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A Strain of Pseudomonas and Its Application

A technology of Pseudomonas and microbial agents, applied in Pseudomonas and its application fields, can solve the problems of fewer types of plant growth-promoting strains, reduced diversification of soil microbial flora, and insufficient chemical fertilizer resources, etc., to achieve Reduce environmental pollution and threats to human health, broad application prospects and market potential, and the effect of high-efficiency plant growth-promoting effects

Active Publication Date: 2019-10-01
江苏联海生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, my country's annual consumption of chemical fertilizers is as high as 90 million tons, but relying on a large amount of chemical fertilizers has also brought about a series of problems: 1) my country's chemical fertilizer resources are insufficient, and it is heavily dependent on imports, which is unsustainable; 2) The excessive use of chemical fertilizers throughout the year has caused The reduction of soil organic matter, soil compaction, and the diversification and function of soil microbial flora have seriously hindered the sustainable development of agriculture in my country; 3) leading to serious environmental pollution (such as soil acidification, groundwater pollution), and the destruction of the ecological environment
[0004] At present, the plant growth-promoting strains have the following problems in the development and application of bio-organic fertilizers: 1) The growth-promoting function is single, and some plant growth-promoting strains only have one growth-promoting function and do not have comprehensive plant growth-promoting characteristics; 2) There are few types of plant growth-promoting strains currently used in the research and development of bio-organic fertilizers, most of which are Bacillus, which are not targeted; 3) The activity of the strains is unstable, and the duration of action is short after mixing with organic fertilizers; 4) The environmental adaptability of the strain is weak, and the strains isolated and screened from different samples and sampling locations do not have wide applicability
Therefore, single-function bio-organic fertilizers can no longer meet the requirements of modern agricultural development, and adding functional microorganisms such as disease resistance, growth promotion, and fertilizer utilization efficiency has become the development trend of new bio-organic fertilizers.

Method used

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  • A Strain of Pseudomonas and Its Application
  • A Strain of Pseudomonas and Its Application
  • A Strain of Pseudomonas and Its Application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1, Pseudomonas Pseudomonas sp . Isolation and Screening of N21

[0023] 1.1. Sampling

[0024] Screening was carried out from the rhizosphere soil of Hainan Fenban, which was coastal sandy loam, and put the Fenba rhizosphere soil into a clean and sterilized kraft paper bag prepared in advance, and brought it back to the laboratory.

[0025] 1.2. Primary screening and purification of strains

[0026] Weigh 1 g of rhizosphere soil sample, place it in a test tube filled with 9 ml of sterile water, shake it evenly to make soil bacteria suspension, and dilute it to 10 -4 、10 -5 、10 -6 The soil suspension was spread on TSA solid medium (tryptone 1.2~1.5%, soybean peptone 0.5~0.8%, sodium chloride 0~0.5%, agar powder 1.5~2.0%, distilled water 1L, pH 7.3±0.2.) placed upside down in a biochemical incubator at 30°C for 3 days. The colonies on the plate were randomly picked, and repeatedly streaked and purified on TSA solid medium until a single colony was obtain...

Embodiment 2

[0049] Embodiment 2, Pseudomonas Pseudomonas sp . Identification of N21

[0050] A series of physiological and biochemical identifications were carried out on the pure culture strains obtained after the above primary screening and purification, and the DNA of the strains was extracted, 16S rDNA sequence was amplified and sequenced.

[0051] Pseudomonas Pseudomonas sp . The main biological traits of N21 are as follows:

[0052] (1) Morphological characteristics of the bacteria: Gram staining is negative, short rod-shaped, without spores.

[0053] (2) Morphological characteristics of colonies: On TSA agar medium, the colonies are round, off-white, raised, and the surface of the colonies is moist and smooth (such as figure 2 shown).

[0054] (3) Physiological and biochemical characteristics: aerobic, positive for catalase, oxidase, nitrate reduction, gelatin, arginine dihydrolase, and indole; citrate, urease, H 2 S reaction negative, can assimilate D-galactose, D-glucuron...

Embodiment 3

[0056] Embodiment 3, Pseudomonas Pseudomonas sp . Determination of Phosphorus and Potassium Solubilizing Activity of N21

[0057] 3.1. Determination of Phosphorus Solubilizing Ability

[0058] Phosphorus dissolved amount: will Pseudomonas sp . N21 was cultured overnight in LB liquid medium at 30°C and 180rpm, and the bacterial concentration was diluted to 1×10 with LB medium 8 Individual / ml, with 10% inoculum size into the inorganic phosphorus liquid medium. Take 10% LB medium as the control, set three repetitions for each treatment, culture on a shaker at 30°C, 180rpm for 3 days, centrifuge the fermentation broth at 12000 rpm for 5min, take the supernatant, and use molybdenum antimony at a wavelength of 700nm Determination of soluble phosphorus content by anti-colorimetric method.

[0059] The specific method is as follows: Take 200 μL of the fermentation supernatant in a colorimetric tube, add 10 mL of distilled water, add two drops of 2,4 (or 2,6 ) dinitrophenol color...

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Abstract

The invention discloses a strain of Pseudomonas, which is preserved in the China Center for Type Culture Collection, with the preservation number CCTCCNO: M2016180, the preservation date is April 7, 2016, and the preservation address is Wuhan-Wuhan University, China. The strain has the characteristics of fast growth, strong strain activity, comprehensive growth-promoting function, and stable growth-promoting effect; the developed bio-organic fertilizer can greatly reduce the environmental pollution caused by chemical fertilizers and the threat to human health; it can not only improve soil Fertility, fertilizer utilization rate, quality of agricultural products, and promote the sustainable development of agriculture, has broad application prospects and market potential.

Description

technical field [0001] The invention relates to the technical field of microorganism application, in particular to a strain of pseudomonas and its application. Background technique [0002] my country's cultivated land area is about 1.82 billion mu, but low-yield fields account for 41.6% of my country's cultivated land. The application of chemical fertilizers is the most important measure to increase the yield of medium-low-yield fields and ensure my country's grain output. At present, my country's annual consumption of chemical fertilizers is as high as 90 million tons, but relying on a large amount of chemical fertilizers has also brought about a series of problems: 1) my country's chemical fertilizer resources are insufficient, and it is heavily dependent on imports, which is unsustainable; 2) The excessive use of chemical fertilizers throughout the year has caused The reduction of soil organic matter, soil compaction, and the diversification and function of soil microbial...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/02A01P21/00C05G3/00C12R1/38
CPCA01N63/10C05F5/008C05G3/00C12N1/205C12R2001/38C05F7/00C05F11/08
Inventor 唐波顾佳宇董存明陈赛王贵云徐清辉
Owner 江苏联海生物科技有限公司
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