Tissue culture method of haricot in situ cluster seedling
A technology of tissue culture and lentils, applied in horticultural methods, botany equipment and methods, plant regeneration, etc., can solve the problems of large differences between genotypes, few adventitious buds of lentils, and low reproduction number, etc., to achieve small regeneration differences and cultivate Shorter time and better repeatability
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Embodiment 1
[0013] The test variety is Jiaoxuan Lentil No. 1, 50 grains;
[0014] Wash the seeds with water and put them in 70% alcohol for 25 seconds, remove them in 10% sodium hyposulfite for 15 minutes, wash them twice with sterile water, put the sterilized seeds in MSB+6-BA 0.1mg / L, pH 5.8, and culture for germination Germinate at 23°C for 5 days; use a scalpel to remove the hypocotyl, the growth points of the terminal buds and lateral buds between the two cotyledons of the germinated lentils under a dissecting microscope, keep the two cotyledons, and put them in MSB+6-BA 2mg / L, pH 5.8, germination at 23°C for 30 days on the bud growth medium, cultured to a height of 2 cm; cut off adventitious buds and put them in MSB+IBA 0.1mg / L, pH 5.8, rooting medium at 23°C to induce rooting for 25 days, Transplant to large pots to obtain regenerated seedlings; obtain 356 regenerated plants of cross-selected lentil No. 1. There are 7-10 clusters of adventitious buds per tree, and the test-tube s...
Embodiment 2
[0016] The test variety is Nicheng No. 1, 50 grains;
[0017] Wash the seeds with water and put them in 70% alcohol for 30 seconds, take them out and put them in 10% sodium hyposulfite for 20 minutes, wash them 3 times with sterile water, put the sterilized seeds in MSB+6-BA 0.5mg / L, pH 5.8, germination medium Germinate at 24°C for 4 days; use a scalpel under a dissecting microscope to remove the hypocotyl, the growth points of the terminal buds between the two cotyledons and the lateral buds, keep the two cotyledons, and put them in MSB+6-BA 3mg / L , pH 5.8, germination at 25°C for 28 days on the bud growth medium, and cultured to a height of 3 cm; cut off adventitious buds and place them in MSB+IBA 0.3mg / L, pH 5.8, induce rooting at 23°C on the rooting medium for 24 days, Transplant to large pots to obtain regenerated seedlings; Obtain Nicheng No. 1, 389 regenerated plants. Produce 7-10 clusters of adventitious buds per tree, cultured for 56 days in test tube seedlings, buds...
Embodiment 3
[0019] The test variety is Nicheng No. 2, 50 grains;
[0020] Wash the seeds with water and put them in 70% alcohol for 35 seconds, take them out and put them in 10% sodium hyposulfite for 25 minutes, wash them with sterile water for 3 times, put the sterilized seeds in MSB+6-BA 1mg / L, pH 5.8, germination medium Germinate at 25°C for 3 days; use a scalpel to remove the hypocotyl, the growth point between the two cotyledons and the growth points of the lateral buds of the germinated lentils under a dissecting microscope, keep the two cotyledons, and put them in MSB+6-BA 4mg / L , pH 5.8, germination at 27°C for 26 days on the long-bud medium, cultured to a height of 4 cm; excised adventitious buds and placed in MSB+IBA 0.5mg / L, pH 5.8, induced rooting at 23°C on the rooting medium for 23 days, transplanted Planted in large pots to obtain regenerated seedlings; obtained 373 regenerated plants of Nicheng No. 2. Produce 7-10 clusters of adventitious buds per plant, cultured for 52 ...
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