Phosphorous-dissolving pseudomonas strain and application thereof

A technology of Pseudomonas and microbial strains, applied in the field of microorganisms, to achieve the effects of promoting seed germination, increasing plant biomass, and promoting plant growth

Active Publication Date: 2019-03-08
四川大宇中和生物质能源科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, research in this area is emerging in an endless stream, but the bottleneck of high-efficiency strain breeding h...

Method used

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  • Phosphorous-dissolving pseudomonas strain and application thereof
  • Phosphorous-dissolving pseudomonas strain and application thereof
  • Phosphorous-dissolving pseudomonas strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] The screening of embodiment 1 target bacterial strain

[0032] (1) Preparation of culture medium

[0033] Beef extract peptone solid medium: peptone 10g, beef extract 3g, sodium chloride 5g, agar 18g, distilled water 1000mL, pH=7.0, sterilized at 121°C for 20min.

[0034] Beef extract peptone liquid medium: peptone 10g, beef extract 3g, sodium chloride 5g, distilled water 1000mL, pH=7.0, sterilized at 121°C for 20min.

[0035] CCM medium: NH 4 NO 3 1g; MgSO 4 ·7H 2 O 0.2g; KH 2 PO 4 0.2g; Mannitol 5.0g; K 2 HPO 4 0.8g; CaCl 2 2H 2 O 0.06g; sucrose 5.0g; NaMoO 4 2H 2 O 2.5mg; yeast powder 0.1g; lactic acid 0.5mL; NaCl 0.1g; 1.64% sodium iron ethylenediaminetetraacetate (Na·Fe·EDTA) 4mL; total volume 1000mL (make up with distilled water); pH=7.0. Note: MgSO 4 ·7H 2 O, CaCl 2 2H 2 O and Na·Fe·EDTA are sterilized separately, otherwise precipitation will occur.

[0036] Salkowski Reagent: Accurately Weigh FeCl 3 4.5g, dissolved in 10.8M H 2 SO 4 Afte...

Embodiment 2

[0048] The identification of embodiment 2 Pseudomonas KSX1-1

[0049] The Pseudomonas KSX1-1 with the strongest IAA secretion ability screened in Example 1 was preserved on the slant of beef extract peptone solid medium, and a series of physiological and biochemical identifications were carried out. The colony morphology of the strain is shown in figure 1 Shown, Gram stain see figure 2 Shown, growth curve see image 3 Shown, SEM see Figure 4 As indicated, DNA extraction, amplification and sequencing of 16SrDNA were performed.

[0050] 16S rDNA was amplified using primers 27F and 1492R, and the primer sequences were as follows:

[0051] 27F: 5-AGAGTTTGATCCTGGCTCAG-3

[0052] 1492R: 5-GGTTACCTTGTTACGACTT-3

[0053] The PCR amplification conditions were 94°C for 3min; 30 cycles of 94°C for 30s, 55°C for 30s, and 72°C for 90s; 72°C for 10min.

[0054] The PCR amplification product is sequenced, and the sequence result (see SEQ ID NO: 1) is used to make a phylogenetic tree,...

Embodiment 3

[0055] The ACC deaminase activity analysis of embodiment 3 Pseudomonas KSX1-1

[0056] (1) Preparation of culture medium

[0057] DF medium: MnSO 4 ·7H 2 O 0.2g, KH 2 PO 4 4.0g, Na 2 HPO 4 6.0g, citric acid 2.0g, glucose 2.0g, sodium gluconate 2.0g, (NH 4 ) 2 SO 4 2.0g, take 0.1mL each of component 1 and component 2 solutions, H 2 O 1000 mL, pH=7.2. (preparation of component one: the CuSO 4 ·5H 2 O 78.22 mg, MoO 3 10 mg, H 3 BO 3 10mg, ZnSO 4 ·7H 2 O124.6mg, MnSO 4 ·H 2 O 11.9mg, dissolved in 100mL sterile distilled water. Preparation of component two: FeSO 4 ·7H 2 O100mg, dissolved in 10mL of sterilized distilled water, fully shaken. Note: Components 1 and 2 are stored at -4°C for future use. )

[0058] ADF medium: Dissolve ACC in ultrapure water, filter and sterilize with a bacterial filter, add to the medium that does not contain (NH 4 ) 2 SO 4 And in the pre-sterilized DF medium, pH=7.2. The final concentration of ACC added was 3.0mmol / L. ...

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Abstract

The invention relates to the field of microbial technology, particularly to a phosphorous-dissolving pseudomonas strain and application thereof. according to the technical scheme, the phosphorous-dissolving pseudomonas strain is preserved in CGMCC (China General Microbiological Culture Collection Center) with a preservation number of CGMCC No. 15915. The phosphorous-dissolving pseudomonas strain is efficient in IAA (indoleacetic acid) secretion to promote growth of crops and increase the biomass of plants, and meanwhile, can grow by taking insoluble phosphates as the phosphorous source and release soluble phosphorous. Besides, the phosphorous-dissolving pseudomonas strain can promote seed germination and improve the root length, stem length, dry weight and wet weight of seedlings, and is applicable to preparing microbial fertilizers.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a strain of Pseudomonas phospholytica and its application. Background technique [0002] Indole-3-acetic acid (IAA) is widely used in agriculture because it participates in the regulation and control of physiological and biochemical processes such as cell growth and division, differentiation of tissues and organs, and transport of assimilates in plants. In addition, indole acetic acid can also directly or indirectly improve the stress resistance of plants, so it has begun to be applied to ecological restoration, but IAA is unstable in the natural environment, easy to degrade, and artificial synthesis is expensive, so adding IAA from exogenous sources is practical. There are certain limitations in application. Several studies have shown that a class of microorganisms that promote plant growth in soil, called plant growth promoting rhizobacteria (PGPR), can secrete IAA, whi...

Claims

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Application Information

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IPC IPC(8): C12N1/20C05G3/00C12R1/38
CPCC05D1/00C05G3/00C12N1/205C12R2001/38C05D9/00C05D9/02C05F3/00C05F5/002C05F11/08
Inventor 刘强
Owner 四川大宇中和生物质能源科技有限公司
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