Cotton endophytic fungus cef-559 and its application in the control of cotton Verticillium wilt
A technology of CEF-559 and cotton verticillium wilt, which is applied to cotton endophytic fungus CEF-559 and its application in the control of cotton verticillium wilt, can solve the problems of poor cotton verticillium wilt and achieve control of verticillium wilt Disease hazards, flexible application methods, and the effect of improving disease resistance
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Embodiment 1
[0022] Embodiment 1: the isolation of bacterial strain
[0023] (1) Sample collection
[0024] From June to August, healthy cotton plants were selected, and the middle and upper parts of the main stem and fruiting branches were taken 15 cm.
[0025] (2) Isolation of strains
[0026] First, the collected samples (cotton stalks) are washed with clear water, peeled off, placed in a sterilized petri dish with a diameter of 9 cm, and sterilized with 75% alcohol for 1 min. Then disinfect with 5.0% sodium hypochlorite for 60 seconds, and wash with sterilized water for 3 times, each time not less than 3 minutes. Then use sterilized absorbent paper to dry the water on the surface, cut into 0.4cm long pieces with sterilized scissors, place them in a 9cm petri dish, and incubate at 25°C. Use the last cleaning solution as a control to check whether the disinfection is thorough. Check the growth of endophytic fungi every day, and find that there are colonies growing out, and transfer t...
Embodiment 2
[0034] Embodiment 2: The inhibitory effect of CEF-559 on Verticillium dahliae of cotton
[0035] 1. Test method
[0036] The inhibitory effect of non-volatile metabolites of CEF-559 on Verticillium dahliae was determined by disc filter method. On a PDA plate with a diameter of 9 cm, spread a double layer of cellophane (nitrocellulose membrane) with a diameter slightly larger than 9 cm, inoculate CEF-559 bacteria blocks (5 mm in diameter) in the center of the plate, and culture in a constant temperature incubator at 25°C for 7 days Remove the cellophane, inoculate Verticillium dahliae Vd080 (5mm in diameter) in the center of the plate, and cultivate in a constant temperature incubator at 25°C for 15 days. The PDA plates without CEF-559 metabolites were inoculated with Verticillium dahliae Vd080 as a control. Each treatment was repeated 4 times, and the diameter of the colony was measured by the cross method, and the inhibition rate was calculated.
[0037] Effects of CEF-559...
Embodiment 3
[0046] Example 3: Controlling effect of CEF-559 on cotton Verticillium wilt
[0047] 1. Test methods Two methods were used to test the control effect of CEF-559 on cotton Verticillium wilt.
[0048] pre-inoculation method. After CEF-559 was shaken and cultivated in Chapic liquid medium at 25°C in the dark for 7 days, it was filtered with four layers of gauze, and then filtered with a 0.22 μm microporous membrane filter to remove bacteria and spores to obtain the culture filtrate of CEF-559 spare.
[0049] Mix the sterilized vermiculite and sand at a ratio of 6:4, then make a bottomless paper bowl with a diameter of 6cm, and place it in a plastic tray of 45cm×35cm. The susceptible variety Jimian 11 was used as the test variety, and the delinted cotton seeds were soaked in clean water for 12 hours, then drenched and ready for sowing. 8 cotton seeds were sown in each pot, and 5 seedlings were retained after emergence. There were 15 pots per treatment, and every 5 pots was a re...
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