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Erwinia carotovora pv. Carotovora strain Ecc 36, plant immune activator protein secreted by same and application of plant immune activator protein

A carrot soft rot and Erwinia technology applied in the biological field to achieve long shelf life, avoid residue and pollution, and facilitate storage and transportation

Active Publication Date: 2017-06-20
BIOLOGY INST OF HEBEI ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are relatively few studies on the use of plant immune activators to biologically control vegetable diseases. Therefore, it is urgent to screen out a microbial strain that can efficiently secrete plant protein activators, so as to reduce the impact of Erwinia soft rot on vegetable production through plant immune activators. Harm

Method used

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  • Erwinia carotovora pv. Carotovora strain Ecc 36, plant immune activator protein secreted by same and application of plant immune activator protein
  • Erwinia carotovora pv. Carotovora strain Ecc 36, plant immune activator protein secreted by same and application of plant immune activator protein
  • Erwinia carotovora pv. Carotovora strain Ecc 36, plant immune activator protein secreted by same and application of plant immune activator protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Isolation and Screening of Bacterial Strains with Activation of Plant Immunity and Disease Resistance

[0027] The rotten Chinese cabbage in the plots with serious incidence of Chinese cabbage was collected as the test material. Bacteria were isolated from the sample in the laboratory, and the bacteria were isolated and purified on LB medium for many times. After the pure cultured bacteria were obtained, the strain identification was carried out.

[0028] Specific steps: the above soil samples come from Chinese cabbage fields infected with Chinese cabbage rot disease in Hebei Province, Shandong, Shanxi and other places. The samples are diluted in a gradient, spread on LB medium, and cultivated in a constant temperature incubator at 28°C. The above-mentioned medium was prepared according to the records in the "Microbial Culture Medium Manual". The formula of the medium is: 5g of beef extract, 10g of peptone, 5g of sodium chloride, 15g of agar, 1000ml of tap wate...

Embodiment 2

[0030] Example 2 Toxicity Test of No. 36 Bacterial Strain with Activation of Plant Immunity and Disease Resistance

[0031] The isolated No. 36 strain was activated in LB liquid medium and cultured at 25°C for 24 hours to prepare a bacterial suspension. After sterilizing the surface of healthy cabbage leaves with alcohol cotton for 3 times, rinse them with sterile water for 3 times, use a sterilized inoculation needle to prick 5 holes on each healthy cabbage leaf, and then add LB liquid bacterial suspension to each hole. Each treatment was repeated 3 times, and the addition of sterile LB medium was used as a control. Put the treated cabbage leaves into a white culture pot, cover the mouth of the pot with plastic wrap to keep moisture, place it in a constant temperature incubator at 25°C, and observe the disease after 24 hours and compare whether it is consistent with the symptoms in the field ( figure 2 ).

[0032] The isolated No. 36 strain was inoculated into a low-salt m...

Embodiment 3

[0034] Example 3 Identification of antagonistic strains

[0035]According to the experimental content and experimental methods recorded in the "Common Bacteria System Identification Manual", the screened preservation number is: CGMCC No.11751 for identification, pick the cultures of the screened soft rot strains, and culture them in LB. After cultured at 28°C for 48 hours, the colony morphology was observed with naked eyes and photographed. The colonies of soft rot pathogens were picked for smear staining, and their characteristics were observed under a 4×100 magnification microscope.

[0036] The observation of colony morphology was carried out on LB medium. After the strains were streaked and inoculated, they were cultured at a constant temperature of 25° C. for 24 hours to observe the colony morphology. The colony of the strain with the preservation number CGMCC No. 11751 is round or irregular, slightly raised, off-white, shiny, translucent, smooth, and not tightly combine...

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Abstract

The invention discloses an Erwinia carotovora pv. Carotovora strain Ecc 36 with the preservation number of CGMCC No. 11751; plant immune activator protein HrpNEccPR secreted by the Erwinia carotovora pv. Carotovora strain Ecc 36 has an effect of preventing and controlling plant soft rot. The invention also discloses a protein sequence of the plant immune activator protein HrpNEccPR and a gene sequence that expresses the protein, and further provides a method for obtaining the plant immune activating protein. According to the invention, an activator protein crude product generated by the selected strain with the preservation number of CGMCC No. 11751 has a good antagonistic effect for plants, especially for Chinese cabbage soft rot, and the activator protein crude product can effectively prevent and control the occurrence of the Chinese cabbage soft rot.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to Erwinia carotovora, the plant immune activation protein secreted by it and its application. Background technique [0002] Erwinia soft rot is a broad pathogen that mainly occurs in Chinese cabbage, cabbage, cauliflower, potato and other crops, and mainly damages leaves, stems and tubers. Leaf infection occurs first on old leaves near the ground, and the diseased part is irregular dark brown lesions, which rot when the humidity is high. In recent years, due to various reasons, the frequency and area of ​​major diseases have increased significantly, and the resulting yield loss is about 50-60%. And these vegetables are indispensable important vegetables in northern my country, but the annual loss due to disease is estimated to reach several billion yuan. In order to deal with severe soft rot diseases of these vegetables, agricultural producers use chemical pesticides in l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C07K14/27C12N15/31C12P21/02A01N47/44A01P3/00C12R1/18
CPCA01N47/44C07K14/27C12P21/02C12N1/205C12R2001/18
Inventor 贾振华宋水山刘方黄亚丽宋聪郝少彦李海利赵芊黄媛媛马宏刘昆昂
Owner BIOLOGY INST OF HEBEI ACAD OF SCI
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