Epiphyte Xylaria sp.FDYS-1, biological agent prepared from the same and application thereof in preventing and curing bursaphenchus xylophilus nickle
A biological agent and fungus technology, which is applied in the application field of controlling pine wood nematodes, and achieves the effects of strong selectivity, good thermal stability and high lethal activity
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Embodiment 1
[0030] Example 1 Isolation of Xyalria sp.FDYS-1 and its activity on pine xylophilus
[0031] Collect the healthy leaves of the Gentleman plant in Guangzhou South China Botanical Garden, rinse them with tap water, dry the water, cut them into small pieces of about 1.5×1.5cm, and carry out surface disinfection. The specific steps are as follows: rinse with 70% alcohol for 30s, and then rinse with 0.1 Rinse with %mercuric chloride for 1min, then rinse with 70% alcohol for 30s, rinse with sterile water for 4 times, and blot dry on sterile filter paper.
[0032] Use the rinsing liquid test method and the tissue imprinting method to test the surface disinfection effect: the rinsing liquid test method is to draw 0.2 mL of the rinsing liquid from the last rinse with sterile water and spread it on the PDA plate; the tissue imprinting method is to use sterilized tweezers to The above-mentioned surface-sterilized materials were transplanted onto the PDA plate and removed after contacting...
Embodiment 2
[0037] The influence of embodiment 2 culture conditions on the production of Xylaria sp.FDYS-1 active substance
[0038] 1. Selection of culture medium
[0039] Inoculate Xylaria sp.FDYS-1 into potato dextrose medium (PD), potato carrot medium (PC), potato sucrose medium (PS), Martin's medium, Czapek medium, Czapek containing 20% potato juice The culture medium and improved Fries No. 3 medium were cultured in 7 kinds of liquid medium including 120r / min shaking at 25°C in the dark for 5 days. The culture filtrate was prepared and the lethal activity of the filtrate against B. xylophilus was tested by the contact method.
[0040] Conclusion: if image 3 As shown, Xylaria sp.FDYS-1 can grow and produce nematocidal active substances in 7 kinds of liquid media, but the culture filtrates obtained from different media have obvious differences in nematocidal activity, and the culture filtrates of PD and PS The nematicide activity of the strains was the strongest, reaching 95.67% a...
Embodiment 3
[0048] Embodiment 3 Xylaria sp.FDYS-1 produces the genetic stability of nematode active substance and the thermostability of active substance
[0049] Xylaria sp.FDYS-1 was continuously subcultured for 6 generations, and then the culture filtrate was prepared and its lethal activity against pine xylophilus was detected.
[0050] Conclusion: by Figure 6 As shown, after continuous subculture of Xylaria sp.FDYS-1 for 6 generations, the nematicidal activity of the culture filtrate remained basically unchanged, and the lethal rate against pine xylophilus was still as high as 90.43%.
[0051] Treat the culture filtrate of Xylaria sp.FDYS-1 in a water bath at 60, 70, 80, 90, and 100°C for 20, 30, 40, 50, 60 minutes and autoclave at 121°C for 20 minutes, and use sterile distilled water to make up the lost water , to detect line killing activity. The test results showed that the nematicidal activity of the culture filtrate of the strain Xylaria sp.FDYS-1 had better thermal stability...
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