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Method for reducing microbiological contamination rate by encapsulating explant by virtue of coating film containing fungicide in plant tissue culture

A technology of plant tissue culture and fungicide, which is applied in the field of plant tissue culture, can solve the problems of pollution, strong human toxicity, and great environmental hazards, and achieve the effects of expanding the contact area and time, facilitating popularization and application, and simple operation

Active Publication Date: 2015-04-22
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, first use 70% Soak in alcohol for 10-30 seconds, soak in 0.1% mercury chloride for 8-12 minutes or soak in 2% sodium hypochlorite (saturated calcium hypochlorite solution) for 15-20 minutes. Salt disinfection effect is 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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  • Method for reducing microbiological contamination rate by encapsulating explant by virtue of coating film containing fungicide in plant tissue culture
  • Method for reducing microbiological contamination rate by encapsulating explant by virtue of coating film containing fungicide in plant tissue culture

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Embodiment 1

[0011] Bougainvillea ( 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% mercury chloride for 10 minutes, rinsed with sterile water for 5 times, and the surface moisture was blotted with filter paper. The medium composition was MS+ 2.0 mg / L 6-BA+1.0 mg / 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%, add nutrients and hormones consistent with the composition of the medium, adjust the pH to 5.8 (after autoclaving to 5.6), cool down after autoclaving for use. Fungicide 3-hydroxy-5-methylisoxazole (hymexazol) mixed with hexamethylenetetramine (urotropine) mother liquor, adding autoclaved sodium alginate, nutrition and hormone mixture, Make the final concentration of hymexazol 0.1%, and the final...

Embodiment 2

[0017] Thin-leaved variegated wood ( variegated laurel ) terminal buds, washed with running water for 2 hours, 75% ethanol for 30 seconds, washed twice with sterile water, soaked with 0.1% mercuric chloride for 12 minutes, rinsed with sterile water for 5 times, the surface moisture was blotted with filter paper, and the control group was inoculated directly , the nutrients and hormones of the medium are MS+6-BA2mg / L+NAA0.1mg / L+sucrose 30g / L, and the pH is 5.8 (down to 5.6 after autoclaving). Coating encapsulated explant method, the concentration of sodium alginate is 3%, add nutrients and hormones consistent with the composition of the medium, adjust the pH to 5.8, and cool down after autoclaving. Bactericide cresolsulfonic 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 cresolsulfonic acid 0.1%, and the final conc...

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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, first use 70% Soak in alcohol for 10-30 seconds, soak in 0.1% mercury chloride for 8-12 minutes or soak in 2% sodium hypochlorite (saturated ...

Claims

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Application Information

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