Bacillus velezensis and application thereof
A technology of Bacillus Velez and Bacillus, applied in the direction of application, bacteria, fungicides, etc., can solve the problems of short duration and low colonization ability, achieve excellent performance, promote plant growth, and have good development and application prospects Effect
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Embodiment 1
[0026] Example 1 Isolation and purification of bacterial strain TYGF10-2F9 and molecular biology identification
[0027] Weigh 10g of wheat rhizosphere soil and add 200mL 0.01M sterile PBS buffer solution, shake at room temperature and 180r / min for 30min to obtain a rhizosphere soil suspension, and gradiently dilute to 10 with sterile water. -1 、10 -2 and 10 -3 , spread evenly on TYG medium (Tryptone Yeast extract Glucose Medium, TYG) with a coating stick, and its composition is: tryptone 1g, yeast extract 1g, glucose 0.5g, KCl 6.34g, NaCl 1.2g, MgSO 4 ·7H 2 O 0.25g, K 2 HPO 4 0.13g, CaCl 2 2H 2 O 0.22g, K 2 SO 4 0.17g, Na 2 SO 4 2.4g, NaHCO 3 0.5g, Na 2 CO 30.09g, FeEDTA 0.07g, agar 15g, distilled water 1000mL, PH=7.2-7.4; after constant temperature cultivation at 28°C for 2 days, pick a single colony for purification.
[0028] DNA extraction was performed in accordance with the procedures of the Column Bacterial DNA Extraction Kit of Shanghai Sangon Bioengi...
Embodiment 2
[0033] Example 2 Determination of Bacteriostasis of Bacterial Strain TYGF10-2F9
[0034] Plate confrontation method was used to control rape sclerotia (Sclerotinia sclerotiorum), rice sheath blight (Rhizoctonia solani), wheat gibberella (Fusarium graminearum), apple ring pattern (Botryospuaeria berengeriana), tobacco red star (Alteraria alternata), pepper phytophthora (Phytophthora capsicici) ), watermelon blight (Fusarium oxysporum f.sp.niveum), cucumber blight (Fusarium oxysporum.sp.cucumebrium Owen), strawberry blight (Fusarium oxysporum f.sp.fragariae), tomato gray mold (Botrytis cinerea), apple rot (Valsa mali Miyabe et Yamada) and wheat total eclipse (Gaeu-mannomyces graminsis (sacc.) Arx & Olivier Var tritici J.walker) a total of 12 plant pathogenic fungi were tested. First, take the center of the petri dish as the center point, draw a cross on the bottom of the petri dish, and inoculate the strains that have been activated on LB medium at a distance of 2.5 cm from the ...
Embodiment 3
[0038] Example 3 Determination of growth-promoting index of bacterial strain TYGF10-2F9
[0039] First, the strains stored at -80°C were streaked on LB medium, and cultured in a 28°C incubator for 24h to activate. Then inoculate a bacterial needle into a 250mL Erlenmeyer flask filled with 100mL LB liquid medium, and add 1mL L-tryptophan to the Erlenmeyer flask to make the final concentration 100μg / mL. Cultivate at 28°C and 180r / min for 5 days, and carry out 2 biological replicates for each strain. Take 8mL strain fermentation broth, centrifuge at 8000r / min, 4°C, absorb supernatant for quantitative determination of strain IAA content. For the quantitative determination of IAA content, 60 μL of supernatant was pipetted into a 2 mL centrifuge tube, and then 120 μL of Salkowski reagent was added to develop color under dark conditions, and the absorbance of each treatment at 530 nm was measured with a UV spectrophotometer. At the same time, a standard curve of IAA content and abs...
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