Composite microbial inoculant containing bacillus subtilis and application of composite microbial inoculant in degrading straws at low temperature
A technology of Bacillus subtilis and compound bacterial agent, applied in the field of microorganisms, can solve the problems of high energy consumption and high operation requirements, and achieve the effects of strong soil purification, enhanced resistance, and obvious effect of increasing production.
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Embodiment 1
[0020] The mutagenic screening of embodiment 1 verrucous Myromyces
[0021] (1) Morphological identification and screening
[0022] The soil in the forest area of Changbai Mountain Nature Reserve was collected and cultivated on PDA medium by conventional methods. The mycelium of the strain was white flocculent at first, and grew divergently towards the periphery. After 8 days of culture, the color of conidia continued to deepen, and glue dots appeared; after 10 days of culture, the colony was in the shape of concentric rings, the conidia showed black glue spots, and the back of the colony appeared Hazel radial folds.
[0023] Use a puncher to get a 1cm-diameter bacterial block, inoculate it on an aniline blue selective medium, and culture it in an aerobic incubator at 30°C for 10 days, observe the fading of aniline blue, and select a strain with a fast fading rate and strong ability as candidate strains.
[0024] At the same time, in order to directional screen lignin-deg...
Embodiment 2
[0030] The low-temperature acclimatization of embodiment 2 bacterial strains
[0031] (1) Enrichment culture strains
[0032] Inoculate the strain into a Erlenmeyer flask filled with 150mL of PDA medium, add filter paper pieces with a length of 5cm and a width of 2cm, incubate at a constant temperature at 30°C for 1 hour and mix evenly, and make 10 -1 -10 -9 For dilution, 1 mL of each dilution was inoculated in PDA medium with filter paper as the only carbon source, and static enrichment culture was carried out at 28°C for 4 generations. Screen out the materials with short breakage time and high degree of ulceration of filter paper. PDA medium: potato 200g / L, glucose 20g / L, agar 15g / L, enrichment culture for 4 generations.
[0033] (2) Preliminary screening and domestication of low-temperature cellulose-degrading composite strains
[0034] Using the filter paper disintegration method, the above-mentioned enriched culture was inoculated into the filter paper strip medium to...
Embodiment 3
[0045] The preparation of embodiment 3 composite bacterial agents
[0046] Cellulase-producing Paenibacillus, xylanase-producing Alternaria, laccase-producing Mycobacterium verrucosus, lignin-peroxidase-producing Bacillus colloid, manganese-peroxidase-producing Lys Bacillus subtilis and Bacillus subtilis were activated in a conventional manner and cultured until the number of viable bacteria in the bacterial liquid reached 2.0×10 8 individual / mL.
[0047] Mix the bacterial solution according to the following mass ratio: 20% of Paenibacillus cellulase-producing bacteria, 10% of Alternaria xylanase-producing bacteria, 20% of Laccase-producing Mycobacterium verrucosus, lignin peroxidase glue 10% of Bacillus plasmodia, 10% of Manganese peroxidase-producing Layseia lanceolata, and 30% of Bacillus subtilis, fully stirred and mixed to obtain a mixed bacterial liquid.
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