Inorganic phosphorus solubilizing plant growth promoting rhizobacteria strain NG-33 and application thereof
A technology of NG-33 and inorganic phosphorus, which is applied in the direction of plant growth regulator, plant growth regulator, application, etc., can solve the problem of reduced utilization rate, and achieve the effects of saving phosphorus fertilizer, improving soil, and promoting plant growth
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Embodiment 1
[0033] Example 1 :Isolation and identification of Enterobactercloacae NG-33 (Enterobactercloacae)
[0034] 1.1 Isolation of Enterobacter cloacae NG-33 (Enterobactercloacae)
[0035] The isolation of Enterobacter cloacae NG-33 (Enterobactercloacae) includes three steps: sampling, enrichment and preliminary screening, and re-screening, as follows:
[0036] 1.1.1 Sampling
[0037] Take samples from the soil in the Nonggang National Nature Reserve of Guangxi Zhuang Autonomous Region, put them into clean sampling bags (bottles), mark them, and store them in a refrigerator at 4°C for later use.
[0038] 1.1.2 Enrichment and preliminary screening
[0039] Weigh 10 g of the collected sample and pour it into 100 mL of 800 mM Luria-Bertani (LB) liquid medium at 37° C., 200 rpm, and shake culture for 24 hours. Then take 1 mL of bacterial suspension and inoculate it into an Erlenmeyer flask containing NBRIP liquid medium. Multiple shake flasks are tested in parallel with a control sample (no bacte...
Embodiment 2
[0053] Example 2 : Determination of the soluble phosphorus content of Enterobactercloacae NG-33 (Enterobactercloacae) shake flask culture
[0054] 2.1 The configuration of molybdenum antimony anti-chromogenic liquid
[0055] ①Take potassium antimony tartrate 〔K(SbO)C 4 H 4 O 6 ] 0.5g, dissolved in 100mL water.
[0056] ②Weigh ammonium molybdate ((NH 4 ) 6 Mo 7 O 24 ·4H 2 O] 10g, dissolved in 450mL water, slowly add 153mL concentrated H 2 SO 4 , Stir while adding.
[0057] ③Add the above ① solution to ② solution, and finally add water to 1L. Shake well and store in a brown bottle. This is a mixture of molybdenum and antimony.
[0058] ④Before use, weigh L-ascorbic acid (C 6 H 8 O 5 , Chemically pure) 1.5g, dissolve in 100mL molybdenum-antimony mixture, mix well, this is the molybdenum-antimony reagent. The validity period is 24 hours, if it is stored in the refrigerator, the validity period is longer.
[0059] The test strains were cultured in 10 mL LB liquid medium at 30° C., 180 rpm,...
Embodiment 3
[0061] Example 3 :Determination of the types and amounts of organic acids produced by Enterobactercloacae NG-33 (Enterobactercloacae)
[0062] The NG-33 strain was fermented in liquid NBRIP liquid medium at 37°C and 180 rpm for 5 days. The culture broth was measured at the same time every day, and then the organic acid in the fermentation broth was analyzed by ion chromatography. Take 1mL fermentation broth and place it in a 2mL plastic centrifuge tube, centrifuge at 12000r / min for 5min, take the supernatant, dilute 10 times with ultrapure water, and filter the diluted solution through a 0.23μm filter membrane to remove micron-level suspended solids. Use ICS-5000 ion chromatography device (Thmero, USA), use IonPacAS11 anion exchange column, IC-RP guard column, analytical column (250mm×4mm), guard column (50mm×4mm). Adopt gradient elution, flow rate 1mL / min, column temperature 30℃, injection volume 25μL. The gradient elution procedure is shown in Table 2. The standard curve is o...
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