Actinomycete strain scaut013 and its application
A technology of actinomycetes and strains, applied in the field of microorganisms, to achieve the effect of promoting nodulation and growth, and good application potential
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Embodiment 1
[0028] Example 1 Isolation and purification of Actinomyces rhizosphere SCAUT013
[0029] 1.1 Isolation of rhizobacterial actinomycetes from A. chinensis in Liangshan
[0030] In 2018, A. chinensis was collected in Huidong County, Liangshan Yi Autonomous Prefecture, Sichuan Province, and the excess soil on the roots was shaken off. The roots were placed in a 50mL sterilized centrifuge tube containing 25mL of phosphate buffer, and treated with ultrasonic waves (150W). After 10 minutes, the roots were taken out under sterile conditions, and the supernatant was removed by centrifugation at 3000g for 10 minutes. -1 , 10 -2 , 10 -3 Gradient dilution, take 100ul of the suspension and spread it evenly on the modified Gau's medium for separation. Each dilution concentration is repeated 3 times, and cultured in a constant temperature incubator at 28°C for 5-7 days. Pick out strains with typical actinomycete colony characteristics, streak with dilution plates, and purify until purifie...
Embodiment 2
[0047] Example 2 Screening of the growth-promoting function of Actinomyces rhizosphere SCAUT013
[0048] 2.1 Screening of growth-promoting functions
[0049] 2.1.1 Determination of ability to produce IAA
[0050] Take 20 μL of bacterial suspension into ISP4 liquid medium containing 0.5mol / L tryptophan, repeat 3 times, set blank control, culture at 28°C for 3 days at 120r / min shaker, centrifuge at 8000r / min for 24h, take 1mL of bacterial solution To the supernatant, add 2 mL of IAA color developing solution, react in the dark at 25 °C for 30 min, measure the color at a wavelength of 530 nm, and record the absorbance value. Take the uninoculated liquid medium as the control for zero adjustment, and use the indole acetic acid standard solution with concentrations of 0, 5, 20, 40, and 60 mg / l as the standard as above, and calculate the concentration of IAA in the assay solution.
[0051] 2.1.2 Determination of siderophore capacity
[0052] Dissolve 0.012g of chrome azure in 10m...
Embodiment 3
[0083] Embodiment 3 The hydroponic experiment of A. chinensis
[0084] 3.1 Hydroponics experiment
[0085] Connect the test actinomycete strain to the ISP first 4 Bevel activation, then transfer to ISP 4 The culture medium was cultured overnight to logarithmic phase, the growth of the strains was detected by the plate method, and the bacterial content was calculated.
[0086] The seeds of A. chinensis were soaked overnight for surface disinfection. The seeds were picked up with sterile tweezers and placed in a petri dish with filter paper and sterile vermiculite. Each petri dish was planted with 10 seeds, and then sterile water was added to maintain moisture. , and maintain air circulation. Sterilize the tweezers, filter paper, petri dish, plastic cup, vermiculite, quartz sand, and low nitrogen nutrient solution required for the experiment.
[0087] Low nitrogen nutrient solution formula: KNO 3 10.1g, KH 2 PO 4 2.2g, MnSO 4 ·H 2 O 100.0mg, KCl 15.5g, ZnSO 4 ·7H 2 ...
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