Siderophore high-yielding strain preparation and application of same in remediation of heavy metal contaminated soil
A bacterial preparation and siderophore technology, applied in the field of bioremediation of environmental pollution, can solve the problems of lack of strains, few strains, and limited strains with high siderophore production
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Embodiment 1
[0029] Example 1: Isolation of Pseudomonas sp.S17 and Bacillus sp.S86 and Siderophore Production Ability Test
[0030] 100g of soil in the experimental field of Zhejiang University of Science and Technology, Hangzhou, Zhejiang Province was collected on-site by the general five-point sampling method, packed in a sterile envelope, and brought back to the laboratory quickly.
[0031] General CAS detection solution and MSA liquid medium are based on the report of Chen Shaoxing et al. ) preparation. The solid detection plate is solidified by adding 5% CAS detection solution and 2% agar powder into MSA liquid medium.
[0032] Dilute the soil sample by 10-fold decreasing method, and spread it on one-fifth LB solid plate (that is, all nutrients are 1 / 5 of the general LB medium). Cultivate at 25°C. After colonies appear, pick out the colonies of different forms one by one with a sterile toothpick and transfer them to a new one-fifth LB solid plate until they are purified into single ...
Embodiment 2
[0038] Example 2: Antibiotic susceptibility test and pathogenic fungus antagonistic ability test of Pseudomonas sp.S17 and Bacillus sp.S86
[0039] The antibiotic susceptibility test adopts the filter paper method, and five commonly used antibiotics such as ampicillin, tetracycline, chloramphenicol, kanamycin and streptomycin are selected, and the antibacterial activity of each antibiotic filter paper on the siderophore-producing bacteria S17 and S86 plates is selected. The size of the ring diameter is used as a criterion for judging sensitive or resistant (resistant) drugs. The test results given in Table 4 and Table 5 show that the strains S17 and S86 are sensitive to the above-mentioned tested antibiotics in varying degrees.
[0040] Table 4. Antibiotic susceptibility test results of bacterial strain S17
[0041]
[0042] Note: If the diameter of the inhibition zone is larger than the upper limit of the moderately sensitive range, it is highly sensitive, and if it is sm...
Embodiment 3
[0051] Embodiment 3: The heavy metal minimum inhibitory concentration (MIC) test of Pseudomonas sp.S17 and Bacillus sp.S86
[0052] The design of the MIC experiment was carried out according to the research of Filali et al. (Filali BK, Taoufik J, Zeroual Y, Dzairi FZ, Talbi M, Blaghen M. Waste water bacterial isolates resistant to heavy metals and antibiotics. Current Microbiology, 2000, 41: 151-156), Single colonies of S17 and S86 of equal size and activation treatment were picked and placed in culture test tubes, and cultured at 30° C. with shaking at 160 r / min for 32 hours. The MIC value was determined according to the growth of the bacteria, and three replicates were set up in each group to test the tolerance of the strains to 12 metal ions. The results are shown in Table 7. The tested strains have multiple heavy metal tolerance (resistance) and excellent properties.
[0053] Table 7. MIC test results
[0054]
[0055]
[0056] Note: The data in the table is the av...
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