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Method for detecting generation of plant immunoreaction

An immune response, plant technology, applied in material excitation analysis, fluorescence/phosphorescence, etc., can solve the problems of poor practicability, complex process, low accuracy, etc., and achieve the effect of strong promotion and application value

Inactive Publication Date: 2013-04-17
YUNNAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The present invention provides a method for detecting the immune response of plants, aiming to solve the problems of complex process, low accuracy and poor practicability of the method for detecting immune response of plants provided by the prior art

Method used

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  • Method for detecting generation of plant immunoreaction

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] 5.0 mg / ml fusion protein was used to treat rice root tissue and leaf sheath for 24 hours, and the callose was directly observed after aniline blue staining and the generation of active oxygen was observed by DAB staining.

[0021] Centrifuge the expression product of the effector protein gene of Magnaporthe grisea to collect the cells, and suspend the cells with freshly prepared buffer solution (20mM Tris-HCl, pH7.4; 200mM NaCl; ImM EDTA; 10mM β-mercaptoethanol) . The supernatant was collected by centrifugation after ultrasonography to disrupt the cells in an ice-water bath. The treatment is to purify the prokaryotic expression product of the gene and treat the root tissues and leaf sheaths of 24 rice monogenic lines with different resistance genes at a concentration of 5.0 mg / ml for 24 hours, and observe the callose after direct staining with aniline blue and DAB staining and active oxygen generation. The results are shown in the table. It was found that the 24 rice ...

Embodiment 2

[0023] 5.0 mg / ml fusion protein was used to treat rice root tissue and leaf sheath for 24 hours, and the callose was directly observed after aniline blue staining and the production of active oxygen was observed by DAB staining.

[0024] Centrifuge the expression product of the effector protein gene of Magnaporthe grisea to collect the cells, and suspend the cells with freshly prepared buffer (20mM Tris-HCl, pH7.4; 200mM NaCl; 1mM EDTA; 10mM β-mercaptoethanol) . The supernatant was collected by centrifugation after ultrasonography to disrupt the cells in an ice-water bath. The treatment is to treat the root tissue and leaf sheath of the susceptible rice variety at a concentration of 5.0 mg / ml after purification of the prokaryotic expression product of the gene for 24 hours, and observe the production of callose and active oxygen after aniline blue direct staining and DAB staining. The results are shown in the table. It was found that the susceptible rice variety Lijiang Xintu...

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Abstract

The invention discloses a method for detecting generation of plant immunoreaction. The existing purification expression through prokaryotic expression products and the callose and reactive oxygen observation method are remained the same; the prokaryotic expression products of rice blast mycoprotein genes are adopted; 24 rice single-gene varieties containing different resistance genes, root organizations and sheaths of rice are processed according to the concentration of 5.0 mg / ml; the DAB dyeing and the aniline blue dyeing are implemented after 24 hours; the forming conditions of callose and reactive oxygen are directly observed through a fluorescence microscope; and the generation of the plant immunoreaction is judged through the forming conditions of the callose and the reactive oxygen. The method uses the prokaryotic expression products of purified effect protein genes to process the rice root organizations, and observes the generation of the callose and the reactive oxygen of the root organizations through the fluorescence microscope, so that the generation condition of the rice immunoreaction can be directly obtained, and the rice blast mycoprotein capable of leading a plurality of rice single-gene varieties containing different resistance genes and susceptible varieties to generate the immunoreaction is synchronously obtained.

Description

technical field [0001] The invention belongs to the field of plant protection and biotechnology, in particular to a method for detecting immune response produced by plants. Background technique [0002] The innate immune mechanism of plants can effectively resist the invasion of pathogenic microorganisms. When a susceptible plant is infected by a specific race of pathogenic bacteria, the plant's own immune mechanism cannot be activated, eventually leading to disease. Plants usually resist the invasion of pathogenic microorganisms through two sets of defense mechanisms, the first is a general mechanism, and the second is a mechanism to resist specific races of pathogens. The general defense mechanism of plants is the immune mechanism triggered by pathogen-associated molecular patterns, namely PTI (PAMP-triggered immunity). PTI is activated by host extracellular surface receptors recognizing pathogen-associated molecular patterns, and is an immune response that occurs in the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
Inventor 杨静李成云刘林朱有勇徐畅施竹凤
Owner YUNNAN AGRICULTURAL UNIVERSITY
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