Endophytic fungi CEF-642 of cotton and application thereof
A technology of CEF-642 and endophytic fungi, applied to cotton endophytic fungi CEF-642 and its application field
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Embodiment 1
[0019] Isolation identification and cultivation of embodiment 1 bacterial strain
[0020] (1) Sample collection
[0021] In June 2012, several true leaves were collected from healthy Cottonia sea-island plants in the experimental field of China Cotton Institute in Anyang City, Henan Province.
[0022] (2) Isolation of strains
[0023] Firstly, the collected sea-island cotton leaves were washed with clean water, peeled off, placed in a sterilized petri dish with a diameter of 9 cm, and sterilized with 75% alcohol for 1 min. Disinfect with 5.0% sodium hypochlorite for 30 seconds, and wash with sterilized water for 3 times. Then use sterilized absorbent paper to dry the surface water, cut into 0.5cm2 long tissue pieces with sterilized scissors, place them in 9cm PDA medium, and culture them at 25°C. Apply the last cleaning solution on the PDA medium to check whether the surface disinfection is thorough. Observe the growth of endophytic fungi in the dish every day, and find th...
Embodiment 2
[0032]Example 2 Antagonistic effect of CEF-642 on cotton Verticillium dahliae (indoor)
[0033] 1. Test method
[0034] Adhere to the training method. Pick a fresh mycelium piece of cotton Verticillium dahliae strong pathogenic strain Vd080 (isolated from a severe disease field in Cangzhou, Hebei) with a diameter of about 5mm, and place it on one side of the PDA medium, and inoculate CEF-642 bacteria of the same size at the same time The silk block was placed on the other side, and placed in a 28°C incubator for dark cultivation. The plate only inoculated with Vd080 was used as the control, and 4 replicates were set for each treatment. After 14 days of co-cultivation, the colony diameter of Verticillium dahliae Vd080 was measured, and the inhibition rate (GI) was calculated using the formula.
[0035] where R 1 is the colony diameter of Vd080 in the control, R 2 is the colony diameter of Vd080 in standoff culture.
[0036] Buckle cultivation method. Pour a certain amo...
Embodiment 3
[0044] Example 3 CEF-642 Control Effects on Cotton Verticillium Wilt (Greenhouse)
[0045] 1. Test method
[0046] The effect of CEF-642 on the control of Verticillium dahliae of cotton was tested in the greenhouse by soaking seeds with endophyte culture filtrate and raising seedlings with corn sand culture matrix. Two batches of experiments were carried out for each method.
[0047] (1) Seed soaking method.
[0048] Inoculate the fresh CEF-642 mycelia into Chapei liquid medium, culture at 28°C and 150 rpm in the dark for 14 days, filter the mycelium with 4 layers of gauze, centrifuge at 1000 rpm for 15 min to remove the spore precipitate, and collect the supernatant culture filtrate. The control strain Vd080 (cultured in Chapic liquid medium at 25°C in dark conditions for 7 days with shaking, filtered through 4 layers of gauze, obtained the spore suspension, and adjusted the spore concentration to 1×10 with a hemocytometer. 7 spores / ml to inoculate cotton seedlings.
[004...
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