Cultivation method for highly effective plant tissue differentiation and regeneration
A technology for inducing plants and culturing methods, applied in the field of plant tissue culture, and can solve the problems of low differentiation and regeneration rate and slow regeneration speed.
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Embodiment 1
[0029] Embodiment 1. The tissue culture of the blade regeneration plant of Seoul loose-leaf lettuce
[0030] 1. Soak Seoul loose-leaf lettuce seeds (purchased from Beijing Jinxiu Dadi Company) with mercury chloride for 5 minutes, put them on MS solid medium, cultivate them at 24°C under light for 12h / d, and subculture on MS solid medium after germination to cultivate.
[0031] 2. Get young and tender lettuce true leaves, leaf diameter 2-3cm, cut into 1cm * 1cm square (explant), place the callus that betaine aldehyde (purchased from U.S. DNA Polymerase Technology company) concentration is 5mM on the back side Induction and differentiation solid medium (callus induction and differentiation medium: MS basic medium + IAA (Sigma) 1.0 mg / L + 6-BA (Sigma) 0.2 mg / L, pH 5.8).
[0032] 3. Cultivate at 24°C and light 12h / d.
[0033] 4. After 11 days, calluses began to grow around the explants, and in about 20 days, they began to differentiate into bud points or sprouts, and in about 30...
Embodiment 2
[0039] Example 2. Effects of Different Concentrations of Betaine Aldehyde on Inducing Plant Tissue Differentiation and Regeneration
[0040]The betaine aldehyde concentration added in callus induction and differentiation medium is 4,6,10mM, carries out the tissue culture of the blade regeneration plant of Seoul loose-leaf lettuce in the same manner as embodiment 1 and comparative example 1, all can be different Promote the differentiation regeneration of lettuce to a certain extent, differentiation rate and regeneration rate are all higher than control example, but lower than embodiment 1 (see Figure 6 , taking 10mM as an example, the rest are similar), therefore, 5mM is the optimal concentration that betaine aldehyde promotes the differentiation and regeneration of lettuce leaves.
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