Preparation method and application of a saline-alkali tolerant air bacillus and its live bacterial preparation
A technology of bacillus and living bacteria preparations, applied in the field of agricultural biology, can solve the problems of failure to spread in large areas and limited control effects, and achieve the effects of improving raw material utilization, scientific production methods, and low production costs
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Embodiment 1
[0021] Example 1: Screening of Aerobacillus strain PAPM-14
[0022] (1) Screening of saline-alkali-tolerant strains
[0023] Aerobacillus PAPM-14 was isolated from maize rhizosphere soil. The soil was sampled in Bincheng District, Binzhou City, Shandong Province in September 2018, which is saline-alkali soil. The specific separation method is as follows: mix the soil sample evenly, weigh 5g, put it into a conical flask filled with 95mL sterile water and 10 glass beads, shake it at 37°C and 180rpm for 30min. Take 1 mL of soil suspension for 10 -1 —10 -7 Serial concentration gradient dilution, and then take 10 -5 、10 -6 、10 -7 The three dilutions were spread on the plate containing the selection medium, and cultured upside down at 37°C for 2 days. Then, different single colonies were picked and streaked onto a plate containing selection medium, and cultured upside down at 37°C for 2 days. Pick a single colony from the plate and transfer it to the slant of the preservatio...
Embodiment 2
[0037] Example 2: Identification of Aerobacillus PAPM-14
[0038] (1) Morphological and physiological and biochemical characteristics
[0039] The morphological characteristics of the PAPM-14 strain are: cultured at 37° C. for 48 h on the NA medium, the colony is round, raised, moist, wrinkled, opaque, and milky white; the thalline is short rod-shaped, such as figure 1 shown. The NA medium is nutrient agar medium, and its formula is: beef extract 3g, peptone 10g, NaCl 5g, agar 20g, water 1000mL, pH 7.0.
[0040] The physiological and biochemical characteristics of the PAPM-14 strain are: Gram-positive rod-shaped bacteria; catalase test positive, citrate utilization positive, malonate utilization negative, nitrate reduction positive, V-P test positive; starch The hydrolysis test was positive, the D-glucose fermentation test was positive, the D-mannitol fermentation test was positive, the lactose fermentation test was positive, and the maltose fermentation test was positive. ...
Embodiment 3
[0044] Example 3: Functional Verification of Airbacillus PAPM-14 Strains
[0045] (1) Ability of producing IAA by Bacillus aerogenes PAPM-14 strain
[0046] Add airbacillus PAPM-14 to the LB liquid medium containing L-tryptophan (200mg / L), culture at 37°C, 180r / min, and shaker for 4 days. Take 50 μL of bacterial suspension and drop it on a white ceramic plate, and add 50 μL of Salkowski colorimetric solution at the same time. Add 50 μL of 50 mg / L IAA colorimetric solution as positive control CK. The white ceramic plate was placed at room temperature in the dark for 30 minutes and then observed. Those that turned red indicated that they could secrete IAA. The color of bacterial strain PAPM-14 suspension was red after the test treatment, indicating that bacterial strain PAPM-14 had a strong ability to secrete IAA ( figure 2 ).
[0047] The formula of the LB liquid medium is: peptone 10.0g, yeast extract 5.0g, NaCl 10.0g, distilled water 1000mL; sterilized at 121°C for 20min...
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