A kind of cryopreservation method of embryogenic callus of masson pine
A technology of embryogenic callus and cryopreservation, which is applied to the preservation of plants, botany equipment and methods, and applications, and can solve the problems of systematic technology, poor stability, low somatic embryo induction rate, and long culture cycle , to achieve the effect of solving the difficulty of induction, solving the serious apoptosis, good economic and social benefits
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Embodiment 1
[0027]Using immature cleaved polyembryonic zygotic embryos of P. massoniana as explants, a large number of vigorous embryogenic cell lines of P. massoniana were obtained through induction of somatic embryos. Among them, after the fast-growing embryogenic cell line SE-17 was maintained and cultured for 8 days, the core part of the embryogenic callus was avoided, and the peripheral embryogenic callus with high viscosity and good transparency was collected as the preservation material.
[0028] Select 2 g of embryogenic callus and put it into a 50 mL sterile Erlenmeyer flask, and add 7 mL of base solution into the sterile Erlenmeyer flask, seal and place the sterile Erlenmeyer flask on a rotary shaker for vibration dispersion Treat for 24 h, and the treatment conditions are: dark culture, no light, temperature 20±0.5°C; rotating shaker speed 80 rpm / min. The raw material and content thereof of the base liquid are: KNO 3 200 mg·L -1 ;KH 2 PO 4 170 mg·L -1 ; KCl 625 mg L -1 ;...
Embodiment 2
[0032] Using immature cleaved polyembryonic zygotic embryos of P. massoniana as explants, a large number of vigorous embryogenic cell lines of P. massoniana were obtained through induction of somatic embryos. Among them, the fast-growing embryogenic cell line SE-17 was proliferated and maintained for 7 days, and the core part of the embryogenic callus was avoided, and the peripheral embryogenic callus with high viscosity and good transparency was collected as preservation materials.
[0033] Select 1g of embryogenic callus and put it into a 25 mL sterile Erlenmeyer flask, add 3 mL of base solution into the sterile Erlenmeyer flask, seal it, place the sterile Erlenmeyer flask on a rotary oscillator for vibration dispersion treatment 24 h, the treatment conditions are: dark culture, no light, temperature 20±0.5°C; rotating shaker speed 80 rpm / min. The raw material and content thereof of the base liquid are: KNO 3 200 mg·L -1 ;KH 2 PO 4 170 mg·L -1 ; KCl 625 mg L -1 ;CaCl ...
Embodiment 3
[0037] Using immature cleaved polyembryonic zygotic embryos of P. massoniana as explants, a large number of vigorous embryogenic cell lines of P. massoniana were obtained through induction of somatic embryos. Among them, after the fast-growing embryogenic cell line SE-22 was maintained and cultured for 7 days, the core part of the embryogenic callus was avoided, and the peripheral embryogenic callus with high viscosity and good transparency was collected as the preservation material.
[0038] Select 1 g of embryogenic callus and put it into a 25 mL sterile Erlenmeyer flask, add 4 mL of base solution into the sterile Erlenmeyer flask, seal it, place the sterile Erlenmeyer flask on a rotary shaker for vibration dispersion Treat for 24 h, and the treatment conditions are: dark culture, no light, temperature 20±0.5°C; rotating shaker speed 80 rpm / min. The raw material and content thereof of the base liquid are: KNO 3 200 mg·L -1 ;KH 2 PO 4 170 mg·L -1 ; KCl 625 mg L -1 ;CaC...
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