Method for culturing apple triploid germ plasm
A triploid and germplasm technology, applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve the problems of low emergence rate of hybrids, achieve good disinfection, facilitate rapid growth, and assist rapid growth Effect
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Embodiment 1
[0033] Aseptic treatment of apple hybrid seeds
[0034] Select the mature fruit of 'Tetraploid Gala'×'Hanfu', remove the pulp, and take out the hybrid seeds, a total of 540 seeds, put the seeds into 100ml triangular flasks, and soak them in 70% (V / V) ethanol for 30s in turn , soak in 0.1% (M / V) mercuric chloride for 5 minutes, and rinse with sterile water for 3 times.
Embodiment 2
[0036] Hybrid Embryo Culture in Vitro
[0037] According to the conventional method, F14 medium, 1g / L activated carbon, hydrolyzed milk 500mg / L protein, 150mg / L ascorbic acid, 15g / L sucrose, 0.4mg / L6-BA, 0.2mg / LNAA, 6g / L agar were prepared into the culture medium , use 0.7% agar strips to solidify, adjust the pH of the medium to 5.8 with 1mol / L NaOH, divide the medium into test tubes, autoclave at 121°C for 20 minutes at 1 atmosphere, and then transfer to sterilized In the triangular flask of bacteria;
[0038] The hybrid embryos stripped of the seed coat were inoculated in an Erlenmeyer flask filled with culture medium, and cultured at a temperature of 25±2°C, a light intensity of 1500 lx, and a light cycle of 14 hours / day.
Embodiment 3
[0040] Ploidy Identification and Multiplication of Hybrid Seedlings
[0041] After 30 days of in vitro culture, hybrid embryos develop into full seedlings with a height of 10 cm. The leaves and root tips are collected under sterile conditions for ploidy identification, and the roots of the plants identified as triploid are removed and cut into 1.5 cm stem segments. Transfer to the subculture medium prepared by MS medium, 0.4mg / L6-BA, 0.2mg / LIAA, 30g / L sucrose, subculture once a month.
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