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Separation method for plant endophyte

A plant endophyte and separation method technology, which is applied in the field of separation of plant endophytes, can solve the problems of incomplete disinfection of plant surfaces, affecting the separation effect of endophytes, and introducing miscellaneous bacteria, so as to reduce damage and reduce damage The effect of increasing the probability of survival

Inactive Publication Date: 2014-12-31
INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, if the existing endophyte isolation methods reduce or do not use disinfectants, it will lead to incomplete disinfection of the plant surface, and it is easy to introduce miscellaneous bacteria during the endophyte isolation, which will affect the isolation effect of endophytes.

Method used

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  • Separation method for plant endophyte
  • Separation method for plant endophyte
  • Separation method for plant endophyte

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] A method for isolating plant endophytes, comprising the following steps:

[0022] (1) Plant tissue pretreatment

[0023] Rinse the plant tissue with tap water and then wash it with sterile water for 2-3 times, dry the surface water, cut it into 0.5cm pieces, and then put the cut material in 70% ethanol for 3-5min. , washed 2-3 times with sterile water, then soaked in 0.1% mercury liter for 1-3 minutes or soaked in 4% sodium hypochlorite for 5-10 minutes, then rinsed with sterile water for 3-5 times, and dried the surface moisture reserve;

[0024] (2) Plant tissue culture

[0025] Place the sterilized plant tissue on MS medium and carry out sterile culture for 1-2 weeks until the plant tissue resumes growth; the culture conditions are temperature 15°C, humidity 20%, light intensity 1000Lux, light time 8-10h / d (3) Isolation of endophytes

[0026] The plant tissue after tissue culture is crushed under aseptic conditions, and then ground in a sterilized mortar, the exu...

Embodiment 2

[0028] A method for isolating plant endophytes, comprising the following steps:

[0029] (1) Plant tissue pretreatment

[0030] Rinse the plant tissue with tap water and then wash it with sterile water for 2-3 times, dry the surface water, cut it into 1.3cm pieces, and then put the cut material in 73% ethanol for 3-5min. , washed 2-3 times with sterile water, then soaked in 0.2% mercury liter for 1-3 minutes or soaked in 5% sodium hypochlorite for 5-10 minutes, then rinsed with sterile water for 3-5 times, and dried the surface moisture reserve;

[0031] (2) Plant tissue culture

[0032] Place the sterilized plant tissue on MS medium, and carry out sterile culture for 1-2 weeks until the plant tissue resumes growth; the culture conditions are temperature 25°C, humidity 50%, light intensity 3000Lux, light time 10-12h / d ;

[0033] (3) Isolation of endophytes

[0034] The plant tissue after tissue culture was crushed under aseptic conditions, and then ground in a sterilized ...

Embodiment 3

[0036] A method for isolating plant endophytes, comprising the following steps:

[0037] (1) Plant tissue pretreatment

[0038] Rinse the plant tissue with tap water and then wash it with sterile water for 2-3 times, dry the surface water, cut it into 2.0cm pieces, and then put the cut material in 75% ethanol for 3-5min. , washed 2-3 times with sterile water, then soaked in 0.3% mercury liter for 1-3 minutes or soaked in 6% sodium hypochlorite for 5-10 minutes, then rinsed with sterile water for 3-5 times, and dried the surface moisture reserve;

[0039] (2) Plant tissue culture

[0040] Place the sterilized plant tissue on MS medium and carry out sterile culture for 1-2 weeks until the plant tissue resumes growth; the culture conditions are temperature 30°C, humidity 70%, light intensity 6000Lux, light time 12-14h / d ;

[0041] (3) Isolation of endophytes

[0042] The plant tissue after tissue culture is crushed under aseptic conditions, and then ground in a sterilized mo...

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Abstract

The invention relates to the technical field of microbes, and discloses a separation method for plant endophyte. The separation method comprises the following steps: (1) pretreating plant tissue; (2) culturing plant tissue; and (3) separating endophyte. The disclosed separation method for plant endophyte is capable of effectively removing surface bacteria, fungi, actinomyces and other microbes, and avoiding endophyte separation effect caused by mixing of mixed bacteria. Endophyte in the plant tissue recovers growth after tissue culture, and disinfection processing does not need performing in operation of separating endophyte, so that the damage effect of a disinfectant on plant tissue and endophyte is substantially reduced, the survival probability of endophyte is increase, and the number of endophyte obtained through separation is obviously improved.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, in particular to a method for isolating plant endophytes. Background technique [0002] At present, most methods for isolating plant endophytes are to directly sterilize plant tissues and then crush them by physical and chemical means, then extract the exudate, and use the liquid as a source of bacteria to cultivate and isolate endophytes. The key to the isolation of endophytes is to eliminate the contamination of microorganisms on the surface of plants. In order to ensure that the isolated microorganisms are plant endophytes, very strict disinfection measures are often adopted. At present, most of the disinfection methods used in the isolation of plant endophytes use plant surface disinfection to remove non-endophyte contamination. For example, the Chinese invention patent CN103122331A discloses a method for isolating endophytic bacteria in plants, including steps such as sampling, surfa...

Claims

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Application Information

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IPC IPC(8): C12N1/00
Inventor 冯仁军卢利方云天艳张银东
Owner INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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