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Pseudomonas and screening method and application thereof

A Pseudomonas and screening method technology, applied in the field of microorganisms, can solve the problem of unsatisfactory phosphorus-dissolving effect of strains, achieve good phosphorus-dissolving ability, improve root growth, and reduce pollution problems

Active Publication Date: 2020-04-24
辽宁土木启生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most of the researches are based on traditional strain screening techniques, and the phosphorus-dissolving effect of the screened strains is not satisfactory.

Method used

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  • Pseudomonas and screening method and application thereof
  • Pseudomonas and screening method and application thereof
  • Pseudomonas and screening method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 Screening and Identification of Pseudomonas sp.

[0054] 1. Screening of Pseudomonas sp. P623-9

[0055] The antagonistic antibacterial Pseudomonas P623-9 of the present invention is screened from rhizosphere soil of continuous cropping tomato seedlings.

[0056] 1.1 Preparation of Pseudomonas culture medium

[0057] 1.1.1 Phosphorous bacteria enrichment medium: take the following components by weighing: (NH 4 ) 2SO 0.5g, NaCl 0.5g, KCl 0.3g, MgSO 4 7H 2 0 0.3g, Ca(PO 4 ) 2 10g, FeS0 4 7H 2 O 0.03g, MnSO 4 4H 2 0 0.03g, 1.0g of phosphate rock powder, 10g of glucose, 1000ml of distilled water, adjust the pH to 7.0-7.5, sterilize under high temperature and high pressure at 121℃ for 20min, and set aside.

[0058] 1.1.2 Separation and screening of solid medium: (NH 4 ) 2 SO 4 0.1g, KCL0.2g, MgSO 4 7H 2 0 0.25g, NaCl 0.5g, glucose 10g, phosphate rock powder 5g, agar powder 18g, dilute to 1000ml, adjust the pH to 7.0-7.5, sterilize under high temper...

Embodiment 2

[0069] Example 2 Pseudomonas P623-9 Phosphorus Solubilizing Ability Experiment

[0070] (1) Qualitative phosphorus-dissolving effect test (primary screening): Determination of phosphorus-dissolving ability under solid culture conditions: phosphorus-dissolving circle method. Pick the purified strains, spot them into the solid medium culture dish, repeat each treatment three times, place them in an incubator at 28°C for 7 days, measure the ratio (D / d) of the diameter of the phosphorus-dissolving circle to the diameter of the colony, and determine Phosphorus dissolving ability of each strain, such as figure 1 shown.

[0071] (2) Quantitative phosphorus-dissolving ability test (molybdenum-antimony anti-colorimetric method): pick the larger phosphorus-dissolving circle in the qualitative phosphorus-dissolving effect test, and the bacterial strains with good bacterial colonies are inoculated into 100mL inorganic phosphorus liquid medium (as mentioned above) respectively. ), shake ...

Embodiment 3

[0072] Example 3 Comparative experiment of Pseudomonas P623-9 on phosphorus-solubilizing ability of facility vegetable tomato

[0073] 1. Test sample: Pseudomonas P623-9

[0074] 2. Field test crops

[0075] Facility Vegetables: Tomato Pepper Cucumber Horned Melon

[0076] 3. Test method and steps

[0077] (1) Activation of strains and preparation of bacterial solution

[0078] Pick a single colony on the Pseudomonas P623-9 plate and inoculate it in 5ml liquid LB medium, culture it with shaking at 30°C for 24h, then inoculate it in 100ml liquid LB medium respectively, culture it with shaking at 30°C for 24h, obtain the culture solution, and count ( The number of bacteria is 1.1×10 8 CFU / ml), spare.

[0079] (2) Field test

[0080] After the four kinds of crops were planted, when the second true leaf unfolded, 5 times diluted bacterial solution was added to the roots respectively, and controls were set up for comparison. At the end of a growing season (about 4 months), rh...

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Abstract

The invention provides a pseudomonas strain, which is named P623-9 and has been currently preserved in Guangdong Microbial Culture Collection Center (GDMCC). The address is Floor 5, NO.59 Building, No.100 Yard, Xianlie Middle Road, Guangzhou, the postal code is 510070, the preservation number is GDMCC NO.60806, and the preservation date is October 14, 2019. The invention also discloses applicationof the pseudomonas in improving the phosphorus utilization rate of greenhouse vegetable soil and promoting growth. According to the pseudomonas, the phosphorus dissolving amount of P623-9 is as highas 496 mg / L; the pseudomonas has good phosphorus dissolving capacity, can convert insoluble phosphatephosphorus such as fixed-state phosphorus in soil into soluble phosphorus which is easily absorbedand utilized by vegetables, reduces the application amount of a phosphate fertilizer, promotes the growth of vegetables, increases the yield by 15% or above in the application process, saves the planting cost and relieves the pollution problem.

Description

technical field [0001] The invention belongs to the technical field of microbes, in particular to pseudomonas and its screening method and application. Background technique [0002] Soil is the material basis for the sustainable development of agriculture and an important nutrient source for crop growth. Phosphorus is one of the important nutrient elements for plant growth. 74% of the cultivated land in my country is deficient in phosphorus, and the phosphorus fertilizer applied to the soil is only 5%-25 % utilization rate, most of the phosphorus has become insoluble phosphate that cannot be absorbed, and it is not easy for plants to directly absorb and utilize it. Bacterial strains, researchers use biotechnology to improve the availability of soil phosphorus to plants by breeding phosphorus-dissolving microorganisms. At present, most studies are based on traditional strain screening techniques, and the phosphorus-dissolving effect of the screened strains is not satisfactory....

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/02A01G7/06C12R1/38
CPCC12N1/20C12N1/02A01G7/06C12R2001/38C12N1/205
Inventor 吴红艳池景良于淼冯健冯敏陈飞郭玲玲王智学
Owner 辽宁土木启生物科技有限公司
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