Preparation method of biocontrol microbial agent for controlling apple tree fungal diseases
A biocontrol agent, fungal technology, applied in the field of biocontrol agent preparation
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Embodiment 1
[0072] Example 1 Isolation, purification and screening of Streptomyces cacaoi FN
[0073] 1. Isolation and purification of Streptomyces cacaoi FN
[0074] Collect soil samples from Funing Vegetable Garden and put them into sterile bags, take 10g and put them into a sterilized triangular flask containing 100mL distilled water and glass beads, place on a shaker at 200rpm for 30min, let stand for 10min, shake well and use a pipette Draw 5mL from the liquid device and add it to a small triangle bottle containing 45mL sterile water, that is, dilute 10 2 times, sequentially serially diluted to 10 -3 、10 -4 、10 -5 、10 -6 Use a pipette to draw 100 μl and spread it on a 2216E plate containing antibiotics (potassium dichromate 50 mg / L, nalidixic acid 15 mg / L and nystatin 50 mg / L), and set 3 replicates for each concentration , placed in a 28°C incubator for 5-10 days. Use a sterilized bamboo stick to pick a single colony of actinomycetes and put it on a freshly prepared 2216E plate...
Embodiment 2
[0082] The growth characteristic of embodiment 2 cocoa Streptomyces FN
[0083] 1. Growth temperature
[0084] Dispense 2216E liquid culture medium (bacteria peptone No.2 5g / L, yeast powder 1g / L, the balance is distilled water, natural pH) into transparent test tubes, and sterilize with damp heat at 121°C for 30 minutes. There should be no precipitation in the culture medium. Streptomyces cocoa FN was inoculated with shaker flask 48 in 2216E liquid medium as an inoculant, and was inoculated into test tubes containing 2216E liquid medium at a rate of 1‰, and the control was not inoculated. Cultivate at 0°C, 7°C, 15°C, 20°C, 28°C, 35°C, 40°C, 45°C, 50°C, 55°C, 60°C incubators for 3d-5d, set 3 repetitions, observe And record the growth (precipitation) situation.
[0085] 2. Acid and alkali resistance
[0086] Adjust the 2216E liquid medium with dilute HCl or NaOH to different pH (1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5 , 10.0, ...
Embodiment 3
[0101] Example 3 Application of Streptomyces cocoa FN to the prevention and treatment of fungal diseases of fruits and vegetables
[0102] Determination of the effect of Streptomyces cocoa FN on apple anthracnose leaf blight, apple leaf spot, apple ring spot, apple rot, apple root rot, apple blight rot, cucumber blight, cucumber blight, tomato early blight, Inhibition rate of watermelon wilt, rape sclerotinia and cabbage black spot pathogens. Draw a straight line through the center of the plate on the back of the PDA plate with a diameter of 90 mm, inoculate 7 mm of probiotic bacteria cake at two points 25 mm away from the center on the line, and inoculate 7 mm of blank PDA medium on the control plate. Inoculate the center of the PDA plate with a 7mm bacterial cake of pathogenic bacteria, and culture it at 28°C for 5-10 days. The radius of the control pathogenic bacteria and the radius of the treated pathogenic bacteria were counted respectively, and the antibacterial rate was...
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