Saline-alkali tolerant growth-promoting bacterium and application of growth-promoting bacterium in improving saline-alkali land

A technology of saline-alkali land and single-cell bacteria, which is applied in the field of environmental microorganisms, can solve problems such as high cost and secondary salinization of land, and achieve the effects of good economic benefits, high treatment efficiency and convenient operation

Active Publication Date: 2020-05-22
SHANGHAI ADVANCED RES INST CHINESE ACADEMY OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition to the naturally formed saline-alkali land, people's abuse of chemical fertilizers and pesticides and improper farming methods will also cause the accumulation of salt in the cultivated layer, leading to the formation of secondary salinization of the land
At present, domestic and foreign research on saline-alkal

Method used

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  • Saline-alkali tolerant growth-promoting bacterium and application of growth-promoting bacterium in improving saline-alkali land
  • Saline-alkali tolerant growth-promoting bacterium and application of growth-promoting bacterium in improving saline-alkali land
  • Saline-alkali tolerant growth-promoting bacterium and application of growth-promoting bacterium in improving saline-alkali land

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preparation example Construction

[0071] The third aspect of the present invention provides a method for preparing a liquid bacterial agent, which at least includes the following steps: after activating and culturing the pure bacteria of Halomonas aquamarina DB01 in seawater, inoculate it in LB liquid medium, and inoculate it at 30°C for 180r / min cultured for 48h to obtain liquid bacterial agent.

[0072] Wherein, the LB liquid medium: peptone 10g / L, yeast extract 5g / L, sodium chloride 10g / L.

[0073] The fourth aspect of the present invention provides a solid bacterial agent, the solid bacterial agent includes Halomonas aquamarina DB01, the concentration of Halomonas aquamarina DB01 is at least 1 × 10 9 cfu / g.

[0074] The fifth aspect of the present invention provides a kind of preparation method of solid bacterial agent, at least comprises the steps: the liquid bacterial agent that comprises Halomonas aquamarina DB01 is obtained solid bacterial agent after drying.

[0075] Furthermore, spray drying, vacu...

Embodiment 1

[0088] Example 1: Isolation, screening and performance determination of saline-alkali-resistant growth-promoting bacteria DB01

[0089] (1) Collect saline-alkali soil from different regions, take 1-5g sample and put it into a conical flask filled with 100mL sterile water, and shake at 180r / min at 30°C for 30min to fully disperse the microorganisms in the tested soil in the sterile water. Bacterial water to obtain a soil mixture.

[0090] (2) The obtained mixed solution is diluted according to the gradient, and the concentration gradient is selected to be 10 -2 、10 -3 、10 -4 、10-5 、10 -6 Soil mixture solution, in high-salt and high-alkali LB solid medium (peptone 10g / L, yeast extract 5g / L, sodium chloride 100g / L, agar 15g / L, adjust the pH to 10 with sodium carbonate after sterilization) Coating on the plate is used for primary screening to isolate saline-alkali-resistant microorganisms, and the plate is inverted and placed in a constant temperature incubator at 30°C for 48h...

Embodiment 2

[0098] Embodiment 2: Molecular biological identification of saline-alkali-tolerant growth-promoting bacteria DB01

[0099] The strains were identified by 16S rDNA sequence alignment. The genomic DNA of strain DB01 was extracted according to the prior art, and using this as a template, a pair of universal primers (27F, 1492R) was used to amplify strain 16S rRNA.

[0100] The upstream primer is 27F (5'-AGAGTTTGATCCTGGCTCA-3'),

[0101] The downstream primer is 1492R (5'-GGTTACCTTGTTACGACTT-3').

[0102] The PCR reaction system (20 μL) is as follows: template DNA 0.5 μL, PCR Taqmix 10 μL, upstream and downstream primers 0.6 μL each, add ddH 2 O to 20 μL of the reaction system.

[0103] PCR program: pre-denaturation at 94°C for 5 min, denaturation at 94°C for 30 s, annealing at 55°C for 30 s, extension at 72°C for 1 min and 30 s, and a total of 30 cycles of the above cycles, extension at 72°C for 10 min, and finally storage at 4°C.

[0104] PCR products were purified and seque...

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Abstract

The invention belongs to the technical field of environmental microorganisms, in particular to a saline-alkali tolerant growth-promoting bacterium and application of the growth-promoting bacterium inimproving saline-alkali land. According to the growth-promoting bacterium being halomonas aquamarina DB01, the collection number of the halomonas aquamarina DB01 is CCTCC NO: M 2019511; moreover, thegrowth-promoting bacterium has high adaptability to high-salinity and high-alkali environments, and can grow well in a saline-alkali soil environment; the growth-promoting bacterium can effectively reduce the content of soluble salt in the saline-alkali land, and increases plant biomass and yield; and the growth-promoting bacterium can be used for improving primary and secondary saline-alkali soilof farmland, forests and the like, reduces the toxic action of saline-alkali stress on plants, and has relatively high application value.

Description

technical field [0001] The invention relates to the technical field of environmental microorganisms, in particular to a saline-alkali-tolerant growth-promoting bacterium and its application in saline-alkali land improvement. Background technique [0002] Land salinization is a worldwide soil degradation problem. Salinization not only reduces the productivity of cultivated land, but also seriously restricts the sustainable use of cultivated land, directly affecting the sustainable development of agriculture. The distribution of saline-alkali land in my country is very wide, widely distributed in North China, Northwest, Northeast West and coastal areas north of the Yangtze River, with a total area of ​​about 1×10 8 hm 2 . In addition to the naturally formed saline-alkali land, the abuse of chemical fertilizers and pesticides and improper farming methods can also cause the accumulation of salt in the cultivated layer, leading to the formation of secondary salinization of the ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C09K17/14A01N63/20A01P21/00C12R1/01C09K101/00
CPCA01N63/00C09K17/14C09K2101/00C12N1/20C12N1/205C12R2001/01
Inventor 刘莉孙雪王娜史吉平崔文会李泳洪
Owner SHANGHAI ADVANCED RES INST CHINESE ACADEMY OF SCI
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