Streptomycete S1-5 and uses thereof
A technology for streptomyces and phytopathogenic fungi, applied in the application, bacteria, biocide and other directions, can solve the problem of no streptomyces, and achieve the effects of not easy drug resistance, fast efficacy, and high killing activity
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Embodiment 1
[0026] Embodiment 1: the acquisition of fermented liquid
[0027] (1) Incline cultivation: adopt PDA medium, apply Streptomyces S1-5 strain on the slope, and cultivate at 30° C. for 2 to 3 days to obtain the strain on the slope;
[0028] (2) Seed culture: inoculate the slant bacteria into Gaoshi No. 1 medium, cultivate at 30° C. for 2 to 3 days, and obtain Streptomyces S1-5 colonies;
[0029] (3) Fermentation culture: original fermentation medium preparation: 20 mL of 1% soybean cake powder infusion, 20 g of glucose, 3.0 g of peptone, 2.0 g of calcium carbonate, 2.5 g of sodium chloride, and 1000 ml of distilled water. Scrape the spores from the colony of Streptomyces S1-5 cultured on Gao’s No. 1 medium, place them in sterile water, shake for 10 minutes, and make a concentration of 1×10 8 The spore suspension of each / ml was inserted in the fermentation medium with 10% (V / V) inoculum, placed on a 250ml Erlenmeyer flask on a shaker, and placed in a constant temperature shaker t...
Embodiment 2
[0030] Embodiment 2: Obtaining and measuring of fermented liquid extract
[0031] Take 50ml of fermentation broth, centrifuge at 5000r / min for 10min, add an equal volume of absolute ethanol to the supernatant to obtain the crude extract of the supernatant of biologically active substances. Wash the filter cake twice with distilled water and centrifuge, discard the supernatant, add absolute ethanol to the filter cake to reduce to the volume of the fermentation broth, soak for 1 hour, centrifuge at 5000r / min for 10 minutes, and take the supernatant, which is the crude mycelium of the biologically active substance. Extract. The two kinds of crude extracts and the centrifuged supernatant of the fermented liquid were used for the determination of antibacterial effect and acaricidal test, and the same ethanol aqueous solution was used as a control at the same time.
[0032] Determination method of antibacterial effect: (four kinds of plant pathogenic fungi are donated by the Plant ...
Embodiment 3
[0041] Embodiment 3: Obtaining and measuring of biologically active substances
[0042] After the Streptomyces S1-5 fermentation broth was centrifuged, the supernatant was taken, added an equal volume of ethyl acetate, extracted, and the upper layer was concentrated and collected under reduced pressure. Collect the lower layer for antibacterial determination, use ethyl acetate as the eluent to walk the column, the eluent is petroleum ether / ethyl acetate, column chromatography silica gel (200-300 mesh) is used as the stationary phase, and the column is packed in a dry method. Use the test tube to collect the eluate at the lower end, collect 10 mL in each tube, and collect 10 tubes in total. Tracking of biological activity: The collected 10 tubes of eluate and the extracted lower layer solution were respectively tested for their antibacterial activity against the tested bacteria (Calleatherium longopodium and Rhizoctonia solani, donated by the Plant Protection Department of Zhej...
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