Sinorhizobium nitrogen-fixing strain H6 and application thereof

A technology of Sinorhizobium and Sinorhizobium, applied in application, bacteria, horticulture, etc., can solve problems such as affecting the yield of alfalfa, and achieve the effect of increasing biomass

Active Publication Date: 2018-11-20
XINJIANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Xinjiang is located in the northwestern border of China, and the unique climatic conditions are suitable for the growth of alfalfa, but the salinization of Xinjiang soil seriously affects the increase of alfalfa yield

Method used

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  • Sinorhizobium nitrogen-fixing strain H6 and application thereof
  • Sinorhizobium nitrogen-fixing strain H6 and application thereof
  • Sinorhizobium nitrogen-fixing strain H6 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Isolation and purification of Azotobacter sinensis strain H6:

[0044] Take fresh root nodules, rinse them with water, blot the surface moisture with filter paper, put them in ethanol with a volume concentration of 95% for 3 minutes, take them out, rinse them with sterile water for 5-6 times, and then put them in with a volume concentration of 0.2% HgCl 2Sterilize for 5 minutes, take it out, rinse it with sterile water for 5-6 times, then cut the fresh root nodule in half on a flame sterilized glass slide, clamp half of the nodule with sterile tweezers, and cut it facing YMA for culture The surface of the substrate was marked, and the formula of the YMA medium was shown in Table 1, and cultured at a temperature of 28 °C after inversion;

[0045] Table 1 YMA (Yeast Mannitol Agar) medium formula ( / L)

[0046]

[0047] After the growth of the bacteria, select the colonies that look like rhizobia from the plate and culture them on the plate. After 3 days, observe the c...

Embodiment 2

[0053] Sequencing of the 16S rDNA sequence of Azotobacter sinensis strain H6:

[0054] Use the kit to extract the total DNA of the strain, and perform PCR-specific amplification. After the PCR amplification product is checked on 1.0% agarose gel electrophoresis, it is sent to Sangon Company for sequence determination. The sequencing result is shown as SEQ ID NO:1 shown;

[0055] Sinorhizobium

[0056] >H6

[0057]

[0058]

[0059] The reference strain was obtained from the GenBank database, and the 16SrDNA partial sequence of the isolated strain and the reference strain was analyzed by using the software MEGA, and a phylogenetic relationship diagram between the isolated strain H6 and the standard strain USDA1002 (X67222.2) was constructed, as shown in image 3 As shown, thus confirming that the isolated Rhizobium H6 is a new strain of Sinorhizobium (Sinorhizobium).

Embodiment 3

[0061] Tieback test:

[0062] The alfalfa varieties used in this experiment are Zhongmi No. 3 (ZM) and Algonquin (GJ), which were selected according to hydroponic experiments. ZM is a salt-tolerant variety in the vegetative growth period, and GJ is a salt-sensitive variety. Alfalfa back-inoculation experimental plant material; set no rhizobia (-Ri), connect rhizobia standard strain USDA1002 and Sinorhizobium strain H6 three kinds of inoculation treatments, salt treatment is divided into two NaCl salt gradients of 0mM and 100mM, test A completely randomized block design was adopted, with 4 repetitions for each treatment;

[0063] The test was carried out in the 811 cultivation room of the Root Biology Center of South China Agricultural University. The cleaned quartz sand was mixed with coarse, medium and fine sand at a ratio of 1:2:1, and then sterilized at high temperature. In the pot, the alfalfa seeds were sterilized according to the above method and placed in a filter pape...

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Abstract

The invention relates to a sinorhizobium nitrogen-fixing strain H6 and application thereof. The sinorhizobium nitrogen-fixing strain H6 was deposited at the China Center for Type Culture Collection ofWuhan University on May 7, 2018, which is referred to as CCTCC for short, and the preservation number is CCTCC NO: M2018255. The sinorhizobium nitrogen-fixing strain H6 is obtained by capturing rhizobia in soil by an alfalfa plant, and isolating and purifying the rhizobia from a fresh root nodule; partial 16S rDNA sequence of the strain is measured, the sinorhizobium is determined as a new strainby phylogenetic analysis, and is named H6. The sinorhizobium nitrogen-fixing strain H6 provided by the invention is proved to have the characteristics of alleviating alfalfa salt damage and fixing nitrogen efficiently by a sand culture back-test; under a salt treatment condition, the biomass of the alfalfa is increased by inoculating the rhizobia, and the nitrogen, phosphorus and potassium contents in a plant are improved; and the sinorhizobium H6 can be applied to salinized areas such as Xinjiang and the like as one of measures for conventional cultivation of the alfalfa.

Description

technical field [0001] The invention relates to a Sinorhizobium nitrogen-fixing bacteria strain H6 (Sinorhizobium sp.) and application thereof. Background technique [0002] There are some microorganisms in nature that can reduce nitrogen in the air to ammonia and provide nitrogen sources for microorganisms and plants. This phenomenon is called biological nitrogen fixation. According to the relationship between higher plants and nitrogen-fixing microorganisms, biological nitrogen fixation can be divided into three systems: self nitrogen fixation, symbiotic nitrogen fixation, and joint nitrogen fixation. [0003] Rhizobia are Gram-negative, non-spore-forming Agrobacteria that can form a mutually beneficial symbiotic relationship with the legume alfalfa. This symbiotic relationship has the characteristics of large nitrogen fixation and strong stress resistance. [0004] Alfalfa is a high-quality forage for livestock. It is widely planted around the world. It is known as a "c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01G22/40A01G7/06C12R1/01
CPCA01G7/06A01G22/40C12N1/20C12N1/205C12R2001/01
Inventor 盛建东王秀荣盛立超李锐
Owner XINJIANG AGRI UNIV
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