Marine-derived strain and application thereof in nematode prevention and control
A marine and nematode technology, applied in the direction of application, nematicides, bacteria, etc., can solve the problems of difficult nematode control and achieve good effect, inhibit nematode oviposition, and strong viability
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Embodiment 1
[0031] Example 1: Isolation and identification of Lysobacterium enzymogenes SYZX2010 strain
[0032] The medium 40% TSA for screening lysobacterium enzymogenes was prepared, and the NaCl concentration was increased to 2%.
[0033] Weigh 1g of marine sediment sample, dissolve it in distilled water, spread it on the prepared TSA plate, and grow it at 28°C for one day. Then pick a single colony on the plate and place it on a new 2% 40% TSA plate, and grow it at 28° C. for one day to obtain a single colony, which is named as SYZX2010 strain.
[0034] Pick a single colony of SYZX2010, dissolve it with 200 μL 1×TE, mix it by pipetting, boil for 10 minutes, and immediately put it in the refrigerator at -20°C to obtain bacterial DNA.
[0035] Using DNA as a template, the primers are 16S rDNA full-length universal primers 8F and 1492R, for 16S rDNA amplification, the amplification system: Premix Taq 15 μL, double distilled water 13.4 μL, 8F 0.3 μL, 1492R 0.3 μL, template 1 μL; the PCR...
Embodiment 2
[0038] Embodiment 2: the growth effect of Lysobacterium enzymogenes SYZX2010 strain under high salt concentration
[0039] The Lysobacter zymogenes SYZX2010 strain and the Lysobacter zymogenes SYZX2023 strain were cultured in 40% TSB containing NaCl with a concentration gradient of 0-2%, and grown at 28°C for 24 hours; then the absorbance at a wavelength of 600nm was measured with an ultraviolet spectrophotometer, The data in Table 1 is the absorbance at a wavelength of 600nm, which is used to detect the concentration of bacteria. It can be seen that when the salt concentration increased to 2%, the growth of the terrestrial Lysobacterium zymosus SYZX2023 strain was more inhibited than that of the Lysobacterium zymosus SYZX2010 strain ( image 3 ).
[0040] Table 1: Absorbance table of Lysobacter zymogenes SYZX2010 under different NaCl concentration gradients
[0041] strain 0% NaCl 0.5% NaCl 1% NaCl 1.5% NaCl 2% NaCl SYZX2010 0.4595 0.475 0.3665 ...
Embodiment 3
[0042] Example 3: Lysobacterium enzymogenes SYZX2010 strain inhibits nematode oviposition experiment
[0043] 1) Cultivate the fermentation broth of Lysobacter zymogenes SYZX2010 strain
[0044] Lysobacter zymogenes SYZX2010 strain was transferred from -20°C refrigerator to a 40% TSA plate with a NaCl concentration of 2% for activation, and after 24 hours of growth, it was transferred to a 40% TSB liquid medium with a NaCl concentration of 2%. ℃ for 3 days to obtain the fermentation broth.
[0045] 2) Cultivate Caenorhabditis elegans
[0046]Prepare the NGM plate, spread Escherichia coli OP50 on the plate, culture at 37°C for one day, transfer 3 to 5 Caenorhabditis elegans to the plate with an insect picker, and culture at 20°C for 6 days;
[0047] 3) Inhibition of nematode oviposition experiment
[0048] The NGM medium with OP50 was irradiated with ultraviolet rays for 30 minutes. The experimental group was coated with 200 μL of the fermentation broth of Lysobacter zymogen...
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