Biocontrol actinomycete strain for preventing and controlling tobacco bacterial wilt and application thereof
A technology of actinomycetes and biocontrol agents, applied in the field of microorganisms, can solve problems such as the lack of efficient strains and the weak research foundation of tobacco bacterial wilt, and achieve the effect of reducing agricultural pollution
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Embodiment 1
[0023] Example 1 Screening of antagonistic actinomycetes
[0024] The specific process is as follows:
[0025] 1. Collection of soil samples
[0026] The rhizosphere soil at a depth of 5-20 cm from different tobacco field plots was collected from Zhenghe County, Fujian Province, a total of 4 parts were packaged and marked and brought back to the laboratory, and separated after natural air-drying.
[0027] 2. Isolation of actinomycetes
[0028] Separation was performed using the plate dilution method. Weigh 10g of the soil sample, pour it into a conical flask equipped with small glass beads and 90ml of sterile water, shake it for 30min, then let it stand for 5 minutes, dilute it by 10 times successively, and prepare 10 -2 、10 -3 、10 -4 、10 -5 、10 -6 0.1ml of the suspension of different concentrations was added to Gao's No. 1 medium (adding K with a final concentration of 50mg / L 2 GrO 7 ) plate, spread evenly and place it at 28°C for observation. After 5-7 days, differe...
Embodiment 2
[0034] The mensuration of embodiment two fermented liquid antibacterial activity
[0035] Six strains of actinomycetes whose inhibition zone diameters were all above 30mm were re-screened by cup and saucer method. The strain was inoculated in the seed medium, and after 2 days of fermentation in shake flasks at 28°C, it was inserted into the fermentation medium with 5% inoculum, cultured at 28°C and 200 r / min for 6 days, and the fermentation broth was centrifuged at 8000 r / min for 5 min , to obtain a sterile fermented liquid after being filtered with a bacterial filter. Tobacco R. solanacearum (10 8 CFU / ml) in the center of the NA plate, 100 μl of sterile fermentation broth was added to the hole, cultured at 28°C for 2 days, and the diameter of the inhibition zone was measured, repeated 3 times. The results showed (Table 2) that the inhibition zones of strains P12 and K4 were both above 20 mm, and the K4 fermentation broth had the strongest antibacterial effect, with the diam...
Embodiment 3
[0039] The identification of embodiment three bacterial strains K4
[0040] 1. Observation of morphological characteristics
[0041] Streak inoculation of the strain K4 on Gao’s No. 1 medium, insert a sterile cover glass at an angle of 45 degrees, culture at 28°C for 7 days, take out the cover glass and observe the growth of hyphae and spores under an optical microscope and an electron microscope. Morphological characteristics. It was found that the spore filaments of the strain K4 were helical, often in clusters, and the spore filaments were broken into spores, spores were oval, and the surface was rough ( figure 1 and figure 2 ).
[0042] 2. Observation of culture characteristics
[0043]Using the 8 kinds of media recommended by the Streptomyces Identification Manual, culture the strain K4 at 28°C for 7-10 days, observe the growth of the bacteria, and record the color of the aerial hyphae and basal hyphae and whether soluble pigments are produced. Strain K4 grew well ...
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