Application of bacillus licheniformis in straw degradation, microbial inoculant containing bacillus licheniformis, and application of microbial inoculant
A technology of Bacillus licheniformis and microbial agent, applied in the field of microorganisms, can solve the problems of slow degradation speed and poor adaptability to low temperature in late autumn
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Embodiment 1
[0038] This Example 1 is used to illustrate the primary screening of Bacillus licheniformis of the present invention.
[0039] Collect 5 samples from broiler chicken manure compost products in Yanqing City, Beijing, use the method of dilution and plating, use LB medium (containing 10g / L tryptone, 5g / L yeast extract and 10g / L NaCl) Bacteria were isolated, and 34 strains with different colony forms were divided into 34 strains. The isolated bacteria were streaked and purified multiple times to obtain pure culture strains and stored. The 34 bacterial strains obtained by purification were respectively spotted on the sodium cellulose-Congo red medium (sodium carboxymethylcellulose 5g, (NH 4 ) 2 SO 4 2g, KH 2 PO 4 1g, MgSO 4 ·7H 2 O 0.5 g, NaCl 0.5 g, NaNO 3 1g, peptone 0.5g, yeast extract 0.5g, Congo red 0.2g, agar 12g, water 1000mL, pH 7.0), cultured at 18°C and 30°C respectively, and measured the colony diameter (d) and transparent circle diameter ( D), calculate the...
Embodiment 2
[0041] This Example 2 is used to illustrate the re-screening of Bacillus licheniformis with straw degrading ability in the present invention.
[0042] A 100mL triangular flask was used, and 2 g of crushed corn stalks, 30 mL of nutrient solution (3.0 g of urea, (NH 4 ) 2 SO 4 6.0 g, peptone 3.0 g, CaCl 2 0.1 g, MgSO 4 ·7H 2 O 0.5 g, K 2 HPO 4 1.0 g, NaCl0.1 g, FeSO 4 ·7H 2 O 0.05 g, MnSO 4 ·7H 2 O 0.016 g, ZnSO 4 ·7H 2 O 0.014 g, CoCl 2 · 6H 2 O0.037g, distilled water 1000mL, natural pH), sterilized at 121℃ for 30min. Inoculate the strains with straw degrading ability obtained through preliminary screening into LB medium for 48 hours of shaking culture, then inoculate 1 mL of bacterial liquid into straw medium, place them in incubators at 18°C and 30°C for cultivation, and shake every day Twice, when culturing for 7d and 15d, the stalks in the bottle were washed 3 times with distilled water and dried to constant weight. The weight of corn stalks in each trea...
Embodiment 3
[0046] This Example 3 is used to illustrate the fermentation culture of Bacillus licheniformis of the present invention.
[0047] The Bacillus licheniformis ( Bacillus licheniformis ) was activated by streaking on LB medium, cultured for 48 hours, picked bacterial lawn and inoculated in LB liquid medium for 24 hours on a shaking table, and inoculated peptone beef extract medium (containing 20g / L peptone, 5g / L of beef extract and 5g / L of NaCl), 10 liters of seed tanks were cultivated, and after 24 hours, they were inoculated into 200 liters of fermentation tanks with a 5% inoculation amount and adopted corn steep liquor medium (corn steep liquor 2.3%, soybean meal Powder 0.6%, MgSO 4 ·7H 2 O 0.2%, K 2 HPO 4 0.05%, CaCO 3 0.05%, pH 7.0) for fermentation, sampling and observation every 4 hours until more than 80% of the bacteria form spores.
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