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Rhodopseudomonas palustris strain, bacterial agent, preparation method of bacterial agent, extracelluar protein as well as extraction method and application of extracelluar protein

A technology of Rhodopseudomonas palustris and Rhodopseudomonas, which is applied to the preparation of its bacterial agent and bacterial agent, and the field of Rhodopseudomonas paluscans strains, which can solve the problems that antiviral agents have not been seen, and achieve bluntness in vitro Good chemical effect, high thermal stability, and short process

Active Publication Date: 2015-03-11
HUNAN PLANT PROTECTION INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, protein-based antiviral agents have yet to be found

Method used

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  • Rhodopseudomonas palustris strain, bacterial agent, preparation method of bacterial agent, extracelluar protein as well as extraction method and application of extracelluar protein
  • Rhodopseudomonas palustris strain, bacterial agent, preparation method of bacterial agent, extracelluar protein as well as extraction method and application of extracelluar protein
  • Rhodopseudomonas palustris strain, bacterial agent, preparation method of bacterial agent, extracelluar protein as well as extraction method and application of extracelluar protein

Examples

Experimental program
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Effect test

Embodiment 1

[0035] A strain of Rhodopseudomonas palustris is Rhodopseudomonas palustris LY-6 ( Rhodopseudomonas palustris LY-6), deposited in the China Type Culture Collection, the deposit number is CCTCC M 2014525, the depositary address is at Wuhan University, China, and the deposit date is October 29, 2014. The strain was named Rhodopseudomonas palustris LY-6 ( Rhodopseudomonas palustris LY-6).

[0036] See figure 1 :The Rhodopseudomonas palustris strain LY-6 of the present invention is a gram-negative bacterium. It is identified as Rhodopseudomonas palustris through physiological and biochemical identification and molecular biology. The main biological characteristics are: double-layer solid plate culture Cultivated on the base for 4 to 8 days to form red round colonies with neat and smooth edges and a diameter of 0.2 mm to 0.60 mm; cultivated in liquid medium for 7 days, it is dark red; it grows well under anaerobic and micro-oxygen conditions; physiologically Characteristics: VP r...

Embodiment 2

[0038] A Rhodopseudomonas palustris bacterial agent, using the Rhodopseudomonas palustris strain LY-6 of Example 1 as a raw material, is prepared by activation, seed culture, production culture, and centrifugation. The specific preparation method includes the following steps:

[0039] (1) Activation: The preserved species of Rhodopseudomonas palustris LY-6 strain is cultivated according to the double-layer plate culture method, and cultivated until a single red colony appears; the medium used is a photosynthetic bacteria solid medium, and the formula is: 0.1 wt% (NH 4 ) 2 SO 4 , 0.02wt% MgSO 4 , 0.5wt% NaHCO 3 , 0.05wt% K 2 HPO 4 , 0.02wt% NaCl, 0.15wt% yeast extract and 1.5wt% agar, pH=7.0.

[0040] (2) Seed culture: Put the red single colony cultured in step (1) into a 120mL serum bottle, and use photosynthetic bacteria liquid at a temperature of 30℃~32℃, light conditions of 2500Lx~4000Lx, and pH=7.0 The culture medium is cultivated to the logarithmic growth phase to obtain the b...

Embodiment 3

[0044] A Rhodopseudomonas palustris extracellular protein is extracted from the Rhodopseudomonas palustris inocula in Example 2. The molecular weight of the protein component is between 15kD and 25kD. The specific extraction method is:

[0045] (1) Ammonium sulfate precipitation-salting out: Add ammonium sulfate to the Rhodopseudomonas palustris inocula to 70% saturation at 4°C; wait for the protein precipitation to precipitate out of the fermentation broth to obtain a mixed liquor.

[0046] (2) Centrifugation and protein precipitation collection: Centrifuge the mixture prepared in step (1) at 11000 rpm for 30 minutes (centrifugation temperature is 4°C), discard the supernatant, and collect the precipitate. The pellet was resuspended in 0.01M, pH=7.0 PBS buffer to obtain a protein resuspension.

[0047] (3) Sephadex G-25 desalting: After freezing and concentrating the protein resuspension prepared in step (2) at low temperature, it is desalted using a Sephadex G-25 column at a loadin...

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Abstract

The invention provides a rhodopseudomonas palustris strain, a bacterial agent, a preparation method of the bacterial agent, extracelluar protein as well as an extraction method and application of the extracelluar protein. The rhodopseudomonas palustris strain is rhodopseudomonas palustris LY-6 which is preserved in China Center for Type Culture Collection; the preservation number is CCTCCM2014525; the bacterial agent is prepared by activating rhodopseudomonas palustris strain, culturing seeds, producing, culturing and centrifuging. The invention also provides extracelluar protein which is extracted from the rhodopseudomonas palustris bacterial agent; the extraction method comprises the steps of precipitating and salting ammonium sulfate out, centrifuging, desalting by SephadexG-25 and carrying out SephadexG-25 column chromatography isolation; an extraction method of the extracelluar protein is simple, short in process and high in extraction efficiency; the extracelluar protein has in-vitro inactivation on tobacco mosaic virus.

Description

technical field [0001] The invention belongs to the technical field of biopesticides, and in particular relates to a strain of Rhodopseudomonas palustris, its bacterial agent and a preparation method of the bacterial agent; it also relates to an extracellular protein extracted from the bacterial agent and a method for extracting the extracellular protein And its application in the control of tobacco mosaic virus. Background technique [0002] Plant viral diseases are "plant cancers" that cause serious losses to agricultural production. Plant viruses generally have systemic infectivity and obligate parasiticity, which makes the research and development of antiviral agents difficult and the stability of the effect is low. Tobacco mosaic virus (TMV) is particularly harmful during tobacco cultivation. Tobacco mosaic virus disease is one of the most important tobacco diseases in the world. It has a wide range of distribution and serious onset hazards, resulting in the decline o...

Claims

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Application Information

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IPC IPC(8): C12N1/20C07K14/195C07K1/36C07K1/30C07K1/16A01N63/02A01P1/00C12R1/01
CPCA01N63/10C07K1/16C07K1/303C07K1/36C12N1/205C12R2001/01
Inventor 刘勇张德咏苏品谭新球张松柏冯推紫彭静张卓
Owner HUNAN PLANT PROTECTION INST
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