The method of inducing tetraploidy of candidum and polyploidy with callus tissue
A callus induction, callus technology, applied in the field of plant biology, can solve the problems of pollution, increased seed contamination rate, test failure, etc.
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Embodiment 1
[0028] A method for inducing white and tetraploidy with callus, comprising the following steps:
[0029] (1) On the ultra-clean workbench, cut the callus into small pieces of 0.3cm×0.3cm, insert them into the proliferation medium and culture them for 10 days; under sterile conditions, add sterilized Chemical inducer, chemical inducer soaked callus for 24 h;
[0030] The chemical inducer is: 0.05% colchicine+3%DMSO;
[0031] The proliferation medium is: N 6 +2,4-D 1.0 mg / L+NAA 0.5 mg / L+TDZ 0.5 mg / L, pH6.0;
[0032] (2) On the ultra-clean workbench, take out the callus in step (1), wash it in sterile water for 5 times, blot the water on sterile filter paper, insert it into the differentiation medium and culture it for 22 days Bud points are produced, buds are formed in 36 days, and 2 to 3 leaves grow from buds in 55 days. The buds are thin and weak, and the leaves are yellow-green;
[0033] The differentiation medium is: MS+6-BA 1.0 mg / L+ZT 0.5 mg / L+NAA 0.5 mg / L, pH6.0;
[...
Embodiment 2
[0041] (1) On the ultra-clean workbench, cut the callus into small pieces of 0.3cm×0.3cm, insert them into the proliferation medium and culture them for 10 days; under sterile conditions, add sterilized Chemical inducer, the chemical inducer soaked the callus for 8 h;
[0042] The chemical inducer is: 0.2% colchicine+3%DMSO;
[0043] The proliferation medium is: N 6 +2,4-D 1.0 mg / L+NAA 0.5 mg / L+TDZ 0.5 mg / L, pH6.0;
[0044] (2) On the ultra-clean workbench, take out the callus in step (1), wash it in sterile water for 5 times, blot the water with sterile filter paper, insert it into the differentiation medium for culture, and partially callus The tissue turns black, and it takes a long time to induce budding. Bud points are formed at 30 days, buds are formed at 43 days, and 2-3 leaves grow from the buds at 65 days. The buds are thin and thick, and the leaves are yellow-green and dark green;
[0045] The differentiation medium is: MS+6-BA 1.0 mg / L+ZT 0.5 mg / L+NAA 0.5 mg / L, p...
Embodiment 3
[0053] (1) On the ultra-clean workbench, cut the callus into small pieces of 0.3cm×0.3cm, insert them into the proliferation medium and culture them for 10 days; under sterile conditions, add sterilized Chemical inducer, the chemical inducer soaked the callus for 16 h;
[0054] The chemical inducer is: 0.1% colchicine+3%DMSO;
[0055] The proliferation medium is: N 6 +2,4-D 1.0 mg / L+NAA 0.5 mg / L+TDZ 0.5 mg / L, pH6.0;
[0056] (2) On the ultra-clean workbench, take out the callus in step (1), wash it in sterile water for 5 times, blot the water with sterile filter paper, insert it into the differentiation medium and culture it, and produce Bud point, 40 d to form buds, 60 d buds to grow 2 to 3 leaves, buds thick, leaves dark green;
[0057] The differentiation medium is: MS+6-BA 1.0 mg / L+ZT 0.5 mg / L+NAA 0.5 mg / L, pH6.0;
[0058] (3) Put the buds obtained in step (2) into the rooting medium on the ultra-clean workbench and cultivate them. On the 7th day, protrusions appeared ...
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