Degrading bacteria for degrading allelochemicals in plant root exudates and application of degrading bacteria
By screening and identifying efficient allelochemical-degrading bacteria Achromobacter xylosoxidans and Pseudomonas nitroreducens, the problem of allelochemical inhibition in peanut continuous cropping was solved, and efficient degradation of a variety of allelochemicals was achieved. , alleviates plant autotoxicity and provides scientific basis for alleviating peanut continuous cropping obstacles.
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Embodiment 1
[0034] Screening of degrading bacteria:
[0035] The screening of degrading bacteria adopts the domestication method of using benzoic acid as the sole carbon source and gradually increasing the concentration of benzoic acid. The specific operation is as follows: Accurately weigh 10g of soil sample, add it into a conical flask filled with 90ml of sterile water, shake it fully in a constant temperature shaker for 30min, and make a soil suspension with a concentration of 1:10. After the soil particles have settled, draw 1ml of the supernatant, transfer it into a test tube filled with 9ml of sterile water, and make 10 -2 Bacterial suspension, and so on, make 10 -3 、10 -4 、10 -5 、10 -6 、10 -7 bacterial suspension. choose 10 -4 、10 -5 、10 -6 、10 -7 Add 100 μl of each of the four concentrations to the benzoic acid selection medium, and the concentration of benzoic acid is 5, 10, 15, and 20 mg·L -1 Increase the gradient acclimatization and enrichment of degrading bacteria in ...
Embodiment 2
[0042] Identification of degrading bacteria:
[0043] (1) Identification of morphological characteristics of degrading bacteria
[0044] After the degrading bacteria were cultured in LB medium for 18 hours, observe the colony shape, size, color, transparency, viscosity, humidity, uplift and edge characteristics of the degrading bacteria, and whether they produce pigment, etc. After the bacteria were stained, observe the bacteria with a high-power microscope Gram stain, flagella, capsule, spore and other structures. After the bacteria solution was fixed with glutaraldehyde, dehydrated with ethanol and freeze-dried, the size and surface structure of the bacteria were observed by scanning electron microscope.
[0045] (2) Determination of physiological and biochemical characteristics of degrading bacteria
[0046] After the degrading bacteria were cultured in LB medium for 18 hours, litmus milk, glucose, methyl red reaction, V-P test, indole reaction, citrate, starch hydrolysis...
Embodiment 3
[0057] Influence of Environmental Factors on the Degradation Characteristics of Degrading Bacteria
[0058] In order to study the degradation of benzoic acid by degrading bacteria HJ-2 and HJ-3 under different environmental conditions, the initial pH, temperature, and NaCl concentration were selected as the research objects, and a single factor experiment was carried out.
[0059] (1) Initial pH of culture
[0060] The degrading bacteria were inoculated with 0.1% (OD600nm=0.8) inoculum until the -1 Benzoic acid and inorganic salt culture solution with pH 4, 5, 6, 7, 8, 9, 10 respectively, 30°C, 180r·min -1 After shaking culture for 48h, measure the growth of thalline and the degradation rate of benzoic acid, the growth of thalline is to take the culture medium without degrading bacteria as a contrast, and every treatment repeats 3 times. Measure the absorbance value at 600nm after the treatment, and the degradation rate of benzoic acid is the percentage of the benzoic acid c...
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