In-vitro sodium azide (NaN3) mutation breeding technology of dendranthema morifolium
A technology for mutation breeding and Chu chrysanthemum, which is applied in the field of chemical mutation breeding technology, can solve the problems of delayed flowering period, inability to apply, increase in the number of chromosomes, etc., and achieves the effects of good effect, simple operation procedure and high frequency of induced mutation.
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Embodiment 1
[0038] ①Wash and rinse the young stems of Chuju chrysanthemum with neutral detergent, then disinfect the surface with 75% ethanol for 30-90sec, and then wash with 0.1%HgCl 2 Sterilize the solution for 6-10 minutes, rinse the sterilized young stems with sterile water 4-5 times, and inoculate the explants of the stems with axillary buds in MS+NAA 0.1mgL -1For the culture medium, 2-3 explants are inoculated in each bottle, and can be subcultured once every 20-30 days to form a large number of sterile seedlings.
[0039] ②Choose aseptic seedlings grown for 15d-20d and cut them into stems with axillary buds and plant them in MS+TDZ 0.1-0.2mgL -1 +NAA.01 -0.2mgL -1 +NaN 3 4-8mgL -1 or MS+KT 3-4mgL -1 +NAA.01-0.4mgL -1 + NaN 3 4-8mgL -1 On the clump bud differentiation medium, 35d-45d part of the shoot tips, leaf axils or stem bases formed cluster buds.
[0040] ③Transfer the cluster buds formed by the mutagenesis treatment to MS+NAA 0.1mgL -1 On the strong seedling medium...
Embodiment 2
[0044] ①Wash and rinse the young stems of Chuju chrysanthemum with neutral detergent, then disinfect the surface with 75% ethanol for 30-90sec, and then wash with 0.1%HgCl 2 Sterilize the solution for 6-10 minutes, rinse the sterilized young stems with sterile water for 4-5 times, peel off the shoot tip explants and inoculate in MS+NAA 0.1mgL -1 For the culture medium, 2-3 explants are inoculated in each bottle, and can be subcultured once every 30-40 days to form a large number of sterile seedlings.
[0045] ②Choose aseptic seedlings grown for 15d-20d and cut them into stems with axillary buds and inoculate them in MS+TDZ 0.1-0.2mgL -1 + NAA.01-0.2mgL -1 +NaN 3 4-8mgL -1 or MS+KT 3-4mgL -1 +NAA.01-0.4mgL -1 + NaN 3 4-8mgL -1 On the clump bud differentiation medium, 35d-45d part of the shoot tips, leaf axils or stem bases formed cluster buds.
[0046] ③Transfer the cluster buds formed by the mutagenesis treatment to MS+NAA 0.1mgL -1 On the strong seedling medium, cu...
Embodiment 3
[0050] a. Wash and rinse the young leaves of Chrysanthemum chrysanthemum with neutral detergent, then disinfect the surface with 75% ethanol for 30-90sec, and then wash with 0.1% HgCl 2 Sterilize the solution for 6-10 minutes, rinse the sterilized young leaves 4-5 times with sterile water, cut the young leaves into small pieces of about 0.5×0.3cm2, and inoculate them into MS+TDZ 0.1-0.2mgL -1 +NAA.01-0.2mgL -1 +NaN 3 4-8mgL -1 or MS+KT 3-4mgL -1 +NAA.01-0.4mgL -1 + NaN 3 4-8mgL -1 On the cluster bud differentiation medium, inoculate 2-3 explants in each bottle, culture for 40-50 days, and 20-30% of the leaf explants form cluster buds.
[0051] b. Cut the aseptic seedlings of cluster buds into stems with axillary buds and inoculate them in MS+TDZ 0.1-0.2mgL -1 +NAA.01-0.2mgL -1 +NaN 3 4-8mgL -1 or MS+KT 3-4mgL -1 +NAA.01-0.4mgL -1 + NaN 3 4-8mgL -1 On the clump bud differentiation medium, 35d-45d part of the shoot tips, leaf axils or stem bases formed cluster ...
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