Betula microphylla var.paludosa embryonic callus induction method
An embryogenic callus and leaflet technology is applied in the field of plant tissue culture to achieve the effect of promoting breeding research
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Embodiment 1
[0013] A method for inducing embryogenic callus of Betula microphylla, the specific steps are as follows: (1) Select rooted seedlings of Betula microphylla tissue-cultured with a seedling age of 28 days and a height of 3 cm; (2) Cut the stem section of the rooted seedlings cultured into (3) Inoculate the explants in a medium containing mother liquor, 0.2mg / L 6-benzylaminoadenine, 0.5mg / L naphthaleneacetic acid and 500mg / L hydrolyzed casein Cultivate in protein dedifferentiation medium for 7 days to dedifferentiate and form callus. The culture temperature is 23°C, the light time is 12h / d, and the light intensity is 40umol / m 2 s; (4) Transfer the callus to the redifferentiation medium including the medium mother solution, 0.5mg / L 6-benzylaminoadenine and 0.02mg / L naphthaleneacetic acid for 10 days, and the culture temperature is 23℃ , the light time is 12h / d, and the light intensity is 40umol / m 2 ·s.
Embodiment 2
[0015] A method for inducing embryogenic callus of Betula microphylla, the specific steps are as follows: (1) select rooted seedlings of Betula microphylla tissue-cultured with a seedling age of 32 days and a seedling height of 5 cm; (2) cut the stem section of the rooted seedlings cultured (3) Inoculate the explants in a medium containing mother liquor, 0.3mg / L 6-benzylaminoadenine, 0.8mg / L naphthaleneacetic acid and 800mg / L hydrolyzed casein Cultivate in protein dedifferentiation medium for 5 days for dedifferentiation to form callus. The culture temperature is 25°C, the light time is 13h / d, and the light intensity is 50umol / m 2 s; (4) Transfer the callus to the redifferentiation medium including the medium mother solution, 0.8mg / L 6-benzylaminoadenine and 0.05mg / L naphthaleneacetic acid for 8 days, and the culture temperature is 25℃ , the light time is 13h / d, and the light intensity is 50umol / m 2 ·s.
Embodiment 3
[0017] A method for inducing embryogenic callus of Betula microphylla, the specific steps are as follows: (1) select rooted seedlings of Betula microphylla tissue-cultured with a seedling age of 35 days and a seedling height of 6 cm; (2) cut the stem section of the rooted plantlets cultured into (3) Inoculate the explants in a medium containing mother liquor, 0.5mg / L 6-benzylaminoadenine, 1.0mg / L naphthaleneacetic acid and 1000mg / L hydrolyzed casein Cultivate in protein dedifferentiation medium for 4 days to dedifferentiate and form callus. The culture temperature is 27°C, the light time is 14h / d, and the light intensity is 60umol / m 2 s; (4) Transfer the callus to the redifferentiation medium containing the medium mother solution, 1.0mg / L 6-benzylaminoadenine and 0.1mg / L naphthaleneacetic acid for 7 days, and the culture temperature is 27℃ , the light time is 14h / d, and the light intensity is 60umol / m 2 ·s.
[0018] In the above examples, the callus induction rate of the tis...
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