Method for inductively producing snow lotus somatic embryo
A technology of snow lotus and selection of snow lotus, applied in the field of bioengineering, can solve problems such as endangered, natural resources of snow lotus cannot reproduce, etc.
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Embodiment 1
[0017] callus induction
[0018] a. Select 200 Xinjiang snow lotus seeds, first sterilize the surface with 75% alcohol for 20 seconds, then soak in 0.1% mercuric chloride solution for 3 minutes, then rinse with sterile water until clean, put them on the culture medium of a sterile culture dish, 8 The daily germination rate reaches 70%;
[0019] b. Take the leaves of just germinated seedlings as explants, cut into 0.5cm in size 2 , put it on the callus induction medium of agar-solidified MS+2,4-D0.05mg / L+BA0.5mg / L, the inoculum size is 250ml triangular flask containing 50ml agar-solidified medium, and then 5 explants Body, solid culture conditions are: fluorescent lamp 2500lx, light 12h / d, temperature 25±1°C, humidity 65±5%, after 20-25 days, callus is obtained; solid medium is added with 3% sucrose, and the pH is adjusted to 5.8 , the sterilization time is 20min, and the temperature is 121℃;
[0020] induction of somatic embryos ...
Embodiment 2
[0023] callus induction
[0024] a. Select 300 Xinjiang snow lotus seeds, first sterilize the surface with 75% alcohol for 25 seconds, then soak in 0.1% mercury chloride solution for 4 minutes, then rinse with sterile water for 3 times, put them on MS medium, and the germination rate after 11 days reach 80%;
[0025] b. Take the leaves of just germinated seedlings as explants, cut into 0.6cm in size 2 , put it into agar-solidified MS+2,4-D0.1mg / L+BA1.0mg / L callus induction medium, 250ml Erlenmeyer flask containing 50ml agar solidified medium, connect 6 explants, solid The culture conditions are as follows: fluorescent lamp 3500lux, light 12h / d, temperature 25±1°C, humidity 65±5%, after 23 days, callus is obtained; solid medium is added with 4% sucrose, pH is adjusted to 5.85, and the sterilization time is uniform. for 20 minutes, and the temperature was 121°C.
[0026] induction of somatic embryos
[0027] c. Then subculture once on t...
Embodiment 3
[0029] callus induction
[0030] a. Select 400 snow lotus seeds, first sterilize the surface with 75% alcohol for 30 seconds, then soak in 0.1% mercuric solution for 5 minutes, then rinse with sterile water for 3 times, put them on a sterile petri dish, and the germination rate after 15 days reach 90%;
[0031] b. Take the leaves of just germinated seedlings as explants, cut into 0.8cm in size 2 , put it into agar-solidified MS+2,4-D2.0mg / L+BA3.0mg / L callus induction medium, 250ml Erlenmeyer flask containing 50ml agar solidified medium, connect 6 explants, solid The culture conditions are: fluorescent lamp 3500 lux, light 12h / d, temperature 25±1°C, humidity 65±5%, after 25 days, callus is obtained; solid medium is added with 5% sucrose, pH is adjusted to 5.9, and the sterilization time is uniform. for 20 minutes, and the temperature was 121°C.
[0032] induction of somatic embryos
[0033] c. Then subculture once on the medium of MS...
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