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A Strain of Marine Streptomyces with Bacteriostatic Activity

A technology of marine streptomyces and streptomyces, applied in the field of microorganisms, can solve the problems of screening out new disease-resistant substrates, difficult problems, etc., and achieve the effect of good market application prospects, low residue, and no pollution to produce drug resistance

Active Publication Date: 2020-06-19
ZHEJIANG CITRUS RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many important agricultural antibiotics widely used at present are isolated from actinomycetes, and these actinomycetes mainly come from terrestrial environments. Due to repeated isolation and screening, it is difficult to screen out actinomycetes with new disease-resistant substrates

Method used

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  • A Strain of Marine Streptomyces with Bacteriostatic Activity
  • A Strain of Marine Streptomyces with Bacteriostatic Activity
  • A Strain of Marine Streptomyces with Bacteriostatic Activity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1: the isolation culture of marine streptomyces

[0028] 1. Biological materials and culture medium: samples were taken from the surrounding sea area of ​​Dachen Island, Taizhou City, Zhejiang Province. The samples included fish, shellfish, shrimp and sediment in the intertidal zone. The collected samples were placed in ice cubes for insulation box, and brought back to the laboratory within 24 hours for the isolation of actinomycetes. A total of 6 kinds of isolation media were selected for the isolation medium, and the composition is shown in Table 3. The artificial seawater was 40 g of sea salt produced by Sigma company and 1000 mL of pure water Dissolved. Nalidixic acid 15 μg / mL, nystatin 50 μg / mL and cycloheximide 50 μg / mL were added to the six media.

[0029] Table 3 Components of 6 isolation media

[0030]

[0031] 2. Sample processing: The collected samples are separated by two processing methods. Treatment method A: Take 1 g sample, put it into 4...

Embodiment 2

[0033] Embodiment 2: Screening of marine Streptomyces antagonistic strains

[0034]1. Preparation of indicator bacteria plate: The pathogenic bacteria of citrus green mold is isolated from the diseased citrus fruit by our laboratory and preserved. Transfer the preserved citrus green mold bacteria to PDA medium (the preparation method is: take 200g of fresh peeled potatoes, chop them, add 1000mL of water to boil for half an hour, filter with gauze, take the filtrate to 1000mL, add 20 g of glucose and 20 g of agar, heated and fully dissolved, then packed into Erlenmeyer flasks or glass test tubes, sterilized at 121°C for 20 minutes by autoclaving) on ​​a flat plate, placed in a microbial incubator at 28°C for cultivation, and after the colonies produced spores, Add 10 mL of sterile water to the petri dish to collect the spores, filter with sterile gauze, and adjust the concentration of the spore suspension to 10 with sterile water 4 spores / mL, use a pipette to draw 20 µL of the...

Embodiment 3

[0037] Embodiment 3: Streptomyces ( Streptomyces chumphonensis ) Preparation of AM-4 Fermented Broth Crude Extract

[0038] 1. Activation culture of the strain: transfer the isolated and preserved Streptomyces AM-4 to Gaoshi Synthetic No. 1 medium plate and culture it in an incubator at 28°C for 7 days.

[0039] 2. Fermentation of bacterial strains: Take the activated Streptomyces AM-4 on Gaoshi Synthetic No. 1 medium and inoculate into Gaoshi Synthetic No. 1 liquid fermentation medium (100 mL medium / 250 mL Erlenmeyer flask, the formula is the same as that of Gaoshi Synthetic No. 1 Synthetic No. 1 solid medium, without agar), cultured on a shaker at 200 rpm / min for 4 days at 28°C.

[0040] 3. Preparation of the crude extract of the fermentation broth: take the fermentation broth and filter it through sterile filter paper to obtain the supernatant and mycelia respectively. The supernatant was taken, extracted three times with ethyl acetate, and the extracts were combined to o...

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Abstract

The invention relates to a strain of marine streptomyces with antibacterial activity, belonging to the technical field of microbes. The marine Streptomyces is Streptomyces ( Streptomyces chumphonensis ) AM‑4, which was registered and preserved in the General Microorganism Center of China Microbiological Culture Collection Management Committee on October 30, 2017, with the preservation number CGMCC No.14843. Streptomyces AM‑4 has good inhibitory effects on citrus anthracnose, Penicillium citri, green mold and brown pedicle rot of citrus, and has the potential to be developed as a biopesticide.

Description

technical field [0001] The invention relates to a strain of marine streptomyces with antibacterial activity, belonging to the technical field of microbes. Background technique [0002] After the citrus fruit is harvested, more than 20 kinds of diseases may occur during storage and transportation. The infectious diseases caused by fungi mainly include penicillium, green mold, anthracnose, pedicle rot, black rot, and sour rot Wait. At present, the effective control of citrus storage diseases is mainly based on the mixture of imidazoles, biguanide salts and 2,4-D three chemical fungicides. As we all know, the long-term improper use of chemical pesticides will cause serious consequences such as the destruction of ecological balance, environmental pollution, drug resistance of pathogenic bacteria, and excessive pesticide residues in fruits. Therefore, it is very necessary to develop new biopesticides for the prevention and treatment of citrus storage diseases. [0003] As an i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/28A01P3/00C12R1/465
CPCA01N63/00C12N1/205C12R2001/465
Inventor 胡秀荣黄振东鹿连明蒲占湑杜丹超陈国庆
Owner ZHEJIANG CITRUS RES INST
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