Pseudomonas potassium-solubilizing bacteria and application thereof
Pseudomonas, a technology for promoting plant growth, applied in the field of microorganisms, can solve problems such as poor colonization ability, unstable yield increase effect, and insignificant potassium solution effect, achieve enhanced soil water retention performance, and be beneficial to crop seedling raising and yield increase , good effect of soil potassium solubilization
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Embodiment 1
[0036] The screening of embodiment 1 potassium solution bacterial strain
[0037] (1) Preparation of culture medium
[0038] 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.
[0039] 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.
[0040] Potassium-solubilizing bacteria screening solid medium: 5g sucrose, MgSO 4 ·7H 2 O 0.5g, FeCl 3 0.005g, CaCO 3 0.1g, potassium feldspar 2g, agar 20g, pH=7.4, deionized water 1000mL.
[0041] Potassium-solubilizing bacteria screening liquid medium: 5g sucrose, MgSO 4 ·7H 2 O 0.5g, FeCl 3 0.005g, CaCO 3 0.1g, potassium feldspar 2g, pH=7.4, deionized water 1000mL.
[0042] (2) Screening of Pseudomonas
[0043] Select the native vegetation in the primordial soil area where the erosion is strong and the nutrients are poor (the ...
Embodiment 2
[0050] Physiological and biochemical identification of embodiment 2 potassium-solubilizing bacterial strains
[0051] The Pseudomonas KSX1-2 with the strongest potassium-dissolving 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 as follows: image 3 As shown, Gram staining as Figure 4 As shown, the scanning electron microscope as Figure 5 As shown, the growth curve is as Figure 6 As indicated, DNA extraction, amplification and sequencing of 16SrDNA were performed.
[0052] 16S rDNA was amplified using primers 27F and 1492R, and the primer sequences were as follows:
[0053] 27F: 5-AGAGTTTGATCCTGGCTCAG-3
[0054] 1492R: 5-GGTTACCTTGTTACGACTT-3
[0055] 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.
[0056] The PCR amplificati...
Embodiment 3
[0057] The ACC deaminase activity assay of embodiment 3 Pseudomonas KSX1-2
[0058] (1) Preparation of culture medium
[0059] 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: CuSO 4 ·5H 2 O 78.22mg, MoO 3 10mg,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 was dissolved in 10mL of sterilized distilled water and shaken fully. Store components 1 and 2 at -4°C for future use.
[0060] 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.
[0061] (2) Determ...
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