Rubber tree embryoid induction culture medium, induction method for embryonic callus and culture method of resistant callus
An embryogenic callus and induction medium technology, applied in the field of plant tissue culture, can solve the problems of low overall efficiency and different tissue transformation rates.
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[0064] The preparation method of the induction medium is not particularly limited in the present invention, and the conventional preparation method of the medium well known in the art can be used.
[0065] The invention provides a method for inducing embryogenic callus based on rubber tree anther somatic embryos, comprising the steps of:
[0066] 1) selecting yellow-green male flowers with a length of 2.81 to 3.20 mm to be sterilized, and separating anthers from the sterilized male flowers to obtain sterilized anther somatic cells;
[0067] 2) Inoculate the sterilized anther somatic cells of the step 1) on the first-generation callus induction medium, culture in dark at 26-28°C for 44-46 days, and transfer the obtained callus to the culture medium described in the above scheme. On the embryoid body induction medium, induce culture at 26-28°C for 40-50 days to obtain spherical or cotyledon-shaped somatic embryos;
[0068] 3) Scratching the somatic embryos in step 2) and inocul...
Embodiment 1
[0097] Embryoid induction medium: modified MS medium as the basic medium, including: Kt 0.55mg / L, NAA0.16mg / L, 6-BA0.04mg / L, salicylic acid 0.06mg / L, spermidine 4.5mg / L, acid hydrolyzed casein 310mg / L, coconut water volume concentration is 7%, activated carbon mass concentration is 0.11%, sucrose 55g / L and plant gel 2.4g / L; The pH value of described induction medium is 5.7.
Embodiment 2
[0099] The material is Hevea brasiliensis Müll.Arg. variety Reyan 7-33-97, and the rubber inflorescences were collected from the experimental farm of the Chinese Academy of Tropical Agricultural Sciences. Select yellow-green male flowers with a length of 2.81 mm, first disinfect them with 75% alcohol for 43 seconds, then disinfect them with 1 mg / mL mercury chloride solution for 13.5 minutes, and wash them four times with sterile water to obtain disinfected anthers. Inoculate the sterilized anthers on the first-generation callus induction medium (the first-generation callus induction medium is based on MS medium, including 2,4-D 1.0mg / L, 6-BA0.2mg / L , coconut water volume concentration 5%, sucrose 70g / L and plant gel 2.2g / L, pH value is 5.8), at 26 ℃, dark culture 46d, the callus that obtains is transferred to the embryoid body induction of embodiment 1 On the culture medium, induce at 26°C for 50 days to obtain somatic embryos with a size of 1-1.2 mm.
[0100] The obtained so...
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