Saline-alkali soil resistant nitrogen fixation rhizobium japonicum and application thereof

A technology for saline-alkali soil and slow rhizobium, which is applied in the field of agricultural microorganisms, can solve problems such as inability to apply to alkaline soil, and achieves the effects of strong nitrogen fixation ability, increased total nitrogen content, and increased nitrogen content.

Active Publication Date: 2022-05-10
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the bradyrhizobia identified so far are only applicable to acidic soil areas in South China, but not to alkaline soils, such as saline-alkali soils in rocky desertification areas

Method used

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  • Saline-alkali soil resistant nitrogen fixation rhizobium japonicum and application thereof
  • Saline-alkali soil resistant nitrogen fixation rhizobium japonicum and application thereof
  • Saline-alkali soil resistant nitrogen fixation rhizobium japonicum and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Isolation and purification of strains

[0039] Collect the nodules on the soybean roots cultivated in the rocky desertification area of ​​Xingyi, Guizhou, select fresh and full nodules, soak the nodules in sterile water for 5-6 minutes in the ultra-clean bench, remove the impurities on the surface, and use 75% anhydrous ethanol. After surface disinfection for 3 to 5 minutes, disinfect with 3.5% NaClO for 8 minutes, and then rinse it with sterile water for 9 to 10 times; then cut a single root nodule with a sterilized knife, and then clamp the root nodule with tweezers. The solid YMA medium was streaked and cultured upside down in a constant temperature incubator at 28°C for 5 to 7 days until clear colonies grew. The obtained rhizobia monoclones were streaked and purified 3-4 times until clear monoclones were grown, single colonies were picked to transfer slants, and stored at 4°C at low temperature, and the obtained strains were named as Dian strains.

[0040...

Embodiment 2

[0042] Example 2 Identification of bacterial strains

[0043] 1. Morphological identification of strains

[0044] The Dian strain isolated in Example 1 was inoculated on YMA solid medium for cultivation, and recorded. After culturing for 5 to 7 days under the optimum growth conditions of pH 7.0 and temperature of 28 °C, the isolated and purified strain Dian was subjected to single colony state observation, mainly including colony size, color, transparency, wetness, colony surface state, and colony edge. state. At the same time, for the Dian strain in the logarithmic growth phase, the morphology of the bacteria was observed by light microscope after smear staining.

[0045] The result is as figure 1 shown, indicating that the Dian strain is Gram-negative, short rod-shaped, without spores, most of which are motile. Grow on solid YMA medium plate, the colony shape is approximately round, milky white, translucent, neat edge, more sticky, colony diameter of 0.8mm ~ 1mm.

[004...

Embodiment 3

[0055] Example 3 Performance identification of soybean rhizobia Dian strain

[0056] Prepare 1L YMA liquid medium, divide it into 3 conical flasks, and sterilize it at 121°C for 20min; pipette 100μL of the rhizobial liquid that has been purified and reach the logarithmic growth phase into the triangle containing YMA medium. In the bottle, shake at 180 r / min overnight on a shaker at 28 °C to the OD of the bacterial solution 600 After the value reaches 0.6±0.05, it is divided into sterilized 50mL centrifuge tubes, centrifuged at 5000rpm in a centrifuge for 10min, taken out, poured off the supernatant, and resuspended with low nitrogen nutrient solution to obtain a bacterial suspension for later use.

[0057] Take 5 μL of the prepared rhizobia suspension on the soluble starch solid medium for spot plate inoculation, incubate in a constant temperature incubator at 28°C for 3 to 5 days until obvious single clones appear, and evenly add iodine solution on the plate, around the colon...

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Abstract

The invention discloses a nitrogen-fixing bradyrhizobium japonicum strain resistant to saline-alkali soil and application of the bradyrhizobium japonicum strain. According to the invention, a strain of bradyrhizobium taihuae Dian strain is obtained through separation and is preserved in Guangdong Microbial Culture Collection Center on January 11, 2022, and the preservation number is GDMCC No: 62202. Researches show that the strain has the characteristics of high nodulation rate and strong nitrogen fixation capability, and can optimize the micro-ecological environment of alkaline soil crop root growth; the Dian strain is adopted to treat plants, soybean nodulation and nitrogen fixation can be effectively promoted, plant nitrogen nutrition is increased, plant growth is promoted, and the biomass, root nodule number and nitrogen content of soybean plants can be remarkably improved. The strain is especially suitable for cultivation and production of soybeans in stony desertification areas, expands a strain resource library of nitrogen-fixing rhizobium, is beneficial to promoting nodulation and nitrogen fixation of plants, and increases nitrogen nutrition of the plants.

Description

technical field [0001] The invention belongs to the technical field of agricultural microorganisms. More specifically, it relates to a saline-alkali-tolerant nitrogen-fixing soybean rhizobia and its application. Background technique [0002] Rhizobium is a class of Gram-negative bacteria widely distributed in soil. The symbiotic nitrogen fixation system between legumes and rhizobia has always been a research hotspot. The physical and chemical properties of soil are important factors affecting the symbiotic nitrogen fixation of rhizobia and leguminous plants. In karst ecosystems, the rhizobia symbiotic with legumes are dominated by Bradyrhizobium (Liu Lu et al., 2018). Studies have shown that the formation of a symbiotic nitrogen fixation system between legumes and rhizobia can improve the problems of soil structure deterioration and soil nutrient deficiency caused by pollution and degradation. For example, Li Jianhua (2010) inoculated Rhizobium bacteria while planting wh...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/20A01P21/00A01G7/06A01G22/40C12R1/01
CPCC12N1/20A01N63/20A01G7/06A01G22/40
Inventor 梁翠月陈倩倩陆星谢宝星刘国选管子迪田江田继辉
Owner SOUTH CHINA AGRI UNIV
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