Ultraviolet mutagenesis type pseudomonas florescens and application thereof
A Pseudomonas fluorescens, application technology, applied in the field of microorganisms, can solve the problems of Pseudomonas fluorescens single biocontrol function, single phosphorus-solubilizing effect, etc.
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Embodiment 1
[0079] Example 1: Isolation, purification and identification of ultraviolet mutagenized Pseudomonas fluorescens F1
[0080] The implementation steps of this embodiment are as follows
[0081] A. Separation
[0082] Select a decaying soil plot with wheat take-all in Zhoukou City, Henan Province, take 100 g of soil from the periphery and the center of the plot within a depth of 10-20 cm, and mix them in equal amounts.
[0083] Dissolve 10g of soil sample in 90ml of water, shake and cultivate for 20min in the QHZ-98A full-temperature shaking incubator produced by Taicang Huamei Biochemical Instrument Factory, and obtain the supernatant according to the dilution of 10 -3 、10 -4 、10 -5 For gradient dilution, 100 μl of the diluted solution was coated on 10 g / L glucose, 0.5 g / L (NH 4 ) 2 SO 4 , 0.3g / L MgSO 4 ·7H 2 O, 0.3g / L NaCl, 0.3g / L KCl, 0.03g / L FeSO 4 ·7H 2 O, 0.03g / L MnSO 4 4H 2 O, 10.0g / L Ca 3 (PO 4 ) 2 On the Montina inorganic phosphorus solid medium (pH7.0-7.5...
Embodiment 2
[0096] Example 2: Determination of Phosphorus Solubilizing Ability of Wild-type Pseudomonas fluorescens F0
[0097] Inoculate the wild-type Pseudomonas fluorescens F0 preserved on the slant medium into conventional LB liquid medium, and cultivate it to the logarithmic phase at a temperature of 30°C and a rotation speed of 180rpm, according to the volume of Montina inorganic phosphorus medium Inoculate 1% of the inoculum amount of the wild-type Pseudomonas fluorescens F0 into 100ml of sterilized Montina inorganic phosphate medium, shake and cultivate for 6 hours under the conditions of temperature 28°C and rotational speed 180rpm. Take 5ml shaking culture solution and centrifuge for 5min under the conditions of rotating speed 12000rpm and temperature 4°C, take 0.1ml supernatant and dilute to 5ml in a graduated test tube, then add 1ml constant volume solution to a 10ml colorimetric tube, and then add 2ml molybdenum-antimony anti-color developer, dilute to the scale with water, p...
Embodiment 3
[0100] Embodiment 3: The ultraviolet mutagenesis test of wild-type Pseudomonas fluorescens
[0101] A. Pick the single colony F0 of wild-type Pseudomonas fluorescens isolated from the decayed soil of wheat take-all, and insert it into NB composed of 5g / L beef extract, 10g / L peptone, and 5g / L sodium chloride In the culture medium (pH7.0-7.2), cultivate at a temperature of 28-30°C and a rotation speed of 180rpm for 16-20 hours; take 2ml of the culture solution and put it in a sterile petri dish with a diameter of 75mm, and then place it in a UV light with a power of 20W. Under the lamp, irradiate for 90s at a distance of 30cm from the ultraviolet lamp, and then dilute the bacterial solution to 10 in the dark. -6 , and then coated with 10.0g / L glucose, 0.5g / L (NH 4 ) 2 SO 4 , 0.3g / L MgSO 4 ·7H 2 O, 0.3g / L NaCl, 0.3g / L KCl, 0.03g / L FeSO 4 ·7H 2 O, 0.03g / L MnSO 4 4H 2 O, 10.0g / L Ca 3 (PO 4 ) 2 On the Montina inorganic phosphorus solid medium (pH7.0~7.5) composed of 20.0...
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