Citrus endophytic actinomycetes with antibacterial activity on various plant pathogens

A technology of endogenous actinomycetes and plant pathogenic bacteria, applied in the field of microorganisms, can solve the problems of chemical pesticide environmental pollution, pesticide resistance of pathogenic bacteria, etc., and achieve the effects of no pollution, less resistance, and low residue

Active Publication Date: 2014-07-30
ZHEJIANG CITRUS RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the prevention and control of crop diseases is mainly based on chemical control. As we all know, the long-term and large-scale use of chemical pesticides often leads to many problems such as environmental pollution, pesticide residues, and drug resistance of pathogenic bacteria that are difficult to solve.

Method used

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  • Citrus endophytic actinomycetes with antibacterial activity on various plant pathogens
  • Citrus endophytic actinomycetes with antibacterial activity on various plant pathogens
  • Citrus endophytic actinomycetes with antibacterial activity on various plant pathogens

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Experimental program
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Effect test

Embodiment 1

[0030] Embodiment 1: Isolation and cultivation of citrus endophytic actinomycetes

[0031] 1. Biological material and culture medium: The biological material used to isolate endophytic actinomycetes was collected from the root samples of mandarin oranges from different orange orchards in Huangyan District, Taizhou City, Zhejiang Province. The isolation medium used for the isolation of endophytic actinomycetes is tap water yeast powder agar (the formula is: 0.25 g of yeast extract, K 2 HPO 4 0.5 g, 18 g agar, 1000 mL tap water).

[0032] 2. Sample treatment: wash the above-mentioned freshly collected samples under tap water to remove the soil on the surface of the sample, and then clean it with ultrasonic waves; 99% ethanol for another 30 s. The sterilized samples were cut into small pieces of about 1 cm × 1 cm and placed on the surface of the separation medium. At the same time, the cleaning solution of the last processed sample was applied to the separation medium to det...

Embodiment 2

[0034] Embodiment 2: Screening of antagonistic strains of citrus endophytic actinomycetes

[0035] 1. Preparation of indicator bacteria plate: prepare potato dextrose agar (PDA) medium plate (preparation method: weigh 200 g of fresh peeled potatoes, chop and add 1000 mL of water to boil for half an hour, filter with gauze, and take the filtrate to volume to 1000 mL, add 20 g of glucose and 20 g of agar, heat to fully dissolve and distribute into Erlenmeyer flasks or glass test tubes, 121 ° C, autoclave for 20 min), and use a sterile puncher with an inner diameter of 5 mm Take freshly cultivated colonies of 3 pathogens of citrus anthracnose, Myrica rubra and Loquat rotifera (these 3 pathogens are indicator bacteria for screening of antagonistic strains), and transfer them to PDA medium plates. Six identical bacteria cakes were placed around the center of the plate about 3 cm away to make an indicator plate.

[0036] 2. Screening of strains with inhibitory activity: Inoculate t...

Embodiment 3

[0038] Embodiment 3: Streptomyces ( Streptomyces sp.) Preparation of R1020 Fermentation Crude Extract

[0039] 1. Activation culture of the strain: transfer the isolated and preserved strain R1020 to Gaoshi Synthetic No. 1 medium plate and culture it in an incubator at 28°C for 7 days.

[0040] 2. Fermentation of bacterial strains: Take the activated strain R1020 on Gaoshi Synthetic No. 1 medium and inoculate it into Gaoshi Synthetic No. 1 liquid fermentation medium (100 mL medium / 250 mL Erlenmeyer flask, the formula is the same as Gaoshi Synthetic 1 No. solid medium without agar), cultured on a shaker at 200 rpm / min for 4 days at 28°C.

[0041] 3. Preparation of the crude extract of the fermentation product: 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 obtain the ethyl acetate extract of the supe...

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Abstract

The invention relates to citrus endophytic actinomycetes with antibacterial activity on various plant pathogens, belonging to the technical field of microbes. The endophytic actinomycetes are Streptomyces sp. R1020 which was registered and collected in China General Microbiological Culture Collection Center on March 17, 2014 with the collection number of CGMCC No. 8923. The strain R1020 has relatively good inhibitory effect on citrus anthrax bacteria, citrus bacterial canker disease bacteria, citrus penicillium italicum, citrus penicillium digitatum, bayberry deadwood disease bacteria, bayberry acicola, bayberry anthrax bacteria, loquat anthrax bacteria, loquat leaf spot fungi, loquat ringspot pathogenic bacteria, tomato alternaria solani, botrytis cinerea, colletotrichum orbiculare, tea tree anthrax bacteria and elsinoeleucospila and has the potential of being developed into biological pesticides.

Description

technical field [0001] The invention relates to a citrus endophytic actinomycete with antibacterial activity to various plant pathogenic bacteria, belonging to the technical field of microbes. Background technique [0002] Plant endophytes are a new class of biocontrol microbial resources, which can colonize and operate in plant tissues and are harmless or even beneficial to plants. Because it has a stable living space in the plant and can produce the same or similar physiologically active substances as the metabolism of the host plant, it can effectively inhibit the infection of pathogenic bacteria or improve the disease resistance of the host plant. Endophytes are distributed inside the host plant tissue, and have a more stable living environment and are easier to function than epiphytes exposed to the external environment. In addition, plant endophytic actinomycetes have a certain host specificity, except for absorbing nutrients from their host plants and acting on other...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01P3/00
Inventor 胡秀荣鹿连明杜丹超陈国庆黄振东蒲占湑
Owner ZHEJIANG CITRUS RES INST
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