Tissue culture method taking masson pine in-vitro mature embryo as explant
A tissue culture and masson pine technology, which is applied in the fields of botany equipment and methods, horticultural methods, plant regeneration, etc., can solve the problems of only slight elongation or stop growth, unable to meet the requirements of production, difficult tissue culture of masson pine, etc. , to achieve consistent and robust growth, fast start-up, and high rooting rate
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Embodiment 1
[0020] The tissue culture method using the isolated mature embryo of Pinus massoniana of the present embodiment as an explant may further comprise the steps:
[0021] The first step is to collect materials: take the mature seeds of Pinus massoniana, soak them overnight, rinse them under running water, and peel off the outer seed shell with tweezers.
[0022] The second step, material disinfection: the shelled seeds are sterilized with 75% ethanol for 30s in the ultra-clean workbench, then sterilized with 0.1% mercury liter for 10-15min, and finally rinsed with sterile water for 3-5 times.
[0023] The third step, inoculation and adventitious bud induction: In the ultra-clean workbench, use the method of conventional aseptic operation, use a scalpel and tweezers to peel off the seed embryos, inoculate them horizontally on the induction medium, and be careful when peeling off the seed embryos Be careful to avoid damage to the embryos as much as possible. The composition of the i...
Embodiment 2
[0032] The difference from the above examples is that the isolated mature embryos of Pinus massoniana are inoculated on the induction medium and cultured for about 20 days. Divided into DCR conventional basic medium, 6-benzylpurine 2.0 mg / L, naphthalene acetic acid 0.15 mg / L, sucrose 30 g / L; transfer the culture to the differentiation medium, cultivate for several days, and wait for the small buds to differentiate After that, the small buds can be inoculated on the bud elongation medium to elongate the buds, cultivated for 30-40 days, and when the young shoots grow to the height required by the conventional rooting procedure, enter the next step, wherein the components of the differentiation medium It is: DCR conventional basic medium, 6-benzylpurine 0.5 mg / L, indolebutyric acid 0.05 mg / L, sucrose 30 g / L, agar 6.0 g / L, and the composition of shoot elongation medium is: DCR Conventional basal medium, naphthalene acetic acid 0.1 mg / liter, sucrose 30 g / liter, this medium promotes...
Embodiment 3
[0034]The difference from the above examples is that the components of the induction medium are DCR conventional minimal medium, 1.0 mg / liter of 6-benzylpurine, 0.10 mg / liter of naphthaleneacetic acid, and 30 g / liter of sucrose; The components are: DCR conventional basic medium, 0.5 mg / L 6-benzylpurine, 0.05 mg / L indolebutyric acid, 30 g / L sucrose, 5.7 g / L agar; the components of the shoot elongation medium are : DCR conventional basic medium, indole butyric acid 0.05 mg / liter, sucrose 30 grams / liter; the components of the rooting medium are: 1 / 2DCR conventional basic medium, indolebutyric acid 2.0 mg / liter, sucrose 20 grams / Lift.
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