Tissue culture and rapid propagation method of Jiuhua Polygonatum sibiricum
A technology of tissue culture rapid propagation and Jiuhua Polygonatum, applied in horticultural methods, botany equipment and methods, horticulture, etc., can solve the problems of low reproductive rate of Polygonatum seedlings and inability to carry out industrial production
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Embodiment 1
[0023] 1. Plant material
[0024] Polygonatum cyrtonema Hua (Polygonatum cyrtonema Hua), belonging to the genus Polygonatum cyrtonema of Liliaceae, is taken from Mount Jiuhua, Anhui Province.
[0025] 2. Induction of Adventitious Buds
[0026] Take the young rhizomes of Polygonatum, and wash off the soil. Cut the rhizomes with buds with a knife, soak them in an aqueous solution of detergent for 5 minutes, and rinse them with tap water. Scrub the surface with 75% ethanol and rinse with tap water. The following is done in an ultra-clean workbench: soak in 75% ethanol for 0.5 min, rinse with sterilized water; soak in mercuric chloride for 10 min, rinse with sterilized water; dry the water with sterile filter paper, cut off the necrotic part of the wound with a knife, and inoculate the induced on the culture medium. The composition of the medium is: MS basic medium + 1.0-6.0mg / L 6-BA + 0.1-0.2mg / L NAA + agar 7g / L + sucrose 30g / L, the pH is 5.8. Hormone composition and content...
Embodiment 2
[0033] 1. Materials
[0034] The adventitious buds of Polygonatum Polygonatum induced by Example 1.
[0035] 2. Proliferation culture of adventitious buds
[0036] The cluster buds obtained in Example 1 are cut into stem segments with a terminal bud or side buds, and are inoculated on the cluster bud proliferation medium respectively. Place them in an environment with a culture temperature of 25±1°C, a light time of 12 hours / day, and a light intensity of 1600 Lx for 40 days. The composition of the culture medium is: MS basic medium+0.6mg / L6-BA+0.2-0.4mg / L IAA+0-0.8mg / L NAA+7g / L agar+30g / L sucrose, pH5.8. Hormone composition and content are shown in Table 2.
[0037] It can be seen from Table 2 that the combination of NAA with a concentration of 0.2 mg / L and IAA with a concentration of 0.2 mg / L has the best effect. Observations also showed that the adventitious buds induced to differentiate on this medium were cut from the base, inoculated on the same medium for subculture,...
Embodiment 3
[0043] 1. Materials
[0044] The differentiated cluster buds obtained in embodiment 2.
[0045] 2. Rooting culture
[0046] The single robust adventitious buds obtained in the differentiation culture obtained in Example 3 were cut from the base, inoculated on the rooting medium, and placed in an environment where the culture temperature was 25±1°C, the light time was 12 hours / day, and the light intensity was 1600Lx cultured for 25 days. The composition of the medium is: 1 / 2 MS basic medium + 0.2-1.0mg / L NAA + agar 7g / L + sucrose 30g / L, the pH is 5.8.
[0047] As can be seen from the results in Table 3, the preferred composition of the rooting medium of the present invention is: 1 / 2MS+0.6mg / L NAA. And the rooting test-tube seedlings cultivated on this medium have strong stems, dark green leaves, and vigorous growth. When cultivating to 20 days, almost all the cultured seedlings will grow roots of varying numbers.
[0048] table 3
[0049]
[0050] 4. Seedling hardening ...
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