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A kind of Pseudomonas phospholytica and application thereof

A technology of Pseudomonas and monas, which is applied to Pseudomonas phosphate and its application field, can solve the problems of increasing phosphorus release, inefficient absorption and utilization, eutrophication of water bodies, etc., and achieves the promotion of plant growth. , the effect of improving the effectiveness

Active Publication Date: 2021-07-27
GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The total phosphorus content in my country's soil is about 40-2500mg / kg, but the average content of available phosphorus that can be directly absorbed by plants is less than 10mg / kg
However, the seasonal utilization rate of the externally applied phosphate fertilizer is only 10% to 15%, and most of the phosphorus is mixed with Ca in the soil. 2+ , Fe 3+ 、Al 3+ and other cations combine to form insoluble orthophosphate compounds, which cannot be absorbed by plants
The long-term large-scale application of phosphorus fertilizer in agricultural production and the inefficient absorption and utilization of it by crops, on the one hand, continuously increase the release of phosphorus to the water environment, resulting in serious water eutrophication. On the other hand, some heavy metal elements in phosphorus fertilizer It also accumulates in the soil and surrounding water with the application, posing a serious threat to the ecological environment and sustainable agricultural development

Method used

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  • A kind of Pseudomonas phospholytica and application thereof
  • A kind of Pseudomonas phospholytica and application thereof
  • A kind of Pseudomonas phospholytica and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] The isolation of embodiment 1 phosphate-dissolving bacterial strain PD171

[0017] Heavy metal-contaminated soil samples were collected from the Dabaoshan mining area in Shaoguan City, Guangdong Province. Weigh 10g of soil sample into 90mL sterilized double-distilled water, shake at 180r / min for 30min, and then use 10-fold gradient to gradually dilute the soil solution to 10 -1 ~10 -5 100 μL of bacterial suspensions with different dilution gradients were taken and spread on different PVK solid medium plates. After culturing at 30°C for 3 days, the strains with different morphologies that produced transparent circles were picked and placed on TSA medium for isolation and purification, so as to finally isolate and obtain strain PD171.

[0018] PVK solid medium: glucose 10g, Ca 3 (PO 4 ) 2 5g, (NH 4 ) 2 SO 4 0.5 g, yeast extract 0.5 g, NaCl 0.3 g, KCl 0.3 g, MgSO 4 ·7H 2 O 0.3g, FeSO 4 ·7H 2 O 0.03g, MnSO 4 ·H 2 O 0.03g, agar 15.0g distilled water 1000mL, pH ...

Embodiment 2

[0020] Classification and Identification of Example 2 Bacterial Strain PD171

[0021] Phenotypic characteristics: The strain PD171 was inoculated on TSA medium by three-section line method, and the colony morphology was observed after culturing at 30°C for 3 days. The cultured colonies were taken for Gram staining, and samples for transmission electron microscopy were prepared, and the cell morphology of the strains was observed with a Hitachi H7650 transmission electron microscope. For the test methods of fluorescein, pyocyanin, oxidase, catalase, methyl red, and hydrolyzed starch, refer to the "Common Bacteria Identification Manual", nitrate reduction, indole, β-galactosidase, hydrolyzed gelatin, hydrolyzed seven Yeling, arginine dihydrolase and other tests use API 20NE reagent strips (Mérieux Bio, France). For the hemolysis test, blood plates were inoculated and cultured at 30°C for 7 days to observe whether there were hemolysis circles.

[0022] Strain PD171 is light yel...

Embodiment 3

[0029] Example 3 Phospholytic ability analysis of Pseudomonas phospholyticum PD171 in NBRIP medium

[0030] Use R2A liquid medium to activate Pseudomonas phospholytica PD171, cultivate and grow to the logarithmic phase at 30°C and inoculate it into an Erlenmeyer flask containing 40mL NBRIP liquid medium at 30°C The shaker was shaken at 180r / min, and the sterilized R2A liquid medium not inoculated with Pseudomonas phospholytica PD171 was added as a control, and 4 replicates were set for each treatment. From the third day, samples were taken every 24 hours and monitored until shaking culture for 7 days. The samples collected at different times were first centrifuged at 12000r / min, and the supernatant was filtered and sterilized with a 0.02 μm sterile filter membrane, and then stored in a freezer at -30°C until determination. The content of water-soluble phosphorus (ws-p) in samples collected at different times was measured by molybdenum-antimony anti-colorimetry, and the pH val...

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Abstract

The invention discloses a pseudomonas phospholytica and application thereof. Pseudomonas phospholytica PD171 was deposited in Guangdong Microbial Culture Collection Center (GDMCC) on October 09, 2020, address: 5th Floor, Building 59, Compound, No. 100 Xianlie Middle Road, Guangzhou City, and the preservation number is: GDMCC No: 61233. The Pseudomonas phosphate-solubilizing bacteria PD171 disclosed by the present invention is a new species of Pseudomonas genus, which can efficiently dissolve insoluble calcium phosphate, and the phosphorus-dissolving amount after 4 days of cultivation is as high as 607.37 mg / L, and the phosphorus-dissolving performance is significantly excellent. in most of the reported phosphate-solubilizing strains. Therefore, the Pseudomonas phospholyticum PD171 disclosed in the present invention can be used to increase the availability of phosphorus in soil, and has important application potential in promoting plant growth, preparing microbial agents and biological fertilizers, and the like.

Description

Technical field: [0001] The invention belongs to the technical field of agricultural microorganisms, and in particular relates to a pseudomonas phospholytica and an application thereof. Background technique: [0002] Phosphorus is an essential macroelement for plant growth and the most important nutrient limiting factor in agricultural production. The total phosphorus content in my country's soil is about 40-2500mg / kg, but the average content of available phosphorus that can be directly absorbed by plants is less than 10mg / kg. However, the seasonal utilization rate of the externally applied phosphate fertilizer is only 10% to 15%, and most of the phosphorus is mixed with Ca in the soil. 2+ , Fe 3+ 、Al 3+ Such cations combine to form insoluble orthophosphate compounds, which cannot be absorbed by plants. The long-term large-scale application of phosphorus fertilizer in agricultural production and the inefficient absorption and utilization of it by crops, on the one hand, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C05F11/08C12R1/38
CPCC05F11/08C12N1/20C12N1/205C12R2001/38
Inventor 冯广达羊宋贞臧毅朱红惠姚青陈家舒
Owner GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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