Bacteriostatic and detoxicating composite biocontrol inoculant for relieving continuous cropping of watermelons
A technology of biocontrol bacteria and watermelon, applied in the field of agricultural microorganisms, can solve the problems of difficulty, inability to stabilize colonization, serious environmental pollution, etc., and achieve the effects of easy industrial production, good application prospects, and stable growth rate
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Embodiment 1
[0023] Embodiment 1 PFC mixture preparation
[0024] (1) Medium preparation: LB solid medium formula (sodium chloride 10g, tryptone 10g, yeast powder 5g, agar powder 15g, distilled water to 1L, pH ≈ 7.2) was sterilized at 121°C for 30min. The LB liquid medium formula (sodium chloride 10g, tryptone 10g, yeast powder 5g, distilled water to 1L, adjust pH ≈ 7.2) was sterilized at 121°C for 30min.
[0025] (2) Strain activation: inoculate three biocontrol strains PNC25 (preservation number: GDMCC NO.60108), FR026 (preservation number: GDMCC NO.60725) and CHS55 (preservation number: CGMCC NO.4738) stored at -80°C It was activated in LB solid medium and placed in a constant temperature incubator at 28°C for 24h.
[0026] (3) Preparation of seed liquid: Inoculate a single colony of the three activated strains into LB liquid medium, and place them in a constant temperature oscillator at 28°C and 200r / min for 12 hours to prepare the seed liquid .
[0027] (4) Fermentation liquid prep...
Embodiment 2
[0029] Example 2 PFC mixture and watermelon Fusarium wilt synergistic / antagonistic effect analysis
[0030] This case explores the following two contents: 1. Comparing the growth and reproduction of three strains of PNC25, FR026 and CHS55 under single culture and co-culture, and exploring whether there is a synergistic growth relationship among the three strains. 2. Compare the growth and reproduction of PFC mixture and Fusarium wilt of watermelon in single culture and co-culture, and explore whether there is an antagonistic relationship between PFC mixture and Fusarium wilt of watermelon. The specific operation is as follows:
[0031] (1) Preparation of PDA culture medium: chop 200 g of potatoes, add water to boil for 20 min, filter, add 20 g of glucose to the filtrate, and dilute to 1 L with distilled water.
[0032] (2) Cultivation of three bacterial strains: Take a 10mL centrifuge tube, add 4mL of PDA culture solution, add 400μL of PNC25, FR026, CHS55 fermentation broth a...
Embodiment 3
[0038] Example 3 Determination of antagonistic activity between PFC mixture and watermelon Fusarium wilt plate
[0039] In this case, the plate confrontation method was used to investigate the inhibitory effect of PFC mixture, PNC25, FR026 and CHS55 strains on Fusarium wilt of watermelon. Specific steps are as follows:
[0040] The activated watermelon wilt bacteria is evenly punched into circular bacterium blocks with a diameter of 5 mm from the outer edge of the colony with a sterilized hole puncher. The bacterial block (5 mm) was inoculated in the center of the PDA medium, and the treatment group was inoculated with 10 μL of PFC mixture, PNC25, FR026 and CHS55 fermentation broth at a distance of about 27 mm from the four directions of the medium center, and the control group added the same amount of no Bacterial water was placed in a constant temperature incubator at 28°C for 10 days. After the control bacteria covered the entire plate, the diameter of the pathogenic bacte...
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