Tissue culture method for inducing immature zygotic embryos of paris polyphylla to generate somatic embryos
A somatic embryo and tissue culture technology, which is applied in horticultural methods, botany equipment and methods, plant regeneration, etc., can solve the problem of lower induction rate of adventitious buds, insignificant tissue culture effect, and low reproduction coefficient of callus of P. problems, to achieve the effect of avoiding endophyte infection, good plasticity, and low infection rate
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Embodiment 1
[0014] 1) Sampling and disinfection: The suitable sampling time for the explants used is August. The best when the fruit is about to mature, but the skin has not cracked. Cut the whole fruit of C. Chonglou and open it, remove the attached fibrous tissue of C. Chonglou seeds with fleshy seed coats, wash in running water, and soak in tap water for 24 hours, and pass 0.1% HgCl under sterile conditions. 2 After 5 minutes of disinfection, wash with sterile water 10 times;
[0015] 2) Peeling of immature zygotic embryos: Use tweezers to peel off the fleshy testa of the sterilized seeds in 1) under aseptic conditions, use ultra-fine tissue culture forceps to fix the peeled endosperm under a stereo microscope, and use surgery The blade cuts the endosperm longitudinally. Because the color of the embryo is the same as that of the endosperm, but the tissue density is different, the vertical light intensity of the microscope can be adjusted to find the shadow-like immature zygotic embryo in ...
Embodiment 2
[0019] 1) Sampling and disinfection: In September each year, remove the attached fibrous tissues of the seeds with fleshy seed coats, wash them in running water, and soak them in tap water for 24 hours, and pass them to 0.1% HgCl under aseptic conditions. 2 After disinfection for 5 minutes, wash with sterile water 10 times;
[0020] 2) Stripping of immature zygotic embryos: peel off the fleshy testa from the sterilized seeds under aseptic conditions, fix the peeled endosperm with tissue culture forceps, and cut the endosperm longitudinally with a scalpel;
[0021] 3) Induction of somatic embryos: inoculate the stripped immature embryos in a pH 5.8 medium (medium is 1 / 2MS, agar powder concentration is 7.5g / L, sucrose concentration is 30g / L), at 19℃ Continuous dark light training;
[0022] 4) Establishment of embryogenic tissue culture system: After 6 months of culture, after 6 months of culture, the somatic embryos obtained by induction are stripped and placed on a new induction mediu...
Embodiment 3
[0024] 1) Sampling and disinfection: In August each year, remove the attached fibrous tissues of the seeds with fleshy seed coats, wash them in running water, and soak them in tap water for 24 hours, and pass them to 0.1% HgCl under aseptic conditions. 2 After disinfection for 5 minutes, wash with sterile water 10 times;
[0025] 2) Stripping of immature zygotic embryos: peel off the fleshy testa from the sterilized seeds under aseptic conditions, fix the peeled endosperm with tissue culture forceps, and cut the endosperm longitudinally with a scalpel;
[0026] 3) Induction of somatic embryos: inoculate the stripped immature embryos in a pH 5.8 medium (medium is 1 / 2MS, agar powder concentration is 7.5g / L, sucrose concentration is 30g / L) at 21℃ Continuous dark light training;
[0027] 4) Establishment of embryogenic tissue culture system: After 6 months of culture, the somatic embryos obtained by induction are stripped and placed on a new induction medium to continue culturing to make...
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