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A kind of antibacterial protein pbr1 and its preparation method and application

An antibacterial protein and aspect technology, applied in the fields of molecular biology and biological pesticides, can solve problems such as affecting human health and ecological balance, easily producing drug resistance, polluting the environment, etc., achieving good development prospects, remarkable control effects, and environmental protection. Effect

Inactive Publication Date: 2016-07-06
YUNNAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Chemical pesticides have played a very important role in agricultural production for a long time in the control of crop diseases and insect pests. However, due to the extensive use of chemical pesticides, it has brought many negative effects, such as polluting the environment, endangering humans and animals, and prone to drug resistance, which seriously affects human health. and ecological balance

Method used

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  • A kind of antibacterial protein pbr1 and its preparation method and application
  • A kind of antibacterial protein pbr1 and its preparation method and application
  • A kind of antibacterial protein pbr1 and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1: Preparation and properties of the antibacterial protein PBR1 of the present invention

[0038] 1. Preparation of antibacterial protein PBR1

[0039] (1) Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) XF1 strain CGMCC NO.2357 was inoculated in LB liquid medium, and cultured at 30° C. and 170 rpm for 48 hours with shaking to obtain a fermentation broth.

[0040] (2) separating and purifying the fermented liquid obtained in step (1) to obtain the target protein; the method for separating and purifying is:

[0041] ① Ammonium sulfate precipitation: take the fermented liquid obtained in step (1), centrifuge at 4°C for 10 minutes at 8000rpm, collect the supernatant, and use the salting-out method to add solid ammonium sulfate with 80% saturation to the fermented liquid, Precipitate for about 24 hours, then freeze and centrifuge at 12,000 rpm for 30 minutes to collect the precipitate.

[0042] ②Dialysis: Dissolve the precipitate collected in the step i...

Embodiment 2

[0061] Embodiment 2: the inhibitory effect of antimicrobial protein PBR1 to Plasmodium spore

[0062] The antibacterial protein PBR1 has a strong lytic effect on the spores of Plasmodium spores. After adding the antibacterial protein PBR1, it can be seen that the spores aggregate into chains or clusters ( Figure 6 B). After 6 hours, it can be seen that the cell wall of the Plasmodium spores is obviously thinner and the shape of the spores begins to become deformed ( Figure 6 C), after 24h, spore lysis has been seen, and the cell contents are extravasated ( Figure 6 D). However, the spores in the control (water, buffer solution) did not change ( Figure 7 ).

Embodiment 3

[0063] Embodiment 3: the mensuration of antibacterial protein PBR1 bacteriostatic spectrum

[0064] According to the inhibitory effect on 11 fungal strains, the isolated antibacterial protein PBR1 has a broad-spectrum inhibitory effect on pathogenic bacteria (see Table 1). It also has a good inhibitory effect on tomato cinerea, wheat take-all, and pomegranate wilt. These pathogens include Fusarium of the subphylum Deuteromycota, genus Ceratocystis of the subphylum Ascomycota, and genus Botrytis of the subphylum Deuteromycota.

[0065] Table 1 The antibacterial effect of antibacterial protein PBR1

[0066]

[0067] Note: +: antibacterial bandwidth ≥ 0.25cm: ++: antibacterial bandwidth ≥ 0.5cm;

[0068] +++ Antibacterial bandwidth ≥ 1cm; ++++ Antibacterial bandwidth ≥ 1.5cm.

[0069] Yunnan Agricultural University

[0070] An antibacterial protein PBR1 and its preparation method and application

[0071] /

[0072] 1

[0073] PatentInversion3.3

[0074] 1

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Abstract

The invention discloses an anti-bacterial protein PBR1 as well as a preparation method and application thereof, and relates to the technical field of molecular biology and biological pesticides. The amino acid sequence of the anti-bacterial protein PBR1 is shown as SEQ ID NO.1. The anti-bacterial protein PBR1 has a superior effect on preventing and curing plant diseases such as cruciferous plasmodiophoromycetes, paddy rice magnaporthe oryzae, curvularia lunata, fusarium oxysporum f. sp. dianthi, fusarium oxysporum vasinfectum, konjac foot rot bacteria, panax notoginseng root rot pathogen, pomegranate wilt pathogen, gaeumannomyces gramini, cylindrosporium pomi brooks, alternaria alternata and / or botrytis cinerea. A pesticide taking the anti-bacterial protein PBR1 as an active ingredient is a novel environment-friendly biological pesticide, is safe to people and livestock, protects the environment, and has a favorable development prospect.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and biological pesticides, in particular to a novel antibacterial protein PBR1 from microorganisms and its application. Background technique [0002] Chemical pesticides have played a very important role in agricultural production for a long time in the control of crop diseases and insect pests. However, due to the extensive use of chemical pesticides, it has brought many negative effects, such as polluting the environment, endangering humans and animals, and prone to drug resistance, which seriously affects human health. and ecological balance. As an important measure to prevent and control plant diseases, biological control has become an inevitable trend of agricultural development. [0003] Bacillus subtilis is a Gram-positive rod-shaped bacterium widely distributed in various living environments. It can produce endophytic spores. It is heat-resistant and stress-resistant. It is ubi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/32A01N47/44A01P1/00A01P3/00C07K1/36C07K1/14C07K1/34C07K1/16
CPCA01N47/44C07K14/32
Inventor 何月秋吴毅歆
Owner YUNNAN AGRICULTURAL UNIVERSITY
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