Dendrobium hirsutum tissue culture and rapid propagation culture medium composition and application of dendrobium hirsutum tissue culture and rapid propagation culture medium composition in dendrobium hirsutum seedling cultivation
A technology of tissue culture rapid propagation and medium, applied in application, plant regeneration, agricultural gas emission reduction, etc., can solve the problems of unreported, low proliferation coefficient, long time consumption, etc.
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Embodiment 1
[0047] 1) Select the Dendrobium capsules, which are healthy, plump, and not yet cracked 150 days after pollination, as explants, and soak them in a detergent solution with a mass concentration of 1% for 10 min. After taking them out, rinse them with running tap water. Put it on the ultra-clean workbench, soak it in 75% ethanol for 60s, take it out and place it in 0.1% HgCl 2 Soak for 8 minutes, take out, and wash 5 times with sterile water.
[0048] 2) Inoculate the sterilized explants in the primary medium, after dark cultivation for a week, the seeds can be observed to germinate, and change from light yellow to light green. Continue the light cultivation for 30 d to observe that the seeds develop and form protocorms. The bulbs are bright green in color, grow well, the pollution rate is 20%, and the seed germination rate is 94.6%;
[0049] The formulation of the primary induction medium used is: 1 / 2MS+6-BA 1.0mg·L -1 +NAA 0.2mg·L -1 +Activated carbon 0.1%+Banana puree 10%+...
Embodiment 2
[0056] Repeat Example 1, except:
[0057] Modify the formula of subculture proliferation and differentiation medium in step 3) to: MS+6-BA 2.0 mg·L -1 +NAA0.4mg·L -1 +Activated carbon 0.1%+Banana puree 10%+Sucrose 30g L -1 + Agar 3.4g L -1 , pH 5.8; the clump seedlings obtained from light culture for 50 d were relatively sturdy, with bright green leaves, and the proliferation coefficient was 5.2. It indicated that the concentration of cytokinin 6-BA was not as high as possible, and the appropriate hormone concentration and ratio were more conducive to the differentiation of protocorm and the reproduction of clump seedlings.
Embodiment 3
[0059] Repeat Example 1, except:
[0060] Modify the strong seedling rooting medium formula in step 4) to: 1 / 2MS+NAA 0.5mg·L -1 +ZnSO 4 1.0mg·L -1 +Activated carbon 0.1%+Banana puree 10%+Sucrose 20g L -1 + Agar 3.5g L -1 , pH 5.8; compared with Example 1, the rooting number, average root length and rooting rate decreased slightly; but its cultured for 50d, the plant grew well, with an average of 5 roots per plant, the average root length 2.2cm, and the rooting rate 93%. During the culture process, it was found that Dendrobium dendrobii has a certain tillering ability while taking root, and sometimes it can also dedifferentiate to form callus. The suitable concentration of ZnSO 4 It can promote the growth and rooting of plants while inhibiting the formation of callus or protocorm; but when ZnSO 4 When the concentration is higher, it may also have a certain inhibitory effect on the growth of the root system.
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