In-vitro sprout quick propagation method of betula microphylla in marsh
An in vitro and marsh technology, applied in the field of plant tissue culture, can solve problems such as pollution, hindering plant tissue culture work, vitrification, etc., and achieve the effect of reducing pollution rate, reducing tissue culture risk, and high rooting rate
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Embodiment 1
[0037] The rapid propagation method of isolated buds of Betula microphylla, using the new shoots of Betula microphylla as explants, first treat them with 70% alcohol for 20s on the ultra-clean workbench, and then use 0.1% HgCl 2 Or 10% NaClO disinfectant adopts different disinfection time (Table 1) after disinfection, rinse 5 times with sterile water. Put the sterilized explants in a petri dish covered with sterile filter paper, blot the moisture on the surface of the material, cut off the part of the base that is in contact with the medicinal solution with a scalpel, and remove the bud scales and unexpanded stipules. Inoculate on MS+6-BA1.0mg / L growth medium, receive 12 bottles for each treatment, and receive 3 explants for each bottle, regularly observe and record, and count the contamination rate, mortality rate and germination rate 25 days after inoculation.
[0038] Table 1 Design of different disinfectants and disinfection time
[0039] Processing number
70...
Embodiment 2
[0045] A method for rapid propagation of isolated buds of birch microphylla, comprising the following steps:
[0046] 1) Establishment of explants: Select ungerminated robust plants with a plant height of about 1-1.5m, and pick new shoots such as main branch terminal buds, lateral buds, and lateral branch terminal buds as plant materials in April to May. Before the new shoots germinate, spray the plants with 50% carbendazim wettable powder twice a week until the plants spit out buds and collect materials. In order to reduce material pollution, pick new shoots from strong branches, use a test tube brush dipped in washing powder to gently scrub, and rinse the surface dirt with running water for 2 to 3 hours.
[0047] 2) Disinfection of explants: with the explants obtained in step 1), on the ultra-clean workbench, first process with 70% alcohol for 20 seconds, then disinfect with 0.1% mercury liter for 10 minutes, and rinse with sterile water for 5 times .
[0048] 3) Primary cul...
Embodiment 3
[0058] A method for the quick propagation of isolated buds of Betula marsh, same as embodiment 2, wherein when the primary generation is cultivated, MS is the basic medium, and 1.0g / L of PVP and 6-BA of different concentrations are added in the medium (Table 4), each handles 10 bottles, 3 explants of every bottle, counts germination rate after 25d.
[0059] Table 4 shows that the germination rate of explants increases with the increase of 6-BA concentration. When no 6-BA was added, the germination rate was the lowest, 16.67%, and there was no callus formation, but the growth was relatively slow; when the addition amount was greater than 0.8mg / L, the germination rate could reach more than 90%, although there was hemispherical callus formation at the base, But the plant grows well; when the addition amount is 1.5mg / L, the germination rate is the highest is 96.67%. figure 1 a), larger and harder, with thinner leaves, yellowish color, and poor growth ( figure 1 b).
[0060] Tab...
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