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A method for alleviating contamination of plant tissue culture explants

A plant tissue culture and explant technology, applied in the field of plant tissue culture, can solve the problems of pollution, strong human toxicity, and great environmental hazards, and achieve the effect of expanding the contact area and time, causing less damage to plant tissues, and facilitating popularization and application

Active Publication Date: 2017-02-22
SUBTROPICAL CROPS INST OF FUJIAN PROVINCE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The microorganisms carried by explants include surface miscellaneous bacteria and endophytic bacteria. Surface miscellaneous bacteria are mainly fungi and bacteria, which are easier to remove by soaking for a short time. The preferred explant disinfection method is the soaking method. Soak in alcohol for 10-30s, soak in 0.1% mercuric chloride for 8-12min or soak in 2% sodium hypochlorite (saturated calcium hypochlorite solution) for 15-20min. Mercury chloride is extremely toxic to humans and harmful to the environment. Salt disinfection is also not satisfactory
Endophytes exist inside the tissue, mainly bacteria, liquid disinfectants are not easy to penetrate and kill in a short time, and endophytes have a long incubation period (more than 5 days), causing large-scale contamination during the culture period. Although antibiotics are added to the culture medium, but Antibiotics have a narrow range of sterilization, and microorganisms are prone to drug resistance
In short, there is still no ideal method for explant sterilization, and the prevention and control of explant contamination is the main difficulty in the success of tissue culture.

Method used

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  • A method for alleviating contamination of plant tissue culture explants
  • A method for alleviating contamination of plant tissue culture explants
  • A method for alleviating contamination of plant tissue culture explants

Examples

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Embodiment 1

[0011] Bougainvillea spectabilis leaves were rinsed with running water for 30 minutes, immersed in 75% ethanol for 30 seconds, washed twice with sterile water, soaked in 0.1% mercuric chloride for 10 minutes, rinsed with sterile water for 5 times, and dried on filter paper. Medium composition MS+2.0mg / L 6-BA+1.0mg / L NAA+30g / L sucrose, pH to 5.8 (down to 5.6 after autoclaving), the leaves were cut into small pieces, and the control group was directly inoculated. Coated explant method, the concentration of sodium alginate is 4%, the nutrients and hormones consistent with the composition of the medium are added, the pH is adjusted to 5.8 (after autoclaving to 5.6), and it is cooled after autoclaving for use. Fungicide 3-hydroxy-5-methylisoxazole (hymexazol) mixed with hexamethylenetetramine (urotropine) mother liquor, adding autoclaved sodium alginate mixed with nutrients and hormones, Make the final concentration of hymexazol 0.1% and the final concentration of urotropine 0.2%, ...

Embodiment 2

[0019] For the terminal buds of Variegated laurel, rinse with running water for 2 hours, wash with 75% ethanol for 30 seconds, wash with sterile water twice, soak with 0.1% mercury chloride for 12 minutes, rinse with sterile water for 5 times, and blot dry with filter paper Surface moisture, the control group was inoculated directly, the nutrition and hormones of the culture medium were MS+6-BA2mg / L+NAA0.1mg / L+sucrose 30g / L, pH to 5.8 (down to 5.6 after autoclaving). The explant method of coating film encapsulation, the concentration of sodium alginate is 3%, and the nutrients and hormones consistent with the composition of the culture medium are added, the pH is adjusted to 5.8, and it is cooled after autoclaving for use. Bactericide cresol sulfonic acid and sodium bisulfite mother liquor were filtered and sterilized respectively, and the mixture of high-pressure sterilized sodium alginate, nutrients and hormones was added to make the final concentration of cresol sulfonic aci...

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Abstract

The invention discloses a method for reducing the microbiological contamination rate by encapsulating an explant by virtue of a coating film containing a fungicide in plant tissue culture, and belongs to plant tissue culture technologies in the fields of agriculture and forestry. The method comprises the following steps: encapsulating a tissue culture explant by using the coating film which has good gas permeability, water permeability and a light transmission performance and has small damages to plant tissues; and selecting a sodium alginate solution as a film-forming agent, adding the fungicide, nutritional components and plant hormones, then encapsulating the explant, feeding a calcium chloride solution for performing crosslinking and solidification, and performing inoculation and culturing. According to a tissue culture explant encapsulated by the coating film containing the fungicide, disclosed by the invention, the contact area between the fungicide and the explant can be expanded, the action time of the fungicide can be prolonged, and surface contamination bacteria and endophytic bacteria of the explant can be effectively killed. The method disclosed by the invention is simple and convenient to operate, has significant effects on reducing the contamination rate of the explant, is low in cost, and is convenient to promote and apply.

Description

technical field [0001] The invention belongs to the plant tissue culture technology in the field of agriculture and forestry, and in particular relates to a method for reducing the pollution rate of plant tissue culture explant culture. Background technique [0002] Plant tissue culture technology has broad market prospects for seedling production, virus removal, and germplasm improvement. At present, in the technical link of plant tissue culture, the prevention and control of explant contamination is the main difficulty in the success of tissue culture. The microorganisms carried by explants include surface miscellaneous bacteria and endophytic bacteria. Surface miscellaneous bacteria are mainly fungi and bacteria, which are easier to remove by soaking for a short time. The preferred explant disinfection method is the soaking method. Soak in alcohol for 10-30s, soak in 0.1% mercuric chloride for 8-12min or soak in 2% sodium hypochlorite (saturated calcium hypochlorite solu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 刘福平
Owner SUBTROPICAL CROPS INST OF FUJIAN PROVINCE