Method for removing endophytic bacteria of uniaxial orchids and application thereof to rapid propagation in tissue culture of vanilla
A technology of tissue culture and endophytes, applied in application, horticultural methods, botanical equipment and methods, etc., can solve the problems affecting the production of tissue culture seedlings and the difficulty of completely removing endophytes, achieve good application value, and avoid toxicity Side effects, high-efficiency bactericidal effect
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Embodiment 1
[0027] 1. Materials: Wild Vietnamese vanilla (V. annamica) was collected from Sanya, Hainan Province and planted in the Orchid Germplasm Resource Nursery of South China Botanical Garden, Chinese Academy of Sciences.
[0028] 2. Preparation of sterile rifampicin solution
[0029] To buy medical rifampicin powder, choose a 250ml drop bottle with a straw, pack it with newspaper after drying, and put it at 1.06kg / cm 2(121° C.) and sterilized for 20 minutes, and another 500 ml of methanol was used to prepare rifampin into a solution with a concentration of 25-50 mg / L and put it into an orchid vase. Place the above medicines, utensils and disposable bacterial filter on the ultra-clean workbench, filter the rifampicin solution into a sterile drop bottle under aseptic conditions, and prepare a sterile solution with a concentration of 25-50mg / L For rifampicin solution, seal the neck of the drop bottle with sterile kraft paper and store it in a refrigerator at 4°C until use.
[0030] ...
Embodiment 2
[0044] 1. Materials: Wild vanilla (V.siamensis) was collected from Sanya, Hainan Province and planted in the Orchid Germplasm Resource Nursery of South China Botanical Garden, Chinese Academy of Sciences.
[0045] 2. Preparation of sterile rifampicin solution
[0046] To buy medical rifampicin powder, choose a 250ml drop bottle with a straw, pack it with newspaper after drying, and put it at 1.06kg / cm 2 (121° C.) and sterilized for 20 minutes, and another 500 ml of methanol was used to prepare rifampin into a solution with a concentration of 25-50 mg / L and put it into an orchid vase. Place the above medicines, utensils and disposable bacterial filter on the ultra-clean workbench, filter the rifampicin solution into a sterile drop bottle under aseptic conditions, and prepare a sterile solution with a concentration of 25-50mg / ml For rifampicin solution, seal the neck of the drop bottle with sterile kraft paper and store it in a refrigerator at 4°C until use.
[0047] 3. Cultur...
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