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Serratia marcescens, and screening method and application thereof

A technology of Serratia marcescens, screening method, applied in the field of microorganisms, can solve problems such as drug resistance, environmental pollution and human health threats

Inactive Publication Date: 2014-10-22
SHAANXI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] Plant diseases have always been a serious threat to agricultural production. The prevention and control of plant diseases by humans can be traced back thousands of years ago. In the early 1940s, a large number of organic synthetic pesticides were produced and the diseases were effectively controlled. However, drug resistance, environmental pollution and human health have been affected. Threat, the three evils

Method used

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  • Serratia marcescens, and screening method and application thereof
  • Serratia marcescens, and screening method and application thereof
  • Serratia marcescens, and screening method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Serratia marcescens named Serratia marcescens subsp.PGPRB, the preservation number is CCTCC NO: M2013617;

[0035] The 16S rDNA nucleotide sequence of Serratia marcescens subsp.PGPRB is:

[0036]

[0037] The rpoB nucleotide sequence of Serratia marcescens subsp.PGPRB is:

[0038]

[0039]

[0040] The screening method of the above-mentioned Serratia marcescens subsp.PGPRB consists of the following steps:

[0041] 1. Strain isolation

[0042] Take 1g of the tobacco rhizosphere soil sample from Yongding County, Fujian Province, put it in a centrifuge tube, add 10g of sterile water to make a soil suspension, place it in a vortex shaker for 15 minutes, let it stand for 5 minutes, and absorb the supernatant Add 10 times, 100 times, 1000 times and 10000 times the volume of sterile water to make a bacterial suspension, and take different concentrations of the bacterial suspension and spread them on the KB solid medium on the flat glass (KB medium 1L contains pepton...

Embodiment 2

[0062] Serratia marcescens subsp.PGPRB that adopts embodiment 1 to screen and isolate is prepared the method steps of prodigiosin prodigiosin as follows:

[0063] 1. Activation of strains

[0064] Use an inoculating loop to dip Serratia marcescens subsp.PGPRB into a line on the LB solid slope, and culture overnight in a 28°C incubator.

[0065] 2. Seed preparation

[0066] Take 50ml of seed medium (peptone 20g / L, MgSO 4 .7H 2 O1.5g / L, K 2 HPO 4 .3H 2 O2g / L, glycerol 10ml / L, pH7.0), put in a 250ml triangular flask, take one slant of the activated strain, add 0.5ml sterile water to make a bacterial suspension, pour it into the seed medium, and keep it at constant temperature Cultivate overnight on a culture shaker with a rotation speed of 200 r / min and a temperature of 28° C. to obtain a seed culture solution.

[0067] 3. Fermentation culture

[0068] Take 200mL of fermentation medium (peptone 20g / L, MgSO 4 .7H 2 O1.5g / L, K 2 HPO 4 .3H 2 O2g / L, glycerol 10ml / L, pH 7...

Embodiment 3

[0078] The method steps for preparing the lipopeptide serrawettin W2 from Serratia marcescens subsp.PGPRB screened and isolated in Example 1 are as follows:

[0079] The strain activation step 1, seed preparation step 2, fermentation and cultivation step 3, lipopeptide serrawettin W2 extraction step 4 are the same as the preparation of prodigiosin.

[0080] 5. Separation and purification of lipopeptide serrawettin W2

[0081] (1) Preliminary separation and purification on silica gel column

[0082] The crude product was dissolved in ethyl acetate, separated by silica gel column chromatography (200-300 mesh), eluted with a 1:1 mixture of petroleum ether and ethyl acetate by volume, and then eluted with ethanol to obtain lipopeptide serrawettin Mixture of W2.

[0083] (2) Purify and identify the mixture containing lipopeptide serrawettin W2 with LC-8A semi-preparative high-performance liquid chromatograph from Japan Shimadzu Company. The semi-preparative column of LC-8A chroma...

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Abstract

A serratia marcescens is named as Serratia marcescens subsp. PGPRB has a preservation number of CCTCC No:M2013617. A screening method of the Serratia marcescens subsp. PGPRB comprises the steps of strain separation and identification. The Serratia marcescens subsp. PGPRB can be used for preparing prodigiosin and can also be used for preparing lipopeptide serrawettin W2.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to Serratia marcescens. Background technique [0002] Plant diseases have always been a serious threat to agricultural production. The prevention and control of plant diseases by humans can be traced back thousands of years ago. In the early 1940s, a large number of organic synthetic pesticides were produced and the diseases were effectively controlled. However, drug resistance, environmental pollution and human health have been affected. Threats are the three evils that people often say. The emergence of the three pests has aroused people's widespread concern, so the research, development and application of biological control has been put on the agenda, and has been valued by governments of all countries. Plant growth promoting rhizobacteria (PGPR) are some microorganisms that can promote and protect plant growth in the rhizosphere of plants. Among them, Pseudo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12Q1/04C12P17/16C12P21/02C12R1/43
Inventor 孙燕向照举李治刘清梅
Owner SHAANXI NORMAL UNIV
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