Tissue culture propagation method of Osmanthus chinensis induced by young sporangia group through ggb pathway
A technology for sporangia and E. japonica, which is applied in the field of tissue culture and reproduction of E. chinensis in South China, can solve the problems of difficulty in obtaining explants and difficulty in induction, and achieve the effects of high induction rate, short induction time, and high environmental requirements.
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Embodiment 1
[0029] (1) Selection of explants and surface disinfection: Using the high-quality Osmanthus chinensis seedlings successfully introduced and cultivated in the school base as materials, on April 20, 2018, healthy incomplete sporangia with young sporangia were cut. Unfold the leaves, gently remove surface dirt with a soft brush, rinse with running water, soak in 75% ethanol for 20 s, then wash with 0.1% mercuric chloride for 4 min, rinse with sterile water 7 times, and obtain Osmanthus chinensis without water. Sporangia explants of young leaves of fungi;
[0030] (2) Induction of green spheroids: The aseptic young leaf sporangia explants of Osmanthus chinensis obtained in step (1) were excised end to end, cut into 2 cm segments, and carefully inoculated into GGB induction medium according to the direction of polarity. The medium is B5 basic medium, with 30 g / L sucrose and 7 g / L agar added, and the pH is 5.8. The number of inoculations is 30 bottles, with 1 explant in each bottle...
Embodiment 2
[0037] (1) Selection of explants and surface disinfection: On June 7, 2018, the high-quality Osmanthus chinensis seedlings successfully introduced and cultivated in the school base were used as materials, and healthy incomplete sporangia with young sporangia were cut. Spread the leaves, gently remove surface dirt with a soft brush, rinse with running water, soak in 75% ethanol for 20 s, then shake and wash with 0.1% mercuric chloride for 3 min, rinse with sterile water 5-7 times, get Huanan Osmanthus aseptic young leaf sporangia explants;
[0038] (2) Induction of green spheroids: The aseptic young leaf sporangia explants of Osmanthus chinensis obtained in step (1) were excised end to end, cut into 2 cm segments, and carefully inoculated into GGB induction medium according to the direction of polarity. The composition of the medium is B5+6-BA1.0 mg / L+NAA0.1 mg / L, 30 g / L sucrose and 7 g / L agar are added, and the pH is 5.8. The number of inoculations and culture conditions are ...
Embodiment 3
[0045] (1) Selection of explants and surface disinfection: On July 4, 2018, the high-quality Osmanthus chinensis seedlings successfully introduced and cultivated in the school base were used as materials, and healthy incomplete explants with young sporangia were cut. Spread the leaves, gently remove surface dirt with a soft brush, rinse with running water, soak in 75% ethanol for 20 s, then wash with 0.1% mercuric chloride for 2 min, rinse with sterile water 5-7 times, get Huanan Osmanthus aseptic young leaf sporangia explants;
[0046] (2) Induction of green spheroids: the inoculation method, culture medium, inoculation quantity and culture conditions are the same as in Example 2, and will not be repeated here.
[0047] (3) Proliferation of green spheroids: On July 10, 2018, it was observed that there were many green spores scattered from the sporangia on the surface of the medium. On July 22, it was observed that the spores gradually expanded to form green spheroids of Osman...
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