Hormone-free tissue culture method for dendrobium candidum
A Dendrobium candidum, tissue culture technology, applied in the field of agriculture, to achieve the effect of reducing production costs, low cost, resource waste and time consumption
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Embodiment 1
[0020] Dendrobium officinale hormone-free tissue culture method:
[0021] a) The mature Dendrobium candidum capsules or Dendrobium candidum seeds collected from the field are sterilized, planted in hormone-free MS medium, and cultivated for 25 days. Seed embryos expand and germinate to form green round bulbs; culture conditions: culture temperature 23~ 27℃, illuminance 1500~2000lx, light time 12h / d;
[0022] b) Transfer the bulbs into hormone-free basic MS basic medium + mashed potato 20g / l medium, and cultivate them for 25 days. The bulbs overlap and pile up in the hormone-free medium, and leaf primordia and differentiated seedlings appear; Culture conditions: culture temperature 23~27℃, light intensity 1500~2000lx, light time 12h / d;
[0023] c) transfer the round corm differentiated seedlings into the hormone-free MS basic medium+mashed banana 15g / l+mashed potato 15g / l medium, after 20 to 30 days of induction, leaf primordia and seedlings reaching 1 to 2 cm in height appear...
Embodiment 2
[0031] Dendrobium officinale hormone-free tissue culture method:
[0032] a) The mature Dendrobium candidum capsules or Dendrobium candidum seeds collected from the field are sterilized, planted in hormone-free MS medium, and cultivated for 30 days. Seed embryos expand and germinate to form green round bulbs; culture conditions: culture temperature 23~ 27℃, illuminance 1500~2000lx, light time 12h / d;
[0033] b) Transfer the bulbs into the hormone-free basic MS basic medium + mashed potato 15g / l medium, and cultivate them for 30 days. The bulbs overlap and pile up in the hormone-free medium, and leaf primordia and differentiated seedlings appear; Culture conditions: culture temperature 23~27℃, light intensity 1500~2000lx, light time 12h / d;
[0034] c) transfer the round corm differentiated seedlings into hormone-free MS basic medium+mashed banana 20g / l+mashed potato 20g / l medium, after 20 to 30 days of induction, leaf primordium and a height of 1 to 2cm seedlings appear, and t...
Embodiment 3
[0042] Hormone culture: The medium used for seed germination into spherical bulbs is MS basic medium + sucrose 30g / l + agar 6g / l + 6-BA (benzylaminopurine) 0.5mg / l + NAA (α-naphthaleneacetic acid) 0.2mg / l , The medium used for proliferation and differentiation from round bulbs is MS basic medium + sucrose 30g / l + agar 6g / l + 6-BA (benzylaminopurine) 0.5mg / l + NAA (α-naphthaleneacetic acid) 0.2mg / l, The culture medium used from the differentiated seedling to the stem section is MS basic medium+sucrose 30g / l+agar 6g / l+6-BA (benzylaminopurine) 0.5mg / l+NAA (α-naphthaleneacetic acid) 0.5mg / l), The medium used for rooting and strong seedlings is 1 / 2 MS basic medium + sucrose 30g / l + agar 6g / l + 6-BA (benzylaminopurine) 2mg / l + NAA (α-naphthaleneacetic acid) 0.5mg / l, and the cultivation method is the same as In Example 1, the culture time is 30-35 days, 30-35 days, 60-65 days, and 35-40 days, respectively.
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