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Method for removing explant endophyte during plant tissue culturing

A plant tissue culture and explant technology is applied in the field of removing explant endophytes during plant tissue culture, which can solve the problem of inability to remove poplar and other explant endophytes, and achieve the effect of maintaining vitality and sufficient vitality.

Inactive Publication Date: 2016-11-16
INST OF FORESTRY CHINESE ACAD OF FORESTRY
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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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  • Method for removing explant endophyte during plant tissue culturing
  • Method for removing explant endophyte during plant tissue culturing
  • Method for removing explant endophyte during plant tissue culturing

Examples

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Embodiment 1

[0028] Embodiment 1: the cultivation of black poplar explant

[0029] 1. Disinfection treatment of cutting soil

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

[0031] 2. Disinfection treatment of cutting branches

[0032] Cut the hibernating black poplar fringes into 15cm lengths, rinse them with tap water, and remove the dirt on the surface with a soft brush. Afterwards, all the spikes were immersed in the antibiotic disinfection mixture (200 mg / L amphotericin B and 100 mg / L streptomycin sulfate sterile aqueous solution) for 24 hours. Use sterile filter paper to absorb the moisture on the surface of the spikes, and immerse the top 0.5 cm of the spikes in melted paraffin for top wax sealing. The treated cuttings are inserted int...

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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, in particular to a method for removing endogenous bacteria in explants during plant tissue culture. Background technique [0002] Poplar belongs to the Salicacae family Populus L. and is a perennial woody tree. Poplar is not only an important tree species for afforestation and industrial timber, but also a model tree species for perennial forest genetic engineering. Therefore, obtaining a large number of sterile explants is the premise and basis for poplar tissue rapid propagation and genetic engineering research. [0003] In conventional plant tissue culture, the disinfection method of explants is generally to wash with tap water to remove 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 a dozen minutes, rinse with sterile water and ...

Claims

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Application Information

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IPC IPC(8): A01H4/00A01G9/10A61L2/18A61L101/44A61L101/34A61L101/06
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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