Low-temperature-resistant efficient crop decomposition bacterial liquid and preparation method and use thereof
A low-temperature-resistant technology for crops, applied in the field of low-temperature-resistant and high-efficiency crop decomposing bacteria solution and its preparation, can solve the problems of limiting the degradation function of cellulose-degrading bacteria, low degradation efficiency, and single strain composition
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Embodiment 1
[0047] The invention provides a method for preparing a low-temperature-resistant and high-efficiency crop straw decomposing bacterial liquid, comprising the following steps:
[0048] Step 1, enrichment acclimation
[0049] Take 10 mL of soil bacterial suspension according to aseptic technique and put it into 90 mL PCS enrichment medium, carry out enrichment culture at 30 ℃, 150r / min shaker shaking conditions, and select filter paper strips from which the color changes obviously or collapses. The solution was transferred to the next-generation enrichment medium for subculture, and the temperature gradient subculture was reduced by 5°C in each generation to enrich and acclimatize the strains that efficiently degrade straw by volume fraction of 5% (V / V) inoculum .
[0050] Step 2, screening and separation
[0051] Dilute the enriched culture solution in step 1 by a 10-fold gradient method to obtain 10 -3 ~10 -8 For each dilution, draw 100 μL of the dilution and spread it on a...
Embodiment 2
[0061] A kind of low-temperature-resistant and high-efficiency agricultural crop straw decomposing bacterial liquid described in the present invention selects the following 6 kinds of effective active microorganisms: Pseudomonas ( Pseudomonas sp.), Acinetobacter ( Acinetobacter sp.), Trichoderma harzianum ( Trichoderma harzianum Rifai ), Penicillium crustacea ( Eupenicillium crustaceum ), Pediococcus pentosacea ( Pediococcus pentosaceus ), the number of effective viable bacteria in the bacterial fertilizer ≥ 1×10 9 cfu / mL.
[0062] Screening of low temperature resistant and efficient crop straw degrading bacteria.
[0063] According to aseptic operation, use sterile water to make 2 strains of Pseudomonas, 1 strain of Acinetobacter, 1 strain of Trichoderma harzianum, and 1 strain of Penicillium clatices to make a bacterial suspension / spore liquid with the same concentration. Take 5mL and inoculate it into a Erlenmeyer flask containing 45mL of enzyme-producing medium, a...
Embodiment 3
[0069] According to aseptic operation, use sterile water to make 2 strains of Pseudomonas and 1 strain of Acinetobacter into a bacterial suspension / spore liquid with the same concentration, and inoculate 5 mL each into the medium with filter paper as the substrate. ~30°C range, 150r / min shaker culture for 10 days to take samples, and add an appropriate amount (5~10mL) of sulfuric acid and sodium nitrate mixed solution to remove bacteria and insoluble calcium carbonate, pass through an 80-mesh sieve, and sieve the remaining bottom The samples were washed with light buffer of distilled water, dried in an oven at 80°C to constant weight, weighed, and the weight loss rate was calculated. Three parallel samples were used in each group.
[0070] Measurement results such as Figure 6 shown.
[0071] Such as Figure 6 As shown, at 5°C, the filter paper weight loss rate of the three low-temperature resistant cellulose-degrading bacteria is at the lowest level of about 7%; at the opti...
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