Tissue culture method and special culture medium of Spanish dagger anther
A culture medium and a technique for anchovy orchid, which are applied in the field of plant tissue culture and crop breeding to achieve the effect of improving selection efficiency
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Embodiment 1
[0029] Embodiment 1, preparation and statistical observation of Pteris orchid haploid plant
[0030] 1. The acquisition of anther-induced callus and its statistical observation
[0031] 1. Pretreatment of anthers
[0032] Collect the flower buds of Phnom Penh orchids (purchased from Beijing Xiaoying Flower Market) that are in the single-nucleus stage, rinse them with tap water 3-5 times, and treat them in the dark at a low temperature of 4°C for 3 days, then use 75% (volume percentage) ) alcohol disinfection for 40 seconds, rinsed with sterile water for 5 times, then sterilized with 0.1% (mass percentage) mercuric chloride for 6 minutes, and rinsed with sterile water for 5 times.
[0033] 2. Acquisition of callus induced by anthers
[0034] The flower bud after the pretreatment in the above-mentioned steps 1 is carefully anther (the anther microspore of uninucleate marginal stage such as figure 1 Shown) were inoculated on the callus induction medium, and under the condition...
Embodiment 2
[0055] Embodiment 2, plant ploidy doubling
[0056] Homozygous diploid plants can be obtained by treating the two haploid Phnom Penh orchid plants detected as haploid obtained in Example 1 above with a concentration of 0.1% colchicine (mass percentage) for 24 hours.
[0057] The treatment method is as follows: firstly filter and sterilize 0.1% colchicine, then use a sterile cotton ball to dip an appropriate amount of the above-mentioned colchicine solution, then place the small cotton ball on the growth point of the plant, and treat for 24 hours.
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