A kind of saline-alkali land improving microorganism and its application
A technology of saline-alkali land and application rate, applied in the field of environmental microorganisms, can solve problems such as affecting the sustainable development of agriculture, restricting the sustainability of cultivated land use, and reducing the productivity of cultivated land.
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[0058] The third aspect of the present invention provides a method for preparing a liquid microbial agent, which at least includes the following steps: activating the pure bacteria of the sea-sourced bacteria Idiomarina sp.YJY02 and then culturing it to obtain the liquid microbial agent.
[0059] Further, after activation and seed culture of the pure sea-source bacteria Idiomarina sp.YJY02, they were inoculated in LB liquid medium for culture.
[0060] Furthermore, the culture temperature is 25°C-35°C, for example, it may be 30°C.
[0061] Furthermore, the rotation speed during the cultivation is 150r / min-200r / min, for example, it can be 180r / min.
[0062] Furthermore, the culture time may be 48 hours.
[0063] Wherein, the LB liquid medium: peptone 10g / L, yeast extract 5g / L, sodium chloride 10g / L.
[0064] The fourth aspect of the present invention provides a solid bacterial agent, the solid bacterial agent includes sea-derived bacteria Idiomarina sp.YJY02, and the concentr...
Embodiment 1
[0079] Example 1: Isolation, screening and performance determination of microbial strains improved in saline-alkali land
[0080] (1)) Collect 5 g of saline-alkali soil, add it to an Erlenmeyer flask filled with 100 mL of sterile water, and incubate at 180 r / min at 30°C for 30 min to fully disperse the microorganisms in the tested soil in the sterile water to obtain a soil mixture.
[0081] (2) Dilute the mixed solution of step (1) according to the gradient, and select the concentration gradient to be 10 -2 、10 -3 、10 -4 、10 -5 、10 -6 The soil mixture solution was spread on LB solid medium containing 100g / L sodium chloride for the isolation of salt-tolerant microorganisms, and the plate was inverted and placed in a constant temperature incubator at 30°C for 48h to 96h. The composition of LB solid medium containing 100g / L sodium chloride is: peptone 10g / L, yeast extract 5g / L, sodium chloride 100g / L, agar 15g / L.
[0082] (3) According to the differences in colony shape, col...
Embodiment 2
[0090] Embodiment 2: the molecular biological identification of bacterial strain YJY02
[0091] The strain YJY02 was identified by the method of 16S rRNA gene sequence analysis. In this experiment, the AxyPrep Bacterial Genomic DNA Miniprep Kit was used to extract the genome, and common primers commonly used in bacteria were used for PCR.
[0092] The upstream primer is 27F: 5'-AGAGTTTGATCCTGGCTCAG-3',
[0093] The downstream primer was 1492R: 5'-GGTTACCTTGTTACGACTT-3'.
[0094] PCR uses 20 μL reaction system: 10 μL Taq enzyme, 0.5 μL template, 0.6 μL each primer, add ddH 2 O to a total system volume of 20 μL.
[0095] PCR reaction program: After pre-denaturation at 94°C for 5 min, start the following cycle program: denaturation at 94°C for 30 s, annealing at 55°C for 30 s, extension at 72°C for 90 s, cycle 30 times, additional extension at 72°C for 10 min, and cooling at 16°C for 5 min.
[0096] The PCR products were detected by 1.5% agarose gel electrophoresis, and sent t...
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