Method for quickly identifying disease resistance of inoculated in vitro branched to pear fire blight

A technology for branch inoculation and E. amylovora, which is applied in horticultural methods, botany equipment and methods, biochemical equipment and methods, etc., can solve the problem of the lack of a unified and clear evaluation method for disease resistance measurement of E. amylovora. Quantitative statistics on the incidence of disease, rating of disease resistance and other issues, to achieve the same inoculation amount, standard grading standard, and consistent wound size

Pending Publication Date: 2019-07-23
XINJIANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is difficult to quantify the incidence of statistics, and cannot accurately rate its disease resistance
So far, there are no unified and clear evaluation methods for the determination of the resistance of different hosts to E. amylovora by various existing methods

Method used

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  • Method for quickly identifying disease resistance of inoculated in vitro branched to pear fire blight
  • Method for quickly identifying disease resistance of inoculated in vitro branched to pear fire blight
  • Method for quickly identifying disease resistance of inoculated in vitro branched to pear fire blight

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1 Determination of pathogenicity of different bacterial strains of E. amylovora

[0027] 1. Preparation of E. amylovora inoculum: the E. amylovora strain E.a 0001, E.a 0017、 E.a 0019, E.a 0033、 E.a 0032 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 to 10 with sterilized water. 8 as an inoculum.

[0028] 2. Preparation of isolated branches: select the new branches of the high-susceptibility pear fire blight variety Xiangli in the same year as the test branches, cut the stems of the test branches to about 50 cm, disinfect with 75% alcohol, and insert 1 / 3 of them In a Erlenmeyer flask filled with 0.5‰NaCl sterile saline.

[0029] 3. Inoculation: Use a sterilized scalpel to cut a wound of about 5 mm at the leaf axil of...

Embodiment 2

[0036] Embodiment 2 Different pear varieties (germplasm) in vitro branch inoculation E. amylovora Classification of disease resistance

[0037] 1. the preparation of E. amylovora inoculum: according to the measurement result of embodiment 1, will E.a The 0001 strain was identified as a highly pathogenic strain, and this strain was used as the test strain for disease resistance determination. Blight blight E.a 0001 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 to 10 with sterilized water. 8 as an inoculum.

[0038] 2. Preparation of isolated branches: collect the young shoots of the 21 tested pear varieties (germplasm) listed in Table 2 as the inoculation material, cut the stems of the tested branches to about 50 cm, and disinfect them...

Embodiment 3

[0046] Example 3 Different apple varieties (germplasm) in vitro branch inoculation E. amylovora Classification of disease resistance

[0047] 1. Preparation of E. amylovora inoculum: the E. amylovora strain E.a 0001 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 to 10 with sterilized water. 8 as an inoculum.

[0048] 2. Preparation of isolated branches: collect the young shoots of the 19 test apple varieties (germplasm) listed in Table 3 as inoculation materials, cut the test branches to a stem segment of about 50 cm, and disinfect them with 75% alcohol , Insert 1 / 3 of it into the Erlenmeyer flask filled with 0.5‰ NaCl sterilized saline.

[0049] 3. Inoculation: Use a sterilized scalpel to cut a wound of about 5mm at the leaf axil of 3~4...

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Abstract

The invention relates to the technical field of plant protection, and particularly discloses a method for quickly identifying disease resistance of inoculated in vitro branched to pear fire blight. The method comprises the following steps of (1) preparing erwinia amylovora inoculating liquid; (2) preparing in vitro branches; (3) performing inoculation; and (4) evaluating variety disease resistance. According to the identifying and evaluating method for the disease resistance of the pear fire blight, provided by the invention, pathogenesis symptoms and pathogeny quantifying through a molecularbiology technique are combined, so that the method is quick, simple, standard and practical, the result is accurate and reliable, and the method is suitable for performing quick identification and screening on disease resistance of bulk materials.

Description

technical field [0001] The invention relates to the technical field of plant protection, in particular, the invention relates to a method for rapid identification of resistance to amyrrhia blight by inoculation of isolated branches. Background technique [0002] Pear fire blight is a devastating disease on pome fruit trees of the Rosaceae family. It is a class of phytosanitary dangerous pests imported into my country. It harms more than 220 kinds of plants belonging to more than 40 genera. Among them, the most susceptible hosts are pears, Apple, hawthorn, etc. Phytophthora amylovora ( Erwinia amylovora ) infects flowers, leaves and new shoots, branches and fruits. When the disease is severe, it quickly spreads from flower vessels and shoots to branches, trunks, and roots, resulting in the death of fruit trees within a few weeks, the destruction of orchards, and heavy economic losses. Pear fire blight has now occurred in more than 60 countries in the world. At present, there...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/18C12Q1/6851C12Q1/06A01G7/06
CPCA01G7/06C12Q1/18C12Q1/6851C12Q2531/113C12Q2563/107
Inventor 罗明李洪涛袁英哲韩剑蒋萍张祥林唐章虎
Owner XINJIANG AGRI UNIV
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