Bacillus aerophilus and application of bacillus aerophilus in killing meloidogyne
A technology of Bacillus aerophilus, ZDXC-01, applied in the field of microorganisms, can solve the problems of rampant occurrence of secondary diseases, pollution, loss of resistance, etc., and achieve good application prospects, fast growth and strong stress resistance
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Embodiment 1
[0022] Example 1 Bacillus aerophilus ( Bacillus aerophilus ) Discovery and Identification of ZDXC-01
[0023] The present inventor isolates monoclonal bacterial strains from seawater by dilution coating method, and the specific separation method is: take 1mL seawater sample and dissolve in 99mL sterile water, vibrate to make it fully mix, promptly obtain 10 -2 seawater dilution. Take 1ml of the above seawater dilution to 9ml sterile water, shake well to obtain 10 -3 seawater dilution. Configure 10 in the same way -4 、10 -5 、10 -6 seawater dilution. take 10 -4 、10 -5 、10 -6 Put 100 μL of seawater dilution solution on the agar-added 2216E medium plate, spread the dilution evenly with a spreader, place the plate in an incubator at 28-30°C for 24 hours, and colonize different forms of bacteria on the medium on the Streak on blank 2216E medium plate, purify the obtained strains, and store them with respective numbers. Among them, the microscopic characteristics of a stra...
Embodiment 2
[0024] Embodiment 2 Preparation of anti-root-knot nematode fermented liquid
[0025] The fermentation process for preparing a large amount of Bacillus aerophilus ZDXC-01 is as follows:
[0026] (1) Streak the Bacillus aerophilus ZDXC-01 stored on the slant of the test tube on the 2216E medium or LB medium with agar for activation, and culture it in a constant temperature incubator at 28-30°C for 3 days;
[0027](2) Transplant the spores of Bacillus aerophilus ZDXC-01 in step (1) into 2216E (yeast extract 1g, peptone 5g, filtered seawater 1000mL, pH 7.2-7.4) or LB liquid medium (tryptone 10g , yeast extract 5g, sodium chloride 10g, distilled water 1000mL, pH adjusted to 7.2-7.4), shake culture at 28-30°C for 3 days to obtain seed liquid;
[0028] (3) Inoculate the seed liquid into the fermentation medium (corn flour 10g, soybean meal 10g, soluble starch 5g, potassium dihydrogen phosphate 1g, sodium chloride 5g, distilled water 1000 mL, pH 7.2- 7.4), shaking culture at 28-30°C...
Embodiment 3
[0029] Example 3 Indoor in vitro test against tomato root-knot nematode
[0030] 1.1 Preparation of test agents
[0031] The anti-root-knot nematode fermentation broth of Bacillus aerophilus ZDXC-01 freshly prepared according to Example 2.
[0032] 1.2 Preparation of test nematodes
[0033] The nematodes tested were obtained from tomato diseased roots in tomato protected areas. Take tomato diseased roots, wash the surface soil with distilled water, pick out the egg mass inside the root knot with a dissecting needle, put it in a petri dish filled with sterile water, and let it stand for incubation at 28°C for 24 hours; collect the hatched root-knot nematode second-instar larvae, Gently mix in the Erlenmeyer flask to obtain nematode suspension (about 100 / mL), and store at 4°C for later use.
[0034] 1.3 Experimental design
[0035] Using the collected tomato root-knot nematode as the test nematode, the anti-root-knot nematode activity of the 24-well plate was tested indoors ...
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