Cotton endophytic fungus cef‑193 and its application
A technology of CEF-193 and cotton verticillium wilt, applied to the endophytic fungus CEF-193 and its application field, can solve the problems of poor cotton verticillium wilt effect and the like, achieve good immune induction, be beneficial to mass reproduction and popularization and application Effect
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Embodiment 1
[0022] Example 1 Isolation of strains
[0023] (1) Sample collection
[0024] From June to August, select healthy cotton plants, and take 15cm from the middle and upper parts of the main stem and fruit branches.
[0025] (2) Isolation of strains
[0026] First, the collected samples (cotton stalks) were washed with clean water, peeled, placed in a sterile petri dish with a diameter of 9 cm, and disinfected with 75% alcohol for 1 min. Then disinfect with 5.0% sodium hypochlorite for 60s, and wash with sterilized water for 3 times, each time not less than 3min. Then use sterilized absorbent paper to dry the water on the surface, use sterilized scissors to cut into 0.4 cm long pieces, put them in a 9 cm petri dish, and cultivate at 25°C. Use the last cleaning solution as a control to check whether the disinfection is complete. The growth of endophytic fungi was checked every day, and if colonies were found to grow, they were promptly transferred to a petri dish with a diamete...
Embodiment 2
[0035] Example 2 Inhibitory effect of CEF-193 on cotton Verticillium wilt
[0036] 1. Test method
[0037] Buckle training method. Pour a certain amount of medium on the lid and bottom of a 90mm diameter petri dish, and the medium on the lid is slightly less. After solidification, 5 μl of Acremonium aromaticum CEF-193 bacterial solution was placed in the center of the lid of the dish (Chappy liquid culture for 7 days), and 5 μl of Verticillium wilt pathogenic bacteria solution was inoculated into the center of the bottom of the dish (Chappy liquid culture for 7 days). Placed in a constant temperature incubator at 25°C and cultured upside down, with the Petri dish only inoculated with pathogenic bacteria as a control, and each treatment was repeated 4 times. The growth status of pathogenic bacteria and Acremonium aromaticum hyphae were observed every day, and the colony diameter of pathogenic bacteria was measured by the cross method after culturing for 7 days, and the inhibi...
Embodiment 3
[0044] Example 3 Induced immunity effect of CEF-193 on cotton Verticillium wilt
[0045] 1. Test method
[0046] CEF-193 was cultured in Chapek liquid medium at 25°C under dark conditions for 7 d with shaking. After filtering through filter paper, the culture filtrate was obtained for seed soaking. The control strain Vd080 (a strong virulent strain of cotton verticillium wilt) was cultured in Chapek liquid medium at 25° C. under dark conditions for 7 days with shaking, and filtered through three layers of gauze to obtain a spore suspension for inoculation. Four treatments were set up in the experiment, which were 25%, 50% and 100% CEF-193 culture filtrate and water soaking. All four treatments were inoculated with Vd080 conidia suspension of cotton seedlings when a true leaf was flat.
[0047] The sterilized vermiculite and sand were mixed at a ratio of 6:4 to make a bottomless paper pot with a diameter of 6 cm, which was placed in a plastic tray of 45 cm × 35 cm. Taking the...
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