Growth-promoting rhizobacteria SXH-2 and application thereof
A technology of SXH-2, rhizosphere growth-promoting bacteria, applied in application, bacteria, biocide, etc., can solve the problems of reduced utilization rate, damage to soil environment, limited yield of chives, etc., to promote phosphorus absorption and reduce massive accumulation. , the effect of improving stress resistance
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Embodiment 1
[0038] Example 1. Isolation and Identification of Klebsiella oxytoca SXH-2
[0039] 1. Isolation of Klebsiella oxytoca SXH-2
[0040] The isolation of Klebsiella oxytoca (Klebsiella oxytoca) SXH-2 includes two steps of sampling, screening and purification, the specific method is as follows:
[0041] 1.1. Sampling
[0042] Screening is carried out from the rhizosphere soil of Chinese chives. The specific chives and soil for testing are taken from Shuangjing Town, Hulan District, Harbin City, Heilongjiang Province (the physical and chemical properties of the soil are as follows: pH 8.15, total salt content 1.83%, alkali-hydrolyzable nitrogen 36mg / kg , available phosphorus 8mg / kg, available phosphorus 132mg / kg, organic matter 2.69%). Put leeks and plant rhizosphere soil into clean fresh-keeping bags prepared in advance, and bring them back to the laboratory for planting for testing.
[0043] Weigh 1g of rhizosphere soil sample into a Erlenmeyer flask containing 50mL of PAF cul...
Embodiment 2
[0061] The ACC deaminase activity determination of embodiment 2, CGMCC No.6297
[0062] The tested strain was cultured overnight in TSB culture solution (tryptone 17.0g, soytone 3.0g, NaCl 5.0g, glucose 2.5g, K2HPO 42.5g, distilled water 1000mL, pH 7.5), and the bacteria were collected by centrifugation at 4°C. The bacterial cells were washed three times with DF medium, resuspended in ADF medium, cultured with shaking at room temperature (21±1°C) for 2 days, collected by centrifugation at 4°C, and washed with 0.1mol·L -1 Wash and centrifuge three times with Tris-HCl buffer (pH=7.6), resuspend in 600 μL 0.1mol L -1 In Tris-HCl buffer (pH=8.0), add 30 μL of toluene and shake rapidly for 30 s to break the cells, transfer 200 μL of toluene-containing cell extract to a 1.5 mL centrifuge tube, add 20 μL of 0.5 mol·L -1 Mix ACC evenly, and do a blank test without adding ACC at the same time, and incubate at 30°C for 15 minutes. Add 1mL 0.56mol·L -1 HCl, centrifuge at 16000g for 5m...
Embodiment 3
[0067] Embodiment 3, the mensuration of CGMCC NO.6297 growth hormone IAA synthetic content
[0068] The test strain Klebsiella oxytoca (Klebsiella oxytoca) SXH-2 was first cultured in DF culture medium for 2 days, and then transferred to DF culture medium with different concentrations of tryptophan (L-Trp) (containing 0, 50, 100, 200 and 500 μg L-Trp mL -1 ) to continue culturing for 2 days, and take a sample to test the OD of the bacterial solution 600 , centrifuge the remaining culture solution at 8000 g at room temperature, take 500 μL supernatant, add 2 mL Salkowski reagent (containing 150 mL H 2 SO 4 , 250mL ddH 2 O and 7.5mL 0.5mol / L FeCl 3 ), after incubating in the dark at room temperature for 20min, measure the absorbance at 535nm (OD 535 ). The sterile medium was treated the same as above and set to zero as a control. With concentrations of 0, 0.01, 0.05, 0.25, 0.5mg·mL -1 The IAA standard solution was used to make a standard curve in the same way. The unit ...
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