KR100817708B1 - Novel Salt-resistant Halomonas Strain with Excellent Degradation of Organic Matter, Soil Reclaimer for Seashore Landfill Containing It, and Method for Promoting Growth of Plant - Google Patents

Abstract

The present invention relates to a halomonas genus strain having excellent organic degradability and flame resistance, a soil improving agent of a seashore landfill or a landfill containing the same, and a method for promoting plant growth using the same, and more specifically, an organic degradability separated from a tidal flat Excellent salt-resistant halomonas genus ( Malomonas sp.) NM5 strain (KACC 91326P), a soil improver of a seashore landfill or landfill containing the same, and a method for promoting plant growth in a seashore landfill or landfill using the same. The use of the strain having excellent organic matter decomposition ability and flame resistance according to the present invention can be usefully used as a biological soil improver or microbial fertilizer in place of the conventional chemical fertilizer-based soil improver.

Description

Novel saline-resistant halomonas strains with excellent organic decomposition, soil reclamation landfill containing the same and methods for promoting plant growth using the same {A novel halophilic Halomonas sp. having an excellent digestive effect of organic material, a soil conditioner for the reclaimed land using said strain and a method for plant growth-promoting

The present invention relates to a salt-tolerant halomonas genus strain having excellent organic degradability, a salt-improving soil improving agent containing the same, and a method for promoting plant growth using the same, and more particularly, to a salt-resistant halomonas genus having excellent organic degradability separated from a tidal flat. sp.) NM5 (KACC 91326P) strain, a soil improver of a seashore landfill or landfill containing the same, and a method for promoting plant growth using the same.

As 65% of the country's land is narrow in land area compared to the population in Korea, it is difficult to supply the land necessary for the development of new industrial complexes, housing complexes, ports and airports due to economic development. A landfill is being built as part of the national land development project. The land use plan of the coastal landfill in Korea was mainly aimed at creating industrial facilities in the early stages of reclamation, but recently, large-scale residential complexes are being constructed. Many trees are planted.

The coastal landfill uses heavy machinery to cut the soil and collect soil to reclaim the sea, or to pump up dredged sand or tidal-flat soil in the sea outside the landfill site. Has chemical properties There are many causes of the death or sluggish growth of plants planted in coastal landfills, but the problems caused by the physical and chemical properties of soils are not suitable for tree growth. It is hard to expect. Therefore, before the tree planting, the soil survey should be carried out to understand the site environment characteristics, soil cross-sectional shape and soil physicochemical characteristics, and then the soil map is prepared. It should be planted after prior review.

Until now, it has been common to cover the soil and plant it in the landfill site, but in this case, if it is not possible to completely eliminate the damage to the trees caused by the rising salinity, The damage is that the salt-containing water from the ground soil Gaepal is moved through the capillary to the ground, losing water on the surface and a large amount of salt accumulates on the surface. Representative obstacles to plants caused by increased salt concentrations are absorption disorders due to a decrease in osmotic pressure, which increases the soil's water retention capacity, causing plant roots to become unable to absorb water, and excessive salt concentrations inhibit absorption of root physiology. It is to accelerate the phenomenon.

Landfill sites are also damaged due to the accumulation of chemicals and heavy metal contamination, and the surrounding environment is polluted by high concentrations of organic matter contained in landfill gas and leachate from landfills.

Seashore landfills and landfills generally have poor plant growth because of the soil conditions that microorganisms cannot live in. Soil microbial habitat environment corresponds to plant growth environment, so it can be said that the basic concept of soil improvement is to correct it as microbial habitat environment. Therefore, it is necessary to stabilize these landfills biologically early for plants to grow.

To date, efforts to restore landfills or landfills have been made mainly through physicochemical methods to improve the physicochemical properties of soil. Republic of Korea Patent Publication No. 199595-14476 describes a salted soil improving agent and a planting method using the same to block salt-containing maternal water and to retain ions required for plant growth, and Patent No. 10-696099 A mixed soil and its manufacturing method for soil improvement of a seaside land, a landfill, a crushed land and a golf course composition land are described. Patent No. 10-697671 improves the physicochemical properties of clay-like particulates and restores soils contaminated with heavy metals. Soil improver, the preparation method thereof and the soil improvement method using the same are described.

However, while these methods mainly aim to improve soil physicochemical properties to stabilize soils and create soils for plant growth, they can grow even under conditions of high microbial or salt concentrations with high resolution of high concentrations of organic matter contained in landfills. Efforts to directly improve the microbiological properties of soils and to create conditions for plant growth by using soil modifiers or microbial fertilizers containing salt-tolerant microorganisms are relatively insignificant.

Accordingly, the present inventors have directly identified a novel Halomonas sp. NM5 strain (KACC 91326P) having excellent organic matter resolving power and flame resistance from a tidal flat having a function of purifying various pollutants discharged from the land. The present invention was successfully completed by finding that the strain has excellent ability to promote the growth of plants by improving contaminated soil such as coastal landfill or landfill, and confirming that it can be effectively applied as a soil improving agent or microbial fertilizer. It was.

Literature Information of the Prior Art

[Document 1] Korean Patent Publication No. 1995-14476 (1995.12.02)

[Document 2] Republic of Korea Patent Publication No. 10-696099 (2007.03.19)

[Document 3] Korean Patent Publication No. 10-697671 (2007.03.21)

The present invention has been proposed to solve the problems of the prior art as described above, to isolate the salt-resistant microorganisms that can be grown in conditions of high salinity of the high concentration of organic matter contained in the coastal landfills or landfills and high salt concentration, and using the same The purpose of the present invention is to provide a method of improving soil physicochemical properties as well as soil properties and using them as soil improving agents or microbial fertilizers to promote plant growth.

In order to achieve the above object, the present invention provides a novel Halomonas sp. NM5 strain (KACC 91326P) having excellent organic resolution and flame resistance.

In addition, the present invention provides a soil improver of a coastal landfill or landfill containing the Halomonas sp. NM5 strain (KACC 91326P) as an active ingredient.

In another aspect, the present invention provides a method for producing a soil modifier by culturing the Halomonas sp. NM5 strain (KACC 91326P).

In addition, the present invention provides a method for promoting the growth of plants in coastal landfills or landfills using the culture medium cultured the strain or strains.

The present invention utilizes a novel Halomonas sp. NM5 strain (KACC 91326P), which has excellent organic resolving power and flame resistance, isolated from a tidal flat having a function of purifying various pollutants, such as a coastal landfill or a landfill. Improving contaminated soil not only promotes plant growth, but also contributes to creating a foundation for ecologically sound growth of the trees planted in the landfill.

The present invention is described in detail as follows.

The present invention provides a novel Halomonas sp. Strain. Specifically, the present invention provides a strain of Halomonas sp. Isolated from a tidal flat having a function of purifying various pollutants.

Halomonas sp. Strain of the present invention is preferably Halomonas sp. ( Mhalomonas sp.) NM5 strain.

The halomonas strain of the present invention was named " halomonas sp. ( Nhalonas sp.) NM5" and was deposited with the deposit number "KACC 91326P" as of August 31, 2007 to the Korea Agricultural Microbiological Conservation Center of the Institute of Agricultural Biotechnology.

The present invention provides a method for separating a Halomonas sp. NM5 strain from a tidal flat.

Soil modifiers containing the Halomonas sp. NM5 strain of the present invention has excellent organic matter resolving power and flame resistance in coastal landfills or landfills. In addition, the NM5 strain or its culture medium has an effect of promoting the growth of plants in the soil.

The present invention can be used as a biological soil improver or microbial fertilizer because the strain improves the biological properties as well as the physicochemical properties of contaminated soil such as coastal landfills or landfills. The biological soil modifier or microbial fertilizer may include the genus Halomonas sp. ( Mhalomonas sp.) NM5 strain as an active ingredient. The biological soil improver or microbial fertilizer according to the present invention may be prepared in the form of a liquid fertilizer, and may be used in the form of a powder powder by adding an extender thereto or granulated by formulating it. However, the formulation is not particularly limited.

The present invention provides a method of using as a microbial fertilizer to promote the growth of plants in soil such as coastal landfills or landfills using Halomonas sp. NM5 strain. Using the culture medium cultured the strain, it can be irrigated as it is in a liquid state, or immersed or sprayed into the seed of the plant.

Hereinafter, the contents of the present invention will be described in more detail with reference to Examples. However, the following examples are only presented to understand the contents of the present invention, but the scope of the present invention is not limited to these examples.

Example 1 Isolation of Halomonas Strains from Tidal Flats

In the present invention, in order to isolate strains that have high salt resistance and salt resistance from tidal flats, soil soil or salt water samples were prepared using Bacto peptone 5g, Bacto yeast extract 1g, Fe (III) citrate 0.1, MgCl ₂ (dried) 5.9g, Na ₂ SO ₄ 3.24g, CaCl ₂ 1.8g, KCl 0.55g, Na ₂ CO ₃ 0.16g, KBr 0.08g, SrCl ₂ 34mg, H ₃ BO ₃ 22mg, Na-silicate 4mg, NaF ₂ 4mg, (NH ₄ ) NO ₃ 1.6mg, Na ₂ HPO ₄ 8mg, NaCl 33g, agar 15g in 1L of distilled water and plated in a solid medium prepared by high temperature sterilization. A total of 25 strains were initially selected by incubating at 37 ° C. for 24 hours.

Five strains capable of using a broad spectrum carbon source were selected through the various carbon source utilization experiments in the above-selected strains, and the temperature, pH, and NaCl concentrations necessary for their growth were all measured. Good growth was observed at ˜42 ° C., pH 6-10 and 1-20% NaCl concentration, confirming the possibility of application in a wide range of environments where these conditions are met.

As a result of testing the degradability of high concentration organic matter in 5 strains selected secondarily, the strain NM5 which was excellent in degrading organic matter was finally selected and the activity of the strain was confirmed through subculture and the glycerol stock was stored for the next experiment. ).

     Example 2 Identification of Isolated Halomonas NM5 Strains

The nucleotide sequence of 16S rDNA of strain NM5 which had the selected flame resistance and excellent organic matter resolution was analyzed. For sequencing ribosomal 16S rDNA, a molecular biological microorganism identification method, genomic DNA was obtained from the strain using the DNeasy Tissue Kit (Qiagen, Valencia, Calif.), And then PCR was performed using the universal primers 9F and 1512R. It was. The PCR result was purified again using the QIAquick PCR purification kit Qiagen, Valencia, Calif., USA), and then the reaction product was prepared using ABI PRISM BigDye Terminator cycle sequencing ready reaction kit (Applied Biosystems, Foster, Calif., USA). Then, the nucleotide sequence was analyzed using an ABI Prism 3700 DNA analyzer. The analysis results are shown in SEQ ID NOs: 1 and 2.

16S rDNA nucleotide sequence (SEQ ID NO: 3) of the NM5 strain obtained through the sequencing method was found to be BLAST (Basic Local Alignment Search Tool). As a result, it was determined that the NM5 strain belongs to the genus Halomonas sp. Halomonas alkaliantartica DSM 15686 ^T and 99.8%, Halomonas neptunia ATCC BAA-805 ^T and 99.6%, and Halomonas boliviensis DSM 15516 ^T , respectively Sequence homology of 99.5% was shown (see FIG. 1).

The NM5 strain was aerobic, gram-negative, heavy basophilic, did not form spores, and had a rod-shaped (0.8-1.0 x 2.0-4.0 μm) with peritrichous flagella as shown in electron micrographs. (See Figure 2). Gram staining and biochemical characteristics of the Halomonas sp. NM5 strain of the present invention showed positive reactions in the oxidase test and the catalase test as shown in Table 1 below. In addition, using the API 20NE / 50CH / ZYM kit (bioMerieux) to test the carbon source and enzyme activity, various carbon sources were used as substrates (see Tables 2 and 3), and also have the ability to produce various enzymes. (See Table 4).

Biochemical Properties of NM5 Strains of Halomonas sp. characteristic NM5 strain Gram Dyeing Gram-negative Colony smooth, translucent and circular with entire edges. Colony Ceamy white Cell morphology  rods Cell size 0.8-1.0 × 2.0-4.0 μm flagellum motile with peritrichous flagella Sporulation non-spore-forming Oxidase + Catalase + Urease + Growth at / in / on: 0 ° C + Optimum 25-30 ° C 45 ° C + pH 6 + Optimum pH 7-8 pH 10 + 0 to 20% NaCl + Optimum 5-15% NaCl NO3-NO2 + NO2-N2 + H2S generation - Voges-Proskauer Test +

Carbon Source Availability of the Halomonas sp. NM5 Strain Substrate NM5  Strain Mannitol + Rhamnose - D-glucose + Mannose - N - acetyl glucosamine frame (N -acetylglusosamine) - Salincin + D-ribose - D-melibiose + Inositol + L-fucose - D-Sucrose + D-sorbitol + D- Maltose + L-arabinose + Adipate + Itaconate - Propionate + Suberate + Caprate - Malate + Malonate + Gluconate + Valerate + Acetate + Citrate + DL-lactate + Phenylacetate - Histidine + L-alanine + 2-ketogluconate + 5-ketogluconate - 3-hydroxybutyrate + Glycogen - 4-hydroxybenzoate + 3-hydroxybenzoate - L-proline + L-serine +

Acid Production Patterns of the NM5 Strain of Halomonas sp. Substrate NM5  Strain Glycerol + Erythritol - D-arabinose - L-arabinose + Ribose - D-xylose + L-xylose - Adonitol - α-methyl-D-xylose - Galactose + Glucose + Fructose + Mannose - Sorbose - Rhamnose - Dulcitol - Inocitol + Mannitol + Sorbitol + α-methyl-D-mannoside - α-methyl-D-glucoside + N - acetyl glucosamine frame (N -acetylglucosamine) - Amigdalin - Arbutin - Esculin + Salincin + Cellobiose + Maltose + Lactose + Melibiose + Sucrose + Trehalose + Inulin - Melezitose + Raffinose + Gentiobiose - D-turanose + D-lyxose - D-tagatose - D-fucose + 2-keto-gluconate - 5-ketogluconate -

Enzymatic Activity of the NM5 Strain of Halomonas sp. ENZYME SUBSTRATE NM5 strain Alkaline phosphatase 2-naphthyl phosphate (pH 8.5) - Esterase [C4] 2-naphthyl butyrate + Esterase Lipase [C8] 2-naphthyl caprylate - Lipase [C14] 2-naphthyl myristate - Leucine arylamidase L-leucyl-2-naphthylamide + Valine arylamidase L-valyl-2-naphthylamide - Cystine arylamidase L-cystyl-2-naphthylamide - Trypsine N-benzoyl-DL-arginine-2-naphthylamide - α-chymotrypsin N-glutaryl-phenylalanine-2-naphthylamide - Acid phosphatase 2-naphthyl phosphate (pH 5.4) - Naphthol-AS-BI-phosphohydrolase Naphthol-AS-BI-phosphate - α-galactosidase 6-Br-2-naphthyl-αD-galactopyranoside - β-galactosidase 2-naphthyl-βD- galactopyranoside - β-glucuronidase Naphthol-AS-BI-βD glucuronide - α- (glucosidase) 2-naphthyl-αD-glucopyranoside + β- (glucosidase) 6-Br-2-naphthyl-βD-glucopyranoside - N-acetyl-N-β-galactosidase (acetyl-β-galactosidase) 1-naphthyl-N-acetyl-βD-glucosaminide - α-mannosidase 6-Br-2-naphthyl-αD-mannopyranoside - α-fucosidase 2-naphthyl-αL- fucopyranoside -

The quinone of the NM5 strain was ubiquinone Q-9, and the representative fatty acids were C18: 1w7c, C16: 0, C16: 1w7c / C15: 0 iso 2-OH and C12: 0 3-OH, and G + C content of DNA Was 56.1 mol%.

Morphological, resulting the strains was analyzed for biochemical and molecular biological classification characteristic of the NM5 strain 5 is halo Pseudomonas genus is identified as a new species belonging to the (Halomonas sp.) "Halo Pseudomonas genus (Halomonas sp.) NM5" strain It was deposited on August 31, 2007 with the deposit number "KACC 91326P" at the Korea Institute of Agricultural Biotechnology.

     Example 3 Degradation of Organic Matter and Salinity of Halomonas sp.

In order to evaluate the effect of reducing salt concentration of Halomonas sp. NM5 strain having excellent organic matter degradability and flame resistance, reclaimed land (sea landfill) was used. Experimental method is to fill the reclaimed soil with Falling Head Method in an acrylic column (inner diameter of 6cm, length of 30cm) to a volume density of 1.25Mg / cm ³ , and then saturate each column with a top volume corresponding to 1 pore volume. Soil and plant concentrations were collected on 10, 20, and 30 days after distilled water (control) and microbial product (500-fold dilution) containing NM5 strain of halomonas were collected. (2000).

Before the test, the reclaimed soil was near-neutral with a pH of 6.9, a salt concentration of 1.25%, and a silty loam with an electrical conductivity (EC) of 25.9 dS / m. The chemical and salinity of reclaimed soils by column test showed a tendency to decrease in electric conductivity (EC) and salinity as the test period passed, and the microbial treatment group showed a greater decrease than the control. Other chemistries resulted in slightly reduced results before the test (see Table 5).

TABLE 5

Degradation of Organic Matter and Salinity of Halomonas sp. NM5 Strains in Reclaimed Soil

division pH (1: 5) EC T-N O.M NaCl Ava .- P ₂ O ₅ Ex.- cations ( cmolc / kg) CEC dS / m % mg / kg Ca ^{2 +} Mg ^{2 +} K ⁺ Na ⁺ cmol _c / kg Before the test  Day 0 6.90 25.9 0.10 0.29 1.25 32 2.1 12.0 3.4 21.4 8.5 Control Day 10 6.85 21.0 0.08 0.18 1.15 24 2.1 10.6 2.6 19.3 9.5 Day 20 6.81 19.3 0.08 0.17 1.05 21 2.0 10.3 2.5 18.1 9.3 Day 30 6.79 18.7 0.07 0.17 0.99 20 2.1 10.0 2.4 17.8 9.4 Treatment Day 10 7.08 16.2 0.10 0.27 0.95 26 2.2 11.7 2.5 16.6 10.1 Day 20 7.12 13.8 0.09 0.25 0.75 24 2.2 11.9 2.6 14.9 9.9 Day 30 7.11 13.0 0.09 0.24 0.62 22 2.2 11.8 2.6 14.0 9.8

Changes in pH of reclaimed soils during the course of the test tended to decrease over time, and microbial treatment tended to increase slightly. EC and salinity changes showed a tendency to decrease in both control and microbial treatments, and the trend and width of the decrease over time was greater in the microbial treatments than the control (see FIG. 3). This effect is believed to be due to the activity of the basophilic microorganism NM5 strain.

The results of the column test using the Falling Head method on the effects of microbial agents containing halomonas NM5 strains on the chemical and salinity changes of reclaimed soils showed that the electrical conductivity (EC) and salinity concentrations (NaCl) were higher than those of the control in the microbial treatments. ), The salinity reduction rate was 14% in the control group and 35% in the microbial treatment group, showing a greater reduction in salt concentration.

     Example 4 Organic Degradation and Salinity Reduction Effects of Halomonas sp. NM5 Strains in Mixed Soils between Landfill and Field Soils

In order to plant plants in coastal landfills, it is often used after mixing soil in the field. Therefore, the salinity reduction effect of Halomonas sp. In order to do so, experiments were carried out using mixed soil of reclaimed land (soil landfill) and field soil.

Experimental method is a pot experiment to mix reclaimed soil and general field soil in a weight ratio of 1: 2, 1: 1 and 2: 1, respectively, and fill it in 1 / 5000a pot, distilled water in the control, and NM5 strain of halomonas in the treatment. Soil was collected on the 10th day after irrigation of the appropriate amount of the microbial agent containing solution (500-fold dilution) in the same amount (500ml), and the soil was collected after 2 times on the 20th day and 3 times on the 30th day. . When collecting soil samples, the soil (10 ~ 20cm), which is the middle part of the pot, was collected and prepared to be 2mm or less after air drying, and chemical and salinity concentrations were carried out according to the Soil and Plant Analysis Method of Rural Development Administration.

The chemical and salinity of the initial soils in which the reclaimed soil and the field soils were mixed in weight ratios of 1: 2, 1: 1, and 2: 1, respectively, differed according to the mixing ratio. The electrical conductivity and salinity increased with increasing soil mixing ratio (see Table 6).

TABLE 6

Results of chemical and salinity analysis of reclaimed soils, field soils and mixed soils before the test

division pH (1: 5) EC T-N O.M NaCl Ava .- P ₂ O ₅ Ex.- cations ( cmolc / kg) CEC dS / m % mg / kg Ca ^{2 +} Mg ^{2 +} K ⁺ Na ⁺ cmol _c / kg Reclaimed soil 6.90 25.9 0.10 0.29 1.25 32 2.1 12.0 3.4 21.4 8.5 Soil 7.25 0.75 0.11 1.55 0.28 278 4.8 2.2 0.8 0.1 10.6 1: 2 7.24 9.1 0.11 1.26 0.55 82 3.9 6.5 1.3 7.4 8.9 1: 1 7.15 11.8 0.10 1.02 0.75 76 3.5 7.6 1.7 10.9 9.3 2: 1 7.05 14.4 0.10 0.87 0.90 64 3.2 7.7 2.2 13.5 10.1

The reclaimed soil and the field soil were mixed at a weight ratio of 1: 2, 1: 1, and 2: 1, respectively. The electrical conductivity and salinity of the control and control groups were lower than before the test, and the reduction of microbial treatments was greater than that of the control (see Table 7). This showed similar results with the column test using the Falling Head method.

TABLE 7

Degradation of Organic Matter and Salinity in Mixed Soils of Reclaimed Soil and Field Soil by Halomonas sp.

division pH (1: 5) EC T-N O.M NaCl Ava .- P ₂ O ₅ Ex.- cations ( cmolc / kg) CEC dS / m % mg / kg Ca ^{2 +} Mg ^{2 +} K ⁺ Na ⁺ cmol _c / kg Control I (1: 2) 10 days 7.16 8.07 0.10 1.18 0.50 76 3.7 4.7 1.3 7.3 8.4 20 days 7.10 7.22 0.09 1.11 0.45 76 3.8 4.5 1.2 6.6 8.2 30 days 7.02 7.01 0.09 1.02 0.41 72 3.6 4.4 1.1 6.1 8.2 Contrast II (1: 1) 10 days 7.01 9.15 0.08 0.94 0.70 71 3.4 6.7 1.7 10.8 9.0 20 days 6.97 8.03 0.08 0.90 0.65 70 3.4 6.4 1.6 9.5 8.7 30 days 6.91 7.83 0.07 0.88 0.58 67 3.3 6.3 1.5 9.0 8.6 Control III (2: 1) 10 days 6.94 11.75 0.07 0.83 0.80 59 3.2 7.0 2.0 12.5 9.2 20 days 6.89 10.45 0.07 0.78 0.75 59 3.2 6.9 1.8 11.2 9.0 30 days 6.82 9.25 0.06 0.73 0.69 55 3.1 6.7 1.9 10.0 8.8

Treatment I (1: 2) 10 days 7.28 6.11 0.11 1.25 0.35 79 3.9 5.1 1.3 6.2 8.7 20 days 7.28 5.20 0.10 1.23 0.25 75 3.5 5.2 1.5 5.1 8.8 30 days 7.31 4.86 0.10 1.21 0.20 73 4.0 5.1 1.6 4.9 8.6 Treatment Zone II (1: 1) 10 days 7.15 7.48 0.10 1.05 0.55 72 3.4 7.3 1.8 9.1 9.2 20 days 7.17 6.21 0.09 1.02 0.38 70 3.7 7.3 1.9 7.9 9.4 30 days 7.21 5.92 0.09 0.98 0.32 67 3.7 7.3 2.0 6.8 9.1 Treatment III (2: 1) 10 days 7.09 9.30 0.08 0.92 0.70 61 3.2 9.3 2.2 10.2 9.8 20 days 7.13 7.30 0.08 0.90 0.51 57 3.1 9.4 2.3 9.1 10.1 30 days 7.16 6.84 0.08 0.87 0.43 54 3.4 9.3 2.4 9.0 9.8

The pH, EC, and salt concentrations of the reclaimed soils and the soils of the soils tended to decrease as the test progressed, but the treatments tended to increase. This is believed to be due to the high pH of the microbial preparation, 7.3. EC and salinity tended to decrease in both control and treatment, but the decrease of treatment with microorganisms was greater, and the decrease was higher in the order of treatment III> treatment II> treatment I (see Figure 4).

The effect of microbial agent on the change of salinity of soil according to the landfill soil conditions is that the pot test showed that the conductivity and salinity of the microorganisms were lower than that of the control. The conductivity and salinity tended to decrease. The decrease in salinity concentration was 14 ~ 18% in the control group, and the salinity reduction effect was more significant in the microbial treatment group at 39 ~ 43%.

In the evaluation of the above results, the salt concentration (NaCl) change according to the microbial agent treatment was examined through the column test and the Pot test. As a result, the electrical conductivity decrease and the salinity of the microbial agent containing the NM5 strain of halomonas in the reclaimed soil It showed the effect of reducing the concentration and showed the same salinity reduction effect as the column test under the landfill soil conditions. Therefore, the microbial agent containing Halomonas genus NM5 strain could be used as a salt reduction material in landfill and reclaimed land. It is expected.

Example 5 Plant Growth Promoting Effect of Halomonas sp. NM5 Strains

In order to evaluate the plant growth promoting effect of Halomonas sp. NM5 strain having excellent organic resolution and flame resistance, the microbial agent containing the Halomonas genus NM5 strain was used as a fertilizer. The effect was observed. The test area per treatment area was 15m ² (1.5m × 10m), and it was carried out in three steps of random placement in a plastic house. Fertilization method of microbial preparations were applied to foliar fertilization and irrigation four times as much as the amount set in each treatment group at a weekly interval after fertilization after flowering of strawberries, and the growth, yield and quality of strawberries were investigated.

Examination of soil physicochemical properties before and after the test showed that pH, effective phosphorus and organic matter tended to decrease after the test, and the electrical conductivity (EC) increased slightly after the test. After the crop cultivation test, the conductivity of microbial treatments tended to be lower than that of conventional treatments, and no significant difference was found between treatments (see Table 8).

TABLE 8

Changes in Physicochemical Properties of Soil Before and After Crop Experiments

process  phrase pH (1: 5) EC T-N O.M Ava .- P ₂ O ₅ Ex.- cations ( cmol _c / kg) CEC dS / m % mg / kg Ca ^{2 +} Mg ^{2 +} K ⁺ Na ⁺ cmol _c / kg Before the test 6.78 0.78 0.22 3.50 500 5.58 2.47 1.34 0.32 14.3 After test Practice 6.55 1.38 0.19 3.19 426 5.12 2.13 1.13 0.38 13.6 Control 6.59 1.32 0.19 3.21 434 5.11 2.10 1.11 0.35 13.6 Half-volume 6.62 1.32 0.20 3.21 438 5.11 2.08 1.10 0.32 13.9 Suitability 6.64 1.22 0.20 3.27 438 5.09 2.06 1.06 0.30 13.9 Drain 6.64 1.20 0.19 3.25 440 5.04 2.03 1.04 0.28 13.6

The effects of microbial agents containing Halomonas sp. NM5 strains on the growth of strawberries showed that the height, leaves, leaf width, tube diameter, and chlorophyll content tended to be higher than the control (sterilized) during the growth period. (See Table 9).

As a result of the quality inspection of the strawberry, the exaggeration of the exaggeration and the exaggeration were increased by 3-7% and 2-9% in the microbial treatments than the control (sterile), and the overweight was 2-6% in the microbial treatments. Sterile spheres). The sugar content was 2-5% higher in the control group than the control (sterile) (see Table 10).

TABLE 9

Effects of Halomonas sp. NM5 Strains on the Growth of Strawberries

process  phrase Candle sheet (cm) Leaf disease (cm) Lobe width (cm) 1st ^* 2nd ^** 1st 2nd 1st 2nd Practice 1.24.2 ^{a #} 28.2 ^a 6.77 ^a 9.67 ^a 4.83 ^a 7.77 ^a Control 24.4 ^a 28.2 ^a 7.05 ^a 9.71 ^a 4.94 ^a 7.83 ^a Half-volume 25.3 ^a 28.4 ^a 7.09 ^a 9.95 ^a 4.97 ^a 7.85 ^a Suitability 25.6 ^a 28.7 ^a 7.16 ^a 9.93 ^a 5.00 ^a 8.01 ^a Drain 25.9 ^a 29.0 ^a 7.35 ^a 10.1 ^a 5.10 ^a 8.11 ^a

process  phrase Conifer (dog) Tube diameter (mm) Chlorophyll (mg / 100 cm ² ) 1st ^* 2nd ^** 1st 2nd 1st 2nd Practice 2. 17.8 ^a 19.0 ^a 5.10 ^a 6.94 ^a 3.94 ^a 3.99 ^a Control 18.0 ^a 19.2 ^a 5.05 ^a 6.94 ^a 4.01 ^a 4.08 ^a Half-volume 19.6 ^a 19.8 ^a 5.10 ^a 7.01 ^a 4.08 ^a 4.14 ^a Suitability 20.0 ^a 21.0 ^a 5.13 ^a 7.09 ^a 4.18 ^a 4.23 ^a Drain 20.0 ^a 21.2 ^a 5.21 ^a 7.13 ^a 4.19 ^a 4.31 ^a

TABLE 10

Effects of Halomonas sp. NM5 Strains on Strawberry Quality

process  phrase Exaggeration And width Evaluation Fungus  medium Sugar mm Indices mm Indices g / ea Indices ^o Brix Indices Practice 42.0 ^{a *} 98 32.8 ^a 98 17.4 ^a 99 11.0 ^a 99 Control 42.7 ^a 100 33.5 ^a 100 17.6 ^a 100 11.1 ^a 100 Half-volume 44.0 ^a 103 34.2 ^a 102 18.0 ^a 102 11.3 ^a 102 Suitability 45.0 ^a 105 35.2 ^a 105 18.2 ^a 103 11.4 ^a 103 Drain 45.9 ^a 107 36.5 ^a 109 18.6 ^a 106 11.6 ^a 105

Fruit yield per week and fruit weight per week were increased by 2 ~ 11% and 6 ~ 18%, respectively, and the yield increased by 6 ~ 18%. Showed. Duncan's new multi-test resulted in a significant 5% level of overweight and yield per control (sterile) and microbial titration and batch treatment (see Table 11).

TABLE 11

Effects of Halomonas sp. NM5 Strains on Yield of Strawberry

process  phrase Per week  Number Per week  medium Yield recast Indices g / week Indices kg / 15m ² Indices Practice 8.60 ^a 98 146.5 ^b 98 20.5 ^b 98 Control 8.80 ^a 100 149.1 ^b 100 20.9 ^b 100 Half-volume 9.00 ^a 102 157.9 ^ab 106 22.1 ^ab 106 Suitability 9.47 ^a 108 167.9 ^a 113 23.5 ^a 113 Drain 9.73 ^a 111 175.5 ^a 118 24.6 ^a 118

In conclusion, there was no damage of strawberry due to fertilization of microbial preparations containing Halomonas sp. NM5 strains throughout the entire test period. Height, leaf number, chlorophyll, etc. were increased according to the throughput, and the fruit yield, fruit weight, and total yield per week were also increased compared to the control (sterile) (see FIG. 5).

Figure 1 shows a phylogenetic analysis using the 16S rDNA gene sequence of the novel Halomonas sp. NM5 strain of the present invention.

Figure 2 shows an electron micrograph of a novel Halomonas sp. ( NM5 ) strain of the present invention.

Figure 3 shows the effects of organic matter degradation and salt concentration reduction in reclaimed soil of microbial preparations containing a novel Halomonas sp. NM5 strain of the present invention.

Figure 4 shows the organic degradation and salt concentration reduction effect in the mixed soil of reclaimed soil and field soil of the microbial agent containing a novel Halomonas sp. NM5 strain of the present invention.

Figure 5 shows the growth promoting effect on the plant of the microbial agent containing a novel Halomonas sp. NM5 strain of the present invention.

Claims (8)

New Halomonas sp. NM5 (KACC 91326P) with excellent organic resolving power and flame resistance. A microbial fertilizer comprising the cells of the genus Halomonas sp. ( Mhalomonas sp.) NM5 or its culture medium as an active ingredient. The microbial fertilizer according to claim 2, which has a plant growth promoting effect. A landfill or landfill modifying agent comprising the cells of Halomonas sp. NM5 strain of claim 1 or a culture medium thereof as an active ingredient. The soil improving agent according to claim 4, which has a growth promoting effect of plants. A method for promoting plant growth using the microbial fertilizer of claim 2. A method of improving the soil of a coastal landfill or landfill using the soil improving agent of claim 4 or 5. A method of promoting plant growth in a landfill or landfill soil using the soil improving agent of claim 4 or 5.

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