KR20190012685A - Microbial agents for promoting plant growth comprising Pseudomonas nitroreducens or culture thereof as active ingredient - Google Patents

Abstract

The present invention relates to a microbial agent for plant growth comprising a Pseudomonas nitroreducens strain or a culture thereof as an active ingredient, a composition for promoting plant growth, a composition to be added to a fertilizer, and a method for promoting plant growth, including a step of treating a plant or soil with the strain or the culture thereof. According to the present invention, the Pseudomonas nitroreducens strain enhances cell division and nitrate intake to promote growth of plants. Accordingly, the Pseudomonas nitroreducens strain or the culture thereof can contribute to a crop quantity increase or quality improvement, and thus can be variously used in agricultural fields such as a microbial agent for promoting plant growth, a composition for promoting plant growth, and a composition to be added to a fertilizer.

Description

Technical Field [0001] The present invention relates to a microbial agent for promoting plant growth comprising Pseudomonas nitroredicine strains or a culture thereof as an active ingredient, and more particularly, to a microbial agent for promoting plant growth comprising Pseudomonas nitroreducens or a culture thereof,

The present invention relates to a plant growth microorganism preparation, a plant growth promoting composition, a composition for adding a fertilizer, and a step of treating the above-mentioned strain or a culture thereof with a plant or soil, comprising a Pseudomonas nitroredicine strain or a culture thereof as an active ingredient Lt; RTI ID = 0.0 > plant growth. ≪ / RTI >

The abuse of chemical fertilizers and pesticides damages not only the consumers but also the health of the growers, resulting in soil degradation. Soil degradation causes soil acidification and disease caused by plant pathogens, so it is repeated in a vicious cycle that is more dependent on chemical fertilizers and pesticides. Specifically, fertilizer in agriculture is an important agricultural material that determines the quantity of crops, but if used too much, it causes eutrophication of river water, water pollution of ground water, imbalance of soil nutrients, and poor growth of crops. Decreases can also cause crop quality deterioration.

Therefore, fertilizer and pesticides, which are used extensively in order to increase agricultural productivity, are the only alternative that can increase the income of the farm household by maintaining the productivity at the current point where the natural environment and human health are greatly threatened, Can be said to be the development of environmentally friendly farming methods.

However, due to the development and diversification of industry, the use of fertilizer is inevitable to increase the productivity of crops because the area of farmland is decreasing. Therefore, there is a growing interest in developing eco - friendly functional farming materials using microorganisms rather than conventional fertilizers.

Because of this interest, the use of chemical fertilizer has been steadily declining with the peak of the early 1990s. Since 1997, the use of organic fertilizer has been increased by induction of fertilizer reduction and composting such as development and supply of low concentration chemical fertilizer, The use of fertilizer is further reduced. The amount of chemical fertilizer used decreased by 1,104,000 tons in the early 1990s, 954,000 tons in 1995 and 801,000 tons in 2000, and decreased further in 2005, while the usage of compost and organic fertilizer is increasing Trend. However, domestic use of chemical fertilizers is still high among OECD countries.

Recently, crop cultivation methods using soil microorganisms have been used as one of eco-friendly agricultural techniques. Currently, cultivation of crops using soil microorganisms is widely performed in soybean plants. Rhizobium is the main microorganism used for the cultivation of these crops. When this strain infects soybean plants, it forms a root nodule, and the gaseous nitrogen is combined with hydrogen Nitrogen fixation that converts ammonia nitrogen (NH ₃ ) to nitrogen is performed. Nitrogen fixation has a very important agricultural significance. Nitrogen depletion is a natural consequence of non-fertilized soils. Under such soil conditions, soybean plants with root nodules resolve the problem of nitrogen depletion by themselves, And it is possible to grow under soil conditions where plants can not grow.

Accordingly, the inventors of the present invention have investigated a method for effectively promoting plant growth, and confirmed that Pseudomonas nitroredicine strain or a culture thereof has an excellent plant growth promoting effect, and completed the present invention.

KR Patent Publication No. 10-2011-0116734 KR Patent Publication No. 10-2011-0102787

It is an object of the present invention to provide a microorganism preparation for promoting plant growth, comprising Pseudomonas nitroreducens strain or a culture thereof as an active ingredient.

Yet another object of the present invention is to provide a composition for promoting plant growth, for enhancing the intake of nitrates of plants or for adding fertilizers, which comprises Pseudomonas nitroreducens strain or a culture thereof as an active ingredient will be.

It is yet another object of the present invention to provide a method for promoting plant growth, which comprises treating Pseudomonas nitroreducens strain or a culture thereof with a plant or soil.

In order to achieve the above object, the present invention provides a microorganism preparation for promoting plant growth, comprising Pseudomonas nitroreducens strain or a culture thereof as an active ingredient.

The present invention also provides a plant growth promoting composition comprising Pseudomonas nitroreducens strain or a culture thereof as an active ingredient.

In addition, the present invention provides a composition for enhancing the intake of nitrate in plants, comprising Pseudomonas nitroreducens strain or a culture thereof as an active ingredient.

The present invention also provides a composition for fertilizer addition comprising Pseudomonas nitroreducens strain or a culture thereof as an active ingredient.

The present invention also provides a method of promoting plant growth, comprising the step of treating a Pseudomonas nitroreducens strain or a culture thereof with a plant or soil.

The Pseudomonas nitroreducus strain of the present invention has an excellent effect of promoting plant growth by promoting cell division and nitrate intake. Accordingly, the Pseudomonas nitroreducus strain or the culture thereof of the present invention can contribute to the increase of the yield or quality of the crop by replacing the chemical fertilizer in the environment friendly organic agriculture in which the use of fertilizer is restricted, , Compositions for promoting plant growth, compositions for adding fertilizer, and the like.

FIG. 1 shows the results of observing GUS expression in DR5: GUS (a) or CycB1pro: GUS (b) transgenic Arabidopsis thaliana via GUS assay.
FIG. 2 is a graph showing the results of q-PCR analysis of changes in the expression level of the NRT2 gene following co-culture with Pseudomonas nitrorrhosis .
FIG. 3 is a view showing the result of confirming the change in nitrate content according to co-cultivation with a Pseudomonas nitroresundans strain.
Fig. 4 is a diagram showing the effect of the Pseudomonas nitrorei strain on the growth of Arabidopsis (Fig. 4a: appearance, Fig. 4b: chlorophyll level, Fig. 4c: root length, side root number and fresh weight).
FIG. 5 is a graph showing the effect of the Pseudomonas nitrorei strain on the growth of lettuce (FIG. 5 a: appearance observation, FIG. 5 b: raw weight).
FIG. 6 is a graph showing the results of observing GUS expression in DR5: GUS (a) or CycB1pro: GUS (b) transgenic Arabidopsis thaliana via GUS assay under low nitrogen conditions.
Fig. 7 is a graph showing the effect of the Pseudomonas nitrileysense strain on the growth of Arabidopsis thaliana under low nitrogen conditions (Fig. 7a: appearance observation, Fig. 7b: root length, side root number and fresh weight).

Hereinafter, the present invention will be described in detail.

For example, the present invention provides a plant growth promoting microorganism preparation comprising Pseudomonas nitroreducens strain or a culture thereof as an active ingredient.

As another example, the present invention provides a plant growth promoting composition comprising Pseudomonas nitroreducens strain or a culture thereof as an active ingredient.

As another example, the present invention provides a composition for enhancing the intake of nitrate in plants, comprising Pseudomonas nitroreducens strain or a culture thereof as an active ingredient.

In the present invention, 'Pseudomonas nitroresistans' is a soil microorganism belonging to anaerobic Gram-negative bacterium and is characterized by being able to synthesize a homopolymer of PHB (polyhydroxybutyrate) from a chain fatty acid. However, it is not known that the plant has yet to be used for promoting plant growth, and further, it is not known that the strain has a plant growth promoting purpose through promotion of nitrate intake.

In one embodiment of the present invention, Pseudomonas nitroresistans having a 16 rRNA gene containing the nucleotide sequence shown in SEQ ID NO: 1 is isolated from a soil sample through analysis and identification by a molecular genetic method by 16 rRNA sequence search This was used for the experiment, and the isolate was identified as a Pseudomonas nitrorrhizal strain IHB B 13561 strain, but this is only an example of Pseudomonas nitroresistans, and thus the present invention is not limited thereto. The Pseudomonas nitroresistans strain according to the present invention can be obtained by directly isolating from the soil, and any commercially available strain can be used.

In the present invention, 'culture' means not only a specific microorganism itself but also a microorganism cultured in a culture medium or a culture medium, and the culture medium may be a medium containing carbon source, nitrogen source, vitamin and mineral.

More specifically, the term "culture medium" means a culture solution inoculated with a strain in a liquid medium. The culture medium may be a culture solution containing the strain, or a culture filtrate obtained by culturing the strain inoculated with the strain and removing only the strain by centrifugation or the like. In addition, the culture medium is a concept including a concentrate, a dried product, and a fraction of a culture, and the formulation is not limited. For example, it may be a liquid or a solid. Herein, the concentrated liquid of the culture liquid means that the culture liquid is concentrated, and the dried material of the culture liquid means that the moisture of the culture liquid is removed. Drying methods include, but are not limited to, air drying, natural drying, spray drying and freeze drying.

In the present invention, " plant growth promotion " includes promoting cell division through plant nitrate intake, promoting lengthy growth of plants, and volume growth, and as a result, (Biomass) expressed in terms of the content of the biomass. In particular, the strain of the present invention has an advantage of promoting excellent plant growth by promoting nitrate ingestion even if a small amount of nitrate is contained in the soil.

In the present invention, 'plant' is a concept excluding bacteria, fungi, and animals, and refers to a creature that has photosynthesis with cell walls and chlorophyll. Thus, the crop to which the microbial agent or composition of the present invention can be used is not particularly limited as long as it is a plant grown using nitrate. In the present invention, the plant includes a plant belonging to the genus Arabidopsis or a plant of the genus Lactuca . In one embodiment of the present invention, Arabidopsis thaliana or Lactuca sativa ( Lactuca sativa ; Lettuce), but the scope of the plant is not limited.

In the present invention, 'microbial agent' is a kind of biological agent, and when used for soil or the like, has an effect described in the application due to the activity of the useful microorganism contained therein, do. Accordingly, the microorganism preparation of the present invention relates to a microorganism preparation that promotes plant growth through the ingestion of nitrate into a plant, and the microorganism preparation containing the Pseudomonas nitroreducus strain of the present invention or a culture thereof is prepared Can be produced in the form of a liquid fertilizer. That is, the microbial agent of the present invention can be formulated as a biological fertilizer to overcome this in environmentally friendly organic farming with limited supply of chemical fertilizer.

In the present invention, the microorganism preparation may be prepared in various forms known in the art. For example, when it is prepared in the form of a dry powder or liquid fertilizer, the Pseudomonas nitroreducus strain may be added per gram of dry powder or More preferably 10 ⁷ to 10 ¹¹ CFU / ml, more preferably 10 ⁹ CFU / ml, more preferably 10 ⁷ CFU / ml or more per 1 ml of the liquid phase. have. In addition, it is better when it is applied to the target crops in the state of live bacteria other than dead bacteria.

As another example, the present invention relates to the use of Pseudomonas nitroreducens ) or a culture thereof as an active ingredient.

In the present invention, 'fertilizer' is also referred to as fertilizer, which means nutrients that promote the growth of vegetation. The fertilizer containing the composition of the present invention can be easily used by mixing with the soil before sowing, and can be fertilized when spraying pesticides and the like. In particular, the composition for fertilizer addition is an environmentally friendly composition containing a component derived from a natural product and can be used more safely than a chemical fertilizer because it is harmless to human beings, livestock and soil.

In the present invention, the composition may provide a fertilizer containing the same, and the fertilizer of the present invention may be prepared by separately preparing a Pseudomonas nitroreducus strain or a culture thereof in the form of a fertilizer additive, mixing the fertilizer with the fertilizer, Can be added directly.

In the present invention, the Pseudomonas nitroresistans in the composition may be in a liquid or dry state, preferably in the form of a dried powder. Drying methods include, but are not limited to, air drying, natural drying, spray drying and freeze drying. The strain of the present invention or a culture solution thereof preferably contains 10 ⁵ CFU / ml or more per weight of fertilizer, but is not limited thereto. The fertilizer may further include conventional additives that can enhance the preservability of the fertilizer in addition to the strain or the culture of the present invention.

In the present invention, the composition can be prepared according to a method generally used in all fertilizer and pesticide chemistry. Preferably, the composition of the present invention is used in combination with a plant growth promoting agent, that is, a conventional fertilizer And the range of application of the composition to a plant or a plant growing soil may vary depending on the type of plant, age, climatic conditions, and the like.

As another example, the present invention relates to the use of Pseudomonas nitroreducens ) or a culture thereof to a plant or soil.

In the present invention, the method is characterized by promoting plant growth through the ingestion of nitrate. The method of promoting the growth of the plant may be a method of easily solubilizing the nitrate present in the soil using the strain of the present invention or a culture thereof.

In the present invention, the method can be carried out by specifically immersing, spraying or crossing the Pseudomonas nitroredicine strain, the cultured product thereof or the microorganism preparation of the present invention directly into a plant or soil, preferably soil, Spraying.

For example, there is a method of immersing the Pseudomonas nitroreducus strain of the present invention or a culture thereof into the soil around the plant, or immersing the seed in the culture medium or preparation by the method of immersion. In addition, there is a method of spraying the Pseudomonas nitroreducus strain of the present invention or a culture thereof with a technique well known in the art in such a manner that the Pseudomonas nitroreducus strain of the present invention is applied to plants in a continuous manner.

The redundant contents are taken into consideration in the complexity of the present specification, and terms not otherwise defined herein have the meanings commonly used in the art to which the present invention belongs.

Hereinafter, the present invention will be described in detail with reference to Examples and Experimental Examples in order to facilitate understanding of the present invention. However, the following examples and experimental examples are intended to illustrate the contents of the present invention, but the scope of the present invention is not limited to the following examples and experimental examples. The embodiments and experimental examples of the present invention are provided to enable those skilled in the art to more fully understand the present invention.

Example  1. Pseudomonas Nitroly Dysense ( Pseudomonas nitroreducens ) Isolation of strain

A soil sample of a conventional lettuce farmer (odus yarn) was added to a 15 ml conical tube containing 10 ml deionized water. Thereafter, the Pseudomonas nitroreducus strain of the present invention was isolated through a smear plate method. Concretely, the conical tube with the soil sample was vortexed for 5 minutes and 900 μl of luria-bertani broth was added to dilute the resultant 100 μl suspension to 10 ^-1 concentration. ≪ / RTI > Thereafter, the dilution was repeated to dilute the suspension to 10 ^-2 , 10 ^-3, and 10 ^-4 concentrations, and the 10 ^-4 concentration suspension was streaked in tryptic soy agar (TSA). This was incubated at 37 DEG C for 24 hours to isolate a single colony of Pseudomonas nitroreducens of the present invention.

Sequence comparison analysis of 16S rRNA was then performed to identify the strain. The isolated strain has the 16S rRNA sequence of SEQ ID NO: 1 and has a 99% or more homology with the 16S rRNA sequence of the Pseudomonas nitroredicine IHB B 13561 (GenBank KJ767371.1), a known strain And it was confirmed that it has the same mycological characteristics as the known strain.

For use in the subsequent experiments, the stock of the isolate was placed in an 80% glycerol solution and stored at 80 ° C.

EXPERIMENTAL EXAMPLE 1 Identification of Plant Growth Promoting Effect of Pseudomonas nitrilexense Strain of the Present Invention

1-1. Identification of Plant Growth Promoting Effect through GUS Assey

(Hereinafter referred to as " PnIHB ") together with a 4-day-old transgenic Arabidopsis thaliana containing DR5: Gus or CycB1pro: GUS responding to an external auxin (DR5: GUS) or a cell division promoting signal (CycB1pro: ) Were co-cultured. One day before co-culture, PnIHB was incubated for 18 hours at 37 DEG C on a shaker. Four days after co-culture, Arabidopsis was harvested and stained with GUS solution in a 37 ° C incubator for 2 hours. Each sample was then treated with 70% EtOH and incubated at 70 ° C until all chlorophyll was removed. As a control group, Arabidopsis cultured in LB medium without strain was used. The results are shown in Fig.

As shown in FIG. 1, it was confirmed that the GUS activity was significantly increased in the transgenic Arabidopsis thaliana cultured with the Pseudomonas nitroreducus strain (PnIHB) according to the present invention, as compared with the control. In particular, the activity of CycB1pro was significantly higher than that of DR5, which means that the function of Pseudomonas nitroreducus strain (PnIHB) according to the present invention is more closely related to the increase of cell division than auxin production.

1-2. Through q-PCR analysis NRT2 (nitrate specific transport2) Confirmation of increased expression of gene

Changes in NRT2 gene expression known to be associated with nitrate intake were analyzed by q-PCR. Specifically, Arabidopsis thaliana was co-cultured with the strain (PnIHB) of Example 1 for 8 days, and then total RNA was isolated from Arabidopsis thaliana and cDNA was synthesized from the RNA. Q-PCR was then performed on the cDNA in a manner commonly used in the art. As a control group, Arabidopsis cultured in LB medium without strain was used. The sequences of the primer sets used for q-PCR are shown in Table 1, and the results of the experiments are shown in Fig.

Gene Primer sequences NRT1.1
( At1g12110 ) F: AACAAACACCCCCAAAAACTGCA
R: CCTTGGAAGTCCCAAGCATC NRT1.2
( At1g69850 ) F: AGGTCTCAAGATGGGAAGGC
R: GGCGAGCATGCCACCGTGA NRT2.1
( At1g08090 ) F: AAAGACAAATTCGGAAAGATTCTG
R: AAGTACTCGGCGATAACATTATCA NRT2.2
( At1g08100 ) F: CCAAAGACAAATTCGGAAAGATTC
R: AAGTACTCGGCGATAACATTGTCT NRT 2.5
( At1g12940 ) F: CTTGTTCGGTCAGGATCTTCCT
R: GTTATCCATCCCCTATAGTTTTTG NRT2.6
( At3g45060 ) F: TGTTCTTGAGGCCATCACAAG
R: CTTCGTGTAGTCCTAAGCTCT

As shown in FIG. 2, expression of six types of NRT2 genes acting as HATS (high-affinity nitrate uptake system) in the Arabidopsis thaliana co-cultured with Pseudomonas nitroreducus strain (PnIHB) according to the present invention is remarkably increased Respectively.

1-3. Check nitrate content

In order to confirm the nitrate content in the Arabidopsis thaliana of Experimental Example 1-2, it was quantified using the conventionally known salicinic acid method (Cataldo et al., 1975; Na Xu et al., 2016; Vendrell and Zupancic, 1990) . Specifically, 50 mg of crushed Arabidopsis samples were added to 1 mL of distilled water and boiled at 100 ° C for 20 minutes. The sample was centrifuged at 15,871 g for 10 minutes and 0.1 mL of supernatant transferred to a new 12 mL tube. Then, 0.4 mL of salicylic acid-sulfuric acid (5 g of salicylic acid in 100 mL of sulfuric acid) was added to initiate the chemical reaction. After 20 min of reaction at room temperature, 9.5 mL of 8% NaOH solution was added. After 5 minutes of reaction at room temperature, the absorbance at OD410 was measured using a multiple detection microplate reader. The nitrate content was calculated using the following equation.

* Nitrate content = C x V / W (C: nitrate concentration calculated by regression equation at OD410, V: volume of extracted sample, W: sample weight).

0.1 mL of distilled water was used instead of 0.1 mL of the supernatant and the standard curve was made using KNO ₃ at a concentration ranging from 0.5 to 2 mg / L and the regression equation was calculated from a standard curve of r 2 ≥ 0.99. The results are shown in Fig.

As shown in Fig. 3, the Arabidopsis thaliana co-incubated with the Pseudomonas nitroredicus strain (PnIHB) according to the present invention showed a slightly lower nitrate content than the control group. This indicates that the consumption of nitrate by the Pseudomonas nitroreducus strain (PnIHB) according to the present invention is increased, which promotes cell division, requires more nitrogen and ATP, and decreases the nitrate content in the plant it means.

1-4. Identify growth promoting effect of Arabidopsis thaliana

The seeds of Arabidopsis thaliana (Col-0 ecotype)) were sterilized with sodium hypochlorite for 5 minutes and then washed 5 times with sterilized water. After storing for 3 days at 4 ° C, the Arabidopsis thaliana seeds were washed with 2% sucrose ≪ / RTI > Germinated Arabidopsis thaliana seeds (Col-0 seedlings) of uniform size were seeded on a plate (2% sucrose and the strain suspension of Example 1 (15 x 10 ⁸ CFU mL ^{- 1} ) containing the strain (PnIHB) , And then cultured for 8 days. Fig. 4A shows changes in appearance for 8 days, Fig. 4B shows the change in chlorophyll after 8 days, and Fig. 4C shows changes in root length, side root number and fresh weight.

As shown in FIG. 4, it was confirmed that the chlorophyll content of the Arabidopsis thaliana co-cultured with the Pseudomonas nitroreducus strain (PnIHB) according to the present invention was not significantly different from that of the control, but the growth of shoots and roots was promoted . In particular, as shown in FIG. 4C, the Arabidopsis thaliana grown in the medium containing Pseudomonas nitroreducens strain (PnIHB) according to the present invention showed an increase of the side root number by about 15% Respectively.

1-5. Confirming the promotion effect of lettuce growth

Lettuce (L. sativa) was commercially available red seeds from seed shops. The lettuce seeds were sterilized with sodium hypochlorite for 5 minutes, washed 5 times with sterilized water, and stored in a refrigerator for 3 days. The sterilized lettuce seeds were inoculated into sterilized soil in a sterile container. At this time, sterilization of the soil was carried out to remove other microorganisms present in the soil. Four days later, uniform sized lettuce seedlings were transferred to a sterile soil container to which 90 ml of distilled water had been added. For inoculation, LB strain (PnIHB) culture solution in Example 1 in 1 ml of different concentrations to ^- was treated directly in the ^{(10 1, 10 3, 10} 5 CFU mL 1) the roots of a plant into. A soil container treated with 1 ml of LB containing no bacteria in plant roots was used as a control. The lettuce treated with the strain of Example 1 (PnIHB) was co-cultured for 10 days at 23 ° C (16-h light / 8-dark) and 60% relative humidity. Fig. 5A shows the appearance change over 10 days, and Fig. 5B shows the change in the weight after 10 days.

As shown in FIG. 5, the lettuce cultured in the medium containing the Pseudomonas nitroreducens strain (PnIHB) according to the present invention was found to have significantly increased leaf growth as compared with the control.

From the above experimental results, it was confirmed that the Pseudomonas nitroreducus strain according to the present invention has an excellent effect of promoting plant growth by increasing cell division and nitrate ingestion.

Experimental Example  2. Low nitrogen  The present invention Pseudomonas Nitroly Dysense  Identification of Plant Growth Promoting Effect of Strain

In order to confirm whether the strain of the present invention has plant growth promoting effect even in a nitrogen-deficient environment, the following experiment was conducted.

2-1. Identification of Plant Growth Promoting Effect through GUS Assey

To conduct experiments under low nitrogen conditions, Hoagland N-free medium supplemented with 1 mM KNO ₃ as a sole nitrogen source was used. A 4-day-old transgenic Arabidopsis thaliana containing DR5: Gus or CycB1pro: GUS responding to external auxin (DR5: GUS) or cell division promoting signal (CycB1pro: GUS) was incubated with the strain of Example 1 under low nitrogen conditions Co-cultured. The specific experimental method is the same as in Experimental Example 1-1. The results are shown in Fig.

As shown in FIG. 6, the transgenic Arabidopsis thaliana co-cultured with the Pseudomonas nitroredicine strain (PnIHB) according to the present invention showed significantly higher GUS activity than the control group even under the low nitrogen condition.

2-2. Identify growth promoting effect of Arabidopsis thaliana

The Arabidopsis thaliana seeds (Col-0 seedlings) germinated in MS agar containing 2% sucrose were seeded onto a plate (1 mM KNO ₃ and 2% water) containing the strain of Example 1 (15 x 10 ⁸ CFU mL- Hoagland N-free agar supplemented with crossover) and cultured for 8 days. Fig. 7 (a) shows changes in appearance after 8 days, and Fig. 7 (b) shows changes in root length, side length and weight after 8 days.

As shown in FIG. 7, it was confirmed that the Arabidopsis thaliana cultivated in the medium containing Pseudomonas nitroreducus strain (PnIHB) according to the present invention showed increased number of side roots and fresh weight in comparison with the control group even under the low nitrogen condition Respectively.

In general, the Pseudomonas nitroreducus strain of the present invention has an excellent effect of promoting plant growth by promoting cell division and nitrate intake. Accordingly, the Pseudomonas nitroreducus strain or the culture thereof of the present invention can contribute to the increase of the yield or quality of the crop by replacing the chemical fertilizer in the environment friendly organic agriculture in which the use of fertilizer is restricted, , Compositions for promoting plant growth, compositions for adding fertilizer, and the like.

<110> Korea University Research and Business Foundation <120> Microbial agents for promoting plant growth comprising          Pseudomonas nitroreducens or culture thereof as active ingredient <130> 1-89 <160> 1 <170> KoPatentin 3.0 <210> 1 <211> 941 <212> DNA <213> Artificial Sequence <220> <223> 16S ribosomal RNA partial sequence <400> 1 atcgggatta ctgggcgtaa gcgcgcgtag gtggtttggt aagatggatg tgaaatcccc 60 gggctcaacc tgggaactgc atccataact gcctgactag agtacggtag agggtggtgg 120 aatttcctgt gtagcggtga aatgcgtaga tataggaagg aacaccagtg gcgaaggcga 180 ccacctggac tgatactgac actgaggtgc gaaagcgtgg ggagcaaaca ggattagata 240 ccctggtagt ccacgccgta aacgatgtcg actagccgtt gggatccttg agatcttagt 300 ggcgcagcta acgcgataag tcgaccgcct ggggagtacg gccgcaaggt taaaactcaa 360 atgaattgac gggggcccgc acaagcggtg gagcatgtgg tttaattcga agcaacgcga 420 agaaccttac ctggccttga catgtccgga accttgcaga gatgcgaggg tgccttcggg 480 aatcggaaca caggtgctgc atggctgtcg tcagctcgtg tcgtgagatg ttgggttaag 540 tcccgtaacg agcgcaaccc ttgtccttag ttaccagcac gttatggtgg gcactctaag 600 ggactgccg gtgacaaacc ggaggaaggt ggggatgacg tcaagtcatc atggccctta 660 cggccagggc tacacacgtg ctacaatggt cggtacagag ggttgccaag ccgcgaggtg 720 gagctaatcc cataaaaccg atcgtagtcc ggatcgcagt ctgcaactcg actgcgtgaa 780 gtcggaatcg ctagtaatcg tgaatcagaa tgtcacggtg aatacgttcc cgggccttgt 840 acacaccgcc cgtcacacca tggggagtgg gttgctccag aagtagctag tctaaccgca 900 agggggacgg ttaccacgga gtgattcatg actggggtga a 941

Claims (7)

Pseudomonas nitroreducens ) or a culture thereof as an active ingredient. The method according to claim 1,
Wherein the 16 rRNA nucleotide sequence of the strain is the nucleotide sequence of SEQ ID NO: 1. The method according to claim 1,
Wherein the microbial agent promotes plant growth through promotion of nitrate intake. Pseudomonas nitroreducens ) or a culture thereof as an active ingredient. Pseudomonas nitroreducens ) or a culture thereof as an active ingredient. Pseudomonas nitroreducens ) or a culture thereof as an active ingredient. Pseudomonas nitroreducens ) or a culture thereof to a plant or soil.

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