KR101922428B1 - New microorganism Bacillus toyonensis SB19 having growth promoting of Leafy vegetables and high temperature tolorance and drought resistance of leafy vegetables and microbial agent containing the same and biofertilizer containing the same - Google Patents

Abstract

The present invention relates to novel microorganism bacillus toyonensis SB19 having growth promotion of leafy vegetables and high temperature tolerance and drought resistance of leafy vegetables, a microbial agent containing the same and a biofertilizer containing the same. The novel microorganism bacillus toyonensis SB19 strain (KACC 81045BP) consists of the base sequence represented by SEQ ID NO: 1, and has growth promotion of leafy vegetables and high temperature tolerance and drought resistance of leafy vegetables. The present invention provides the novel microorganism bacillus toyonensis SB19, the microbial agent containing the same and the biofertilizer containing the same to overcome a water deficit environment, i.e., a drought condition, and reduce drought damage of plants to promote the growth of leafy vegetables vulnerable to drought in comparison to other crops for cultivating roots or fruits even under a weak drought condition to be effectively used in eco-friendly leafy vegetable production.

Description

TECHNICAL FIELD The present invention relates to a novel microorganism, Bacillus toxinensis SB19, which is capable of promoting the growth of safflowers and having high temperature resistance and resistance to wrapping, and a microorganism preparation containing the microorganism, and a biological fertilizer containing the microorganism Bacillus toyonensis SB19 having growth promoting properties of Leafy vegetables and high temperature tolerance drought resistance of leafy vegetables and microbial agents containing the same and biofertilizer containing the same}

The present invention relates to a novel microorganism, Bacillus toxinensis SB19, a microorganism preparation containing the microorganism, and a biological fertilizer containing the microorganism. More particularly, the present invention relates to a microorganism, A new microorganism, Bacillus toxinensis SB19, which enables the growth of safflower plants to be achieved, a microbial agent containing the same, and a biocidal fertilizer containing the same.

Drought is one of the abiotic stresses that hinders the growth and development of plants, and is a major reason for limiting agricultural productivity. It is known that drought changes not only plant metabolism and growth but also plant gene expression pattern, and it affects many physiochemical activities and survival. Plant damage caused by drought does not occur suddenly like frost injury, but progresses slowly and gradually increases intensity and lasts. This is because the plants are gradually exposed to drought stress and induce leaf senescence. In other words, due to drought stress, the endogenous cytokinin content of plant leaves is decreased and abscisic acid content is increased, so that pore closure occurs rapidly and the loss of plant water loss by the action of vaporization is reduced. In addition, the use of carbon dioxide is reduced by pore closure, and photosynthesis is suppressed. When water stress is persistent and deteriorates, the leaves are removed because they are only nonproductive sites that do not undergo photosynthesis. In plants where photosynthesis is suppressed, oxygen reacts with electrons to form excessive reactive oxygen species (ROS), which have a strong toxicity. ROS is an oxidative stress such as lipid peroxidation in the cell membrane and inhibition of nitrate anabolism in plant roots Cause.

Especially, in the case of leafy vegetables which are ingested with fresh leaves, this action due to drought causes decrease of productivity. Especially in the summer, high insolation and high temperatures exacerbate the water stress stress, so the vegetables may be more fatal to drought than other crops that grow roots or fruit.

Recently, many studies have been carried out on methods that can increase the plant nutrition through the interaction of beneficial microorganisms and plants and increase the resistance of plants to abiotic stresses such as drought, low temperature, salt and heavy metal pollution It is known. Among them, plant growth-promoting bacteria (PGPB) are useful microorganisms that exist in the rhizosphere and soil of plants. They are nitrogen fixation, solubilization of insoluble phosphorus, plant hormone synthesis, siderophore production, antibiotics, lytic enzymes, hydrogen cyanide production , Control of plant pathogens through competition for nutrients, and so on.

In the present study, PGPB increased the growth of plants in the drought condition compared to the untreated group, suggesting that PGPB could be a good way to overcome drought stress (Mayak et al., 2004; Kasim et al. al., 2013; Seo and Song, 2013; Vurukonda et al., 2016). Various methods for mitigating the drought stress effect of plants caused by PGPB have been known.

Specifically, a useful substance produced by a microorganism induces stress tolerance in a plant, thereby forming a biofilm containing a sugar and an oligopolysaccharide, so that the root of the microorganism- And the water availability at the surface is enhanced. It is also known that microorganisms produce plant hormones such as abscisic acid, gibberellic acid, cytokinin, auxin and the like, or ACC (1- aminocyclopropane-1-carboxylate) deaminase, where ethylene is produced during the maturation of plants, such as accelerating ripening of fruit, but is also caused by external stresses such as drought and high temperature, This study was conducted to investigate the effects of PGPB on the growth of plants under stress conditions by lowering the level of ethylene in plant roots by PGPB. , It was confirmed that drought tolerance was improved when the plants were inoculated. This is due to the action of ACC deaminase produced by microorganisms, (Kiyosue et al., 1994; Glick et al., 1998; Timmusk and Wagner, 1999) have shown that the production of ress ethylene is decreased and the expression of ERD15 ( early response to dehydration 15 ) is increased.

Although the above-mentioned various studies have been carried out, it has not yet been found that strains having both high-temperature and high-temperature resistant strains have not yet been found. Therefore, it is possible to overcome the water-poor environment under the drought condition and alleviate the drought damage of the plants The development of new strains representing such strains is required.

1. Mayak, S., T. Tirosh, and B. R. Glick. 2004. Plant growth-promoting bacteria that confer resistance to water stress in tomatoes and peppers. Plant Sci. 166 (2): 525-530. 2. Kasim, W. A., M. E. Osman, M. N. Omar, I. A. A. El-Daim, S. Bejai, and J. Meijer. 2013. Control of drought stress in wheat using plant-growth-promoting bacteria. J. Plant Growth Regul. 32 (1): 122-130. 3. Seo, M.-S. and H.-G. Song. 2013. Growth promotion of tomato plant under drought conditions by treatment of rhizobacteria producing ACC deaminase and phytohormones. Korean J. Microbiol. 49 (1): 46-50. 4. Vurukonda, S. S. K. P., S. Vardharajula, M. Shrivastava, and A. SkZ. 2016. Enhancement of drought stress tolerance in crops by plant growth promoting rhizobacteria. Microbiol. Res. 184: 13-24. 5. Kiyosue, T., K. Yamaguchi-Shinozaki, and K. Shinozaki. 1994. ERD15, a cDNA for a dehydration-induced gene from Arabidopsis thaliana. Plant Physiol. 106 (4): 1707. 6. Glick, B. R., D. M. Penrose, and J. Li. 1998. A model for plant growth-promoting bacteria. J. Theor. Biol. 190 (1): 63-68. 7. Timmusk, S. and E. G. H. Wagner. 1999. The plant-growth-promoting rhizobacterium Paenibacillus polymyxa induces changes in Arabidopsis thaliana gene expression: a possible connection between biotic and abiotic stress responses. Mol. Plant Microbe Interact. 12 (11): 951-959.

The object of the present invention is to provide a new microorganism, Bacillus toenansis ( Bacillus sp.), Which promotes the growth of sweetened vegetables, toyonensis SB19, a microbial agent containing the same, and a biological fertilizer containing the microbial agent.

In order to achieve the above object, the novel microorganism Bacillus toyonensis SB19 strain (KACC 81045BP) of the present invention comprises the nucleotide sequence represented by SEQ ID NO: 1, and is characterized in that it promotes growth of the transplanting plants, . Such a strain is characterized by having root-fixing ability of wrapping.

It is characterized in that it is applied to any one of kale and red mustard as the wrapping and is used as an active ingredient of any one of microbial pesticides, seed coating agents, soil improving agents, composting homework agents, leaf side spraying agents and rooting agent spraying agents.

The microorganism preparation for enhancing resistance to high temperature or drying stress of wrapping vegetables of the present invention is characterized by containing the above-mentioned new microorganism Bacillus toyonensis strain SB19 (KACC 81045BP) as an active ingredient.

The method comprises the steps of preparing the new microorganism Bacillus toyonensis strain SB19 (KACC 81045BP), and inoculating the strain into TSB medium, followed by incubation at 20 to 35 ° C at 150 to 200 rpm Under conditions of 40 to 50 hours.

The biocidal fertilizer for enhancing resistance to high temperature or drying stress of wrapping vegetables according to the present invention is characterized by containing at least one of the microorganism Bacillus toyonensis strain SB19 (KACC 81045BP) or the microorganism preparation as an active ingredient .

The present invention is characterized in that the biocidal fertilizer is applied to the soil in such a manner as to increase tolerance to high temperature or drying stress of wrapping vegetables.

The technical objects to be achieved by the present invention are not limited to the above-mentioned technical problems.

The present invention provides a novel microorganism Bacillus toxinensis SB19 that promotes the growth of wrapping vegetables and has high temperature resistance and resistance to wrapping, a microorganism preparation containing the microorganism, and a biocide fertilizer containing the same, It is expected that it will be useful for the production of environment friendly safflower because it makes it possible to grow the fried vegetable which is more fatal to the drought than other crops that grow root or fruit, .

In addition, it is expected to support the value of microbiological methods to overcome environmental stress.

FIG. 1A is a diagram showing the growth promoting effect of kale by treatment with Bacillus toyonensis strain SB19, which is a starting strain of the present invention.
A: Control (water treated control)
B: SB19 of the present invention
FIG. 1B is a graph showing the growth promoting effect of kale seedlings by treatment with Bacillus toyonensis strain SB19 as a starting strain of the present invention.
FIG. 2 is a graph showing the results of confirming the molecular system of the selected strain of FIG. 1 based on the 16S rRNA gene sequence.
FIG. 3 is a graph showing a dry stress-relieving effect of kale on days 7 and 14 after treatment with Bacillus toyonensis strain SB19 as a starting strain of the present invention.
A: control (water treated control) -7 days
B: Treatment of SB19 10 ⁶ cell mL < ^-1 >
C: On treatment with SB19 10 ⁷ cell mL < ^-1 >
D: control (water treated control) -14 days
E: Treatment of SB19 10 ⁶ cell mL- ¹ of the present invention -14 days
F: -14 days after SB19 10 ⁷ cell mL- ¹ treatment of the present invention
G: Graphing
FIG. 4 is a graph showing a dry stress-relieving effect of red mustard on the 7th day after the treatment with the strain Bacillus toyonensis SB19 as a starting strain of the present invention.
A: Control (water treated control)
B: SB19 of the present invention
C: Graphing
FIG. 5 is a graph showing a high-temperature stress-relieving effect of an enemy mustard for 7 days after the treatment of the strain Bacillus toyonensis SB19 as a starting strain of the present invention.
A: Control (water treated control)
B: SB19 of the present invention
C: Comparative strain CeR16-2
D: Graphing
FIG. 6 is a graph showing a high-temperature stress-relieving effect of kale on the 7th day after the treatment of the strain Bacillus toyonensis SB19 as a starting strain of the present invention.
A: Control (water treated control)
B: SB19 of the present invention
C: Comparative strain CeR16-2
D: Graphing
FIG. 7 is a graph showing the change in the number of general bacteria in the soil and root of mustard root by the soil treatment of the strain Bacillus toyonensis SB19 as a starting strain of the present invention.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, the present invention will be described in detail, and a detailed description of known functions and configurations that may unnecessarily obscure the gist of the present invention will be omitted.

The inventors of the present invention cultured each strain purely isolated from the cultivated soil of the cabbage cultivated in kale, red cabbage, and rhizosphere soil, collected the bacterial cells, prepared a suspension of the strain, and treated the pots directly with the kale, , Leaf area, biomass, and moisture content (the port showing the growth promoting effect) are finally selected.

Subsequently, the SB19 strain, which was introduced here, was confirmed based on the 16S rRNA gene sequence. As a result, the SB19 strain was closely related to various strains of Bacillus sp. B. toyonensis BCT-7112T strains showed the highest homology (99.93%, respectively), indicating a novel strain of B. toyonensis .

Therefore, the present invention named this strain as Bacillus toyonensis strain SB19, which was deposited with the National Institute of Agricultural Science and Technology , National Academy of Agricultural Science and received the accession number KACC 81045BP on Apr. 3, 2017, It was confirmed that it comprises all or a part of the nucleotide sequence shown in SEQ ID NO: 1 and that the sequence consists of an open reading frame (ORF) of 1,480 bp.

In order to confirm that the SB19 strain having the growth promoting effect of the above-described structured sweet potatoes was able to grow even in the drought condition, the resistance enhancement effect of kale and red mustard on the high temperature or the dry stress was examined. (water treated control), and it was confirmed that the effect was shown up to 7 days in the case of red mustard and up to 14 days in case of kale.

Further, the optimal growth range was examined by examining the physiological characteristics of the new strain. It was confirmed that the optimum growth range was 20 to 35 ° C, the optimum growth rpm was 150 to 200 rpm, and the optimum growth time was 40 to 50 hours.

In particular, the root colonization of the SB19 strain using the rhizosphere soil and root dilution plating method was investigated to confirm that the high temperature or dry stress mushrooming effect was caused by the mechanism of the new strain As a result, it was confirmed that the SB19 strain, which was the new strain, effectively settled on the enemy root after 5 weeks of treatment and persisted. This demonstrates microbial-plant interaction by plant root establishment of the strain SB19, which is the novel strain, and thus the SB19 strain, which is the novel strain, plays an important role in promoting water availability of plants, thereby proving that it is a strain that helps overcome drought stress .

In addition, the microorganism preparation (culture medium) of the present invention includes the novel strain, and any condition such as the growth temperature, rpm, and time of the strain exhibits the resistance enhancement effect of the wrapping to high temperature or dry stress, However, based on the above experimental results, it can be seen that the strain activity most preferably occurs when the temperature is from 180 to 200 rpm at 20 to 35 ° C.

The biological fertilizer of the present invention is characterized in that it comprises at least one of the above-mentioned microbial cells or the microbial cells.

The bioprotective agent of the present invention may further contain glycerol, milk powder, milk, skim milk powder, animal or vegetable oil, etc. suitable as a microorganism protecting agent, and the nutrient source may include fibrinogen powder, soybean meal powder, cottonseed powder, The present invention further includes the carbon source and nitrogen source such as powder, corn extract, yeast extract, molasses, glucose, sugar, corn syrup and the like in addition to the usual amount so as to further promote the efficacy of the strain.

The biological fertilizer obtained by the above method can be formulated into various forms such as powders, pellets or granules of appropriate size using a general formulator.

The biological fertilizer of the present invention formulated in the above may be used as it is or may be dried by air drying at room temperature or by lyophilization or high temperature drying method.

By treating the biological fertilizer according to the present invention with the soil, it promotes the growth of the wrapping plants and improves resistance to high temperature or drying stress of the wrapping itself, thereby enhancing the drought stress inhibition, .

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for illustrating the present invention and that the scope of the present invention is not construed as being limited by these embodiments.

≪ Example 1 > Separation of strains and selection of strains promoting growth of kale

1. Experimental Method

1) Strain isolation and culture method

The isolates were isolated from the soil of Kale, Redsuckle, Chicory and Celery cultivated soil and rhizosphere soil. 100 μL of a suspension (10 ^-1 to 10 ^-6 g mL ^-1 ) diluted with 1 g of soil sample in 9 mL of sterilized water was plated on a Tryptic Soy Agar (TSA; Difco, USA) medium and cultured at 28 ° C. for 48 hours While isolating single colonies of strains showing differences in color and morphology. Forty-six strains isolated purely were suspended in 20% glycerol and stored at -70 ° C for growth-promoting bioassay and identification. The isolated strains were inoculated into Tryptic Soy Broth (TSB; Difco, USA) and cultured at 28 ° C and 180 rpm for 48 hours. The culture was centrifuged at 8000 rpm for 15 minutes and the supernatant was removed to collect the cells. The collected cells were diluted with sterilized water to a concentration of 10 ⁷ cell mL ^-1 .

2) Selection method of strain promoting growth of kale

BM6 Sao (Berger Peat Moss, Canada) containing a square hole 32 ports (one per ball Sao requirements; 125 mL) were inoculated the kale (Brassica oleracea var alboglabra Bailey.) A (n = 8). The suspension of the strain prepared 7 days after kale sowing was treated once per 50 mL per pot, and treated with 50 mL of distilled water. The growth of kale was investigated after two weeks of strain treatment. Growth investigations were carried out on the basis of plant height (㎝), root length (㎝), leaf width (㎝), leaf length (㎝), maximum leaf area (㎠), root weight (㎎) (㎎) were used as subjects. Leaf length, leaf width, and maximum leaf area were investigated for leaf length and leaf width. In the selection of the second strain, the pot was directly assayed by the same method as that of the first strain selection using KARCOTO (Seoul Bio, Korea).

2. Experimental results

The results of the experiment are shown in Table 1 below.

Selection of primary strain plant height
(cm) leaf width
(cm) leaf length
(cm) shoot fresh weight
(mg) root fresh weight
(mg) shoot dry weight
(mg) root dry weight
(mg) control 8.7 2.5 3.7 419.8 307.1 65.3 39.5 SB3 10.2 3.4 4.4 926.3 456.6 125.5 52.0 SEL5 10.2 3.2 4.4 824.8 313.5 132.0 40.3 SB22 9.3 3.1 3.9 565.3 265.8 78.0 48.0 SRS25 12.5 4.2 5.0 1373.5 410.0 160.3 55.3 SEL2 11.1 3.2 4.1 785.3 443.0 95.0 55.3 KR14 11.9 4.2 5.0 1226.3 432.8 137.5 57.3 KR16 14.0 5.0 6.1 1585.0 232.0 142.4 37.5 KR31 13.5 4.5 6.0 1899.8 300.3 190.8 42.8 SB19 14.5 5.8 6.9 2725.3 475.5 275.0 45.5 RbR16 11.5 4.2 4.9 1250.0 370.5 144.5 52.3 CeR16-2 11.5 4.5 5.3 1728.0 435.0 210.3 57.8

As shown in Table 1, the plant height, leaf length, leaf width and fresh weight of the 12 strains were higher than those of the untreated control, , 6 strains SRS25, KR16, KR31, SB19, RbR16 and CeR16-2 were selected in order of greatest live weight gain.

After the above experiment, it was judged that it is preferable to select the soil with various physical and chemical properties when selecting the plant growth promoting strains by the bioassay. Thus, The strains were selected.

The results are shown in Table 2 below.

Secondary strain selection plant height (cm) Shoot fresh weight (mg) BM6 Baroker BM6 Baroker control 8.7 9.9 419.8 698.8 SRS25 12.5 12.6 1373.5 993.8 KR16 14.0 11.7 1585.0 837.5 KR31 13.5 13.8 1899.8 1318.8 SB19 14.5 14.1 2725.3 1662.5 RbR16 11.5 11.9 1250.0 877.5 CeR16-2 11.5 13.3 1728.0 991.3

As shown in Table 2 above, the control showed better growth of kale on the baroque soil than on the BM6 soil used in the primary strain selection, while in the case of the strain treated with BM6, And SB19 was the most effective for promoting the growth of kale on the BM6 soil.

≪ Example 2 > Effect of promoting growth of kale by the finally selected strain SB19

1. Experimental Method

The strains SB19 finally selected in Example 1 were cultured and bacterial cells were collected to prepare a strain suspension. After 7 days of kale sowing, the suspension of the prepared strains was treated once per 50 mL per pot, and the same amount of distilled water was applied to the control. The growth of kale was investigated after two weeks of strain treatment. Plant height, maximum leaf area, and number of leaves were measured in the isolated strains to confirm the growth promotion effect on the surface.

2. Experimental results

As a result of the above experiment, as shown in Figs. 1a and 1b, it was finally confirmed that the SB19 strain treatment showed the effect of enhancing the growth of excellent kale seedlings.

Example 3 DNA Extraction of SB19 Strain and Analysis of 16S rRNA Gene

1. Experimental Method

Based on the 16S rRNA gene sequence of strain SB19, which was finally selected through Examples 1 and 2, molecular schematics were created.

2. Experimental results

As shown in FIG. 2, the SB19 strain is closely related to the Bacillus sp . Strains and has the highest homology (99.93%) with B. toyonensis BCT-7112T strain. The bootstrap value, which indicates the stability of the system, is also relatively high at 77%.

Thus, it was confirmed that the new strain selected in Example 1 was identified as a final new microorganism B. toyonensis strain. As described in the deposit document attached thereto, the SB19 strain is referred to as ' Bacillus toyonensis SB19' , And this strain was deposited with the National Institute of Agricultural Science and Technology, National Institute of Agricultural Science and Technology and received the accession number KACC 81045BP on April 3, 2017.

<Experimental Example 1> Drying stress relieving effect of kale by treatment with strain SB19

1. Experimental Method

After 7 days of sowing, the strain suspension was treated with 10 ⁶ cell mL ^-1 and 10 ⁷ cell mL ^-1 for 1 time and the same amount of distilled water was applied to the control. Water stress was given without irrigation for 7 days from 10 days after the strain treatment, and the degree of damage was examined on the 7th day. Immediately after the irradiation, 50 mL of distilled water per pot was administered to all treatments, and water stress was again given for 7 days, and then the degree of damage was examined 14 days after the treatment. The extent of damage was divided into 0 ~ 5 stages based on Table 3 below.

Severity scale of drought stress injury Extent of damage 0 healthy One 1 ~ 10% damage 2 11 ~ 25% damage 3 26 ~ 50% damage 4 51 ~ 80% damage 5 > 80% damage or death

The results of the survey were expressed as% by the following equation (1), and the damage of healthy plants was 0% and the damage of dead plants was 100% without drought damage.

(1)

(One)

2. Experimental results

As shown in FIG. 3, when the suspension of SB19 strain was treated, there was an improvement in resistance to dryness compared to the untreated solution. Specifically, the degree of damage in the untreated area reached 82.5% on the 7th day after the treatment, while the damage rates of the 10 ⁶ and 10 ⁷ cell mL ^-1 SB19 strains were 65% and 30%, respectively, . On the 14th day after the treatment, the damage level of untreated control reached 87.5% and the damage of 10 ⁶ and 10 ⁷ cell mL ^-1 SB19 strains were 71.3% and 47.5%, respectively, It was observed that drought damage to kale was alleviated. The effect of treatment concentration on drought mitigation was better at 10 ⁷ cell mL ^-1 than 10 ⁶ cell mL ^-1 at 7 and 14 days after dry stress induction. This, initially 10 ⁷ more effective in overcoming dry stress in the cell mL ^-1 concentration because until the 7th day after the drought stress induced damage due to water shortage in the concentration of 10 ⁶ cell mL ^-1 SB19 strains faster generation Seems to be. Furthermore, since significantly enemy to all the road damage due to drought in a concentration of 10 ⁶ cell mL ^-1 when viewed findings day 7 and day 14 compared with no treatment, 10 ⁷ cell mL ^-1 concentration but less effective than 10 ⁶ cell mL- ¹ concentration was effective in overcoming the water stress stress. Therefore, it was concluded that the treatment of strain SB19 was effective in reducing plant stress and drying stress by inhibiting the aging of kale induced by moisture deficiency.

<Experimental Example 2> Effect of SB19 strain treatment on drying stress (weather resistance) of enemy mustard

1. Experimental Method

Experiments were carried out in the same manner as in Experimental Example 1, except that dry stress relieving effect was confirmed on red mustard in place of kale, and the effect was confirmed on day 7.

2. Experimental results

As a result, as shown in FIG. 4, as shown in FIG. 3, it was confirmed that dry stress relieving effect was not exerted even in red mustard seeds, while drying stress relieving effect was observed in SB19 seedlings.

<Experimental Example 3> Effect of SB19 strain treatment on the reduction of hot stress (hypertonicity) of red mustard and kale

1. Experimental Method

Kale and red mustard seeds, which had passed seven days after sowing, were treated once in the same manner as in Experimental Example 1. After 10 days from the day of the strain treatment, high temperature treatment was carried out at 40 ° C in a thermostat. After 24 hours, the damage was investigated. After the irradiation, 50 ml of distilled water per port was used in all treatments. After that, high temperature treatment was performed for 24 hours and the degree of damage was investigated at 48 hours after the high temperature treatment.

2. Experimental results

As shown in FIG. 5 and FIG. 6, it was confirmed that both the red mustard and kale had no effect of reducing the stress at high temperatures, whereas the SB19 treatment showed high stress relief effect. In particular, the CeR16-2 strain shown in Table 2 is a Bacillus velezensis CeR16-2 strain belonging to the genus Bacillus such as the SB19 strain of the present invention, and exhibits a somewhat enhanced promoting effect on growth of the transplant, as shown in Table 2, The results of the high temperature tolerance test of FIG. 6 show that the resistance to high temperature is not higher than that of the control. As a result, the same Bacillus sp. And SB19 strain according to the present invention were found to be capable of growth of the pickled vegetables in the drought condition.

<Experimental Example 4> Determination of plant root rooting ability of SB19 strain

There are many ways to mitigate the effects of drought stress on useful microorganisms. a) an induced systemic tolerance (ISR) is induced in a plant by a useful substance produced by the microorganism, b) a biofilm containing a sugar and an oligo-polysaccharide is formed, This is done in a way that water availability at the roots surface by interaction is enhanced. C) the microorganism produces a plant hormone such as abscisic acid, gibberellic acid, cytokinin, auxin, or the like; or d) (1-aminocyclopropane-1-carboxylate deaminase).

Thus, in order to determine whether microorganisms settle in plant roots for microbial-plant interaction, root colonization of SB19 strain using rhizosphere soil and root dilution plating method was investigated .

1. Experimental Method

After 1 day of sowing, the suspension of SB19 strain was treated once with kale and red mustard, and after 5 weeks from the date of the strain treatment, the soil and roots of the red mustard root were collected. Sterilized water (total volume 10 ml) was added to 1 g of rhizosphere soil and allowed to stand at room temperature for 3 hours. The roots were surface-sterilized and then ground and cultured on TSA medium by grinding stepwise. Colony forming units were investigated per 1 g of rhizosphere soil and roots.

2. Experimental results

As a result of the soil- borne treatment of the strain Bacillus toyonensis SB19 of the present invention, As shown in Table 1, SB19 bacterial counts (7.92 × 10 ⁷ CFU / ㎖) and root endogenous bacterial counts (4.83 × 10 ⁶ CFU / ㎖) were untreated (rhizospheric soil; 1.68 × 10 ⁷ CFU / And 3.33 × 10 ⁴ CFU / ml, respectively.

In other words, SB19 strain was found to be persistent even after 5 weeks of treatment, effectively settling on enemy roots. Therefore, microbial - plant interaction by plant root establishment of SB19 strain is considered to help overcome drought stress by playing an important role in enhancing water availability of plants.

As described above, in the present invention, SB19 strain and plant interaction play an important role in promoting the water utilization rate of plants, and it is expected to improve the quality of cabbage such as kale and mustard in weak drought conditions, The SB19 strain is expected to be useful for production.

It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the invention as defined by the appended claims. It will be possible. The scope of the present invention is defined by the appended claims, and all differences within the scope of the claims are to be construed as being included in the present invention.

Depositor Name: National Institute of Agricultural Science

Accession number: KACC 81045BP

Funding date: 04/03/2013

<110> REPUBLIC OF KOREA (MANAGEMENT: RURAL DEVELOPMENT ADMINISTRATION) <120> New microorganism Bacillus toyonensis SB19 having growth          promoting of Leafy vegetables and high temperature tolorance and          drought resistance of leafy vegetables and microbial agent          containing the same and biofertilizer containing the same <130> p2017-0126 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 1480 <212> DNA <213> Bacillus toyonensis SB19 <400> 1 ttttgaagtt tggaattcgc tcaggatgaa cgctggcggc gtgcctaata catgcaagtc 60 gagcgaatgg attgagagct tgctctcaag aagttagcgg cggacgggtg agtaacacgt 120 gggtaacctg cccataagac tgggataact ccgggaaacc ggggctaata ccggataaca 180 ttttgaactg catggttcga aattgaaagg cggcttcggc tgtcacttat ggatggaccc 240 gcgtcgcatt agctagttgg tgaggtaacg gctcaccaag gcaacgatgc gtagccgacc 300 tgagagggtg atcggccaca ctgggactga gacacggccc agactcctac gggaggcagc 360 agtagggaat cttccgcaat ggacgaaagt ctgacggagc aacgccgcgt gagtgatgaa 420 ggctttcggg tcgtaaaact ctgttgttag ggaagaacaa gtgctagttg aataagctgg 480 caccttgacg gtacctaacc agaaagccac ggctaactac gtgccagcag ccgcggtaat 540 acgtaggtgg caagcgttat ccggaattat tgggcgtaaa gcgcgcgcag gtggtttctt 600 aagtctgatg tgaaagccca cggctcaacc gtggagggtc attggaaact gggagacttg 660 agtgcagaag aggaaagtgg aattccatgt gtagcggtga aatgcgtaga gatatggagg 720 aacaccagtg gcgaaggcga ctttctggtc tgtaactgac actgaggcgc gaaagcgtgg 780 ggagcaaaca ggattagata ccctggtagt ccacgccgta aacgatgagt gctaagtgtt 840 agagggtttc cgccctttag tgctgaagtt aacgcattaa gcactccgcc tggggagtac 900 ggccgcaagg ctgaaactca aaggaattga cgggggcccg cacaagcggt ggagcatgtg 960 gtttaattcg aagacacgcg aagaacctta ccaggtcttg acatcctctg aaaaccctag 1020 agatagggct tctccttcgg gagcagagtg acaggtggtg catggttgtc gtcagctcgt 1080 gtcgtgagat gttgggttaa gtcccgcaac gagcgcaacc cttgatctta gttgccatca 1140 ttaagttggg cactctaagg tgactgccgg tgacaaaccg gaggaaggtg gggatgacgt 1200 caaatcatca tgccccttat gacctgggct acacacgtgc tacaatggac ggtacaaaga 1260 gctgcaagac cgcgaggtgg agctaatctc ataaaaccgt tctcagttcg gattgtaggc 1320 tgcaactcgc ctacatgaag ctggaatcgc tagtaatcgc ggatcagcat gccgcggtga 1380 atacgttccc gggccttgta cacaccgccc gtcacaccac gagagtttgt aacacccgaa 1440 gtcggtgggg aacctttgga gccgccgcta agtgacatgt 1480

Claims (8)

1 &lt; / RTI &gt; comprising the nucleotide sequence represented by SEQ ID NO: 1,
Promoting the growth of wrapping vegetables, and imparting high temperature resistance or resistance to wrapping,
New microorganism Bacillus toyonensis strain SB19 (KACC 81045BP).
The method according to claim 1,
Wherein the strain has root-fixing ability of wrapping vegetables.
New microorganism Bacillus toyonensis strain SB19 (KACC 81045BP).
The method according to claim 1,
Wherein the safflower is one of kale and red mustard,
New microorganism Bacillus toyonensis strain SB19 (KACC 81045BP).
The method according to claim 1,
A microbial pesticide, a seed coating agent, a soil improving agent, a compostant homework agent, a foliar spraying agent, or a root spraying agent,
New microorganism Bacillus toyonensis strain SB19 (KACC 81045BP).
A novel microorganism Bacillus toyonensis strain SB19 (KACC 81045BP) according to claim 1 as an active ingredient.
Microbiological preparation for enhancing tolerance to hot or dry stress of wrapping vegetables.
Preparing a new microorganism Bacillus toyonensis strain SB19 (KACC 81045BP) according to any one of claims 1 to 4,
Culturing the strains in a TSB (Tryptic Soy Broth) medium at 20 to 35 DEG C and 150 to 200 rpm for 40 to 50 hours;
A method for producing a microorganism preparation for enhancing tolerance to hot or dry stress of wrapping vegetables.
A microorganism according to any one of claims 1 to 4, which contains as an active ingredient at least one of the microorganism Bacillus toyonensis SB19 strain (KACC 81045BP) or the microorganism preparation according to the above-mentioned 5,
Biological fertilizer for improving tolerance to high temperature or drying stress of wrapping vegetables.
A method for increasing the tolerance to hot or dry stress of wrapping vegetables by treating the biocidal fertilizer according to claim 7 with the soil.

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