Method of propagation of Tilia miqueliana by tissue culture
A technology of linden tissue and culture medium, applied in the field of woody plant seedling propagation
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Embodiment 1
[0014] 1. On April 15, 2006, 50 robust annual seedlings of Nanjing Tilia were selected. Cut the aerial parts, remove the leaves, and use the stem segments as explants, each stem segment has an axillary bud, 2 cm long. Rinse the explants with clean water, rinse once with washing powder, and sterilize with 84 disinfectant on a shaker for 15-20 minutes. After rinsing with running water for 20-30 minutes, sterilize with 75% ethanol for 50-60 seconds on a sterile operating table, immediately pour 0.1% mercuric chloride for 10 minutes, and finally wash 4 times with sterile water, shaking for 3-5 minutes each time. Both ends of the sterilized explant were excised by 0.5 cm to remove the injured part. Access induction medium MS+6-BA0.2mg / L+KT0.05mg / L+IBA0.02mg / L+GA 3 0.1mg / L, add agar 6.5g / l, sucrose 30g / l. The medium pH is 5.8-6.0. Each bottle received 3 plants, a total of 70 bottles were connected. Under the conditions of temperature 20-26°C, light time 10h / day, and light inten...
Embodiment 2
[0018] 1. On April 25, 2007, 30 robust annual seedlings of Nanjing Tilia were selected. Cut the aerial parts, remove the leaves, and use the stem segments as explants, each stem segment has an axillary bud, 2 cm long. Rinse the explants with clean water, rinse once with washing powder, and sterilize with 84 disinfectant on a shaker for 15-20 minutes. After rinsing with running water for 20-30 minutes, sterilize with 75% ethanol for 50-60 seconds on a sterile operating table, immediately pour 0.1% mercuric chloride for 10 minutes, and finally wash 4 times with sterile water, shaking for 3-5 minutes each time. Both ends of the sterilized explant were excised by 0.5 cm to remove the injured part. Access induction medium MS+6-BA0.2mg / L+KT0.05mg / L+IBA0.02mg / L+GA 3 0.1mg / L, add agar 6.5g / l, sucrose 30g / l. The medium pH is 5.8-6.0. Each bottle received 3 plants, a total of 50 bottles were connected. Under the conditions of temperature 20-26°C, light time 10h / day, and light inten...
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