Shewanella piezotolerans 34# and application thereof to algae inhibition
A Shewanella, pressure-resistant technology, applied in marine pressure-resistant Shewanella bacteria and its application in inhibiting the growth of red tide algae, can solve the lack of pertinence of strains, and the lack of strict distinction between red tide occurrence period and non-occurrence period And other issues
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Embodiment 1
[0045] Embodiment 1, screening, identification and characteristics of pressure resistant bacterium (Shewanella piezotolerans) 34#
[0046] 1. Screening of Shewanella piezotolerans 34#
[0047] On April 21, 2011, a localized red tide occurred near the coast of the South China Sea (Shenzhen Shekou Port) (the surface layer of the red tide occurrence area of Shenzhen Shekou Port). Phylum Bacillariophyceae, Fragilariaceae Schroder, genus Thalassionema Grunow. Retrieve the water sample from this area, bring the water sample back to the laboratory and filter it through gauze to remove impurities and large particles in the water; then filter it through a 100μm metal mesh to remove debris in the water; finally filter it through a 10μm filter membrane to remove algae cells , and the filtrate was coated and screened for bacteria.
[0048] Dilute the filtrate prepared above, take 30 μL and spread it on the 2216E seawater solid medium (recipe: peptone 5.0g, yeast extract 1.0g, iron pho...
Embodiment 2
[0062] Embodiment 2: the detection of the QS ability of Shewanella piezotolerans (Shewanella piezotolerans) 34# production
[0063] 1. Preparation of Shewanella. The 34# bacterial strain of identification is cultivated in the eppendof tube of 1.5 milliliters for 6-8 hours, and the liquid volume of the eppendof tube is 0.5 milliliters, and the substratum is LB liquid medium (medium formula: tryptone 10g, yeast extract 5g, NaCl 10g, adjust pH to 7.0. Dilute to 1L with deionized water. Steam sterilize at 15psi high pressure for 20min). The concentration of the cultured Shewanella liquid was detected by a visible light spectrophotometer, and the concentration value was about OD 600 = 0.35. The prepared bacterial solution was stored in a 4°C refrigerator for later use.
[0064] 2. Preparation and detection of test strains. The QS produced by the 34# strain was detected with the reporter strains CV026 and A136 that specifically detect QS, wherein the reporter strain CV026 is sui...
Embodiment 3
[0066] Embodiment 3, the application of Shewanella piezotolerans (Shewanella piezotolerans) 34# in inhibiting algae
[0067] 1. The method of inhibiting algae by Shewanella piezotolerans 34#
[0068] 1. High-density culture of Shewanella piezotolerans 34#
[0069] Carry out primary culture (2216E liquid culture medium, formula: peptone 5.0g, yeast extract 1.0g, ferric phosphate 0.01g, old seawater 1000mL, adjusted by the pressure-resistant Shewanella 34# obtained by embodiment 1 in a 10mL conical flask. ph to 7.8), the culture condition is 30°C, the rotation speed is 200r / min, and the time is 6h, when the strain density is close to OD 600 = 0.5, transfer to a 100mL Erlenmeyer flask for secondary culture. Continue to cultivate for 18 hours until the strain density reaches 1×10 9 Individual / ML concentration, spare.
[0070] 2. Cultivation of algae cells for experiments
[0071] The algal species used in the experiment was Alexandrium catenella (Hong Kong strain). Take the ...
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