Screening method and application of Raoultella planticola strain SRPG-4 producing ACC (1-aminocyclopropane-1-carboxylic acid) deaminase activity

A deaminase and strain technology, which is applied in the field of agricultural biology to achieve the effects of strong environmental adaptability, stable effect, good ecological benefits and social benefits

Inactive Publication Date: 2015-03-25
SHIHEZI UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no report of a fast and operable screening metho

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  • Screening method and application of Raoultella planticola strain SRPG-4 producing ACC (1-aminocyclopropane-1-carboxylic acid) deaminase activity
  • Screening method and application of Raoultella planticola strain SRPG-4 producing ACC (1-aminocyclopropane-1-carboxylic acid) deaminase activity
  • Screening method and application of Raoultella planticola strain SRPG-4 producing ACC (1-aminocyclopropane-1-carboxylic acid) deaminase activity

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Experimental program
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Effect test

Embodiment 1

[0020] Embodiment 1: have the screening and identification of ACC deaminase bacterial strain

[0021] 1. DF liquid medium:

[0022] Component one: H 3 BO 3 10mg, MnSO 4 ·H 2 O 11.19 mg, ZnSO 4 ·7H 2 O 124.6 mg, CuSO 4 ·5H 2 O 78.22 mg, MoO 3 Dissolve more than 10mg in 100mL sterilized distilled water and store at -4°C;

[0023] Component two: FeSO 4 ·7H 2 O (100mg) dissolved in 10ml sterilized distilled water, stored at -4°C;

[0024] Take 0.1 mL each of the above component 1 and component 2 solutions, and then add KH 2 PO 4 4.0g, Na 2 HPO 4 6.0g, MgSO 4 ·7H 2O 0.2g, glucose 2.0g, gluconic acid 2.0g, citric acid 2.0g, (NH 4 ) 2 SO 4 2.0g, H 2 O 1000ml.

[0025] 2. ADF medium:

[0026] The (NH 4 ) 2 SO 4 Replace it with ACC. ACC cannot be sterilized by high temperature. When using it, first make ACC into a mother solution with a concentration of 0.5M, sterilize it through a 0.2μm bacterial filter, and add the mother solution at 6mL / L to the DF aft...

Embodiment 2

[0055] Example 2: Culture characteristics of R. planticola SRPG-4

[0056] Pick a single colony from the ADF plate, re-inoculate the screened strains in DF and ADF culture medium, measure the optical density at 600nm, and draw the growth curve of each strain.

[0057] figure 1 It shows that the strain SRPG-4 can grow normally in both DF and ADF medium. At the same time, the cell density of SRPG-4 in ADF medium is lower than that in DF medium, but the growth trend is basically the same, which also shows that the strain SRPG -4 can make good use of ACC as nutrients for growth.

Embodiment 3

[0058] Embodiment 3: the salt tolerance test of R.planticola SRPG-4

[0059] Using the shake flask culture growth curve method, different NaCl concentrations (0.5%, 1.0%, 2.0%, 4.0%, 6.0% and 8.0%) were set to measure the salt tolerance of the SRPG-4 strain. image 3 As shown, the results show that when the salt concentration does not exceed 4.0%, the time to enter the plateau is more consistent, indicating that the strain SRPG-4 has good tolerance, but when the salt concentration exceeds 2%, the bacterial biomass lower. SRPG-4 grew very slowly when the salt concentration was 6.0%, and the bacterial concentration was also very low, and it entered a plateau after 24 hours. When the salt concentration was 8.0%, the bacteria almost stopped growing. It shows that the strain SRPG-4 will not affect the survival of the strain when the soil salinity does not exceed 2.0%. Because the salt content of secondary salinized soil or salinized soil capable of growing crops is generally wit...

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Abstract

The invention provides a screening method (comprising four steps of gathering, preliminary screening, secondary screening and identifying) and an application of a Raoultella planticola strain SRPG-4 producing 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity, wherein the strain is identified as Raoultella planticola SRPG-4 with a preservation number of CGMCC No.9392. The strain SRPG-4 obtained by the method provided by the invention can grow well in an ADF (Augmented Dickey-Fuller) culture medium, and the ACC specific enzymatic activity of the strain reaches 0.832+/-0.032U/mg. Under a salinization condition, the application of an SRPG-4 bacterium solution has an obvious effect of improving the germination rate, the planting percent and the biomass accumulation of cotton, and can adjust the output of an auxin (IAA namely indole-3-acetic acid) and relieving the generation of ethylene (Eth) and a cytokinin (ABA), thereby relieving salt stress. The strain screened by the method provided by the invention is derived from northwest arid areas, has strong adaptive capacity to environment and has advantages of stable application effect and the like, so that the strain used for developing a microorganism bacterium agent has a wide application prospect to solve the problem of soil salinization.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology. More specifically, the present invention relates to a screening method and application of a vegetative Raoult strain SRPG-4 producing ACC deaminase activity and the salt-dissolving and growth-promoting effect of the strain, which was preserved on June 30, 2014 In the General Microbiology Center of China Microbiological Culture Collection Management Committee, the preservation number is CGMCC No.9392, and the preservation address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing. Background technique [0002] Northwest China is the largest cotton production area and high-quality commercial cotton base in China. However, in order to solve the secondary salinization of soil caused by the long-term use of drip irrigation under plastic film in arid and semi-arid areas of cotton cultivation, crops lack seedlings, broken ridges, and yields The decline has seriously restricted...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/02A01P21/00C12R1/01
CPCC12N1/02C12N1/205C12R2001/01
Inventor 武占省何艳慧李春凃亮樊艳爽
Owner SHIHEZI UNIVERSITY
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