Method for propagating betula luminifera aseptic seedlings by using tissue culture technology
A technology of tissue culture and sterile seedlings, applied in horticultural methods, botanical equipment and methods, applications, etc., can solve problems such as tight supply and demand, long growth cycle of forest trees, etc., to shorten germination time, improve emergence efficiency, and increase the number of seeds Effect
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Embodiment 1
[0042] A kind of method utilizing tissue culture technology to propagate the aseptic seedling of Betula glabra, comprising the following steps:
[0043] 1. Collection of explants: Collect mature seeds on sunny days, and cut the tender stems of Birch glabra with axillary buds growing well.
[0044] 2. Disinfection of explants:
[0045] a. Cut the collected tender stems into 4cm stem segments, peel off the seed coats of the seeds, place them in a clean beaker, soak them in detergent water for 30 minutes, and then rinse them with running water;
[0046] b. The washed stems and stems are treated with PEF with an electric field strength of 10Kv / cm, a pulse width of 35μs, and a pulse frequency of 700Hz, wherein the stems are treated for 15s and the seeds are treated for 22s;
[0047] c. Place the treated stems and seeds on an ultra-clean workbench, first soak them with 75% alcohol for 45 seconds, rinse them with sterile water for 5 times, then soak them with 0.1% mercury liter for ...
Embodiment 2
[0069] Under the same conditions as in Example 1, each culture medium was replaced with:
[0070] (1) Adventitious bud induction medium: its composition is: MS+6-BA 2.0mg / L+2,4-D 0.05mg / L+NAA0.4mg / L+agar 6.5g / L+sucrose 25g / L; pH condition It is 5.8; it is prepared by sterilizing at 121℃101KPa for 20min.
[0071] (2) Proliferation medium: its composition is: MS+2,4-D 3.0mg / L+NAA 2.0mg / L+IAA 1.0mg / L+agar 6.5g / L+sucrose 30g / L; pH condition is 6.0; Sterilized at 121°C and 101KPa for 20 minutes.
[0072] (3) Seed germination induction medium, which consists of: MS+6-BA 3.0mg / L+NAA 0.25mg / L+GA3 2.0mg / L+6.5g / L carrageenan+25g / L sucrose; the pH condition is 6.0; prepared by sterilizing at 121°C and 101KPa for 20 minutes.
[0073] (4) Callus induction medium: its composition is: MS+6-BA 2.0mg / L+NAA 0.5mg / L+TDZ 1.0mg / L+6.5g / L carrageenan+25g / L sucrose; pH condition It is 5.9; it is prepared by sterilizing at 121℃101KPa for 20min.
[0074] (5) Cluster bud induction medium: its compo...
Embodiment 3
[0084] Under the same conditions as in Example 1, each culture medium was replaced with:
[0085] (1) Adventitious bud induction medium: its composition is: MS+6-BA 1.5mg / L+2,4-D 0.1mg / L+NAA 0.3mg / L+agar 6.5g / L+sucrose 30g / L; pH conditions It is 5.8; it is prepared by sterilizing at 121℃101KPa for 20min.
[0086] (2) Proliferation medium: its composition is: MS+2,4-D 4.0mg / L+NAA 1.5mg / L+IAA 2.0mg / L+agar 6.5g / L+sucrose 25g / L; the pH condition is 6.0; Sterilized at 121°C and 101KPa for 20 minutes.
[0087] (3) Seed germination induction medium, which consists of: MS+6-BA 4.0mg / L+NAA 0.4mg / L+GA3 3.0mg / L+6.5g / L carrageenan+25g / L sucrose; the pH condition is 6.0; prepared by sterilizing at 121°C and 101KPa for 20 minutes.
[0088] (4) Callus induction medium: its composition is: MS+6-BA 1.5mg / L+NAA 0.3mg / L+TDZ0.75mg / L+6.5g / L carrageenan+30g / L sucrose; pH condition It is 5.9; it is prepared by sterilizing at 121℃101KPa for 20min.
[0089] (5) Cluster bud induction medium: its c...
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