Xylariaceae fungus sj18 and application thereof in prevention and treatment of bursaphelenchus xylophilus
A sj18, pine wood nematode technology, applied in the fields of application, nematicides, chemicals for biological control, etc., can solve problems such as insufficient attention, inability to answer, and inability to prevent and control pine wood nematode, and achieve Unique control, environmentally friendly and compatible effects
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Embodiment 1
[0018] Embodiment 1: the isolation of Xylariaceae fungus
[0019] Take the root of a wild hickory tree, wash it, cut it into small pieces of about 2cm, first sterilize the surface with 70% alcohol, then soak it in 50% antifomin (sodium hypochlorite solution) for half an hour, and finally soak it in sterile water Rinse 4-5 times;
[0020] Cut the sterilized material on the ultra-clean bench, inoculate it on the PDA medium (solid potato medium) plate and cultivate it. When bacteria grow, they are respectively streaked and purified, and the DNA is extracted for molecular identification, so as to obtain And confirm the strain. The specific steps to confirm the strain are as follows:
[0021] 1) Pick the bacteria to be tested from the PDA plate and add them to the suspension solution A, add a sterilized tungsten carbide bead with a diameter of 3-5mm to the bacteria solution, put it in a boiling water bath for 5 minutes, and then place it in liquid nitrogen Under quick freezing, ...
Embodiment 2
[0037] Example 2: Morphological and biological characteristics of bacterial strains
[0038] Cyclobacteriaceae fungi ( Metarhizium anisopliae ) sj18 grows well on PDA medium, and the optimum temperature is 32-34°C. At 32°C, a 0.5cm block can grow to 8cm in 4 days. It can grow normally at pH5-11, and the growth rate is not affected. The mycelium is white when it first grows, and after 1 week of cultivation, the fungus turns dark brown and does not produce spores. figure 1 It is the morphology of sj18 bacteria under the microscope.
Embodiment 3
[0039] Embodiment 3: the effect of bacterial strain fermented liquid killing pine xylophilus
[0040] The specific implementation steps are as follows:
[0041] 1. Pine xylophilus collection
[0042] 1) Take a spare PDA plate, inoculate it with Botrytis cinerea, incubate at 25°C until the entire plate is covered, then store it upside down at 4°C for later use;
[0043] 2) Wrap the pine wood block with gauze, put it in a sealed plastic bag, add sterile water, soak in a water bath at 20-25°C overnight, centrifuge the soaking solution at 2000rpm for 3-5min, remove the supernatant, and leave 1-5mL, that is, for insect fluid;
[0044] 3) Take 20-50 μL of insect liquid and add it to the Botrytis cinerea plate covered with mycelia in step 1) in the form of a streak, grow at 25°C until the area of the insect reaches more than 2 / 3 of the petri dish, and store in a 4-degree refrigerator for later use;
[0045] 2. nematode screening
[0046] 1) Take the nematode plate out of the ...
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