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Pharming bacteriophage PSA-Pe and application thereof

A bacteriophage, potent technology, applied in the field of microbiology, can solve the problems of low development speed of new antibiotics and resistance, and achieve the effect of low cost, strong specificity and remarkable effect.

Pending Publication Date: 2022-07-29
苍溪县兴科现代农业科技研究院有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

According to reports, 75% of bacterial infections in the United States are resistant to one or more antibiotics. Due to the continuous emergence of drug-resistant bacteria due to the abuse of antibiotics, the therapeutic effect of antibiotics is declining, especially the "super bacteria" However, the development speed of new antibiotics is far lower than the speed of bacteria overcoming antibiotic resistance, making the application of phages come back to the stage of history

Method used

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  • Pharming bacteriophage PSA-Pe and application thereof
  • Pharming bacteriophage PSA-Pe and application thereof
  • Pharming bacteriophage PSA-Pe and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The present embodiment is the preparation method of the potent bacteriophage PSA-Pe of the present invention, comprising the following steps:

[0037] (1) Isolation, purification and titer determination of kiwifruit canker phage

[0038] The isolated PSA bacterial liquid was transferred to 20 mL of LB medium as the host, and cultured with shaking at 150 rpm / min for 16 h at 25°C. Take the above shaken PSA culture medium, mix it with sewage or soil samples, take 200uL of the mixture and add 10mL of semi-solid medium, put the semi-solid medium in a water bath in advance, and keep its temperature at 34-36°C. It was quickly poured onto the thin-layer solid medium cooled in advance, that is, the phage was separated by the double-layer plate method, and the formation of plaque was observed after culturing at 25 °C for 16 h.

[0039] If clear plaques are observed on the double-layer plate after culturing for 16 hours, pick a single plaque and inoculate it into 200uL log-phase ho...

Embodiment 2

[0051] The present embodiment is the stability test of the potent phage PSA-Pe of the present invention, including the following steps:

[0052] (1) Thermal stability

[0053] To evaluate the stability of phage at different temperatures, experiments were performed in sterile LB at pH 7.0. Put the phages to be tested in a water bath at 4°C, 25°C, 37°C and 50°C, respectively, and take samples at 0h, 3h, 6h, 12h, and 24h. The culture solution was serially diluted, and the phage titer was measured by the double-layer plate method. The phage was cultured in a 25°C incubator for 16 hours, and three replicates were performed at each time point to obtain the average value.

[0054] The phage titer is obtained by counting the plaques on the plate, such as Figure 4 As shown, the phage was stored at 4 °C and 25 °C for 24 hours, and the concentration decreased by less than 0.16 lg PFU / mL. When the temperature was increased to 37 °C, the maximum inactivation of the experimental phage wa...

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Abstract

The invention discloses a virulent bacteriophage PSA-Pe and application thereof, the virulent bacteriophage PSA-Pe is preserved in China General Microbiological Culture Collection Center (CGMCC) on July 14, 2020, and the preservation number is CGMCC No.18920. The virulent bacteriophage PSA-Pe has the advantages that the virulent bacteriophage PSA-Pe can be applied to the field The virulent bacteriophage PSA-Pe provided by the invention has the remarkable advantages that (1) pathogenic bacteria can be specifically killed, the acting time is short, and the lasting time is long; (2) the drug resistance of pathogenic bacteria is not caused; (3) normal physiological metabolism of plants is not influenced, normal florae on soil and the plants are not influenced, and the fertilizer is nontoxic, harmless, eco-friendly and green and environment-friendly; (4) as a specific bacteriophage of pseudomonas syringae macaque pathotype, the bacteriophage has strong specificity, can be applied to prevention and treatment of kiwifruit canker caused by pseudomonas syringae macaque pathotype (PSA), and can be used for rapid detection of pathogenic bacteria in plants, food and environment; and (5) self-copying can be realized, and the research and development production cycle is short.

Description

technical field [0001] The invention relates to the field of microbiology, in particular to a potent bacteriophage PSA-Pe and an application thereof, which is a potent bacteriophage taking the pathogenic type of Pseudomonas syringae kiwifruit as a host. Background technique [0002] Kiwifruit canker disease occurs in all kiwifruit producing areas of the world. Although the origin of the pathogens varies from place to place, genome sequence analysis shows that the pathogens are all Gram-negative bacteria of the genus Pseudomonas, and the pathogenesis of the pathogens is closely related to their host plants. Therefore, it is urgent and necessary to study the incidence of canker in different planting areas and develop high-efficiency antibacterial agents, especially biological agents, to effectively control the occurrence of canker. [0003] The control of plant bacterial diseases is a worldwide problem. Traditionally, the use of chemical pesticides, together with crop rotatio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A01N63/40A01P1/00C12Q1/04C12R1/92
CPCC12N7/00A01N63/40C12Q1/04C12N2795/00021G01N2333/21G01N2333/005Y02A50/30
Inventor 夏勉李刚贾燕涛金一白珺刘成刘彦希张继贤杨长铜
Owner 苍溪县兴科现代农业科技研究院有限公司