Multifunctional oligotrophic cellulose-degrading bacterium compound bacterial agent and application thereof
A cellulose-degrading bacteria and oligotrophic technology, applied in application, fungicide, bacteria and other directions, can solve the problems of lack of high temperature and salt resistance, eutrophication of water quality and deterioration of soil physical and chemical properties, inability of phosphorus and potassium elements, etc.
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Embodiment 1
[0029] Screening and Identification of Example 1 Bacterial Strains Lzh-X14 and Lzh-10
[0030] Strains Lzh-X14 and Lzh-10 are two oligotrophic strains isolated from Datian, Dezhou City, Shandong Province.
[0031] (1) Primary screening of cellulose degrading bacteria
[0032] Use the dilution coating method to apply the dilution to the oligotrophic bacteria screening medium (peptone 1g; yeast extract 0.5g; NaCl 1g; agar 20g, dissolved in deionized water 1000ml. Sterilize at 121°C for 30min.) and the resulting colonies Use a pick loop to pick and inoculate in oligonutrient cellulose Congo red medium (sodium carboxymethylcellulose 1.88g; Congo red 0.25g; MgSO 4 0.025g; K 2 HPO 4 0.05g; mix 20g of agar, dissolve in 1000ml of deionized water. Sterilize at 121°C for 30min. ), select the colony that can produce a transparent circle on this medium and use the three-section line method to purify the bacterial species.
[0033] (2) Screen high temperature and saline-alkali toler...
Embodiment 2
[0044] Antagonism of Bacillus amyloliquefaciens Lzh-X14 and Streptomyces Lzh-10 in embodiment 2 of the present invention
[0045] Bacillus amyloliquefaciens Lzh-X14 and Streptomyces Lzh-10 were inoculated on Gaoshi No. 1 solid medium in a criss-cross pattern, and cultured at 30°C for 24 hours. If there is a bacteriostatic zone, it means that the two strains have antagonistic effect. If the two strains grow cross-growth, the two strains have no antagonistic effect. figure 1 It shows that Bacillus amyloliquefaciens Lzh-X14 and Streptomyces Lzh-10 have no mutual antagonism, and can be used to prepare compound bacterial agents.
[0046] Adopt plate confrontation method to detect the antagonism of Bacillus amyloliquefaciens Lzh-X14 and Streptomyces Lzh-10, inoculate pathogenic bacteria Fusarium moniliforme on the PDA solid medium (200g of peeled potatoes, 20g of sucrose, 20g of agar, 1000mL of distilled water, natural pH), cultured at 30°C for 48 hours and then used for later use....
Embodiment 3
[0047] Example 3 Cellulose Degrading Bacteria Enzyme Activity Determination
[0048] The enzyme activity of cellulase was measured by DNS method, and the strain with higher enzyme activity was selected.
[0049] Experimental method: DNS method to measure cellulase enzyme activity: use the inoculation loop to inoculate the obtained strain into the general liquid medium at 120r / min and cultivate for 12-24h, until the bacterial liquid is turbid, centrifuge at 4000r / min for 10min and take the supernatant as crude Enzyme solution, take 1.0mL enzyme solution and put it into a 15mL test tube, add sodium carboxymethyl cellulose solution at 37°C for 30min, shake evenly, then react in a water bath at 45°C for 30min, add 3.0mL DNS reagent accurately, boil in water for 7min and flush After cooling and oscillating evenly, use a 722 spectrophotometer to compare color at a wavelength of 540nm, record the absorbance against the standard curve, and calculate the enzyme activity. Select the str...
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