all show that the effect i...
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