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Guava leaf endophyte separation method

A separation method, the technology of guava leaves, applied in the field of separation of endophytes, can solve the problems of not being suitable for the endophytes of guava leaves, etc., achieve the effect and efficiency of wall breaking, promote leaching, and facilitate operation Effect

Inactive Publication Date: 2018-12-07
THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The isolation method of endophytes announced in the prior art is not suitable for the isolation of endophytes from guava leaves. Research and development of a method for efficiently isolating endophytes from guava leaves can significantly expand the application of guava range, with broad market value

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0029] (1) Collection and low-temperature preservation of guava leaves: in summer, collect fresh and healthy old guava leaves with a tree age of more than 10 years, and place the picked guava leaves at 4°C for 24 hours;

[0030] (2) Pretreatment and surface disinfection of guava leaves: get the guava leaves collected, rinse them with tap water, and gently brush away impurities such as soil on the surface of the leaves with a hairbrush, soak the leaves in sterile water, and add 2 - 3 drops of detergent, stir for 30 minutes with a magnetic stirring device, remove the damaged leaves, rinse 5 times with sterile water, remove the residual detergent, then soak the leaves in 0.2% chlorine dioxide solution for 3 minutes, use sterile Rinse 5 times with sterile water; put the guava leaves in a sealed glass container, immerse them in sterile water, use an ultrasonic sterilizer at 10KHz, 80W, sterilize ...

Embodiment 2

[0035] A method for isolating guava leaf endophytes, comprising the following steps:

[0036] (1) Collection and low-temperature preservation of guava leaves: in summer, collect fresh and healthy old guava leaves with a tree age of more than 10 years, and place the picked guava leaves at 4°C for 24 hours;

[0037] (2) Pretreatment and surface disinfection of guava leaves: get the guava leaves collected, rinse them with tap water, and gently brush away impurities such as soil on the surface of the leaves with a hairbrush, soak the leaves in sterile water, and add 2 - 3 drops of detergent, stir for 30 minutes with a magnetic stirring device, remove the damaged leaves, rinse 5 times with sterile water, remove the residual detergent, and then soak the leaves in 0.3% chlorine dioxide solution for 1 min, use sterile Rinse 5 times with sterile water; put the guava leaves in a sealed glass container, immerse them in sterile water, use an ultrasonic sterilizer at 10KHz, 80W, sterilize fo...

Embodiment 3

[0042] A method for isolating guava leaf endophytes, comprising the following steps:

[0043] (1) Collection and low-temperature preservation of guava leaves: in summer, collect fresh and healthy old guava leaves with a tree age of more than 10 years, and place the picked guava leaves at 4°C for 24 hours;

[0044] (2) Pretreatment and surface disinfection of guava leaves: get the guava leaves collected, rinse them with tap water, and gently brush away impurities such as soil on the surface of the leaves with a hairbrush, soak the leaves in sterile water, and add 2 - 3 drops of detergent, stir for 30 minutes with a magnetic stirring device, remove the damaged leaves, rinse 5 times with sterile water, remove the residual detergent, and then soak the leaves in 0.2% chlorine dioxide solution for 1 min, use sterile Rinse 5 times with sterile water; put the guava leaves in a sealed glass container, immerse them in sterile water, use an ultrasonic sterilizer at 10KHz, 80W, sterilize ...

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PUM

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Abstract

The invention discloses a guava leaf endophyte separation method, which comprises the following steps: (1) acquiring fresh old guava leaves of more than 10 years in summer, and storing at low temperature; (2) performing surface sterilization treatment on old guava leaves to obtain standby leaves; (3) shearing the standby leaves obtained in the step (2), adding normal saline and cellulase, and thengrinding into homogenate, diluting the homogenate and then coating onto a culture plate for culturing; and (4) after colonies grow on the culture plate, selecting single colony, and performing streaking and purification, to obtain guava leaf endophyte. The method is especially suitable for separating the endophyte from guava leaves, and is simple in technological steps and convenient to operate.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for isolating endophytes. Background technique [0002] Endophytes refer to microorganisms that live in living plant tissues at all or part of their life cycle without causing obvious plant disease symptoms. The "endosymbiotic theory" believes that endophytes co-evolving with the host may obtain the transfer of related genes from the host, and have the ability to synthesize the same or similar metabolites as the host. In 1993, for the first time, an endophytic fungus capable of producing the anticancer substance paclitaxel was isolated from Taxus brevifolia, which further confirmed the "endosymbiotic theory". Secondary metabolites produced by plant endophytes are an important source of natural medicinal active substances, and a large part of natural products are new bioactive substances, which have broad application prospects. Due to the fast propagation speed of plant end...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/02C12N1/20C12N1/14
CPCC12N1/02C12N1/14C12N1/20
Inventor 谢志国吴学玲罗奎曾伟民周智广胡芳周翔宇李交昆申丽余润兰刘元东刘阿娟邱冠周
Owner THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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