A Leptinia scsio T-2 and its application

A technology of scsiot-2 and sheath filaments, applied in the biological field, to achieve good nitrogen fixation activity, broad application prospects, and high value effects

Active Publication Date: 2021-07-20
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

[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 the emergence rate in the field (Wang Shaomei et al., 1991), but there are few reports on the application of nitrogen-fixing cyanobacteria as bacterial fertilizers in marine ecosystems

Method used

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  • A Leptinia scsio T-2 and its application
  • A Leptinia scsio T-2 and its application
  • A Leptinia scsio T-2 and its application

Examples

Experimental program
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Embodiment 1

[0019] Example 1: Isolation, purification and identification of Leptolyngbya sp. scsio T-2

[0020] 1. Isolation and purification of Leptolyngbya sp. scsio T-2

[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 T-2 was obtained by separation and purification.

[0022] 2. Identification of Leptolyngbya sp. s...

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 cyanobacteria scsio T-2 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 Dessaux Y.A).

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

[0039] In the...

Embodiment 3

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

[0045] Select the pure bacterial strain cyanobacteria scsio T-2 obtained by separation and purification in Example 1 to carry out DNA extraction and purification. For the extraction and purification of DNA in this embodiment, its operation method refers to Dong Xiuzhu's "Common Bacteria System Identification Manual" P 409 . 1SF / 16SR (Weisenburg et al., 1991, 16S ribosomal DNA amplification forphylogenetic study., Journal of bacteriology, 1991 173(2): 697-703 and nifH gene general primer PolF / PolR (reference Poly et al. , 2001, Comparison of nifH 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 5min; denaturation at 95°C fo...

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Abstract

The invention discloses a scsio T‑2 strain and application thereof. Leptolyngbya sp. scsio T‑2, deposit number: CCTCC NO: M 2015061. The present invention isolates and screens a new species of cyanobacteria Leptolyngbya sp. scsio T-2 from the leaves of Thalassia hemperichii, which has good nitrogen fixation activity and can secrete and produce 3-indoleacetic acid Therefore, the Leptolyngbya sp. scsio T‑2 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 scsio T-2 and application thereof. Background technique: [0002] Seagrass is capable of photosynthetic autotrophy and is widely distributed in temperate, tropical and subtropical coastal ecosystems around the world. It is an important component of typical tropical marine ecosystems and one of the most productive ecosystems in nature. [0003] Seagrass is a flowering plant that grows in the ocean. Compared with terrestrial higher plants, its species are extremely rare. There are only 72 known species in the world, of which 15 species are endangered or extinct; Waycott et al. (2009) analyzed 215 species in the world. According to the analysis of the data of monitoring points since 1879, it is found that about 58% of the seagrasses are in a state of decline. 7%, habitat loss, biodiversity loss and global climate change are all important reasons, so it is particularly im...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/12C12P17/10C05G3/00C12R1/89
Inventor 凌娟董俊德张渊洲张燕英杨清松江玉凤曾思泉
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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