Method for separating cadmium-tolerant iron-generating carrier bacterial strain and application
A separation method and siderophore technology are applied in the field of separation of cadmium-resistant siderophore-producing strains, which can solve the problems of ecological environment impact, single function, difficult strains and the like, and achieve fast growth rate, strong environmental adaptability and improved germination rate. Effect
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[0026] The preparation method of the reagent required for the test is as follows:
[0027] (1) LB medium: tryptone 10.0g, yeast extract 5.0g, NaCl 10.0g, agar 20.0g, distilled water 1000mL, pH 7.0, sterilized by autoclaving at 121°C for 20min;
[0028] (2) SMA medium: 20.0g sucrose, 2.0g L-aspartic acid, K 2 HPO 4 0.5g, MgSO 4 ·7H 2 O 0.5g, 8-hydroxyquinoline, distilled water 1000mL, pH 7.0, sterilized by autoclaving at 121°C for 15min;
[0029] (3) Inorganic phosphorus medium (NBRIP): glucose 10g, (NH 4 ) 2 SO 4 0.5g, NaCl 0.3g, KCl 0.3g, MgSO 4 ·7H 2 O 0.3g, Ca 3 (PO 4 ) 2 5g, 1000mL distilled water, pH 7.0-7.5, 121℃, 15min high pressure steam sterilization;
[0030] (4) Salkowski'S chromogen: 150mL concentrated sulfuric acid was dissolved in 250mL deionized water, and 7.5mL 0.5molL was added -1 FeCl 3 ·6H 2 O solution;
[0031] (5) CAS detection solution: dissolve 0.079g CAS in 50mL deionized water, then add 10mL 1mmol L -1 FeCl 3 solution (prepared wit...
Embodiment 1
[0034]Take a ryegrass plant with good growth, shake off the soil around the ryegrass root system, leaving the rhizosphere soil closely attached to the root system, remove the rhizosphere soil, put it in a sterilization bag for later use, take an appropriate amount of root Add sterile water to the soil, shake it and let it stand still, absorb the upper layer of soil suspension, and apply it to LB+2mg / L Cd 2+ On the solid medium plate, invert culture at a constant temperature for 2-5 days, pick a single colony that grows well, streak and purify, and obtain a cadmium-resistant siderophore strain.
[0035] The preferred solution, the weight ratio of root soil to sterile water is 1:10, after adding sterile water to the root soil, shake at 150r / min for 30 minutes and then let it stand for 10 minutes, absorb the upper layer of soil suspension, and apply it on LB+2mg / L On a Cd2+ solid medium plate, culture upside down at 25-30°C for 3-4 days, pick a well-growing single colony and stre...
Embodiment 2
[0037] Take a ryegrass plant with better growth, shake off the soil around the ryegrass root system, leaving the rhizosphere soil closely attached to the root system, remove the rhizosphere soil, put it in a sterilization bag for later use, weigh the root Put 10g of soil into the Erlenmeyer flask and add 100mL of sterile distilled water, shake at 150r / min for 30min and let it stand for 10min, absorb 1mL of the upper layer of soil suspension, the dilution of the soil suspension is 10 -1 , and then serially diluted 10 times with sterile distilled water to prepare 10 -2 、10 -3 、10 -4 、10 -5 、10 -6 100 μL of each diluted suspension was spread on a solid medium plate of LB+2mg / L Cd2+, and cultured upside down at 28°C for 3 days, wherein the dilution of the soil suspension was 10 -1 、10 -2 and 10 -6 The single colony grows well, and the dilution of the soil suspension is picked up at 10 -1 、10 -2 and 10 -6 The well-growing single colonies were streaked and purified to obtai...
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