Burkholderia sp. resistant to heavy metal copper and application
A technology of Burkholderia and Kholderia, applied in application, bacteria, restoration of contaminated soil, etc., to achieve broad application prospects and the effect of promoting stem length and plant height
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Embodiment 1
[0044] The isolation and preservation of embodiment 1 Burkholderia HXC-1
[0045] 1. Isolation and cultivation of Burkholderia HXC-1
[0046]Take Mimosa pudica Linn. collected from the suburbs of Chaozhou City, Guangdong Province, remove the stems and leaves, wash the roots with water, choose a large, plump, fresh root nodule from the roots, wash away impurities, rinse, and rinse with water. After soaking in 95% ethanol for 5 min, use 0.1% HgCl 2 The surface was sterilized for 5 minutes, and then rinsed 10 times with sterile water under sterile conditions. On the ultra-clean workbench after ultraviolet light sterilization, shake the sterile water flowing down from the root nodules for the last time, take a small amount to spread evenly in YMA medium, and incubate upside down at 28°C for 3 to 5 days. Observe the growth of the strain on the culture medium. If no bacterial colonies appear, it proves that the root nodules have been cleaned and disinfected; otherwise, the cause ...
Embodiment 2
[0071] Physiological and biochemical characteristics of embodiment 2 Burkholderia HXC-1
[0072] 1.H 2 o 2 Enzyme activity assay
[0073] The Burkholderia HXC-1 strain was inoculated on YMA solid medium, cultured at 28°C for 3 days, and 1 mL of 3% H 2 o 2 Drop it on the bacterial lawn, and repeat each treatment three times; if bubbles appear within 5 minutes, it is positive, and if there are no bubbles, it is negative. The result is as image 3 As shown, the bacterial lawn added H 2 o 2 After that, more air bubbles were produced rapidly, indicating that Burkholderia HXC-1 strain H 2 o 2 Enzyme activity is high.
[0074] 2. Congo red experiment
[0075] The Congo red assay is used to detect the presence of cellulose in cell surface components. Congo red can dye cellulose into a red compound, so the colony turns red, which is a positive reaction, indicating that there is cellulose on the cell surface; if the color of the colony remains unchanged, it is a negative reac...
Embodiment 3
[0092] Example 3 Burkholderia HXC-1 Tolerance Determination of Metallic Copper
[0093] (1) Activation of Burkholderia HXC-1: first inoculate the Burkholderia HXC-1 strain on the solid YMA medium, and then inoculate it in an incubator with a temperature of 28°C. Cultivate for 3 days until the lawn is plump;
[0094] (2) Preparation of bacterial suspension: in the cultured plentiful bacterial lawn, use an inoculation loop to select a ring of bacterial colonies in 80mL of YMA liquid medium (250mL Erlenmeyer flask), and use a shaker (rotating speed is 120r / min, temperature is 28 ℃) fully shaken for 15 hours to obtain a bacterial suspension;
[0095] (3) Determination of the highest tolerance concentration of copper
[0096] Configure the YMA solid medium containing copper (0, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600mg / L), for subsequent use;
[0097] Get 1mL of the bacterial suspension prepared in step (2) and add it dropwise to the above-mentioned YMA solid m...
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