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Burkholderia sp. Strain ES-89 for preventing and treating fungal diseases of plants as well as culture method and application

A technology of Burkholderia and Kholderia, which is applied in the field of plant disease biological control to achieve the effects of good control effect, reduced pollution and low production cost

Active Publication Date: 2017-03-15
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the late 1980s, several strains of Burkholderia biocontrol bacteria have been submitted to the U.S. Environmental Protection Agency (EPA) for registration as biopesticides, but no Burkholderia strains have been seen in China. Report on Prevention and Control of Coptidis Diseases

Method used

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  • Burkholderia sp. Strain ES-89 for preventing and treating fungal diseases of plants as well as culture method and application
  • Burkholderia sp. Strain ES-89 for preventing and treating fungal diseases of plants as well as culture method and application
  • Burkholderia sp. Strain ES-89 for preventing and treating fungal diseases of plants as well as culture method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1: Screening of Burkholderia strain ES-89

[0048] Screening of strains:

[0049] (1) Collection of samples: The soil for this experiment was collected from the Coptidis GAP planting base in Taishan Temple, Enshi Prefecture, Hubei Province, China. The five-point sampling method was used for sampling. The depth of sampling was around the rhizosphere of Coptis chinensis and 15 or 17 or 19 or 20 cm underground. , take a sample of 200 or 240 or 260 or 280 or 300g at each point, and then put the sample into a sterilized sealed bag and bring it back to the laboratory for bacterial screening.

[0050] (2) Preparation of sample suspension: Grind the collected soil sample (through 80-mesh sieve) and add 5 g into a conical flask filled with 45 ml of sterile water. Shake fully at 180 rpm for 20 min at 28 ° C, and then make 1 :10(V / V) of soil suspension. After the soil particles are left to settle, absorb 1ml of the supernatant and transfer it into a test tube filled with...

Embodiment 2

[0053] Example 2: Identification of Burkholderia strain ES-89

[0054] Identification of strains:

[0055] The applicant isolated and screened a Burkholderia strain with strong control effect on Coptis chinensis from the rhizosphere soil of Coptis chinensis GAP planting base in Taishan Temple, Enshi Prefecture, Hubei Province, China. Named ES-89. The isolation and identification method of ES-89 bacterial strain of the present invention, with reference to plant pathological research method (Fang Zhongda, 1998), Berger Bacteria Identification Handbook (eighth edition) and Common Bacteria System Identification Handbook (Dong Xiuzhu and Cai Miaoying, 2001) Test methods in other monographs. The ES-89 strain was placed in NA culture based on overnight culture at 28°C, and the genomic DNA was extracted using the Bacterial Genomic DNA Extraction Kit (TIANGEN). -TACGGYTACCTTGTTACGAC TT-3') to amplify its 16S rDNA region.

[0056] Morphological identification results of ES-89 strain...

Embodiment 3

[0059] A Burkholderia ES-89 culture method for preventing and treating plant fungal diseases, the steps are as follows:

[0060] A. Inoculate 1ml of ES-89 strain into 100ml NB culture solution and activate for 22 or 23 or 24 or 25 or 26h;

[0061] B. Inoculate 1ml of ES-89 bacterial classification activated in step (A) into a 250ml Erlenmeyer flask containing 100ml NB culture solution, 26 or 27 or 28 or 29 or 30°C, 160rpm, shaking culture 46 or 47 or 48 or 49 or 50h;

[0062] C. Place the fermented liquid obtained after shaking culture in step (B) at 4°C, centrifuge at 8000rpm for 18 or 19 or 20 or 21 or 22min, and filter the upper layer of the fermented liquid with a bacterial filter (0.22um) to obtain sterile fermentation supernatant;

[0063] D. Wash the thalline with sterile water for 2 or 3 or 4 times to make its thalline suspension. Store the fermentation supernatant and cell suspension at 4°C for later use;

[0064] The composition and formula of the nutrient broth ...

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Abstract

The invention discloses a Burkholderia sp. Strain ES-89 for preventing and treating fungal diseases of plants as well as a culture method and application. The culture method comprises the following steps: A, inoculating an ES-89 strain into a nutrient broth culture liquid for activation; B, inoculating the activated ES-89 into a triangular flask with the nutrient broth culture liquid, and performing shake cultivation; C, centrifuging fermentation broth obtained from shake cultivation under a certain condition, and filtering an upper layer of fermentation broth by a bacterial filter to obtain sterile supernate; D, washing the bacterium with sterile water to prepare a bacterial suspension, and storing the fermentation broth, the fermentation supernate and the bacterial suspension. The strain is preserved with the preservation number of CCTCC, CCTCC NO: M 2016665, and the strain ES-89 can be applied to preparation of medicines for treating or preventing plant pathogenic fungi. The strain has an inhibition function on coptis chinensis leaf spot diseases and the like, and can be applied to prevention and treatment on multiple plant diseases. The prevention and treatment effect of the strain on the coptis chinensis leaf spot diseases can be 88% or greater, the culture method is feasible, the operation is simple and convenient, the pollution in the production process is reduced, solvent poisoning is avoided, and the application prospect is relatively good.

Description

technical field [0001] The invention belongs to the technical field of biological control of plant diseases, and in particular relates to a Burkholderia sp. The cultivation method of the Germania strain ES-89 also relates to a use of the Burkholderia strain ES-89 for preventing and treating plant fungal diseases. Background technique [0002] Coptis chinensis is a commonly used traditional Chinese medicine, and it is mostly planted in the mountains of Sichuan, Hubei, Hunan and Guizhou, the mountains of southwestern Sichuan, the mountains of southern Sichuan, Yunnan and Tibet, and the mountains of Qinba and other regions. Coptis chinensis has high medicinal value, and it has been reported that it has good curative effects in anticancer and hypoglycemic aspects, which has attracted widespread attention. With the increasing interest in the development of natural medicines in the international community, especially the in-depth research and extensive application of Coptis chine...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/02A01P3/00C12R1/01
CPCA01N63/10C12N1/20C12N1/205C12R2001/01
Inventor 郑露席中刚黄俊斌刘浩游景茂
Owner HUAZHONG AGRI UNIV
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