Tissue culture regeneration method of paeonia suffruticosa
A technology of tissue culture and peony, which is applied in plant regeneration, horticultural methods, botanical equipment and methods, etc., can solve the problems that peony traits are difficult to make a breakthrough and is in its infancy, and achieve the convenience of large-scale seedling planting, season and the effect of small environmental impact and fast seedling growth
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Embodiment 1
[0035] The collection and processing of embodiment 1 explant material
[0036] Choose lush and disease-free plants, collect crab-yellow fruits in July, and let them dry in the shade indoors for 7 days to collect seeds, remove the shriveled seeds and impurities, and leave plump seeds. Put the seeds at 4°C for 90 days, break the dormant state of the epicotyls of the seeds, soak them in concentrated sulfuric acid for 2-3 minutes at 25°C, wash them with running water for 3-4 times, and then disinfect them with 20% sodium hypochlorite solution for 20 days Minutes, then wash 3-4 times with sterile water under sterile conditions, and dry the water with sterile filter paper for later use.
Embodiment 2
[0037] Example 2 Induction and differentiation of callus, regeneration of somatic embryos, differentiation of cotyledon embryos and induction of adventitious roots
[0038] 1) Induction and differentiation of callus:
[0039] The sterilized peony seeds obtained in Example 1 were inoculated in MSB medium, the culture temperature was 20-25° C., the light intensity was 20000 lux, and the light was 16 hours per day. Radicles appeared in 10 days. After 14 days, cut the radicle and place it in the induction medium (30g / L sucrose, 4.44g / L MSB, 40mg / L 2,4-D, 2g / L gel, pH 7.0), and the callus will appear after 15 days of culture Tissues, after 30 days of culture, large groups of callus and somatic embryos appeared.
[0040] 2) Somatic embryo regeneration:
[0041] Remove the somatic embryos with a smooth surface and yellow-green color from the callus mass to ensure that the bottom of each somatic embryo is connected with as little tissue as possible to minimize the differentiation of...
Embodiment 3
[0046] Embodiment 3 complete plant regeneration and seedling hardening
[0047]When the differentiated rooted explant grows to a plant body with a height of about 6 cm, the seedling is exposed to the air for hardening for 5-7 days. Then take it out, rinse the medium of the root, transfer it to a flowerpot containing moist vermiculite, pour nitrogen fertilizer once, and add water to moisturize every day. The control culture conditions are temperature 20-25°C, light intensity 20000 lux, and 16 hours of light per day. One week later, the plants were moved into the greenhouse field for cultivation. The cultivation conditions were: temperature 20-25°C, air humidity 85%, spraying a broad-spectrum fungicide every 7 days, and the survival rate reached 95% after hardening for 1 month. 10 pieces, 10cm high.
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