KR101624628B1 - Novel bacillus vallismortis bs07m with promoting effect of plant growth and improving effect of cold-tolerance, and microbial agent containing the same - Google Patents

Abstract

The present invention relates to a novel strain of Bacillus balisumotis BS07M having the effect of promoting growth and cold tolerance of crops and a microorganism preparation containing the same.
Novel microorganism Bacillus of the present invention balise motiseu (Bacillus vallismortis) BS07M KCTC11991BP strain is represented by SEQ ID NO: 1, and is characterized with a cold resistance enhancing effect of promoting the growth of plants and crops, the microorganism preparation of the present invention of the novel microorganism Bacillus balise motiseu (Bacillus vallismortis) it is characterized by containing the bacterial cells or the culture solution of the strain BS07M KCTC11991BP.
According to the present invention, a new microorganism Bacillus balismotis BS07M, which is a novel microorganism that not only has a direct inhibitory effect on plant pathogens but also has resistance to plant disease and has enhanced immune function activity and promotes the growth of crops, And a microbial agent containing the same.

Description

TECHNICAL FIELD [0001] The present invention relates to a novel strain of Bacillus balisumotis BS07M having an effect of promoting growth and cold resistance of a crop, and a microorganism preparation containing the same. [0002] The present invention relates to a novel strain of Bacillus balisumotis BS07M,

The present invention relates to a novel strain of Bacillus balisumotis BS07M having the effect of promoting growth and cold resistance of crops and a microorganism preparation containing the same. The present invention relates to a novel strain of Bacillus balisumotis BS07M having an effect of suppressing the growth of a plant pathogenic bacteria, To promote the growth of crops and to increase the cold resistance of the crops, and a microbial preparation containing the same.

Recently, agricultural ecosystem pollution and safety due to the abuse of chemical pesticides have become more societal concern for agricultural products, and thus environmentally friendly biocontrol methods using microorganisms have been attempted for various diseases which are difficult to control with chemical agents.

In recent years, in order to induce disease resistance to crops, inoculation of microorganisms in advance in a crop has resulted in a resistance reaction induced in the crop body (Induced Systemic Resistance, "ISR").

Microorganism to induce resistance to such plant disease is one or more different types of microorganisms belonging to the bacteria involved, and that, among the Bacillus (Bacillus) in such a known strain which is put to practical use is to only a few, and the strains belonging to the same genus The immunological activity of the inducible resistance to the crops varies considerably depending on species, and selection of excellent strains is necessary.

For this purpose, it is required to select a strain having excellent function to exhibit immune function until the late stage of crop growth by effectively treating the plant disease stall while stably expressing the induction resistance to the plant disease.

It is an object of the present invention to provide a novel microorganism, Bacillus balisumotis BS07M, which not only has a direct inhibitory effect on plant pathogens but also induces resistance to plant diseases, enhances immune function activity and promotes the growth of crops, And to provide a microbial preparation containing the same.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are not intended to limit the invention to the particular embodiments that are described. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are not restrictive of the invention, There will be.

Novel microorganism Bacillus of the present invention balise motiseu (Bacillus vallismortis) BS07M strain KCTC11991BP is represented by SEQ ID NO: 1, and is characterized for promoting the growth of crops, especially characterized with the immune active performance, cold resistance increase performance of the crop for the antimicrobial active performance, plant diseases of the plant pathogen .

The crop is characterized in that it is any one selected from pepper, cucumber and tobacco.

It said plant pathogen is a collation Sat tree Colchicum Aku tatum (Colletotrichum acutatum), and Leah Alter Alter appear (Alternaria alternata , Botrytis Cinerea , Penicillium < RTI ID = 0.0 > italicum ), Penicillium expansum), Fusarium oxy Spokane Room (Fusarium oxysporum , Rhizoctonia solani , Sclerotinia sclerotiorum , and the like.

The plant disease is caused by Phytophthora capsici , Pectobacterium carotovora , red pepper anthracis ( Colletotrichum acutatum ), cucumber mosaic virus ( cucumber mosaic virus) .

In addition, the novel microorganism Bacillus vallismortis BS07M strain KCTC11991BP of the present invention is represented by SEQ ID NO: 1 and is characterized in that it has a cold tolerance enhancing ability of a crop.

The new microorganism is characterized by being used as an effective ingredient of any one of microbial pesticide, microbial fertilizer, seed coating agent, soil improving agent, composting homework, foliar spraying agent, or traditional spraying agent.

Microorganism preparation of the present invention is characterized by comprising the novel microorganism Bacillus balise motiseu (Bacillus vallismortis) cells or a culture broth of strains BS07M of KCTC11991BP.

The method for promoting crop growth, method for controlling plant pathogenic bacteria, and method for enhancing cold tolerance of a crop according to the present invention is characterized in that the microorganism preparation containing the above-mentioned new microorganism is sprayed or treated with soil.

According to the present invention, a new microorganism Bacillus balisumotis BS07M is provided, which has a feature that the cold resistance of a crop is enhanced to withstand cold weather.

The new microorganism Bacillus balisumotis BS07M is also significantly superior to the conventional effect of suppressing plant disease and promoting plant growth. Thus, the novel microorganisms and microorganism preparations containing the microorganisms of the present invention not only effectively inhibit plant pathogens but also significantly reduce the disease caused by an increase in the activity of the plant immunity function, .

Figure 1 is a novel Bacillus balise motiseu BS07M (Bacillus vallismortis BS07M) strains using the 16S rRNA gene sequence.
Fig. 2 is a diagram showing the growth promotion effect of the red pepper seedlings obtained by the soil cultivation treatment of the novel strain of Bacillus balismotis BS07M.
Fig. 3 is a diagram showing the effect of the growth of the pepper fruit by the new strain of Bacillus balismotis BS07M
4 is a view showing the antimicrobial activity of a novel strain of Bacillus balismotis BS07M against a plant pathogenic bacterium.
FIG. 5 is a diagram showing disease resistance gene expression by a novel strain of Bacillus subtilis BS07M.
1. Col O: Arabidopsis wild type
2. etr3-5: Arabidopsis transformed with the gene
3. nah G: Transgenic Arabidopsis unable to express PR gene
4. npr1: Arabidopsis transformed with the gene
Fig. 6 is a diagram showing the lignin accumulation of cucumber seedlings root by treatment with a novel strain of Bacillus balismotis BS07M.
FIG. 7 is a graph showing the effect of the Pectobacterium ( Bacillus thuringiensis) carotovora ) SCC1. < / RTI >
Fig. 8 is a graph showing an effect of suppressing anthrax anthracis in pepper plants treated with a new strain of Bacillus balismotis strain BS07M under the condition of cultivation in a non-growing condition.
Fig. 9 is a graph showing the inhibitory effect of the cucumber mosaic virus in the treatment of the novel strain of Bacillus balismotis BS07M.
10 is a view showing the effect of increasing the cold tolerance of cucumber by treatment with a new strain of Bacillus valis smothus BS07M.
Fig. 11 is a graph showing the effect of increasing the cold tolerance of tobacco seedlings by the novel strain of Bacillus balismotis BS07M.

The terms, techniques, and the like described in this specification are used in the meaning commonly used in the technical field to which the present invention belongs, unless otherwise specified.

Hereinafter, the present invention will be described in detail.

In the present invention, a novel Bacillus Vermis BS07M ( Bacillus sp. Bacillus sp. vallismortis BS07M) were selected.

The obtained from the starting strain comparing the homology of the 16S rRNA analysis bar, been identified as a novel strain of Bacillus Barris.And motiseu (Bacillus vallismortis), this Bacillus Barris.And motiseu BS07M (Bacillus vallismortis BS07M), and deposited this strain on the Korea Research Institute of Bioscience and Biotechnology on July 28, 2011 under the accession number KCTC11991BP.

Novel microorganism of the present invention, the Bacillus Barris.And motiseu BS07M (Bacillus vallismortis BS07M) consists of the entire nucleotide sequence shown in SEQ ID NO: 1 or a part thereof and is composed of an open reading frame (ORF) of 1478 bp.

As a result of confirming the growth of the crop and the enhancement of the immune function of the microorganism preparation containing the microorganism of the new strain or the microorganism culture of the microorganism as described above, it was confirmed that all of the crops, regardless of the kind of the crop, Immunity enhancement effect and cold tolerance enhancing effect can be obtained. Especially, pepper, cucumber and tobacco showed the best growth promotion, immune function enhancement and cold tolerance enhancement effects. That is, when the new strain is treated with pepper, the yield of pepper is increased by 30% or more.

It also produces powerful antimicrobials that inhibit major plant pathogens such as anthrax, and is also known as Phytophthora capsici , Pectobacterium carotovora , red pepper anthracis ( Colletotrichum acutatum ), cucumber mosaic virus ( cucumber mosaic virus , and the like.

In addition, the disease resistance gene is expressed and the immune function through the salicylic acid pathway is enhanced.

In addition, since the new strain is treated with a crop, the survival rate is 100% even when it is exposed at a low temperature of 4 캜 or lower for 24 hours or more, thereby increasing the cold tolerance of the crop.

Hereinafter, the content of the present invention will be described in more detail with reference to Examples and Experimental Examples.

< Example  1> Isolation and selection of strains

In order to isolate the excellent bacteria that activate the immune function against the plant disease, the plant rhizosphere soil was collected from farmland of all over the country.

The collected samples were placed in sterilized water, shaken, and diluted by a dilution plate method to prepare a stepwise diluted solution, which was spread on a solid medium TSA medium.

After incubation at 30 ° C for 2 days, the bacteria Bacillus valismortis strain BS07M was isolated pure.

The pure isolates were tested for inducible resistance to Colletotrichum orbiculare .

The seeds of the varieties of Oryza sativa L. cucumber were surface sterilized with 1% sodium chloride, and then seeded on plastic pots. After 2 weeks, the suspension of the isolates was subjected to a cross treatment.

After 1 week, the spores suspension of cucumber anthracnose was sprayed on the treated leaves to investigate the lesion formed, and the most excellent strain resistant to plant disease, Bacillus valismortis ) BS07M were selected.

< Example  2> Identification of the novel strain of the present invention

The identification of the strain selected in Example 1 was carried out by 16S rRNA gene sequencing.

(1) 16S rRNA sequencing

For the identification of bacteria, 16S rRNA gene sequencing was used.

The 16S rDNA gene was amplified by PCR using the universal primers fD1 (5'-AGAGTTTGATCCTGGCTCAG-3 ') and rP2 (5'-ACGGCTACCTTGTTACGACTT-3') after DNA extraction with a DNA extraction kit Lt; / RTI &gt;

The resulting PCR product was reacted using a DNA sequencing kit and sequenced with a genetic analyzer (3100 Genetic Analyzer). The nucleotide sequence was identified to the genus through the BLAST program of the NCBI server.

The 16S rDNA sequence was aligned with the nucleotide sequence of the standard strain using the CLUSTAL W program. The evolution of the data set was made using the MEGA version program. The bootstrap value of the branch was researched through 1,000 resampling.

Molecular phylogenetic analysis As shown in Fig. 1, the Bacillus balismotis strain BS07M was identified as Bacillus subtilis subtilis , Bacillus mojavensis , Bacillus vallismortis ) , And Bacillus It was identified as Bacillus vallismortis because it has the highest affinity with vallismortis . On July 28, 2011, KCTC11991BP was deposited with the Korea Research Institute of Bioscience and the new microorganism was deposited with Bacillus vallismortis ) BS07M.

< Example  3> New Bacillus Balis Motis BS07M  Culture of strain

The above-named Bacillus balis malthus BS07M is used, which is obtained by spreading Bacillus balis malthus BS07M stored in a cryogenic tank (-80 ° C) on sterilized TSA (Tryptic soy agar) medium and incubating at 30 ° C for 48 hours Cultured and reactivated. (At this time, the TSB liquid medium is inoculated with Bacillus virulis mothis BS07M and then reactivated by culturing at 30 ° C for 48 hours.)

The cultured cells were collected and put into 1 L of sterilized water or 0.2 M phosphate buffer to prepare 10 ^{9-10 10} cfu / ml cell suspension.

The cell suspension was diluted to a concentration of 10 ⁶ to 10 ⁷ cfu / and used in the following soil treatment and plant treatment experiments.

< Experimental Example  1> New Bacillus Balis Motis BS07M  Strain Promotion ability of pepper growth  Confirm

1) vinyl house condition

A cell suspension of the novel strain of Bacillus balismotis BS07M of Example 3 was prepared.

In the vinyl house condition, the above cell suspension was treated in the rhizosphere soil during the raising period.

Specifically, first, the suspension of the Bacillus valis smothis BS07M cells (10 ⁹ cfu / ml) was diluted 100 times with the second to third leaf spot of the main leaf. Then, it ran to the rhyolite soil about twice every 10 days.

The results are shown in Fig. 2 and Tables 1 and 2 below.

That is, as shown in FIG. 2, it was confirmed that the growth promoting effect of the new Bacillus ballis motif BS07M strain was remarkably improved in the seedling growth promoting effect, and the growth rate thereof was also increased in the subsequent growth.

process Length (cm) Stem diameter
(mm) fruit tree
(recast) 5.28 6.18 6.28 7.19 7.19 7.19 Control 23.70 50.50 70.83 101.90 11.9 41.4 Disease resistance inducer
(0.1 mM-BTH) 20.90 58.08 76.17 116.20 11.0 33.8 Invention
(BS07M) 27.10 73.50 102.50 132.80 13.9 53.0 LSD ( p = 0.05) 1.20 5.71 5.71 8.50 1.7 14.8

* Least significant difference (LSD): the least significant difference

In addition, as shown in Table 1, the growth rate of the plant length and stem diameter was remarkably superior to that of the control and the 0.1 mM BTH treatment as the disease resistance inducer, as well as the novel strain of Bacillus balismotis BS07M of the present invention, It was confirmed that the new strain of Bacillus balismotis BS07M strain of the present invention was significantly increased in the fruit juice per serving.

process
Quantity (kg / plot) With guns
(kg) 5 days 17th 30 days contrast 16.0 35.8 42.0 93.8 Bacillus subtillis S08 22.0 40.4 34.0 96.4 Bacillus vallismortis BS07M (invention) 20.0 45.4 37.0 102.4 Bacillus subtillis BS5b 20.0 38.6 40.0 98.4

Also, as shown in Table 2, it was confirmed that Bacillus strains were also significantly different. That is, in the total fruit weight of pepper are different Bacillus (Bacillus) the inventors novel Bacillus strain balise motiseu BS07M treatment than the strain appears the highest was found to indicate the excellent effect of flooding.

2) Nogi condition

During the incubation period, the cell suspension was treated in the same manner as in the above-mentioned vinyl house condition to the rhizosphere soil in the vinyl house. Thereafter, the plant was planted at the end of the growing season and compared with the control.

The results are shown in Table 3 below.

process Length (cm) Quantity (g) / week Number of fruits Fruit inside diameter (mm) Percussion thickness (mm) Conventional (control) 127.1 795.0 31.1 21.58 2.34 BS07M (invention) 131.5 929.0 40.1 26.08 3.3 LSD ( p = 0.05) 16.0 29.0 10.8 2.1 0.5

* Least significant difference (LSD): the least significant difference

As shown in Table 3, in the new strain of Bacillus balismotis strain BS07M, the growth rate of the pepper seedlings such as the length of the fruit, the borehole diameter and the thickness of the pericarp were significantly superior to those of the control (control), and the fruit number and yield were increased.

In addition, as shown in FIG. 3, the size of the fruit of the pepper fruits was larger and longer than that of the control (control), and the growth of Bacillus vulnificus BS07M strain was increased.

< Experimental Example  2> New Bacillus Balis Motis BS07M  Strain On plant pathogens  About Identification of antimicrobial activity

The antimicrobial activity effect of the novel strain of Bacillus balismotis BS07M on major plant pathogens is shown in Fig.

That is, the novel strain of Bacillus balisumotis BS07M of the present invention is a plant pathogenic fungus, Colletotrichum &lt; RTI ID = 0.0 &gt; acutatum , Alternaria alternata), Botrytis cine Liao (Botrytis cinerea), Penny room Solarium itaconic rikum (Penicillium italicum ), Penicillium expansum KC08001), Penicillium expansum JVV81), Fusarium oxy Spokane Room (Fusarium oxysporum , Rhizoctonia &lt; RTI ID = 0.0 &gt; solani , and Sclerotinia sclerotirum , respectively. The results are shown in FIG.

Among them, Colletotrichum acutatum , Alternaria alternata , Botrytis cinerea , and Sclerotinia sclerotirum . The antimicrobial activity of these plant pathogens was shown to be high.

< Experimental Example  3> New Bacillus Balis Malthus BS07M Identification of the induction resistance promoting activity by the strain treatment

1) Experimental method

(1) Cultivation of Arabidopsis Plants

In order to confirm the expression of the antifungal gene by the induction resistance promoting activity in the treatment of the new strain Bacillus balismotis BS07M, expression of PR gene derived from Arabidopsis wild type (Col 0) and transgenic strain (parent Arabidopsis ecotype) Transgenic Arabidopsis thaliana (nah G: SA), transgenic Arabidopsis thaliana (etr3-5, npr1) were used.

At this time, Arabidopsis wild type (Col 0) was obtained from the center, and the nah G transgenic plant (Arabidopsis thaliana) is known to encode a salicylate dehydrogenase and degrade SA.

Arabidopsis seeds were surface-sterilized (dipped in 70% ethanol for 2 minutes, immersed in 1% sodium hypochlorite for 20 minutes), washed three times in sterilized distilled water, then 0.8% agar and 1.5% sucrose, pH 5.7 After germination of the seeds was promoted for 2 days at 4 ° C in which the light was blocked, the seedlings were allowed to stand for 12 hours - light / 12 hours - placed on a plant growth plant set to a dark cycle and left under a 40W fluorescent lamp.

At a relative humidity of 50 to 60%, the temperature was maintained at 22 +/- 1 deg. Two weeks later, the seedlings were transplanted into 60 ml pots containing a potting soil mixture for 2 hours, autoclaved sterilized pollen for 1 hour at intervals of 24 hours.

Prior to transferring the seedlings to the pots, the potting soil was treated with either the fresh Bacillus balisumotis strain BS07M suspension or the same volume of sterile distilled water. The grown plants were grown under plant growth conditions at a relative humidity of 70% for 9 hours (200 μE / m ² s, 24 ° C) and 15 hours at night (20 ° C).

The plants were irrigated every other day and treated with Hoagland nutrient solution at half concentration once a week. Two weeks later, the bacterial suspension of the novel strain of Bacillus balismotis BS07M of Example 3 was transplanted to the Arabidopsis thaliana plants.

(2) Confirmation of the defense gene expression by the treatment of the novel Bacillus balisumothis BS07M strain

To the thus prepared Arabidopsis thaliana plants, a bacterial suspension of the novel Bacillus balisumotis BS07M strain of Example 3 was inoculated in a soil at a concentration of 10 ⁸ cfu / ml, and after one week, leaf tissue was obtained from the plant for RNA analysis Respectively.

Arabidopsis total RNA was extracted by homogenizing at least 2 g of frozen tissue in equal volume of extraction buffer (0.35 M Glycine, 0.048 M NaOH, 0.34 M NaCl, 0.04 M EDTA, 4% (w / v) SDS) . The homogeneous suspension was then extracted with phenol and chloroform, and the RNA was precipitated with LiCl.

RT-PCR was performed using Ex Taq polymerase.

The reaction mixture was prepared to contain 0.1 μg of cDNA, 10 pMol of forward and reverse primers, 250 nM of dNTP, and 0.5 U of Ex Taq polymerase in 20 μl of the buffer solution.

The PCR was carried out at 94 ° C. for 5 minutes, followed by treatment at 94 ° C. for 1 minute and 57 ° C. for 1 minute under 25 cycles, followed by extension at 72 ° C. for 10 minutes.

The primer for the defective gene was a forward primer 5'-TGCGGTAACACCGAACCATAC-3 '(SEQ ID NO: 1) and a reverse primer 5'-CGACAGTTGCATTGGTCCTCT-3' (SEQ ID NO: 2) (SEQ ID NO: 3) and the reverse primer 5'-TCACGGAGGCACAACCAAGTC-3 '(SEQ ID NO: 4) were used as a primer for the Gene-1 gene.

The amplified PCR product was analyzed using 1.2% agarose gel to record the gel.

When the induction resistance of a plant by infection with a microorganism or an inducible resistance is expressed, an hypersensitive reaction occurs, followed by pathogenesis related protein (PR) proteins such as glucanase, chitinase, , Lignification of plant cells is induced, and resistant substances such as phytoalexin and phenol compounds accumulate and a resistive reaction appears.

Similarly, the rhizosphere treatment of a new non-pathogenic strain of Bacillus balis malthus BS07M induces the immune function of the plant.

2. Experimental results

As shown in FIG. 5, the expression of the PR1 gene, which is a disease-resistant gene in the wild-type Arabidopsis thaliana prepared as described above, was confirmed by the novel Bacillus valisymotis BS07M strain-treated plant, and it was not expressed in the NahG mutant The disease resistance pathway by Boa SA is thought to increase the plant immunity activity.

In order to prove the above experiment, a new strain of Bacillus balisumotis BS07M was treated to examine the reaction of roots. As a result, the induction of ligninization was confirmed in the plant root tissue as shown in Fig. In other words, the red lignin formation represents one of the disease-resistant reactions, which means that the plant cell wall thickens. Therefore, it has been found that the cell wall of the cucumber root is thickened by the treatment of the novel strain of Bacillus balisumotis BS07M of the present invention, and plant pathogens can not enter.

< Experimental Example  4> New Bacillus Balis Motis BS07M By induction resistance of the strain Plant disease  Inhibitory effect

In order to examine the inhibitory effect of the novel Bacillus valis smothis strain BS07M on the induction resistance,

The expression of the disease resistance of the red pepper by the treatment of the new Bacillus varios motif BS07M strain was tested by inoculating pathogenic bacteria into the red pepper leaves and fruits collected from the stools and packaging.

In case of rotting, in the indoor test, the leaves of fresh mushroom and the leaves of fresh mushroom were collected by separating the treated and untreated rice cultivars treated with the new strain of Bacillus balismotis BS07M.

And Pectobacterium induced resistance resistance to carotovora SCC1 was tested.

As a result, the leaves of the pepper seedlings appeared as shown in Table 4 and FIG.

process Lesion area ratio (%) leaf BS07M
(Invention) 0 Control (control) 42 No treatment (superior above ground) 32 LSD ( p = 0.05) 37.23

* Least significant difference (LSD): the least significant difference

As shown in Table 4 and FIG. 7, the control area showed a 42% lesion area ratio, whereas the new Bacillus valis mottus BS07M strain treatment effectively inhibited the lesion from occurring.

Table 5 shows the leaves of the leaves in an open state.

process Inflammation rate (%) Anthrax infection rate (%) Primary Secondary Conventional (control) 71.7 53.7 15 0.1 mM-BTH 16.7 75.7 8 BS07M (invention) 20.0 0.0 One LSD ( p = 0.05) 20.62 23.5 19.1

* Least significant difference (LSD): the least significant difference

As shown in Table 5, the infection rate was 71.7% and 53.7% in the control, respectively, but the treatment was significantly suppressed to 20.0% and 0.0% in the control and the control, respectively.

In the case of red pepper anthracnose, fruit of red pepper cultivated in greenhouse was collected and tested for inhibition of red pepper anthracnose.

And suspended in sterilized water to obtain a pepper anthracnose ( Colletotrichum acutatum ) spores were adjusted to a concentration of 10 ⁵ spores / and a sufficient amount of an electrodepositive agent was added thereto.

Then, the sample was placed in a greenhouse at a relative humidity of 100% for 24 hours, removed from the greenhouse, and examined for disease occurrence on the seventh day. The fresh fruit of the Bacillus balisumotis BS07M strain treated with the red pepper cultivar and the untreated cultivar were examined separately.

As a result, as shown in Table 5, the infection rate was 15% in the conventional strain, but the lowest infection rate was 1% in the Bacillus balismotis BS07M treatment.

Also, in the case of pepper anthracnose, it was carried out under the condition of cultivation in the field of packaging.

Colletotrichum acutatum spores were suspended in sterilized water, adjusted to a concentration of 10 ⁵ spores / and added to the leaves and fruits of the pepper. Then, the occurrence of anthrax was examined four times at intervals of about 15 days.

As shown in Fig. 8, the result showed that the inhibition effect of anthracnose was greatest by the treatment with the new strain Bacillus balismotis BS07M even under the packaging conditions.

In addition, cucumber mosaic virus ( cucumber mosaic virus , and CMV).

Cucumber mushroom virus (CMV) cucumber leaf was crushed and rubbed together with carborandum powder in the main leaf 1 cucumber (Cryptomeria japonica) seedlings after 2 weeks of sowing treated with spore suspension of the strain BS07M The mosaic formed in the greenhouse at room temperature for one week after the incubation was examined for virus infection. Untreated was treated with sterilized distilled water and CMV virus was inoculated by the same method.

As a result, as shown in FIG. 9, the cucumber seedlings treated with BS07M showed no symptoms of infection due to CMV virus, which proved that the virus control effect was excellent

<Experiment 5> The novel Bacillus Balis Motis Expression of water cold resistance increasing effect by the treatment of plant strains BS07M

The cold tolerance of cucumber and tobacco seedlings was examined after treatment with Bacillus balismotis BS07M.

After 24 hours treatment at 4 ℃, the leaves were allowed to stand for 6 hours in the greenhouse.

As shown in FIGS. 10 and 11, the chlorophyll was destroyed by cold treatment due to the low temperature treatment, and the surface of the leaf was shrunken and discolored to an off-white color. On the other hand, treatment with the new strain Bacillus valis smothis BS07M And the cold resistance was increased without cold weather.

Therefore, the novel microorganism of the present invention has an excellent characteristic that the cold resistance is enhanced by the treatment of crops and endures cold weather even at a low temperature condition of 4 캜. Using this, the plant disease prevention and plant growth promoting effect is remarkably superior to the conventional effect, Can be used.

In addition, it not only effectively inhibits plant pathogens but also greatly reduces disease caused by increased activity of plant immunity, and is also effective for promoting crop growth, thus being useful for environmentally friendly agriculture.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments. You can implement the examples. 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.

Korea Biotechnology Research Institute KCTC11991BP 20110728

<110> REPUBLIC OF KOREA (MANAGEMENT: RURAL DEVELOPMENT ADMINISTRATION) <120> NOVEL BACILLUS VALLISMORTIS BS07M WITH PROMOTING PLANT GROWTH AND          IMPROVED COLD-TOLERANCE, AND MICROBIAL AGENT CONTAINING THE SAME <130> P2011-2074 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 1478 <212> DNA <213> Bacillus vallismortis <400> 1 cactcagacg aacgctggcg gcgtgcctaa tacatgcaag tcgagcggac agatgggagc 60 ttgctccctg atgttagcgg cggacgggtg agtaacacgt gggtaacctg cctgtaagac 120 tgggataact ccgggaaacc ggggctaata ccggatggtt gtctgaaccg catggttcag 180 acataaaagg tggcttcggc taccacttac agatggaccc gcggcgcatt agctagttgg 240 tgaggtaacg gctcaccaag gcgacgatgc gtagccgacc tgagagggtg atcggccaca 300 ctgggactga gacacggccc agactcctac gggaggcagc agtagggaat cttccgcaat 360 ggacgaaagt ctgacggagc aacgccgcgt gagtgatgaa ggttttcgga tcgtaaagct 420 ctgttgttag ggaagaacaa gtgccgttca aatagggcgg caccttgacg gtacctaacc 480 agaaagccac ggctaactac gtgccagcag ccgcggtaat acgtaggtgg caagcgttgt 540 ccggaattat tgggcgtaaa gggctcgcag gcggtttctt aagtctgatg tgaaagcccc 600 cggctcaacc ggggagggtc attggaaact ggggaacttg agtgcagaag aggagagtgg 660 aattccacgt gtagcggtga aatgcgtaga gatgtggagg aacaccagtg gcgaaggcga 720 ctctctggtc tgtaactgac gctgaggagc gaaagcgtgg ggagcgaaca ggattagata 780 ccctggtagt ccacgccgta aacgatgagt gctaagtgtt agggggtttc cgccccttag 840 tgctgcagct aacgcattaa gcactccgcc tggggagtac ggtcgcaaga ctgaaactca 900 aaggaattga cgggggcccg cacaagcggt ggagcatgtg gtttaattcg aagcaacgcg 960 aagaacctta ccaggtcttg acatcctctg acaatcctag agataggacg tccccttcgg 1020 gggcagagtg acaggtggtg catggttgtc gtcagctcgt gtcgtgagat gttgggttaa 1080 gtcccgcaac gagcgcaacc cttgatctta gttgccagca ttcagttggg cactctaagg 1140 tgactgccgg tgacaaaccg gaggaaggtg gggatgacgt caaatcatca tgccccttat 1200 gacctgggct acacacgtgc tacaatggac agaacaaagg gcagcgaaac cgcgaggtta 1260 agccaatccc acaaatctgt tctcagttcg gatcgcagtc tgcaactcga ctgcgtgaag 1320 ctggaatcgc tagtaatcgc ggatcagcat gccgcggtga atacgttccc gggccttgta 1380 cacaccgccc gtcacaccac gagagtttgt aacacccgaa gtcggtgagg taacctttta 1440 ggagccagcc gccgaagtgg gacagatgat tgggtgan 1478

Claims (12)

delete 1 < / RTI > of SEQ ID NO: &lt; RTI ID = 0.0 &gt; 1,
( Pectobacterium carotovora ) caused by the red pepper anthracnose fungus caused by Colletotrichum acutatum Malarial, caused by Cucumber mosaic virus Cucumber mosaic virus. The present invention also provides a method for inhibiting the growth of a plant,
The new microorganism Bacillus vallismortis BS07M strain KCTC11991BP.
&Lt; SEQ ID NO: 1:
cactcagacg aacgctggcg gcgtgcctaa tacatgcaag tcgagcggac agatgggagc 60
ttgctccctg atgttagcgg cggacgggtg agtaacacgt gggtaacctg cctgtaagac 120
tgggataact ccgggaaacc ggggctaata ccggatggtt gtctgaaccg catggttcag 180
acataaaagg tggcttcggc taccacttac agatggaccc gcggcgcatt agctagttgg 240
tgaggtaacg gctcaccaag gcgacgatgc gtagccgacc tgagagggtg atcggccaca 300
ctgggactga gacacggccc agactcctac gggaggcagc agtagggaat cttccgcaat 360
ggacgaaagt ctgacggagc aacgccgcgt gagtgatgaa ggttttcgga tcgtaaagct 420
ctgttgttag ggaagaacaa gtgccgttca aatagggcgg caccttgacg gtacctaacc 480
agaaagccac ggctaactac gtgccagcag ccgcggtaat acgtaggtgg caagcgttgt 540
ccggaattat tgggcgtaaa gggctcgcag gcggtttctt aagtctgatg tgaaagcccc 600
cggctcaacc ggggagggtc attggaaact ggggaacttg agtgcagaag aggagagtgg 660
aattccacgt gtagcggtga aatgcgtaga gatgtggagg aacaccagtg gcgaaggcga 720
ctctctggtc tgtaactgac gctgaggagc gaaagcgtgg ggagcgaaca ggattagata 780
ccctggtagt ccacgccgta aacgatgagt gctaagtgtt agggggtttc cgccccttag 840
tgctgcagct aacgcattaa gcactccgcc tggggagtac ggtcgcaaga ctgaaactca 900
aaggaattga cgggggcccg cacaagcggt ggagcatgtg gtttaattcg aagcaacgcg 960
aagaacctta ccaggtcttg acatcctctg acaatcctag agataggacg tccccttcgg 1020
gggcagagtg acaggtggtg catggttgtc gtcagctcgt gtcgtgagat gttgggttaa 1080
gtcccgcaac gagcgcaacc cttgatctta gttgccagca ttcagttggg cactctaagg 1140
tgactgccgg tgacaaaccg gaggaaggtg gggatgacgt caaatcatca tgccccttat 1200
gacctgggct acacacgtgc tacaatggac agaacaaagg gcagcgaaac cgcgaggtta 1260
agccaatccc acaaatctgt tctcagttcg gatcgcagtc tgcaactcga ctgcgtgaag 1320
ctggaatcgc tagtaatcgc ggatcagcat gccgcggtga atacgttccc gggccttgta 1380
cacaccgccc gtcacaccac gagagtttgt aacacccgaa gtcggtgagg taacctttta 1440
ggagccagcc gccgaagtgg gacagatgat tgggtgan 1478
delete delete delete delete delete 3. The method of claim 2,
Wherein the strain is used as an active ingredient of any one of a microbial pesticide, a microbial fertilizer, a seed coating agent, a soil improving agent, a compostant homework agent, a foliar spraying agent,
The new microorganism Bacillus vallismortis BS07M strain KCTC11991BP.
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