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Bacillus megaterium strain X3 and preparation method and application thereof

A technology related to Bacillus megaterium, applied in the field of Bacillus megaterium X3 and its preparation, to achieve the effects of promoting plant growth, good antagonistic effect, and increasing yield

Active Publication Date: 2015-09-23
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These research results are expected to speculate that the Bacillus megaterium agent also has the effect of improving soil microbial diversity and optimizing soil micro-ecological structure, which remains to be studied

Method used

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  • Bacillus megaterium strain X3 and preparation method and application thereof
  • Bacillus megaterium strain X3 and preparation method and application thereof
  • Bacillus megaterium strain X3 and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 Isolation and purification of bacterial strains

[0052] (1) Separation

[0053] Prepare beef extract peptone medium: peptone 10.0g, beef extract powder 3g, sodium chloride 5g, distilled water 1000mL, pH7.0~7.5.

[0054] Taking the soil from the farm of South China Agricultural University in Tianhe District, Guangzhou City, Guangdong Province as the screened soil sample, 10.0 g of the screened soil sample was accurately weighed by the flat plate coating method, and put into a 250 mL triangular flask filled with 90 mL of sterile water (with glass beads) , shaking on a shaker for 30 minutes to fully disperse the cells, let it stand for 20-30 seconds, and take the supernatant for 10-fold decreasing dilution, using 10 3 ~10 5 Dilution: use a pipette to absorb 0.1mL of the dilution respectively, spread it on a beef extract peptone plate, and incubate it upside down at 28°C for 48h. Select a suitable concentration plate, pick typical single colonies of different ...

Embodiment 2

[0060] Example 2 Characterization

[0061] (1) Morphological characteristics of bacteria

[0062] Gram-positive bacteria with spores. The bacteria are rod-shaped, motile, and obligately aerobic. Gram-stained micrographs of bacteria figure 1 , see photos of spore staining figure 2 .

[0063] (2) Colony Morphological Characteristics

[0064] After 24 hours of cultivation on the beef extract peptone medium, the colonies formed are round or irregular. After 48 hours, the colonies are all round, milky white, about 2 to 3 mm in diameter, with irregular edges, flat and moist.

[0065] (3) Growth characteristics

[0066] In the liquid medium of beef extract 5g, peptone 5g, sodium chloride 5g, and water 1L, the rotation speed was 113rpm, the temperature was 30°C, and the initial pH value was 7.0. After culturing for 18 hours, the number of viable bacteria was measured as 2.91±0.60 ×10 8 cfu mL -1 .

[0067] (4) Molecular biological characteristics

[0068] The total DNA of ...

Embodiment 3

[0069] Example 3 Test of siderophores produced by bacterial strains

[0070] Qualitative: transfer the strain to the LB plate and culture it for 24-48 hours, use the spot inoculation method, use a sterilized toothpick to inoculate the strain on the CAS solid detection plate, spot 3 replicates, and culture it at 28°C for 48 hours. On the CAS assay plate, a distinct orange siderophore halo will appear around the siderophore-secreting bacterial colony. The results showed that the strain X3 had the largest orange halo, indicating that the strain had the strongest ability to produce siderophore ( Figure 5 ).

[0071] Quantitative: Inoculate the strains in iron-limited SA liquid medium (sucrose 20.0g, L-aspartic acid 2.0g, K 2 HPO 4 0.5g, MgSO 4 ·7H 2 O 0.5g, distilled water 1000mL, pH7.0), cultured on a shaker at 28°C (150rpm) for 48h; the bacterial suspension was centrifuged at 10,000rpm for 15min, the supernatant was taken, and the bacterial liquid and CAS detection soluti...

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Abstract

The invention discloses a bacillus megaterium strain X3 and application thereof to plant growth promotion. The bacillus megaterium strain X3 is preserved in the China General Microbiological Culture Collection Center (CGMCC) on May 11, 2015 with the preservation number of CGMCC No.10803, is obtained through separation of indoleacetic acid, has the characteristics of siderophore production, phosphate solubilizing, ammonia producing, salt tolerance and disease suppression, can remarkably promote crop growth and improve corn yield, and meanwhile, can increase the diversity and the abundance of microbes in soil. The bacillus megaterium strain X3 is quick in reproduction, simple in production process, strong in adversity ability, easy to preserve and conductive to industrial production, and can not only prevent and cure soil-borne diseases but also promote plant growth when utilized to prepare a bio-fertilizer, thereby having a wide application prospect.

Description

technical field [0001] The invention relates to a microorganism and its application, in particular to a strain of Bacillus megaterium strain X3 (Bacillus megaterium strain) and its preparation method and application. Background technique [0002] Indole acetic acid (IAA) is a signal substance that regulates auxin. It is an auxin that is ubiquitous in plants and plays a key role in plant growth. Bacteria not only secrete IAA, but also achieve the growth-promoting effect on plants through various ways such as nitrogen-fixing and phosphorus-dissolving, siderophore production, inhibition of soil pathogenic bacteria and increase of soil beneficial microorganisms. Relevant patent documents show that the Bacillus megaterium SJ-7 strain has the functions of dissolving phosphorus and potassium and preventing and controlling fungal diseases. Bacillus megaterium JX15 has the characteristics of indole acetic acid production, nitrogen fixation and phosphorus dissolution, which can impro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/08C12N9/54C09K17/00A01C1/00A01N63/00A01P21/00A01P3/00C12R1/11C09K101/00
Inventor 蔡燕飞张娅娅李永涛熊汉琴曹禺郭真真
Owner SOUTH CHINA AGRI UNIV
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