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Root nodule azotobacter strain BXBL9 and uses thereof

A slow rhizobium, nitrogen fixation technology, applied in applications, bacteria, animal repellents, etc., to achieve the effect of improving nitrogen nutrition

Inactive Publication Date: 2008-05-21
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 BXBL9 and uses thereof
  • Root nodule azotobacter strain BXBL9 and uses thereof
  • Root nodule azotobacter strain BXBL9 and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Isolation and purification of nitrogen-fixing bacteria strain BXBL9

[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 Azotobacter tumefaciens strain BXBL9

[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 Figure 3. Therefore, it was determined that the isolated nitrogen-fixing bacteria strain BXBL9 was a new strain of Bradyrhizobium, and it was named Huagu No. 2.

Embodiment 3

[0045] Embodiment 3 tie-back test

[0046] The above-mentioned large test tube paper culture device (Fig. 4) was used for the verification of the back connection test, and the treatment of soybean seeds was 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 raw paper and place it in a 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] Reagent name

Dosage

Reagent name

Dosage

tryptone

5g

Yeast Enzyme Powder

3g

CaCl 2 2H 2 o

0.7g

[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 t...

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Abstract

The invention discloses a Nodule Azotobacter strain BXBL9 and the application. The strain is obtained by a way that the Rhizobia strains in the soil are respectively captured through different soybean varieties; the Rhizobia strains are separated and purified in fresh root nodules; part 16S rDNA of the strain is sequenced; the phylogenetic analysis determines that the strain is a novel Bradyrhizobium strain, which is named as Huagu 2. The Nodule Azotobacter strain BXBL9 of the invention has the characteristics of acid and aluminum tolerance and high level of nodules, which are proved by laboratory pot culture and field experiment. The inoculation of the Rhizobia strain can improve the yield of soybean more than 20 percent to 30 percent. The invention suits for the acid soil area in South China, which can be used as one of the measures for the conventional cultivation of soybean.

Description

technical field [0001] The invention relates to a nitrogen-fixing bacterial strain BXBL9 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 Applications(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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