Tissue culture and rapid propagation method for manglietiastrum sinicum
A culture medium and subculture technology, applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve the problems of narrow distribution area of canopy wood, scarce population, insufficient seedlings, etc.
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Embodiment 1
[0045] Screening method for inducing differentiation medium
[0046] Take the young terminal buds and side buds of Canopy wood as explants, soak them in 1% soapy water for 10 minutes, rinse them with running water, put them on the ultra-clean table, cut them into stems with single nodes, and use 75% Disinfect with alcohol for 10 seconds, rinse with sterile water for 3 times, disinfect the surface with 0.1% mercuric chloride solution for 10 minutes, wash with sterile water for 3 to 5 times, inoculate in the prepared canopy wood induction medium, 1 node per bottle.
[0047] Inoculate the sterilized stem segments in MS+3.0mg / l 6-BA+1.0mg / L IAA, MS+2.5mg / l 6-BA+1.0mg / L IAA, MS+2.0mg / l 6- BA+1.0mg / L IAA, MS+1.0mg / l 6-BA+1.0mg / L IAA, MS+0.5mg / l 6-BA+1.0mg / L IAA culture medium to induce differentiation, the MS The medium also includes 30 g / L sucrose, 5 g / L agar, and a pH value of 5.8. The culture temperature is 23-30° C., and the artificial light is assisted during the culture peri...
Embodiment 2
[0052] Proliferation and Subculture Media Screening Methods
[0053] Using MS medium as the basic medium for proliferation and subculture hormone screening, the cytokinin is 6-BA, and the concentration range is (0mg / L, 0.1mg / L, 0.5mg / L, 1.0mg / L, 1.5mg / L, 2mg / L, 6 concentration gradients), the auxins are all IAA, the concentrations are (0.1mg / L, 0.5mg / L, 1.0mg / L, 3 concentration gradients), and the uniform design method is used. The culture temperature is 23-30° C., the light time is 10 hours, and the light intensity is 1500 Lux.
[0054] The results are shown in Table 2.
[0055] Table 2 Proliferation and Subculture Medium Hormone Screening
[0056]
[0057] It can be seen from Table 2 that MS medium+1.0mg / L 6-BA+0.5mg / L IAA+sucrose 30g / L+agar 5g / L, pH value 5.8, culture period 60 days, light intensity 1500Lux, temperature 23-30 ℃. Count the side buds and terminal buds quantity that each propagule produces, the average value of bud node quantity (comprising terminal bu...
Embodiment 3
[0059] Screening method for strong seedling rooting medium
[0060] Using MS medium as the basic medium for hormone screening of strong seedlings and rooting medium, the concentration range of IBA is 0.1mg / L, 0.5mg / L, 1.0mg / L, 2mg / L 4 concentration gradients, and the concentration of IAA is 0.1 mg / L, 0.5mg / L, 1.0mg / L, 2.0mg / L 4 concentration gradients, using the uniform design method. The culture temperature is 23-30° C., the light time is 10 hours, and the light intensity is 1500 Lux.
[0061] The results are shown in Table 3.
[0062] Table 3 Rooting Medium Hormone Screening Results
[0063]
[0064] As can be seen from Table 3, MS medium+1.0mg / L IAA+1.0mg / L IBA+sucrose 30g / L+agar 5g / L, pH value 5.8, culture period 30 days, rooting rate reached 81%.
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