Bacillus megaterium having nitrogen fixing capacity and application of bacillus megaterium

A Bacillus megaterium, nitrogen fixation technology, applied in application, bacteria, microorganism-based methods, etc., can solve problems such as soil environmental hazards, and achieve the effect of improving soil, promoting growth and root development, and promoting the increase of nutrients

Active Publication Date: 2019-07-16
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But now most of the nitrogen is applied through the application of chemical fertilizers, which will bring a series of harm to the soil, crops and the environment

Method used

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  • Bacillus megaterium having nitrogen fixing capacity and application of bacillus megaterium
  • Bacillus megaterium having nitrogen fixing capacity and application of bacillus megaterium
  • Bacillus megaterium having nitrogen fixing capacity and application of bacillus megaterium

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1: Screening and identification of nitrogen-fixing bacteria

[0021] 1.1 Screening and purification

[0022] The medium for screening nitrogen-fixing bacteria is Ashby nitrogen-free solid medium (Ashby), glucose 10g, potassium dihydrogen phosphate 0.2g, magnesium sulfate heptahydrate 0.2g, sodium chloride 0.2g, calcium carbonate 5g, calcium sulfate dihydrate 0.1g, 18g agar, pH 6.8-7.0, add water to 1000ml. Sterilize at 121°C for 20 minutes.

[0023] Weigh 10g soil samples (collected from the rhizosphere of plants in Tai’an, Shandong), put them in a 250ml Erlenmeyer flask filled with an appropriate amount of glass beads and 90ml sterile water, at 30°C, 180r min -1 Oscillate for 20 minutes and let stand for 10 minutes to obtain a soil suspension. Dilute to 10 by serial dilution -6 After that, draw 0.1ml and spread it on the Ashby solid medium, place the plate in an incubator at 30°C and incubate it upside down for 24 hours, pick out different types of typic...

Embodiment 2

[0028] Embodiment 2: Determination of growth-promoting properties of N3 bacterial strain

[0029] 2.1 Determination of nitrogenase activity

[0030] Put the isolated and purified strains into LB liquid medium, collect the bacterial cells by centrifugation after culturing for 24 hours, and then culture the resuspended bacteria in nitrogen-free liquid to make 10 8 CFU / ml of nitrogen-fixing bacteria suspension, as the bacteria solution to be inoculated.

[0031] Inoculate the prepared bacterial solution to be inoculated in a penicillin vial containing 2ml of nitrogen-free liquid medium, and incubate at 30°C for 24 hours; use a well-sealed syringe to draw 1mL of air from the cultured penicillin vial, and then inject 1ml of acetylene. Seal the eye of the needle with adhesive tape; continue to cultivate for 24h, take 100ul gas sample and measure the peak value of ethylene on the gas chromatograph.

[0032] 2.2 Determination of IAA production capacity

[0033] The isolated and pur...

Embodiment 3

[0039] Embodiment 3: the application of bacterial strain on the growth promotion of green manure crop Er Yuelan

[0040] 3.1 Bacteria preparation

[0041] Inoculate the isolated and purified nitrogen-fixing bacteria into LB liquid medium, culture on a shaker at 30°C and 180rpm for 24 hours, pour the bacterial solution into a sterilized centrifuge tube under aseptic conditions, centrifuge at 8000rpm for 5min, remove the supernatant, Resuspend the bacteria with sterile water to make a concentration of approximately 10 8 CFU / ml bacterial suspension, ready for use.

[0042] 3.2 Bacteria inoculation of Er Yuelan

[0043] Two treatments were set up to carry out the pot experiment of Erchi orchid, the control treatment (CK) was inoculated with the same amount of sterile water, and the N3 bacterial agent inoculation treatment (N3). Soak the seeds of Erchi orchid in 95% alcohol for 5 minutes, then sterilize the surface with 3% NaClO for 2 minutes, and then rinse with sterile water r...

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Abstract

The invention discloses bacillus megaterium having nitrogen fixing capacity and an application of the bacillus megaterium. The bacillus megaterium(Bacillus megaterium) N3 having nitrogen fixing capacity is preserved in the China General Microbiological Culture Collection Center on January 14th, 2019, with the preservation number being CGMCCNo.17170. The invention discloses an application of microbial agents prepared from N3 to promotion of growth of orychophragmus violaceus and nutrient accumulation, and an application of microbial agents prepared from the N3 to soil organism repair. The bacillus megaterium (Bacillus megaterium N3) has good IAA production capacity, nitrogen fixing capacity and phosphorus solubilizing capacity, and besides, microbial agents prepared from the bacillus megaterium can significantly promote growth of orychophragmus violaceus plants, promote increase of nutrients in the orychophragmus violaceus plant bodies, significantly improve soil, and increase soil organism activity and content of available nutrients.

Description

technical field [0001] The invention belongs to the technical field of plant growth-promoting bacteria, and in particular relates to a bacterial strain with nitrogen-fixing ability, and its growth-promoting and soil-improving application to Erchis orchids. Background technique [0002] Orychophragmus violaceus is a cruciferous plant with ornamental and greening value. It is a rare ground cover plant in northern China for viewing flowers in early spring and viewing green in winter. It is also a kind of practical winter green fertilizer. It is rich in nitrogen, phosphorus, potassium and other mineral elements in the full flowering stage, which can increase the content of soil organic carbon, promote the activities of soil microorganisms, help maintain soil fertility, and increase the yield and quality of main crops. quality. Green manure biomass is an important basis for exerting its farmland ecological service functions such as ground cover, soil residual nitrogen intercepti...

Claims

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Application Information

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IPC IPC(8): C12N1/20B09C1/10A01N63/00A01P21/00C12R1/11
CPCC12N1/20B09C1/10A01N63/00C12R2001/11C12N1/205
Inventor 徐莉魏志敏孙斌方成代子雯刘满强焦加国李辉信胡锋
Owner NANJING AGRICULTURAL UNIVERSITY
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