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Root nodule azotobacter strain BXYD3 and uses thereof

A technology of bradyrhizobium and nitrogen fixation, applied in the direction of application, bacteria, animal repellent, etc., to achieve the effect of improving nitrogen nutrition

Inactive Publication Date: 2009-11-04
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For a long time, although soybeans have a certain planting area in the acidic phosphorus-deficient red soil area, common rhizobia are rarely used in the southern area due to the acidic soil in the south.

Method used

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  • Root nodule azotobacter strain BXYD3 and uses thereof
  • Root nodule azotobacter strain BXYD3 and uses thereof
  • Root nodule azotobacter strain BXYD3 and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Isolation and purification of nitrogen-fixing bacteria strain BXYD3

[0038] Take fresh root nodules, rinse them with water, and dry the surface moisture with filter paper. Put it in 95% ethanol for 3 minutes, take it out and wash it with sterile water for 5-6 times, then put it in 0.2% HgCl 2 Sterilize for 5 minutes, take out and rinse with sterile water 5-6 times. Then cut in half on a flame sterilized glass slide, clamp the half tumor with sterile tweezers, mark the incision facing the surface of the YMA (Table 1) medium, incubate at 28°C after inversion.

[0039] After the thalli grow out, select the rhizobia-like colony from the plate and streak on the plate for culture. After 3 days, observe the colony situation, and observe until 15 days, because Bradyrhizobium needs 7 to 15 days to appear colonies. According to the following two aspects, it can be preliminarily judged whether it is rhizobia: ①Colony shape: the colony of rhizobia is round, milky whit...

Embodiment 2

[0042] Example 2 Sequencing of the 16S rDNA sequence of nitrogen-fixing bacteria strain BXYD3

[0043] The total DNA of the strain was extracted, and PCR-specific amplification was carried out. After the PCR amplification product was checked on 1.0% agarose gel electrophoresis, it was sent to Yingjun Company for sequence determination. The sequencing result is shown in SEQ ID NO:1.

[0044] Eight reference strains were obtained from the GenBank database, and the 16S rDNA partial sequences of the isolated strains and reference strains were analyzed using the software GeneDoc and TREECONW, and a phylogenetic relationship diagram between the isolated strains and the standard strains was constructed, as shown in image 3 As shown, the isolated nitrogen-fixing bacteria strain BXYD3 was determined to be a new strain of Bradyrhizobium (Bradyrhizobium), and it was named Huagu No. 3.

Embodiment 3

[0045] Embodiment 3 tie-back test

[0046] The recertification of the tie-back test adopts the above-mentioned large test tube paper culture device ( Figure 4 ), soybean seed treatment is also as described above.

[0047]Bacterial liquid culture: use an inoculation loop to pick a ring of rhizobia in the test tube and put it into the TY liquid medium. The formula of the TY liquid medium is shown in Table 2. After plugging it with a cotton plug, seal it with kraft paper and put it on the shaker Cultivate until the logarithmic growth phase (about 4 to 6 days), and the culture temperature is 28°C.

[0048] Table 2TY liquid medium ( / L)

[0049]

[0050] Inoculation treatment: Put the germinated seeds in a petri dish and soak them in the bacterial solution for 15 minutes, plant the seedlings between the filter paper and the tube wall with tweezers, add 1-2mL of the bacterial solution to each seedling to ensure that each seedling The inoculation amount of seedlings reached 1.0...

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Abstract

The invention discloses a nitrogen-fixing bacterial strain BXYD3 and its application. The bacterial strain captures the rhizobia in the soil through different soybean varieties, separates and purifies the rhizobia from fresh nodules, and extracts the 16S rDNA part of the strain. The sequence was determined, and through phylogenetic analysis, it was determined to be a new strain of Bradyrhizobium (Bradyrhizobium), and it was named Huagu No. 3. The nitrogen-fixing bacterium rhizobium strain BXYD3 of the present invention has been proved by laboratory potting and field back-inoculation tests that it has the characteristics of acid-resistant aluminum and high-efficiency nodulation. Inoculation of the rhizobia can increase soybean yield by more than 20-30%, and it can be applied to acidic soil areas in South China , as one of the measures of conventional soybean cultivation.

Description

technical field [0001] The invention relates to a nitrogen-fixing bacterial strain BXYD3 and its application. Background technique [0002] Biological nitrogen fixation is one of the major basic research topics in life sciences, and it plays an important role in production practice. Symbiotic nitrogen fixation is the most important type of biological nitrogen fixation. The symbiosis between legumes and rhizobia has become the focus of biological nitrogen fixation research because of its strong nitrogen fixation capacity, large nitrogen fixation capacity, and strong stress resistance. A large number of experiments at home and abroad have proved that inoculating rhizobia in most soils can improve the yield and quality of legume crops. Inoculation of high-efficiency rhizobia, as the main measure to increase soybean yield, has been widely used in northern my country. [0003] At present, soybean has become one of the most valuable leguminous crops in the vast acidic phosphoru...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/00
Inventor 曹桂芹廖红严小龙
Owner SOUTH CHINA AGRI UNIV
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