A method for removing endogenous bacteria in explants during plant tissue culture

A tissue culture and explant technology, applied in plant tissue culture and biological fields, can solve the problem of inability to remove bacteria in explants such as poplar, and achieve the effect of maintaining vitality and sufficient vitality

Inactive Publication Date: 2018-11-06
INST OF FORESTRY CHINESE ACAD OF FORESTRY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The present invention aims at the above defects existing in the prior art, and provides a new method for removing endogenous bacteria in explants during plant tissue culture in order to improve the deficiency that conventional disinfection and sterilization methods cannot remove endogenous bacteria in explants such as poplar , this method can effectively remove bacteria and fungi on the surface and inside of poplar and other explants, so that the explants can maintain sufficient vitality

Method used

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  • A method for removing endogenous bacteria in explants during plant tissue culture
  • A method for removing endogenous bacteria in explants during plant tissue culture
  • A method for removing endogenous bacteria in explants during plant tissue culture

Examples

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Embodiment 1

[0028] Example 1: Cultivation of explants of Populus americana

[0029] 1. Disinfection of cutting soil

[0030] Mix turf soil, perlite and river sand in a ratio of 6:2:2 as cutting soil, and apply 50% carbendazim (30-40g / m 3 ), mix well, put it into a plastic bag, airtight for 5 days, open it, and dry it in the sun for 2-3 days, then put it into a culture bowl with a diameter of 6 cm.

[0031] 2. Disinfection of cuttings

[0032] Cut the hibernating Populus nigra fringe into 15cm length, rinse with tap water, and remove surface dirt with a soft brush. Afterwards, all the cuttings were immersed in the antibiotic disinfection mixture (200 mg / L amphotericin B and 100 mg / L streptomycin sulfate sterile aqueous solution) and treated for 24 hours. Use sterile filter paper to absorb moisture on the surface of the cuttings, and immerse 0.5 cm of the top of the cuttings in melted paraffin, and then seal the top with wax. The processed cuttings are placed in the nutrient bowl and placed in a ...

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Abstract

The invention belongs to the technical field of organisms and discloses a method for removing explant endophyte during plant tissue culturing. The cuttage soil can be disinfected by 50% of carbendazim, cuttage twigs are disinfected by sterile water solution containing 200 mg / L of amphotericin and 100 mg / L of streptomycin sulfate, and new semi-lignified twigs can be treated in advance by sterile water solution containing 100 mg / L of amphotericin and 50 mg / L of streptomycin sulfate. Accordingly, bacterial and fungi on the surface of and inside the explant such as aspen can be effectively removed, and sufficient vigor of the explant can be kept.

Description

Technical field [0001] The invention relates to the field of biotechnology, in particular to the technical field of plant tissue culture, and specifically relates to a method for removing internal bacteria from explants during plant tissue culture. Background technique [0002] Poplars belong to the Populus L. plants of the Salicacae family and are perennial woody trees. Poplar is not only an important afforestation tree species and industrial timber species, but also a model tree species for genetic engineering of perennial trees. Therefore, obtaining a large number of sterile explants is the prerequisite and basis for the rapid propagation and genetic engineering of poplar tissues. [0003] In conventional plant tissue culture, the method of disinfection of explants is generally to rinse with tap water to remove the surface soil, disinfect with alcohol for 30 seconds to 1 minute, and then disinfect with 10% sodium hypochlorite or 0.1% raw mercury for a few minutes to more than t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00A01G2/10A61L2/18A61L101/44A61L101/34A61L101/06
CPCA01G9/0299A01H4/005A61L2/18
Inventor 张冰玉苏晓华张伟溪高亚楠
Owner INST OF FORESTRY CHINESE ACAD OF FORESTRY
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