Method for measuring pathogenicity of pear fire blight bacteria by means of in-vitro water culture approach for fragrant pear branches

A pear fire blight and pathogenicity technology, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, and microbial determination/inspection, etc. Reliable, easy to operate, stable in pathogenicity

Pending Publication Date: 2019-07-09
XINJIANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method can only qualitatively identify the pathogenicity of E. amylovora, but does not propose to classify the pathogenicity
I

Method used

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  • Method for measuring pathogenicity of pear fire blight bacteria by means of in-vitro water culture approach for fragrant pear branches
  • Method for measuring pathogenicity of pear fire blight bacteria by means of in-vitro water culture approach for fragrant pear branches
  • Method for measuring pathogenicity of pear fire blight bacteria by means of in-vitro water culture approach for fragrant pear branches

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 The impact of different inoculation methods on isolated branches on the pathogenicity evaluation of pathogenic bacteria

[0026] 1. Preparation of E. amylovora inoculum: the E. amylovora strain E.a 0001, E.a 0019 and E.a 0087 was inoculated on NA+5% sucrose medium, activated at 28.5°C for 48 hours, picked a single colony and inoculated in NA+5% sucrose medium, cultured with shaking at 28.5°C and 160 rpm / min for 12 hours, and measured OD 600 The bacterial solution with a value of about 1.2 was diluted with sterile water to a concentration of 10 8CFU / mL was used as the inoculum.

[0027] 2. Preparation of isolated branches: Select the newborn young branches of the Korla fragrant pear, which is extremely susceptible to pear fire blight, as the test material, cut the test branches up to the stem section of about 30 cm, disinfect it with 75% alcohol, and then 1 / 3 into a conical flask filled with 0.5‰NaCl sterilized saline.

[0028] 3. Inoculation: In this...

Embodiment 2

[0040] Example 2 Determination of pathogenicity of E. amylovora pathogenicity and identification of host selection

[0041] 1. Preparation of E. amylovora inoculum: the E. amylovora strain E.a 0001, E.a 0017、 E.a 0055 were respectively inoculated on NA + 5% sucrose medium, activated at 28.5°C for 48 hours, picked a single colony and inoculated in NA + 5% sucrose medium, cultured with shaking at 28.5°C and 160 rpm / min for 12 hours, and measured OD 600 The bacterial solution with a value of about 1.2 was diluted with sterile water to a concentration of 10 8 CFU / ml was used as the inoculum.

[0042] 2. Preparation of isolated branches: choose pear fire blight susceptible varieties Xiangli, Dangshan pear, black sour pear, Du pear, Xinli No.7, yellow sour pear, brown Juju pear, Hong'anjiu, Huochengdong yellow pear, The newborn young shoots of Kuqa Amut were used as the test materials. The stems of the test branches up to about 30cm were sterilized with 75% alcohol, and 1 / 3...

Embodiment 3

[0049] Embodiment 3: the pathogenicity determination of different bacterial strains of amygdalus blight

[0050] 1. Preparation of E. amylovora inoculum: Inoculate 10 strains of E. amylovora from different sources in Table 1 on NA+5% sucrose medium, culture at 28.5°C for 48 hours to activate, pick a single colony and inoculate on NA +5% sucrose culture solution, shake culture at 28.5℃, 160rpm / min for 12h, measure OD 600 The bacterial solution with a value of about 1.2 was diluted with sterile water to a concentration of 10 8 CFU / ml was used as the inoculum.

[0051] 2. Preparation of isolated branches: select the new branches of the Korla fragrant pear, a high-susceptibility variety of pear fire blight, as the test branches, cut the test branches to a stem section of about 30 cm, disinfect with 75% alcohol, and insert 1 / 3 of them In a Erlenmeyer flask filled with 0.5‰NaCl sterilized saline.

[0052] 3. Inoculation: Use a sterilized scalpel to cut a 5mm hole in the leaf axil...

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Abstract

The invention relates to the technical field of plant protection and particularly discloses a method for measuring the pathogenicity of pear fire blight bacteria by means of an in-vitro water cultureapproach for fragrant pear branches. The method comprises the following steps of (1) preparation of a pear fire blight bacterium inoculation solution; (2) preparation of in-vitro branches; (3) inoculation; (4) observation of an inoculation result and evaluation of the pathogenicity. The method is simple and feasible, the pathogenicity of pathogenic bacteria is stably expressed, the repeatability is high, and the result is reliable; the pathogenicity and pathogenicity intensity of the pear fire blight bacteria can be accurately reflected. In actual operation, the method can be carried out in batches and has the advantages of small occupied space, lower labor cost and shorter experimental cycle. Therefore, the method is suitable for application and popularization.

Description

technical field [0001] The invention relates to the technical field of plant protection. Specifically, the invention relates to a method for measuring the pathogenicity of Phytophthora amylovora by in vitro hydroponics of fragrant pear branches. Background technique [0002] Amylovora blight is caused by Erwinia amylovora ( Erwinia amylovora ) is the most devastating bacterial disease caused by infecting a variety of Rosaceae plants, and is a quarantine pest for imported plants in my country. The disease first occurred in New York State, USA in 1780, and has now spread to nearly 60 countries and regions in the world. Phytophthora amylovora has a wide host range and can harm more than 220 kinds of plants belonging to more than 40 genera of Rosaceae, among which the most susceptible are fruit trees such as pears, apples, and hawthorns. In the trunk, the pathogen can quickly spread from the diseased tip to the branches and trunk, until the root, causing the inflorescence to d...

Claims

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Application Information

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IPC IPC(8): C12Q1/02C12R1/18
CPCC12Q1/02G01N2333/27
Inventor 罗明袁英哲韩剑李洪涛张静文张祥林
Owner XINJIANG AGRI UNIV
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