A method for improving the rate of induction of somatic embryos of Pinus massoniana
A technology of induction rate and masson pine, which is applied in the field of plant reproduction, can solve the problems of low induction rate of embryogenic callus, difficulty in obtaining regenerated plants, and difficulty in differentiation, and achieves the ability to maintain normal proliferation, less browning, and acquisition rate of sub-generation. high effect
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Embodiment 1
[0025]On June 25, 2015, the immature cones of Tongmian pine excellent tree that were healthy, free from diseases and insect pests, and the zygotic embryo was in the stage of fission and multiple embryos were collected as explants in the excellent stand of Tongmian pine in the state-owned Paiyangshan Forest Farm in Guangxi. Use volume concentration 45% alcohol + volume concentration 0.03% streptomycin + 100 mg·L -1 After the nicotinic acid mixture was wiped and disinfected on the surface of the explants, put every 4-5 explants into a plastic ziplock bag with a size of 20 mm×30 mm, seal the bag mouth by 80%, fold the bag mouth down Store in a refrigerator at 8°C for 14 days.
[0026] The pretreated explants were taken out of the refrigerator and rinsed with running water for 2 h, soaked in a broad-spectrum fungicide with a volume concentration of 0.1-0.3 % for 1 h, took out the explants and placed them in a volume concentration of 60 % alcohol for 3-5 min, then transferred to 8...
Embodiment 2
[0030] On July 15, 2016, the immature cones of the Tongmian pine excellent tree that were healthy, free from diseases and insect pests, and the zygotic embryos were in the stage of fission and multiple embryos were collected as explants in the excellent stand of Tongmian pine in the state-owned Paiyangshan Forest Farm in Guangxi. Use volume concentration 45% alcohol + volume concentration 0.03% streptomycin + 100 mg·L -1 After the nicotinic acid mixture was wiped and disinfected on the surface of the explants, put every 4-5 explants into a plastic ziplock bag with a size of 20 mm×30 mm, seal the bag mouth by 80%, fold the bag mouth down Store in a refrigerator at 6°C for 12 days.
[0031] The pretreated explants were taken out from the refrigerator and rinsed with running water for 2 h, soaked in a broad-spectrum fungicide with a volume concentration of 0.1-0.3 % for 1.5 h, took out the explants and placed them in a volume concentration of 60 % alcohol for 3-5 min, then trans...
Embodiment 3
[0035] On July 8, 2017, the immature cones of Tongmian pine excellent tree that were healthy, free from diseases and insect pests, and the zygotic embryo was in the stage of fission and polyembryony were collected as explants in the excellent stand of Tongmian pine in the state-owned Paiyangshan Forest Farm in Guangxi. Use volume concentration 45% alcohol + volume concentration 0.03% streptomycin + 100 mg·L -1 After the nicotinic acid mixture was wiped and disinfected on the surface of the explants, put every 4-5 explants into a plastic ziplock bag with a size of 20 mm×30 mm, seal the bag mouth by 80%, fold the bag mouth down Store in a refrigerator at 7°C for 14 days.
[0036]The pretreated explants were taken out of the refrigerator and rinsed with running water for 2 h, soaked in a broad-spectrum fungicide with a volume concentration of 0.1-0.3 % for 1 h, took out the explants and placed them in a volume concentration of 60 % alcohol for 3-5 min, then transferred to a sodi...
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