Bacillus P75 for improving Pisha sandstone soil fertility and application thereof
A bacillus and soil fertility technology, applied in the direction of application, bacteria, fertilization device, etc., can solve the problems of water shortage, difficult vegetation growth, thin slope soil layer, etc., to improve soil fertility, promote growth, increase plant biological volume effect
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Embodiment 1
[0030] The plant growth-promoting bacterium P75 strain (CCTCC NO: M2018690) of the present invention for improving the fertility of the sandstone soil in Inner Mongolia is isolated and purified from the roots of alfalfa growing in the sandstone area of Inner Mongolia. The isolation and identification method is as follows:
[0031] Rinse the alfalfa root with hot deionized water, disinfect it with 75% alcohol and 2.5% NaClO, and soak it several times with sterile water. Put 0.1 g of the surface-sterilized root into a sterile mortar and grind until homogenized, add 1 mL of sterile water to mix, and then put it in a constant temperature water bath at 85°C for 15 minutes. Take 0.1mL of the suspension and spread it on the LB solid medium containing 5mg / L Cd, and at the same time take 0.1mL of the sterile water from the last immersion and spread it on the LB solid medium to test whether the surface of the sample is thoroughly disinfected. After culturing at 28°C for 72 hours, if n...
Embodiment 2
[0033] Activation of embodiment 2 bacterial strain P75 and bacterial suspension preparation
[0034] The P75 (CCTCC NO: M 2018690) slant strain was inoculated in LB solid medium (tryptone 10.0 g, yeast extract 5.0 g, Nacl 10.0 g, distilled water 1000 ml, agar 20 g, pH 7.0), and cultured at 30°C for 1 day. Then select a plump, viscous P75 single colony and inoculate it in LB liquid medium (tryptone 10.0g, yeast extract 5.0g, Nacl 10.0g, distilled water 1000ml, pH 7.0), 30°C, 160rmp shaking culture for 18-20h, so that the cells The number reaches more than 1 billion CFU / ml.
Embodiment 3
[0035] The ability of embodiment 3 bacterial strain P75 to produce IAA
[0036] According to the determination method of Gordon and Weber (1951), LB liquid medium was divided into test tubes, 4 mL per tube, and after high-temperature sterilization at 121 ° C, 1 mL of 2.5 mg / mL tryptophan solution sterilized by filtration was added to make the color in the medium The final concentration of amino acid was 0.5 mg / mL. The strains were inoculated in the above-mentioned medium, and cultured in a shaker at 30°C for 3 days. The fermentation broth was centrifuged at 12000r / min for 5min, 1mL of the supernatant was taken, 50μL of 10mM orthophosphoric acid was added, and 2mL of Sackowski's chromogenic reagent was added, mixed thoroughly, the color was developed at 25°C for 30min in the dark, and the absorbance value was measured at a wavelength of 530nm. The sterile medium was treated the same as above and set to zero as a control. Use the IAA standard solutions with concentrations of 0...
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