Strong bacillus G25-1-2 and applications thereof

A technology of bacillus and seaweed, applied in the field of microorganisms, can solve the problems of undiscovered nitrogen fixation function, achieve the effects of nitrogen cycle and maintenance of higher productivity levels, high nitrogenase activity, and wide adaptability

Inactive Publication Date: 2012-01-04
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The bacterium of Paenibacillus validus found in existing research has PAHs degradation function (Daane LL, 2002, PAH-degradation by Paenibacillus spp. and description of Paenibacillus naphthalenovorans sp nov., a naphthalene-degrading bacterium from the rhizosphere of Salt marsh plants, International journal of systematic and evolutionary microbiology, 2002, 52(1), 131-139) and growth-promoting function (Ulrich Hildebrandt, 2006, The bacterium Paenibacillus validus stimulates growth of the arbuscular mycorrhizal fungus Glomus to the intraradices formation of fertile spores, FEMS Microbiology Letters, 254(2), 258-267), etc., but no nitrogen fixation function has been found

Method used

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  • Strong bacillus G25-1-2 and applications thereof
  • Strong bacillus G25-1-2 and applications thereof
  • Strong bacillus G25-1-2 and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The separation and purification of embodiment 1 bacterial strain

[0029] The sampling site of this embodiment is the seaweed (18°15′08.6″N; 109°30′51.1″E) that grows after coral degradation in Sanya Bay Coral Reef Reserve, Sanya City, Hainan Province, China. The main seaweed species is Thalassia hemprichii), after the samples were collected, each sample was classified and numbered separately, put into a sterilized sealed polyethylene bag, placed in an ice box and brought back to the laboratory, and stored at -20°C.

[0030] The seaweed samples collected above are taken from the rhizosphere (i.e. leaf sheaths and rhizomes), cut into small pieces of about 2 cm with sterilized scissors, and put into selective nitrogen-fixing liquid medium (this medium contains NaCl 25g, KCl 0.56 g. MgSO 4 -7H 2 O 4.8g, MgCl 2 -6H 2 O 4.0g, K 2 HPO 4 0.01g, FeSO 4 -7H 2 00.001g, Tris 0.48g, peptone 4.0g, yeast extract 2.0g, glycerin 2.0ml and distilled water 1L, the final pH is 8.2...

Embodiment 2

[0033] Nitrogenase activity assay of embodiment 2 bacterial classification

[0034] The nitrogenase activity of the strains in this example was measured by gas chromatography for the acetylene-reducing activity (Acetylene-reducing Activity, ARA) of nitrogen-fixing bacteria (with reference to Capone 1993, Capone DG (1993) Determination of nitrogenase activity in aquatic samples using the acetylenereduction procedure. Aquatic Microbial Ecology, 621-631).

[0035] Take the above 40 μL of the reacted gas to measure the peak value of ethylene on a GC-17A gas chromatograph (Shimadzu), and the concentration of ethylene in the standard gas is 138 ppm. The working conditions of the gas chromatograph are set as follows: the temperature of the detection chamber is 100°C, the temperature of the column is 60°C, the injection port is 60°C, the column length is 1m, and the carrier gas pressure at the head is 0.95kg·cm -2 , hydrogen is 0.8kg·cm -2 , the air is 0.6kg·cm -2 . The calculatio...

Embodiment 3

[0044] The sequence identification of embodiment 3 bacterial strains

[0045] Template preparation: select the pure bacterial strain obtained by separation and purification in Example 1 for DNA extraction and purification. For the extraction and purification of the DNA, refer to Dong Xiuzhu's "Common Bacteria System Identification Manual" P 409 , the extracted and purified DNA is the template used in the subsequent PCR amplification of this embodiment.

[0046] 1. Amplification of nifH gene

[0047] The nifH gene universal primer PolF / PolR (refer to Poly et al., 2001) of nitrogen-fixing bacteria was used for PCR amplification:

[0048] PCR reaction system: the total system is 25uL, 1×PCR Buffer, 200umol L -1 dNTPs, 2.0 mmol L -1 Mg 2+ , 1umol L -1 Primers, 4ug of BSA and 2.0U of Taq DNA polymerase, template 1ul.

[0049] PCR amplification conditions: pre-denaturation at 94°C for 5 min, denaturation at 94°C for 30 s, annealing temperature at 55°C for 30 s, extension at 72...

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Abstract

The invention discloses a strong bacillus G25-1-2 and applications thereof. The bacillus has been preserved by China Center for Type Culture Collection in Wuhan University (short for CCTCC) on 4th, June, 2009, and the preservation number is CCTCC NO:M209122. The bacillus is a pure bacillus with high-efficiency nitrogen-fixing activity, and is separated and sieved from a seaweed ecological system. When the strong bacillus G25-1-2 is used for processing seaweed, compared with a test control group, the processed seaweed has higher plant height, stem diameter average value, carotene content, carotenoid content and soluble saccharide content, thereby the result shows that the bacillus has good growth promotion effect on the seaweed. The strong bacillus G25-1-2 can be used for seaweed nursing and restoration of the whole seaweed ecological system, and has good promotion functions on the nitrogen circulation of the coral reef-seaweed bed composite ecological system and the maintenance of high production level.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to nitrogen fixation by microorganisms, in particular to a Paenibacillus carterus G25-1-2 and application thereof. Background technique [0002] The wetland ecosystem is a special ecosystem with an average annual protein production of 9g / m 2 , is 3.5 times that of terrestrial ecosystems, and also has the functions of regulating and storing water sources, stabilizing coastlines, regulating climate, purifying water quality and fixing nutrients, preserving species, providing wild animal habitats and other basic ecological benefits. , energy, medical industry, etc. provide a large number of raw materials for production. [0003] Coral reefs and seagrass beds are important components of tropical marine wetland ecosystems. High productivity levels and important ecological significance are their basic characteristics, but they also have the vulnerability and variability of wetland ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/02A01P21/00C05F11/08C12R1/01
Inventor 董俊德凌娟张燕英李丽璇王友绍张偲
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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