Method for rapidly propagating seedlings by using leaves of codonopsis lanceolata
A four-leaf ginseng and leaf technology, applied in the field of plant tissue culture, can solve the problems of slow growth, not long, unsatisfactory effect of stem segment and stem tip induction, etc.
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Embodiment 1
[0031] 1. Leaf induced differentiation of adventitious buds
[0032] Take young ginseng leaves, wash and disinfect them, blot dry with sterile paper, and inoculate the whole piece in induction medium (MS+6-BA 0.8mg / L+NAA0.5 mg / L+sucrose 30g / L+Kara Glue 6.5g / L, medium pH value is 5.8) to induce differentiation of adventitious buds. Culture conditions: culture temperature 24~26℃, light culture, light time 10~12h / d, light intensity 20~30μmol·m -2 ·s -1 . After culturing for about 10 days, adventitious buds began to differentiate at the leaf incision or on the surface of the leaf (see figure 1 ). After 20 days of culture, cut into small pieces and inoculate on fresh induction medium for subculture.
[0033] 2. Strong seedling cultivation process
[0034] Cut the adventitious buds after subculture into small clumps (blocks) and transfer them to strong seedling medium (MS+6-BA 0.5mg / L+NAA 0.1mg / L+sucrose 30g / L+carrageenan 6.5g / L+polypeptide 3ml / L) for strong seedling culture...
Embodiment 2
[0044] 1. Leaf induced differentiation of adventitious buds
[0045]Take young ginseng leaves, wash and sterilize, blot dry with sterile paper, then cut into small pieces about 0.5cm×1.0cm perpendicular to the veins and inoculate them in induction medium (MS+6-BA 2.5mg / L+NAA0.1mg / L+sucrose 30g / L+carrageenan 6.5g / L, medium pH value is 5.8) to induce differentiation of adventitious buds. Culture conditions: culture temperature 24~26℃, light culture, light time 10~12h / d, light intensity 20~30μmol·m -2 ·s -1 . After culturing for about 10 days, adventitious buds began to differentiate at the cuts of leaves or on the surface of leaves. After 20 days of culture, cut into small pieces and inoculate on fresh induction medium for subculture.
[0046] 2. Strong seedling cultivation process
[0047] Cut the adventitious buds after subculture into small clumps (blocks) and transfer them to strong seedling medium (MS+6-BA 0.3mg / L+NAA 0.5mg / L+sucrose 30g / L+carrageenan 6.5g / L+polypepti...
Embodiment 3
[0057] 1. Leaf induced differentiation of adventitious buds
[0058] Take young ginseng leaves, wash and sterilize, blot dry with sterile paper, then cut into small pieces of about 0.5cm×1.0cm perpendicular to the veins and inoculate them in induction medium (MS+6-BA 2.0mg / L+NAA0.3mg / L+sucrose 30g / L+carrageenan 6.5g / L, medium pH value is 5.8) to induce differentiation of adventitious buds. Culture conditions: culture temperature 24~26℃, light culture, light time 10~12h / d, light intensity 20~30μmol·m -2 ·s -1 . After culturing for about 10 days, adventitious buds began to differentiate at the cuts of leaves or on the surface of leaves. After 20 days of culture, cut into small pieces and inoculate on fresh induction medium for subculture.
[0059] 2. Strong seedling cultivation process
[0060] Cut the adventitious buds after subculture into small clumps (blocks) and transfer them to strong seedling medium (MS+6-BA 0.2mg / L+NAA 0.25mg / L+sucrose 30g / L+carrageenan 6.5g / L+poly...
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