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Surface sterilizing method of explant

A surface sterilization and explant technology, applied in the field of plant tissue culture, can solve the problems of pollution, tissue culture operation failure, difficult to kill microorganisms, etc., and achieve the effect of improving the sterilization effect

Inactive Publication Date: 2009-06-24
LINGBAO HUABAO IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In this way, even if the follow-up soaking and disinfection work is done well, it is difficult to kill the microorganisms entering the interior, which will eventually lead to contamination and tissue culture operation failure

Method used

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  • Surface sterilizing method of explant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] Take jujube stems, scrub the surface dirt with 75% alcohol solution, soak the stems in 70% alcohol solution for 15 minutes, and then soak them in disinfectant containing 0.1% mercury chloride and 0.5% Tween 5min. During the soaking and disinfection process, gently stir with a glass rod from time to time to promote full contact between all parts of the material and the disinfection solution, drive out air bubbles, and make the disinfection thorough. Pour out the disinfectant solution 1-2 minutes before the time is up, and immediately pour sterile water into it and shake it gently. Rinse with sterile water for about 3 minutes each time, 8-10 times. The entire sterilization process is performed under sterile conditions.

Embodiment 2

[0013] Pick Phalaenopsis peduncles and cut off unnecessary parts, then scrub with 70% alcohol solution to remove surface dirt. After cleaning, soak the pedicel in 75% alcohol solution for 1 min, and then soak it in a disinfection solution with a sodium hypochlorite concentration of 6% and a Tween concentration of 0.1% for 10 min. During the soaking process, use a glass rod to gently stir from time to time to promote full contact between all parts of the material and the disinfection solution, to drive out air bubbles, and to make the disinfection thorough. 1-2 minutes before the time is up, after pouring out the sodium hypochlorite solution, immediately pour in sterile water and shake gently. Rinse with sterile water for about 3 minutes each time, 3-5 times. The entire sterilization process is performed under sterile conditions.

Embodiment 3

[0015] Take the stem tip of the cherry blossoms, wash off the dirt on the surface with 73% alcohol solution, and then put it into a disinfection solution with a sodium hypochlorite concentration of 4% and a Tween concentration of 0.3% for 15 minutes. During the soaking process, use a glass rod to gently stir from time to time to promote full contact between all parts of the material and the disinfection solution, to drive out air bubbles, and to make the disinfection thorough. Pour out the sodium hypochlorite disinfectant 1-2 minutes before the time is up, and immediately pour sterile water into it and shake it gently. Rinse with sterile water for about 3 minutes each time, 3-5 times. The entire sterilization process is performed under sterile conditions.

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Abstract

An exophytic surface sterilization method belongs to the technical field of plant tissue culture, comprising three steps of cleaning explant, soaking sterilization of disinfector solution, aseptic water flushing, characterized by removing an unused part of the explant, directly cleaning using 70-75% alcohol solution rather than tap water, putting into the disinfector solution for explant sterilization after cleaning. In the inventive technical scheme, cleaning and sterilization are developed at the same time, thereby avoiding animalcule flushed in the tap water or the explant enter the inside of vegetable trachea along capillary of the explant, greatly reducing pollution ratio of plant tissue culture.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a method for surface sterilization of explants. Background technique [0002] Whether the explants used for plant tissue culture are taken from the field or indoors, they usually have microorganisms on their surface. Once these pollution sources are brought into the medium, they will cause medium contamination, which is a major obstacle to sterile tissue culture. Therefore, the explants must be subjected to strict surface sterilization, and then inoculated on the culture medium through aseptic procedures. According to the traditional surface sterilization treatment technology, after the unused parts of the explants are removed, the following steps need to be carried out: (1) rinse under tap water for a long time, and scrub with an appropriate brush if necessary, Hard materials can be scraped with a knife, and then rinsed under tap water for a few minute...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 胡士敬赵莉王芳许朝斌王肖东邢红梅郭勇任兵茹赵烈平谷磊
Owner LINGBAO HUABAO IND
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