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A strain of pseudoantaba scsio S3 and its application

A technology of SCSIOS3 and Pseudo-Anabaena, applied in the direction of microorganisms, fermentation, bacteria, etc., to achieve good nitrogen fixation activity, broad application prospects, and high-value effects

Active Publication Date: 2018-07-27
海南三友环境与生态修复有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In 1939, De India used nitrogen-fixing cyanobacteria as a fertilizer in paddy rice. After that, more than 10 countries including the United States, Britain, Japan, and Egypt also carried out related research. Li Shanghao et al. (1959), who first started research in this area, confirmed that Inoculation of nitrogen-fixing cyanobacteria in paddy fields can effectively provide nitrogen fertilizer for rice expenditure, fertilize the soil, and increase rice yield by 10%-30%; application of nitrogen-fixing cyanobacteria can effectively promote the growth of wheat roots and shoots and field emergence rate (Wang Shaomei et al., 1991), but there are few relevant reports on the application of nitrogen-fixing cyanobacteria as a growth-promoting bacterial fertilizer in marine wetlands and other ecosystems

Method used

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  • A strain of pseudoantaba scsio S3 and its application
  • A strain of pseudoantaba scsio S3 and its application
  • A strain of pseudoantaba scsio S3 and its application

Examples

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Embodiment 1

[0019] Example 1: Isolation, purification and identification of Pseudoantabaena sp. SCSIO S3

[0020] 1. Separation and purification of Pseudanabaena sp. SCSIO S3

[0021] Collect the seaweed Thalias algae collected from Xincun Bay, Lingshui, Hainan Province, China, add seawater to scrub, and filter to obtain the cyanobacteria liquid. Pick cyanobacteria hyphae and culture them in marine cyanobacteria medium ATCC Medium 957, nitrogen-fixing cyanobacteria medium ATC Medium 819, marine nitrogen-fixing cyanobacteria medium ATCC Medium 1077 and BG-11Medium, and the continuous light intensity is 150μE / m 2 / s, the light cycle is 12:12h (light: dark cycle), the temperature is 23-27°C, in the light incubator (the model is LRH-250-GH microcomputer-controlled light incubator), every two weeks into fresh medium until the microscopic examination shows a single mycelium. The cyanobacteria SCSIO S3 was obtained by separation and purification.

[0022] 2. Identification of Pseudanabaena sp...

Embodiment 2

[0036] The mensuration of embodiment 2 cyanobacteria indole acetic acid production

[0037] The salkowski colorimetric method was used to determine the ability of the isolate to produce plant growth hormone substances, and the standard curve was prepared using pure 3-IAA (3-indole acetic acid). Inoculate the cyanobacteria SCSIO S3 in 100mL selective liquid nitrogen-free liquid modified medium (formula is the same as in Example 1), and the continuous light intensity is 150 μE / m 2 / s, the light cycle is 12:12h (light: dark cycle), the temperature is 25-28°C, cultivate for 120h, take 5mL of the cyanobacteria culture supernatant every 24h, centrifuge at 10000r / min for 10min, take the supernatant and add colorimetric The solution was left to stand in the dark for 0.5h, taken out, and measured immediately with a spectrophotometer at a wavelength of 530nm (Glickmann and DessauxY.A).

[0038] Indole acetic acid content in the sample (μg / mg) = (A × V1) / (W × V2) × 1000

[0039] In t...

Embodiment 3

[0044] 16SRNA and nitrogen fixation gene amplification of the strain of embodiment 3

[0045] Select the pure bacterial strain cyanobacteria SCSIO S3 that the separation and purification of embodiment 1 obtains to carry out DNA extraction and purification, the extraction and purification of DNA in this embodiment, its operation method refers to Dong Xiuzhu "Common Bacteria System Identification Manual" P 409 . 16S 27F / 1492R of nitrogen-fixing bacteria (Weisenburg et al., 1991, 16S ribosomal DNA amplification for phylogenetic study., Journal of bacteria, 1991173 (2): 697-703 and nifH gene general primer PolF / PolR (refer to Poly et al., 2001, Comparison of nif (Gene Pools in Soils and Soil Microenvironments with Contrasting Properties, Applied and Environmental Microbiology, 2001, 67 (5): 2255-2262) PCR amplification: PCR reaction system as shown in Table 1.

[0046] Table 1 PCR reaction system

[0047]

[0048] The PCR reaction conditions were: pre-denaturation at 95°C for...

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Abstract

The invention discloses a pseudoantaba SCSIO S3 and an application thereof. The present invention provides a new species of cyanobacteria SCSIO S3 isolated from the leaves of seagrass Thaleisis in Xincun Bay, Lingshui, Hainan Province, China, which was preserved in China Center for Type Culture Collection (CCTCC) on May 29, 2015. It has A new species of cyanobacteria with high nitrogen-fixing biological activity and the ability to produce the plant growth hormone indole acetic acid, so the Pseudanabaena sp. SCSIO S3 has high value and broad application prospects in the preparation of marine microbial fertility .

Description

Technical field: [0001] The invention belongs to the field of biotechnology, and in particular relates to a strain of pseudoantaba SCSIO S3 and its application. Background technique: [0002] Prokaryotes can reduce nitrogen in the air, which most organisms cannot directly use, to nitrogen forms that can be used by organisms. This process is biological nitrogen fixation. Nitrogen-fixing organisms mainly include cyanobacteria (cyanobacteria), actinomycetes, proteobacteria and other bacteria and methanogenic archaea, covering about 1500 species of bacteria in 38 genera, 87 species of cyanobacteria in 20 genera and 2 genera of archaea (Dixon and Wheeler, 1986) . In addition to providing new nitrogen sources for the ecosystem through biological nitrogen fixation, cyanobacteria can also produce a series of active metabolites such as plant growth regulators, amino acids and polysaccharides to regulate plant growth (Ahmed et al., 2010; Mandal et al ., 2011). In the rhizosphere, c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C05F11/08C12P17/10C12R1/89
Inventor 凌娟张燕英张渊洲杨清松董俊德江玉凤曾思泉
Owner 海南三友环境与生态修复有限公司
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