Tissue culture method and special culture medium of Spanish dagger anther
A culture medium and technology of Pteris orchids, 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] Example 1. Preparation and statistical observation of haploid plants
[0030] 1. Obtainment and statistical observation of callus induced by anthers
[0031] 1. Pretreatment of anthers
[0032] Collect the flower buds of Phnom Penh Phoenix orchid (purchased from Beijing Xiaoying Flower Market) in the single-core stage, first rinse with tap water 3-5 times, dark treatment at 4 ℃ low temperature for 3 days, and then use 75% (volume percentage ) Alcohol sterilization for 40 seconds, rinse with sterile water 5 times, then sterilize with 0.1% (mass percentage) mercury liters for 6 minutes, and rinse 5 times with sterile water.
[0033] 2. Anther-induced callus acquisition
[0034] Use tweezers to carefully remove the anthers (microspores of anthers in the mononuclear side stage, such as figure 1 (Shown) inoculated on callus induction medium, and cultured in the dark for one week at 25°C, and then cultivated at a temperature of 23°C, an illumination intensity of 2000 Lx, and an illumi...
Embodiment 2
[0055] Example 2. Plant ploidy doubled
[0056] The 2 haploid Phnom Penh chinensis plants that were detected as haploid obtained in Example 1 were treated with 0.1% colchicine (mass percentage) for 24 hours to obtain homozygous diploid plants.
[0057] The treatment method is: first filter and sterilize 0.1% colchicine, then dip a sterile cotton ball into an appropriate amount of the above-mentioned colchicine solution, and then place the small cotton ball on the growth point of the plant for 24 hours.
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