Marine streptomyces with antibacterial activity

A technology of marine streptomyces and streptomyces, applied in the field of microorganisms, can solve difficult problems, screen out new disease-resistant substrates, etc., and achieve the effects of low residue, good market application prospects, and easy cultivation

Active Publication Date: 2018-05-01
ZHEJIANG CITRUS RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Many important agricultural antibiotics widely used at present are isolated from actinomycetes, and these actinomycetes mainly come from te...

Method used

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  • Marine streptomyces with antibacterial activity
  • Marine streptomyces with antibacterial activity
  • Marine streptomyces with antibacterial activity

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Experimental program
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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 200 g of fresh peeled potatoes, chop them, add 1000 mL of water to boil for half an hour, filter with gauze, take the filtrate to 1000 mL, and then Add 20 g of glucose and 20 g of agar, heat to fully dissolve, then dispense into Erlenmeyer flasks or glass test tubes, sterilize at 121°C for 20 minutes by autoclaving) on ​​a flat plate, and culture in a microbial incubator at 28°C until the colonies produce spores Finally, 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 th...

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 marine streptomyces with antibacterial activity and belongs to the technical field of microbes. The marine streptomyces is streptomyces chumphonensis AM-4, is registered and collected in China General Microbiological Culture Collection Center on October 30, 2017 and has a collection number of CGMCC No.14843. The streptomyces chumphonensis AM-4 has an excellent inhibitory effect on ccolletotrichum glecosporioides, penicillium italicum, penicillium digitatum and pathogen of brown stem-end rot, and has the potential of being developed into 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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IPC IPC(8): C12N1/20A01N63/00A01P3/00C12R1/465
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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