Pseudomonas strain and applications thereof
A technology for pseudomonas and microbial inoculants, which is applied to pseudomonas and its application fields, can solve the problems of few types of plant growth-promoting strains, reduced functions of soil microbial flora diversification, and insufficient chemical fertilizer resources, etc. Reduce environmental pollution and threats to human health, broad application prospects and market potential, and the effect of high-efficiency plant growth-promoting effects
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Embodiment 1
[0022] Embodiment 1, Pseudomonas Pseudomonas sp . Isolation and Screening of N21
[0023] 1.1. Sampling
[0024] The rhizosphere soil of Hainan Fenban was screened, and the soil was coastal sandy loam, and the rhizosphere soil was put into a clean and sterilized kraft paper bag prepared in advance, and brought 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 obtained.
[0027] 1....
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 the 10% LB medium as the control, set three repetitions for each treatment, culture on a shaker at 30°C and 180rpm for 3 days, centrifuge the fermentation broth at 12000rpm for 5min, take the supernatant, and use molybdenum-antimony antimony at a wavelength of 700nm Determination of soluble phosphorus content by colorimetry.
[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 col...
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