Tissue cultivating method of dendrobium officinale growing in Yunnan
A technology of tinplate and tissue cultivation in Yunnan, applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve the problems of restricting the production and development of Dendrobium candidum test-tube seedlings, shortage of raw materials, long breeding time, etc., to achieve sustainable development. The effect of utilization and development, ensuring the quality of seedlings, and shortening the growth cycle
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Embodiment 1
[0015] Embodiment 1: explant culture
[0016] The robust dormant buds of Dendrobium candidum were inoculated on MS sucrose agar medium containing auxin, cytokinin, 200ml / L coconut milk and 3% activated carbon after being sterilized by alcohol and mercuric chloride, and cultured in a light culture room for 2~ After 3 weeks, most of them can germinate and grow new shoots. They have a higher germination rate and good growth in the medium of NAA0.5mg / L+6-BA 1.0-2.0mg / L. After 2 months, the seedlings Up to 1.5cm-3.0cm.
Embodiment 2
[0017] Embodiment 2: protocorm induction and cultivation thereof
[0018] Use the tissue culture seedlings that have established a sterile system, cut the stems into small pieces and continue to induce the differentiation of protocorms, add 100ml / L coconut milk in the range of 0.2-0.5mg / L NAA and 1.0-2.0mg / L 6-BA hormone On the MS sucrose agar medium, after being cultured in a light culture room for 1 month, protocorms grow from around the mother body, the differentiation frequency is 100%, and the proliferation coefficient is more than 10 times. According to this method, subculture every one month, the annual proliferation coefficient of each bud is theoretically 1.0×10 10 above.
Embodiment 3
[0019] Embodiment 3: lateral bud differentiation and subculture thereof
[0020] Cut the above-mentioned seedlings into leafy stems of 1~2cm in length, and inoculate them on MS sucrose agar medium containing auxin 0.2-0.8mg / L NAA, cytokinin 1.0-2.5mg / L BA, and 100ml / L coconut milk Above, after one and a half months of cultivation in the light incubator, axillary buds grow from the leaf axils of the stem segment, the differentiation frequency is 100%, and the proliferation coefficient is more than 2 times. According to this method, subculture every one and a half months, the annual proliferation coefficient of each bud is theoretically 1.9×10 4 above.
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