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Anti-bacterial protein PBR1 as well as preparation method and application thereof

A kind of antibacterial protein and technology, applied in the fields of molecular biology and biopesticides, can solve problems that affect human health and ecological balance, easily produce drug resistance, pollute the environment, etc., achieve good development prospects, significant control effects, and protect the environment. Effect

Inactive Publication Date: 2014-04-16
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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  • Anti-bacterial protein PBR1 as well as preparation method and application thereof
  • Anti-bacterial protein PBR1 as well as preparation method and application thereof
  • Anti-bacterial protein PBR1 as well as preparation method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1: antibacterial protein of the present invention PBR1 preparation and properties of

[0040] 1. Antibacterial protein PBR1 preparation of

[0041] (1) Take Bacillus amyloliquefaciens ( Bacillus amyloliquefaciens ) XF1 strain CGMCC NO.2357 was inoculated in LB liquid medium, and cultured at 30°C and 170rpm for 48 hours with shaking to obtain a fermentation broth.

[0042] (2) Separating and purifying the fermentation broth obtained in step (1) to obtain the target protein; the separation and purification method is:

[0043] ① Ammonium sulfate precipitation: take the fermentation broth obtained in step (1), centrifuge it at 4°C and 8000rpm for 10 minutes, collect the supernatant, and use the salting-out method, add solid ammonium sulfate with 80% saturation to the fermentation broth, and put it at 4°C Precipitate for about 24 hours, and then refrigerated and centrifuged at 12000 rpm for 30 minutes to collect the precipitate.

[0044] ②Dialysis: Dissolv...

Embodiment 2

[0063] Embodiment 2: antibacterial protein PBR1 Inhibitory effect on Plasmodium spores

[0064] antimicrobial protein PBR1 It has a strong lytic effect on the spores of Pluzobacterium spores, adding antibacterial proteins PBR1 Afterwards, the spores can be seen to aggregate into chains or clusters ( Figure 6 B). After 6 hours, it can be seen that the cell wall of Plasmodium spores is obviously thinner and the shape of the spores begins to become deformed ( Figure 6 C), after 24 hours, the spores were lysed, and the cell content was extravasated ( Figure 6 D). However, the spores in the control (water, buffer) did not change ( Figure 7 ).

Embodiment 3

[0065] Embodiment 3: antibacterial protein PBR 1 Determination of antibacterial spectrum

[0066] Judging by the inhibitory effect on 11 fungal strains, the isolated antimicrobial protein PBR 1 It has a broad-spectrum inhibitory effect on pathogenic bacteria (see Table 1). It not only has a good inhibitory effect on rice blast fungus, a major airborne disease, but also has a good inhibitory effect on tomato Botrytis cinerea, wheat take-all, and pomegranate wilt. Good inhibitory effect. These pathogens include the Fusarium genus of the subphylum Deuteromycetes ( Fusarium ), the Ascomycota genus Longorosonia ( Ceratocystis ), and the genus Botrytis ( Botrytis ).

[0067]

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[0069] Yunnan Agricultural University

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

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[0076] PatentIn version 3.3

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[0078] 1

[0079] 239

[0080] PRT

[0081] Bacillus am...

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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 biopesticides, in particular to a new antibacterial protein from microorganisms PBR1 and apply. 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 ( Bacillus subtilis ) is a Gram-positive rod-shaped bacterium widely distributed in various living environments. It can produce endophytic spores and has strong heat and stress resistance. It is...

Claims

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Application Information

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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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