Multi-functional Streptomyces amritsarensis and application thereof
A multifunctional technology of Streptomyces alginolyticus, applied in the field of microorganisms, can solve the problems of pathogenic bacteria production, reduced control effect, and reduced wheat yield, and achieve the effect of promoting synthesis
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Embodiment 1
[0038] Isolation and screening of embodiment 1 bacterial strain
[0039] (1) Isolation of actinomycetes: Collect 200g of red soil from the Chongqing Purple Soil Monitoring Station, take 10g of soil and resuspend it in 90ml of sterile water, apply Gao’s No. 1 medium after gradient dilution, and cultivate at 28°C until a single colony grows According to the difference in the color and colony shape of sporocydia and hyphae in the base, different actinomycetes were purified and preserved.
[0040] (2) Screening of actinomycetes that antagonize Fusarium graminearum: Inoculate the bacterium block of Fusarium graminearum in the center of the PDA solid medium, inoculate actinomycetes around it, observe the antagonistic circle after cultivating at 28° C. for 3 days, and select cells that can Actinomycetes that antagonize Fusarium graminearum actinomycetes to produce antagonistic circles are screened for alginolytic bacteria.
[0041] (3) Screening of alginolytic actinomycetes: Inocula...
Embodiment 2
[0045] The cultivation of embodiment 2 bacterial strains
[0046] Seed medium, Gao's No. 1 medium: soluble starch 20g, KNO 3 1.0g, NaCl 0.5g, K 2 HPO 4 0.5g, MgSO 4 ·7H 2 O 0.5g, FeSO 4 ·7H 2 O 0.01g, add deionized water to 1L, agar powder 2% (m / v, add when solid), pH7.4-7.6.
[0047] Fermentation medium, Luria-Bertani (LB) medium diluted 5 times: tryptone 2g, yeast extract 1g, NaCl 1g, agar powder 2% (m / v, add when solid), deionized water to 1L , pH6.8-7.0.
[0048] Antagonist PDA medium: 200g potatoes, add water and boil for 30 minutes, filter with 2 layers of gauze, take the filtrate for medium preparation, glucose 20g, agar powder 2% (m / v, add when it is solid), add deionized water to 1L , pH natural.
[0049] Algae medium BG11 medium (Blue-Green Medium): NaNO 3 1.5g, K 2 HPO 4 0.04g, MgSO 4 ·7H 2 O 0.075g, CaCl 2 ·7H 2 O 0.036g, Na 2 CO 3 0.02g, citric acid 0.006g, ferric citrate 0.006g, EDTANa 2 .001g, trace element solution A5 1ml (A5:H 3 BO 3 ...
Embodiment 3
[0054] The identification of embodiment 3 bacterial strains
[0055] The strain is characterized as follows:
[0056] Colony morphology: The strain HG-16 grows on the modified Gaoshi No. 1 medium, producing white aerial hyphae and gray-pink spores. After prolonged culture, the colony turns yellow when it ages, and does not produce water-soluble pigments. Microscopic observation The spores are linear rods (see figure 1 a); On LB medium and PDA and other nutrient-rich medium, the colony is wrinkled, light yellow, does not form white aerial hyphae, does not produce spores, and forms basal hyphae closely connected with the medium , all the colonies could not be picked.
[0057] Cultivate Streptomyces alginolyticus with the above-mentioned seed liquid medium for 3 days, then collect the thalline by centrifugation at 10000×g, discard the supernatant, extract the total DNA of the thalline cells with a kit (Fast DNA SPIN kit, USA), and then use primers for PCR Amplified sequence, f...
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