Aspergillus sepultus and application thereof
A technology of Aspergillus and seeds, applied in the field of microorganisms, can solve the problems of Aspergillus tubingensis CT1's phosphorus-solubilizing ability and inapplicability, and achieve the effects of increased content, enhanced utilization rate, and enhanced root system
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Embodiment 1
[0048] Example 1 Acquisition and morphological identification of buried Aspergillus sepultus MN114011
[0049] (1) Strain acquisition
[0050] Aspergillus sepultus MN114011 was isolated from agricultural soil collected from a peanut planting site in Jiayi Town, Yanshan County, Wenshan Prefecture, Yunnan Province (23°43′17.07″, 104°02′5.77″).
[0051] The separation method is as follows:
[0052] Weigh 5g of soil, add 45mL of sterile water, shake with a vortex shaker for 5min, and then dilute to 10 -1 ~10 -5 , take 10 -3 、10 -4 and 10 -5 Three gradient bacterial solutions were spread on the flat plate, spread on the organophosphate solid medium, each coating was repeated three times, and cultured upside down in a constant temperature incubator at 28°C for 4 days, after 4 days, the circles with phosphate solution were picked out The strains were purified and stored in glycerol tubes for the next step of re-screening;
[0053] The formula of organic phosphorus solid medium...
Embodiment 2
[0058] Example 2 Molecular identification of buried Aspergillus sepultus MN114011
[0059] The strain was streaked on a plate to obtain a single colony, and the single colony was picked and placed in a shaker tube containing 2mL TSB, and placed at 28°C for shaking culture until turbid; Take 1mL of the bacterial solution in a centrifuge tube, centrifuge at 10000rpm for 3min, discard the supernatant, add 50 μL of sterile water, shake well, incubate at 95°C for 2 minutes, and place in a refrigerator at 0°C for later use;
[0060] Use a toothpick to pick a small amount of hyphae into 20 μL lysate, and lyse at 95°C for 30 minutes; pipette 2 μL lysate into the PCR system, which includes 12.5 μL premix TaqMaster Mix, 10 μL ddH 2 O, 0.7 μ L ITS4 (SEQ ID NO: 2), 0.7 μ L ITS5 (SEQ ID NO: 3), 0.6 μ L DMSO, carry out PCR amplification;
[0061] Take 5 μL of the amplification product, add 1 μL of loading buffer, mix well, perform agarose gel electrophoresis for 45 min, and detect the ampl...
Embodiment 3
[0065] Example 3 strain preservation
[0066] Based on the morphological observation and molecular identification results of Examples 1-2, the strain MN114011 belongs to Aspergillus sepultus, and Aspergillus sp. MN114011 was preserved in Guangdong Microbial Culture Collection Center (GDMCC) on May 14, 2019 , the address is Guangdong Institute of Microbiology, Building 59, Compound, No. 100 Xianlie Middle Road, Guangzhou City, postcode 510070, and the deposit number is GDMCC 60666.
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