Biocontrol actinomycete and application
A technology of actinomycetes and biocontrol agents, applied in the direction of application, bacteria, biocide, etc., can solve the problem of limited microorganisms, achieve the effects of simple cultivation conditions, resistance to ultraviolet radiation, and good application prospects
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Embodiment 1
[0038] Example 1 Isolation of antagonistic actinomycetes
[0039] Separation was performed using the plate dilution method. The specific method includes: taking 3g of soil samples collected from different areas of Jiangsu Province and putting them into sterile Erlenmeyer flasks filled with 27ml of 0.85% sodium chloride solution in mass volume ratio, placing them on a shaker at 150rpm, and cultivating them with shaking at 28°C for 30min After standing still for 5min, the suspension at this time is considered to be 10 -1 diluent, and then diluted 10 times with a mass volume ratio of 0.85% sodium chloride solution, respectively prepared into 10 -2 、10 -3 、10 -4 、10 -5 、10 -6 diluent, draw 10 -4 、10 -5 、10 -6Spread 0.1ml of the dilutions of the three gradients on Gao’s No. 1 medium (add 1ml of 3% potassium dichromate per 300ml of medium), repeat each gradient 3 times, and place at 28°C After culturing for about 3 days, single colonies with different colony shapes were sel...
Embodiment 2
[0040] Identification of embodiment 2 actinomycetes strain JK1
[0041] The plate confrontation test was used to screen the actinomycetes that had an inhibitory effect on the mycelia growth of Pythium conglomerate or Pythium melon fruit among the above 143 strains of actinomycetes, and after using the plate confrontation test to verify the selected bacteria again, a strain against The strain with the best inhibitory effect on Pythium massiferous and Pythium melon fruit was named JK1.
[0042] 1. Observation of morphological characteristics
[0043] Streak inoculation of actinomycete strain JK1 on Gao’s No. 1 medium, insert it obliquely at 45° on a sterile cover glass, culture it at 28°C for 10 days, take out the cover glass and observe the morphological characteristics of the hyphae under an optical microscope .
[0044] The results showed that the aerial hyphae were abundant and luxuriant, dry and velvety, opaque, powdery on the surface, showing a slightly white squid gray ...
Embodiment 3
[0067] The plate antagonistic effect determination of the fermentation liquid of embodiment 3 bacterial strain JK1 and ethyl acetate extract
[0068] (1) Actinomycetes JK1 was inoculated in Gao's No. 1 medium for activation, cultured in a 28°C constant temperature incubator in the dark, and cultured for 2-4 days before use. Gaoshi No. 1 medium is composed of 20g of soluble starch, 0.5g of sodium chloride, 1g of potassium nitrate, 0.5g of dipotassium hydrogen phosphate, 0.5g of magnesium sulfate, 0.01g of ferrous sulfate, and 15-20g of agar, with a pH of 7.2. Distilled water to 1000ml, sterilized at 121°C for 20min;
[0069](2) Inoculate the activated actinomycetes into 1L of fermentation medium for fermentation, shake and culture for 96 hours at a speed of 160-180r / min and a temperature of 28°C in the dark. After the fermentation is over, one of the methods is to The fermented product was centrifuged at 5000r / min for 10min, and the supernatant was filtered with a 0.22μm bacte...
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