Plant tissue culture rapid propagation method taking stem tip as explant
A plant tissue culture and explant technology, applied in the field of fast propagation of plant tissue culture, can solve the problems of unstable germplasm resources, long plant breeding time, and easy aging of "ling seeds", and achieve rapid propagation and tissue culture cycle. Short, well-rooted effect
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Embodiment 1
[0034] Embodiment 1, the influence of different hormone ratios on the shoot tip regeneration of Rhizoma Chuanxiong
[0035] (1) Material collection: The young stem tips of Rhizoma Chuanxiong at the mature stage, about 2 cm to 4 cm in length, are directly picked as explants, without the need to peel off young leaves and leaf primordia or cut out the shoot tips with dissecting needles under a dissecting microscope. In the prior art, the stem tip of Rhizoma Chuanxiong is used as the explant to propagate and cultivate. The stem tip explant of Rhizoma Chuanxiong with 1 to 2 true leaves needs to be peeled off the shoot tip meristem under a microscope, and 2-3 mm The explants are then propagated through callus induction and differentiation culture, which is a cumbersome process and takes a long time.
[0036] (2) Disinfection treatment of explants: Rinse the stem tip of Rhizoma Chuanxiong carefully under running water for 1 hour, then wash it 3 times with sterile water in an ultra-cl...
Embodiment 2
[0050] Embodiment 2, Rhizoma Chuanxiong stem tip tissue culture rapid propagation method
[0051] According to the optimal rooting medium of the shoot apex screened in the previous stage, the method of tissue culture and rapid propagation of the shoot apex of Ligusticum chuanxiong was established by using the direct organogenesis method. The operation steps are as follows:
[0052] (1) Material collection: select young shoot tips of Rhizoma Chuanxiong in the mature stage, about 2 cm to 4 cm in length, as explants.
[0053] (2) Disinfection treatment of explants: Rinse the stem tip of Rhizoma Chuanxiong carefully under running water for 1-2 hours, then wash it with sterile water for 3 times in an ultra-clean bench, then treat it with 70% or 75% alcohol for 30 seconds, and then wash it with 2 % (W / V) sodium hypochlorite solution for 4 minutes, and finally washed 3 times with sterile water.
[0054] (3) Stem tip rooting medium: Transfer the sterilized Rhizoma Chuanxiong stem tip...
Embodiment 3
[0057] Embodiment 3, Ligusticum chuanxiong stem node tissue culture rapid propagation system
[0058] Learn from the tissue culture and rapid propagation system of Rhizoma Chuanxiong stem tip, and use the best rooting medium screened in the previous stage to establish the tissue culture and rapid propagation system of Rhizoma Chuanxiong stem nodes. The operation steps are as follows:
[0059] (1) The explants of Rhizoma Chuanxiong stem nodes near the tip of the stem were selected, and the stem nodes near the underground part were difficult to sterilize and had a high pollution rate.
[0060] (2) Disinfect and sterilize the stem node explants of Rhizoma Chuanxiong. The disinfection method adopted is: first rinse the stem nodes under running water for about 3 hours, then treat them with 75% alcohol for 45 seconds and 2% sodium hypochlorite for 8 minutes under sterile conditions, and finally wash the explants with sterile water for 3-3 hours. 5 times.
[0061] (3) Transfer the ...
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