Sapium discolor endophyte burkholderia cenocepacia WJ-19 with efficient tolerance to multiple heavy metals
A technology of Burkholderia and Holderella, applied in the field of discovery and characteristics of Burkholderia, to achieve the effect of high-efficiency removal of heavy metals
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Embodiment 1
[0020] Example 1: Screening of heavy metal-tolerant Sapium arvensis endophytic bacteria
[0021] 1. Isolation and purification of endophytic bacteria from plants:
[0022] The fresh Sapium aconitifolia samples were sterilized on the plant surface, dried and placed on mannitol soybean medium (soybean powder 20g / L, mannitol 20g / L, agar powder 20g / L, pH value 7.2±0.2), and cultured at 30°C , when the endophytic bacteria grow out, combined with streak separation method in LB solid medium (peptone 10g / L, yeast extract 5g / L, sodium chloride 10g / L, agar powder 18g / L, pH5.6-5.8 ), the temperature is 30°C, the culture time is 1-2 days, and the separation is carried out until the pure culture is obtained.
[0023] 2. The verification of the experimental effect of the surface disinfection of Chinese tallow tree, the specific implementation method is (one of the following two methods can be selected):
[0024] Method 1: Take 200 μL of washing water from the last step of surface disinfec...
Embodiment 2
[0026] Example 2: Biological identification and strain morphological characteristics of bacterial strains
[0027] 1. Morphological characteristics of the strain
[0028] (1) Morphological characteristics of colony population: Streak inoculation of WJ-19 monoclonal colony into LB solid medium, place the plate upside down in a constant temperature incubator, and incubate at 35°C for 18-24h, forming medium-sized, opaque, moist, raised Milky white colonies. Under the conditions of LB liquid culture (temperature 30°C, pH=6, shaker 170r / min), WJ-19 was in the lag phase at 0-4h, entered the logarithmic growth phase at 4-10h, and then entered the stable phase and decay phase.
[0029] (2) Individual morphological characteristics of the bacteria: Gram staining was carried out on the WJ-19 strain, and observation by an optical microscope showed that the strain was a rod-shaped, Gram-negative bacterium.
[0030] 2. Strain 16S rRNA analysis
[0031] Bacterial Genomic DNA Extraction Ki...
Embodiment 3
[0038] Example 3: detection of strain heavy metal tolerance
[0039] The dilution quantitative spotting method and plate coating method were used to detect the tolerance level of the strain to heavy metals such as cadmium, lead and zinc, and the minimum inhibitory concentration of the strain was obtained.
[0040] 1. Colony dilution method to screen heavy metal-tolerant bacteria: OD 600 ≈1 strain fermentation broth, dilute it to OD with saline 600 ≈0.1, 0.01, 0.001, spot the same amount of fermented broth of these four concentration gradients on the solid medium of LB strains added with heavy metal lead nitrate, zinc chloride, and cadmium chloride respectively, and screen to obtain The colony WJ-19 that can grow on both.
[0041] 2. Determination of the minimum inhibitory concentration by plate coating: the microbial plate coating method was implemented on the solid medium of LB strains added with different concentrations of heavy metal lead nitrate, zinc chloride, and cadmi...
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