Efficient phosphorus-solubilizing aspergillus niger strain and application thereof
A technology of Aspergillus niger and phosphate-solubilizing bacteria, which is applied in the field of highly efficient phosphorus-solubilizing Aspergillus niger strains, can solve the problems of low grade, increased agricultural production costs, and large energy consumption, so as to increase the content of available phosphorus, achieve remarkable phosphorus-solubilizing effect, The effect of increasing yield
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[0018] Example 1 Preliminary screening of microorganisms in soil
[0019] Preparation of soil diluent: take calcareous soil from the mountainous area of southern Jinan, weigh 0.5g soil sample and dissolve it in 4.5ml sterile water to make a 1:10 soil solution, then draw 0.5ml soil solution and place it in 4.5ml without In the bacteria water, make a 1:100 soil solution, and by analogy with this method, prepare 1:10 6 -10 7 Dilute the soil solution.
[0020] Take 0.1ml of the soil dilution solution and evenly spread it on the insoluble inorganic phosphorus solid medium, and invert it in a 30℃ incubator for 2-3 days. Observe the colonies grown on the medium. Among them, 11 colonies have obvious surrounding The color changes and there are transparent circles, which are named YW-1, YW-2, YW-3,..., YW-11.
Example Embodiment
[0021] Example 2 Re-screening of Phosphate Solubilizing Microorganisms
[0022] The 11 strains obtained from the preliminary screening in Example 1 were respectively inoculated on a poorly soluble inorganic phosphorus solid medium and cultured at 30°C for 3 days. The size of the transparent circle around the colony was observed. It was found that the five bacteria with the largest transparent circle were YW- 2. YW-3, YW-7, YW-9, YW-10.
[0023] Inoculate the five bacteria with the largest transparent circle into 50 mL of insoluble inorganic phosphorus liquid medium, culture at 30°C, 200 rpm, for 6 days, and use the liquid phosphorus solubilizing medium without any bacteria as the control group; test and culture separately The number of viable bacteria in the liquid, the specific results are shown in Table 1.
[0024] 2.1 Plotting of phosphorus standard curve
[0025] Take 5mg / l of phosphorus standard solution 0.0, 0.2, 0.4, 0.8, 1.6, 2.0, 3.2, 4.0ml in the test tube in turn, the...
Example Embodiment
[0031] Example 3 Identification of YW-9 strain
[0032] 1) Colony morphology of strain YW-9:
[0033] The colony is dark brown, thick and velvety, with round and neat edges; the back of the colony is colorless or the center is slightly yellowish brown. There are a large number of conidia on the front of the colony, brown and black, without exudate; the flora is dark brown, with developed hyphae, and the conidia head is like a "chrysanthemum", with two layers of spore small stems, and the conidia are spherical. It is black, dark brown.
[0034] 2) Molecular biological identification of YW-9 strain:
[0035] The YW-9 strain obtained by the above screening was identified by molecular biology method, and its 18s rDNA sequence was measured, and the blast comparison was performed in the GenBank nucleic acid database. Based on the colony morphology of the YW-9 strain and the 18srDNA comparison result, the applicant confirmed that the YW-9 strain is Aspergillus niger (Aspergillus niger ),...
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