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Method for detecting rice blast pathogenic protein

A technology of rice blast fungus and protein, which is applied in the field of detection of blast fungus pathogenic protein, can solve the problem of inability to quantitatively detect the pathogenicity of blast fungus pathogenic protein, and achieve the effect of effective and simple distinction

Inactive Publication Date: 2011-04-20
YUNNAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The invention overcomes the deficiency of the inability to quantitatively detect the pathogenicity of the rice blast fungus pathogenic protein, and provides a method for detecting the pathogenicity of the rice blast fungus pathogenic protein on rice leaves

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] The susceptible rice variety Lijiang Xintuan Heigu was inoculated when the fourth leaf was fully developed. Select the widest part of the middle part of the inoculated rice leaf, and use a wounding device to form a pressure wound spot with a diameter of 1 mm. After culturing in a complete medium, transfer it to a medium that does not contain a nitrogen source. After nitrogen starvation culture The weakly pathogenic Magnaporthe oryzae protein was obtained and inoculated at the wound site, and inoculated with deionized water as a control. The inoculation concentration of protein was 1 μg / μl, and the inoculation volume was 3 μl. The lesion formed on the rice leaf 72 hours after inoculation was oval, and the longest value extending toward the leaf tip and leaf base was measured as the length of the lesion. According to the classification criteria of lesion length between 1.5mm and 2mm as weak pathogenicity; lesion length between 2.1mm and 3mm as moderate pathogenicity; les...

Embodiment 2

[0011] The susceptible rice variety Lijiang Xintuan Heigu was inoculated when the fourth leaf was fully developed. Select the widest part of the middle part of the inoculated rice leaf, and use a wounding device to form a pressure wound spot with a diameter of 1 mm. After culturing in a complete medium, transfer it to a medium that does not contain a nitrogen source. After nitrogen starvation culture The obtained medium-pathogenic rice blast fungus protein was inoculated on the wounded site, and deionized water was used as a control. The inoculation concentration of protein was 1 μg / μl, and the inoculation volume was 3 μl. The lesion formed on the rice leaf 72 hours after inoculation was oval, and the longest value extending toward the leaf tip and leaf base was measured as the length of the lesion. According to the classification criteria of lesion length between 1.5mm and 2mm as weak pathogenicity; lesion length between 2.1mm and 3mm as moderate pathogenicity; lesion length...

Embodiment 3

[0013] The susceptible rice variety Lijiang Xintuan Heigu was inoculated when the fourth leaf was fully developed. Select the widest part of the middle part of the inoculated rice leaf, and use a wounding device to form a pressure wound spot with a diameter of 1 mm. After culturing in a complete medium, transfer it to a medium that does not contain a nitrogen source. After nitrogen starvation culture The obtained strong pathogenic rice blast fungus protein was inoculated on the wound site, and deionized water was used as a control. The inoculation concentration of protein was 1 μg / μl, and the inoculation volume was 3 μl. The lesion formed on the rice leaf 72 hours after inoculation was oval, and the longest value extending toward the leaf tip and leaf base was measured as the length of the lesion. According to the classification criteria of lesion length between 1.5mm and 2mm as weak pathogenicity; lesion length between 2.1mm and 3mm as moderate pathogenicity; lesion length g...

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Abstract

The invention provides a method for detecting pathogenic proteins of rice blast fungi, wherein a susceptible rice cultivar LTH is inoculated when a fourth leaf is completely unfolded; a compression injury patch with a diameter of 1 millimeter is formed on a widest position in the middle of the inoculated rice leaf by an injury device; proteins obtained after cultivation of a rice blast fungus - nitrogen hunger or recombinant expression of pathogenic genes are inoculated on the injury position, wherein the inoculation concentration of the proteins is 1 microgram per microliter, and the inoculation volume of the proteins is 3 microliters; and a scab formed on the rice leaf in 72 hours after inoculation is elliptical, and the longest value extended to the direction of the leaf apex and the leaf base is measured to be the length of the scab and is the basis for comparing the degree of pathogenicity of different proteins. The classification criteria of the proteins is as follows: the proteins have weak pathogenicity when the length of the scab is between 1.5 and 2 millimeters, have intermediate pathogenicity when the length of the scab is between 2.1 and 3 millimeters, and have strong pathogenicity when the length of the scab is more than 3.1 millimeters. The method has significance in research of the functions of pathogenic genes and the pathogenic proteins of the rice blast fungi.

Description

Technical field: [0001] The invention relates to a method for detecting the pathogenic protein of rice blast fungus, which belongs to the field of plant protection. Background technique: [0002] The rice blast caused by Magnaporthe grisea is one of the most serious diseases that harm rice production in the world. In the study of the pathogenicity of rice blast fungus to rice, usually the spores of different physiological races of blast fungus are used to inoculate the leaves of different strains of rice and observe the symptoms to determine whether the result of the interaction between the two is pathogenic or not. disease resistance. The size of lesions produced by spores inoculated on rice leaves has become the main method for identifying the difference in pathogenicity of different strains of Magnaporthe grisea. Recent studies have shown that the molecular interaction between pathogenic bacteria-produced proteins and host resistance gene protein products is the core of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/02
Inventor 苏源李成云朱有勇杨静刘林王云月孔垂思常明蔡永占
Owner YUNNAN AGRICULTURAL UNIVERSITY
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