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

Active Publication Date: 2018-01-16
SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to: overcome the problems that existing uniaxial orchid endophytes are difficult to completely remove, and then affect the production of tissue cultured seedlings, and provide a method for thoroughly removing uniaxial orchid endophytes by using rifampicin. And this method was used in the rapid propagation of vanilla tissue culture

Method used

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Examples

Experimental program
Comparison scheme
Effect test

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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PUM

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Abstract

The invention discloses a method for removing endophytic bacteria of uniaxial orchids and application thereof to rapid propagation in the tissue culture of vanilla. The method is characterized in thatthe endophytic bacteria existing in an explant material in the tissue culture process of the uniaxial orchids are removed through rifampicin treatment in combination with stem top culture, and the method is combined with the rapid propagation in the tissue culture of the vanilla. The method and the application thereof disclosed by the invention have the beneficial effects that rifampicin with specific concentration and broad spectrum sterilization ability is added into a uniaxial orchid culture medium, so that the efficient bactericidal and bacteriostatic effects of the rifampicin can be realized, the endophytic bacteria in the culture material can be completely eliminated through subculture for four times, and meanwhile, the toxic and side effects of the high-concentration rifampicin onthe uniaxial orchids are avoided; the mass propagation of the uniaxial orchids is promoted through rational combination of all components in the culture medium. The method disclosed by the invention successfully solves the bottleneck problem that the endophytic bacteria is hard to remove in the production processes of tissue culture seedlings of the vanilla and the other uniaxial orchids, lays a foundation for the rapid propagation in the tissue culture and has good application value.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture rapid propagation, more specifically, the invention relates to a method for thoroughly removing endophytic bacteria of uniaxial orchids, and the method is applied to the rapid propagation of vanilla tissue culture. Background technique [0002] Vanilla, also known as vanilla, is a perennial tropical vine spice crop, distributed in tropical regions of the world, within 27° north and south latitude, there are nearly 110 species in the world, and currently 4 species are found in my country, distributed in Guangxi and Guizhou , Yunnan, Hainan, Guangdong and Taiwan and other places in humid tropical valley forests. The commercially cultivated vanilla is primarily the Mexican vanilla (Vanilla planifolia Ames), known for producing aromatic pods. Vanilla is a precious perennial tropical vine spice plant. It is mainly used as a fragrance raw material in the food industry, and can also be used ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 吴坤林郑枫曾宋君张新华马国华简曙光
Owner SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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