A method for tissue culture and rapid propagation of willow stem segments
A technology of tissue culture rapid propagation and willow drilling, applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve the problems of low operability and repeatability, general growth status, etc., and improve the efficiency of breeding seedlings Effect
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Embodiment 1
[0031] Example 1 Screening the medium for inducing clumping buds
[0032] Four mediums, WPM, DKW, 2 / 3DKW and 1 / 2NRM were used as the basic medium to select the basic medium suitable for the induction culture of the stem segments of Dianthus spp. Prepare a medium with the same types of hormones and the same concentration with the above four basic mediums. The hormone used is 6-BA (6-benzylaminmopurine) with a final concentration of 1.0 mg / L plus NAA with a final concentration of 0.1 mg / L (Naphthaleneacetic acid, 1-Naphthaleneacetic acid), that is, prepare the following medium (pH value is 5.80):
[0033] 1. WPM+6-BA1.0mg / L+NAA0.1mg / L medium
[0034] 2. DKW+6-BA1.0mg / L+NAA0.1mg / L medium
[0035] 3. 2 / 3DKW+6-BA1.0mg / L+NAA0.1mg / L medium
[0036] 4. 1 / 2NRM+6-BA1.0mg / L+NAA0.1mg / L medium
[0037] Cultivation of cluster buds was carried out on the stem segment (3cm in length, with one axillary bud) of Dianthus sibiricum that year. The culture conditions were 10h-12h of light per day and temper...
Embodiment 2
[0041] Example 2 Screening of Dedifferentiation Medium
[0042] Four mediums, WPM, DKW, 2 / 3DKW, and 1 / 2NRM were used as the basic medium to select the basic medium suitable for the dedifferentiation culture of the stems of the willow. Prepare the medium with the same types and concentrations of the above four basic mediums. The additives used are the final concentration of 2.0mg / L 6-BA, 0.1mg / L NAA, 4.5g / L agar and 20g / L glucose , That is, prepare the following medium (pH value is 5.80):
[0043] 1. WPM+6-BA2.0mg / L+NAA0.1mg / L+agar 4.5g / L+glucose 20g / L medium
[0044] 2. DKW+6-BA2.0mg / L+NAA0.1mg / L+agar 4.5g / L+glucose 20g / L medium
[0045] 3. 2 / 3DKW+6-BA2.0mg / L+NAA0.1mg / L+agar 4.5g / L+glucose 20g / L medium
[0046] 4. 1 / 2NRM+6-BA2.0mg / L+NAA0.1mg / L+agar 4.5g / L+glucose 20g / L medium
[0047] The clumping buds obtained in Example 1 were inoculated on the above 4 kinds of culture mediums for dedifferentiation culture, and the culture conditions were 10h-12h light per day, and the temperature wa...
Embodiment 3
[0051] Example 3 Using the method of the present invention to carry out tissue culture and rapid propagation of the stems of Willow
[0052] On April 20, 2014, in Huanren County, Benxi City, the original branches of Dianthus sibiricum were collected. The diameter of the branches should be more than 3mm. The collected branches should be cut into 3cm stems without pests and diseases. There is an axillary bud, and the tissue culture and rapid propagation of the stems of the willow will begin:
[0053] (The pH values of the clumping bud induction medium, dedifferentiation medium, differentiation medium and rooting medium used below are all 5.80.)
[0054] 1) Disinfection of explants: Disinfect the collected stems for 7 minutes with a mercury solution with a mass percentage of 0.3%.
[0055] 2) Cluster bud induction culture: 100 sterilized stem segments are inoculated on the cluster bud induction medium, which is based on DKW medium and contains 1.0 mg / L (final concentration, The same b...
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