KR101922410B1 - Novel compounds produced by Bacillus oryzicola YC7011 with activities of induced resistance against plant pathogens and insect and plant growth promotion - Google Patents

Abstract

Plants endogenous bacteria Bacillus originality Cola (Bacillus oryzicola ) The YC7011 culture medium and the substances isolated therefrom are treated to the rhizosphere and seeds of the plant to induce the resistance of the plant to control the plant diseases and pests, and to promote the growth of plants. It also has a growth inhibitory effect on plant and human pathogenic fungi. Therefore, the present invention relates to six resistance-inducing novel cyclic lipopeptides and three basic substances (Bacillopeptin A, B, C, and B), which are metabolites of Bacillus oryzae Cola YC7011 and have plant pest control, ). Bacillus oryzae cola YC7011 is cultured in large quantities to separate and purify the culture medium to provide environmentally friendly multifunctional natural plant protection agents as well as new materials that can be used in various industrial fields.

Description

{Novel compounds produced by Bacillus oryzicola YC7011 with activity against induced plant pathogens and insect and plant growth promoting effect of plant pest control,

The present invention is Bacillus originality Cola (Bacillus oryzicola ) YC7011, and to a host resistance-inducing novel substance having an antibacterial and plant growth promoting effect.

The present invention is derived from the research carried out as part of the "Industrial Convergence Source Technology Development Project" (Project No. 10044909, Title: Development of a Prolonged Effective Probiotic Crop Protection System) supported by the Ministry of Industry and Trade in 2014.

As the problems of environmental pollution, destruction of ecosystem, toxicity of pesticides, resistance to pests and weeds caused by misuse of chemical pesticides all over the world have arisen, many efforts have been made to provide environmentally friendly and sustainable agricultural production system that minimizes safe food production and environmental pollution . Since 2001, Korea has established a 5-year plan to cultivate eco-friendly agriculture in order to reduce the use of conventional chemical pesticides and chemical fertilizers to less than half, increase the number of eco-certified farmers, and increase the size of eco- . As the size of environmentally friendly agriculture gradually increases and consumer demand for agricultural products that do not use chemical pesticides or synthetic fertilizers increases, the environmentally friendly agricultural products market is steadily increasing. As a result, the use of organic synthetic pesticides and chemical fertilizers has been decreasing, and the use of natural plant protection agents for pest control has been increasing.

Natural plant protection products are largely divided into natural product extracts and agro-microbial products. In Korea, the former is mainly used as an insecticide and the latter as a fungicide. Microorganisms useful for agriculture are widely distributed in the natural environment such as symbiosis with soil, plant rhizosphere and plant tissue. Because they are various and secrete various metabolites, they are developed as natural plant protection agents and are effectively used in environmentally friendly agriculture have. Natural plant protection agents using microorganisms can be largely used in three ways, depending on the main constituents used: a method using microorganisms themselves, a method using metabolic antibiotics produced by fermentation of microorganisms, a method of using microbial metabolism And methods of using materials. Among them, methods using microorganisms themselves are widely used. Antagonistic microorganisms capable of directly inhibiting plant pathogenic bacteria and plant growth promoting rhizobacteria (PGPR) promoting plant growth are developed and used as products most domestically and internationally have. The microorganisms used are the fungi Trichoderma harzianum , the bacterium Bacillus subtilis , Bacillus amyloliquefaciens , Bacillus cereus , Bacillus pasteurii , Bacillus methylotrophicus , Pseudomonas fluorescens , Paenibacillus spp., Streptomyces spp. Lysobacter spp. Are well known as the main component. These microorganisms can directly or indirectly supply nutrients to the plants, stimulate the growth of plants through various antagonistic actions such as secretion of various hormones and antimicrobial substances that promote root growth, or induce the defense mechanism of plants before they become infected with pathogens, (Non-patent papers 1-4)

Various useful microorganisms have been used as a natural plant protection agent for controlling pests that cause problems in many crops, but the results of studies on controlling pests of rice have been rare (Non-Patent Document 2). Several studies on rice disease have shown that antagonism of Streptomyces sp. And Bacillus sp. May be used for control of rice sheath blight, or Bacillus vallismortis EXTN-1 and Pseudomonas spp. There is a report on the effective control of rice blast and rice sheath blight by using fluorescens mc75 and pc78. Fusarium causing rice blight and rice grain blight To inhibit moniliforme and Fusarium fujikuroi , P. fluorescens and Bacillus cereus (Non-Patent Documents 4-8).

Recently, it has been reported that treatment of Bacillus oryzicola YC7007, a new bacterial strain isolated from rice seedlings, on rice seeds or roots induces host resistance against bacterial egg wilt , blight of white blotch and Kidari disease in rice, (Korean Patent Publication No. 9-12).

To date, the development or control of natural plant protection products using microorganisms has been based almost exclusively on the direct inhibition of plant pathogens or the promotion of growth. However, when these microorganisms are treated in seeds or rhizosphere, there is little worldwide research on the effect of controlling insect pests or the mechanism of inhibition (Non-Patent Documents 11-12).

The Bacillus sp. Strain, which is widely used in natural plant protection agents, is most characteristic of promoting growth and resistance of antifungal, antibacterial and host plants and forming endospore, which is stable to heat and survive for a long time in a poor environment And is easy to formulate and is the most commercially used microorganism. Some strains are known to directly or indirectly supply nutrients to plants, promote hormonal secretion and various antagonistic actions that promote root growth, promote plant growth and induce defense mechanisms of plants before they become infected with pathogens (Non-Patent Document 2 -3, 12). Bacillus thuringiensis among many Bacillus has been known for a long time and has already been developed as a product since it has shown efficacy of producing toxins and killing pests directly. In contrast, several species of Bacillus strains exhibiting plant disease control and promoting plant growth have been reported to produce a variety of metabolites.

The antimicrobial substances isolated from various Bacillus strains are most well known as cyclic lipopeptides such as Iturins, Surfactins and Fengycins, And lipoic acid sequences which are linked to the ring-form amino acids and the length and characteristics of the lipopeptides have been disclosed (Non-Patent Document 14-17). These anticancer substances of the cyclic lipopeptide series are produced in B. subtilis strains. Iturins AE, Bacillomycines D, F, L, Lc, Mycosubtilin, Bacillopeptins A, B and C. The first three amino acids ( _L- Asx (x = p, n), _D- Tyr and _D- Asn) of this turin family are well conserved and are known to play an important role in antimicrobial activity. Iturin C is the cost of the _L -Asn-1 Iturin A _L substituted with -Asp-1 structure was on antibiotic Iturin A is not on the Iturin C. In addition, methylation of the phenyl group of _D- Tyr-2 of Iturin A resulted in decreased activity. Bacillopeptins are structurally similar to Bacillomycin L, but are linked by _L- Asn and _L- Glu instead of _L- Asp and _L- Gln of Bacillomycin L. Bacillopeptin C has only a slight structural difference from Bacillopeptin A or B, but the inhibitory effect against mold and yeast can be seen only in Bacillopeptin C. Mycosubtilin has a structure in which iturin A and two amino acids ( _D- Ser 6. _L- Asn 7) are changed, but the antimicrobial activity is more potent than Iturin A (Non-Patent Documents 14, 17 and 19). In recent years, studies have been conducted to identify lipoproteins from microbial strains that exhibit antibacterial activity against plant diseases, identify structures, identify structural genes, and identify the actions of these substances in plants through gene manipulation Non-Patent Document 15, 18-22).

Thus, various lipopeptides produced by Bacillus are known to be environmentally friendly and harmless to humans as well as biosurfactant activity that helps antagonism, antibacterial and antifungal action, and establishment of resistance to microorganisms. have. Particularly, as the effects of these effects are enhanced, these substances are known to be applicable not only to natural plant protection agents but also to various industrial fields such as biotechnology, food and pharmaceuticals (Non-Patent Document 15). However, there has been no report that these substances induce host plant resistance to pests.

Bacillus oryzae cola YC7011 isolated from the rice seedlings during the pot experiment in the first green industry of the present invention is an endogenous bacterium that not only inhibits the growth of important bacterial pathogens and fungi in rice but also induces rice resistance and promotes plant growth . In this patent, for the first time in the world, it has been confirmed that this multifunctional microorganism YC7011 strain has a resistance inducing effect on insect pests by rhizosphere treatment or seed treatment. Therefore, the active substances exhibiting the effect are isolated and purified and their structure is revealed. It also reveals the function of these new substances and can be used to develop methods for use in natural plant protection products and bio industries in various fields.

Non-Patent Document 1: Ryu, C.M. (2013). Promoting plant protection by root-associated microbes. Plant Pathol J 29: 123-124. Non-Patent Document 2: McSpadden Gardener B. (2010). Biocontrol of plant pathogens and plant growth promotion by Bacillus. In: Recent Developments in Plant Disease, Plant Pathology in the 21st Century. Eds. By U. Gisi, I. Chet and M.L. Gullino, Chapt. 6, pp. 71-79. Springer-Amsterdam. Non-Patent Document 3: Lee, S.K., Sohn, H. B., Kim, G.G., and Chung, Y.R. (2006). Enhancement of biological control of Botrytis cinerea on cucumber by foliar sprays and bed potting mixes of Trichoderma harzianum YC459 and its application on tomato in the greenhouse. Plant Pathol J 22 (3): 283-288. Non-Patent Document 4: Sung, K.C. and Chung, Y.R. (1997). Enhanced suppression of chitinases or antibiotics. In: Proceedings of the 4th International Workshop on Plant Growth Promoting Rhizobacteria Present Status and Future Prospects, eds. By A. Ogoshi, K. Kobayashi, Y. Homma, F. Kodama, N. Konodo and S. Akino. pp.370-373. OECD, Paris. Non-Patent Document 5: Choi, G.J., Kim, J.C., Park, E.J., Choi, Y.H., Jang, K.S., Lim, H.K., Cho, K.Y. and Lee, S.W. (2006) Biological control activity of two isolates of Pseudomonas fluorescens against rice sheath blight. Plant Pathol J 22: 289-294. Non-Patent Document 6: Park, K.S., Paul, D. and Yeh, W.H. (2006). Bacillus vallismortis EXTN-1 mediated growth promotion and disease suppression in rice. Plant Pathol J 22: 278-282. Non-Patent Document 7: Kazempour, M.N. and Elahinia, S.A. (2007). Biological control of Fusarium fujikuroi, the causal agent of bakanae disease by rice associated antagonistic bacteria. Bulg J Agric Sci 13: 393-408. Non-Patent Document 8: Rosales, A.M. and Mew, T.W. (1997). Suppression of Fusarium moniliforme in rice by rice-associated antagonistic bacteria. Plant Dis. 81: 49-52. Non-Patent Document 9: Chung, E.J., Hossain, M.T., Khan, A., Kim, K.H., Jeon, C.O., and Chung, Y.R. (2015). Bacillus oryzicola sp. nov., and endophytic bacterium isolated from the roots of rice with antimicrobial, plant growth promoting, and systemic resistance inducing activities in rice. Plant Pathol J 31 (2): 152-164. Non-Patent Document 10: Hossain, MT, Khan, A., Rashid, M. and Chung, E. J., Chung, Y.R. (2015). Development of a novel endophytic Bacillus species as a microbial inoculant to control seed-borne rice diseases and brown plant hopper. In: 10th International Plant Growth Promoting Rhizobacteria Workshop. Program and Abstract book. p.47. Non-Patent Document 11: Zebelo, S., Song, Y., Kloepper, J.W. and Fadamiro, H. (2016). Rhizobacteria activates (+) -? - cadinene synthase genes and induces systemic resistance to cotton against beet armyworm (Spodoptera exigua). Plant Cell and Environment 39: 935? 943. Non-Patent Document 12: Hossain, MT, Khan, A., Chung, E.J., Rashid, M.H., and Chung, Y.R. (2016). Biological control of rice bakanae by an endophytic Bacillus oryzicola YC7007. Plant Pathol J 32 (3): 228-241. Non-Patent Document 13: Kwon, Y.S., Lee, D. Y., Rakwal, R., Baek, S. B., Lee, J. H., Kwak, YS, Seo, J. S., Chung, W.S., Bae, D.W., and Kim, S.G. (2016). Proteomic analysis of the interaction between plant-growth promoting rhizobacterium Paenibacillus polymyxa E681 and Arabidopsis thaliana. Proteomics 16: 122-135. Non-Patent Document 14: Govindasamy V., Senthilkumar M., Magheshwaran V., Kumar U., Bose P., Sharma V., and Annapurna K. (2010). Bacillus and Paenibacillus spp .: Potential PGPR for Sustainable Agriculture. Plant Growth and Health Promoting Bacteria. Microbiology Monographs V18: 333-364. Non-patent document 15: Ongena M., and Jacques P. (2008). Bacillus lipopeptides: versatile weapons for plant disease biocontrol. Trends Microbiol 16 (3): 115-125. Non-Patent Document 16: Luo C., Liu X., Zhou X., Guo J., Truong J., Wang X., Zhou H. Li X., and Chen Z. (2015). Unusual biosynthesis and structure of Locillomycins from Bacillus subtilis 916. Appl Environ Microbiol 81 (19): 6601-9. Non-Patent Document 17: Kajimura Y., Sugiyama M., and Kaneda M. (1995). Bacillopeptins, New cyclic lipopeptide antibiotics from Bacillus subtilis FR-2. J Antibiot (Tokyo) 48 (10): 1095-103. Non-Patent Document 18: Chen XH, Koumoutsi A., Scholz R., Eisenreich A., Schneider K., Heinemeyer I., Morgenstern B., Voss B., Hess WR, Reva O., Junge H., Voigt B. , Jungblut PR, Vater J., Sussmuth R., Liesegang H., Strittmatter A., Gottschalk G., and Borriss R. (2007). Comparative analysis of the complete genome sequence of the plant growth-promoting bacteria Bacillus amyloloquefaciens FZB42. Nat Biotechnol. 25 (9): 1007-14. Non-Patent Document 19: Thasana N., Prapagdee B., Rangkadilok N., Sallabhan R., Aye S. L., Ruchirawat S., and Loprasert S. (2010). Bacillus subtilis SSE4 produces subtulene A, a novel lipopeptide antibiotic possessing an unusual C15 unsaturated b-amino acid. FEBS Lett 584 (14): 3209-14. Non-patent document 20: Ma Z., Hu J., Wang X., and Wang S. (2013). NMR spectroscopic and MS / MS spectrometric characterization of a novel lipopeptide antibiotic bacillopeptin B1 produced by a marine sediment-derived Bacillus amyloliquefaciens SH-B74. J Antibiot (Tokyo) 67 (2): 175-178. Non-Patent Document 21: Yamamota S., Shiraishi S., and Suzuki S. (2015). Are cyclic lipopeptides produced by Bacillus amyloliquefaciens S13-3 responsible for the plant response to strawberry against Colletotrichum gloeosporidides Lett Appl Microbiol 60 (4): 379-86. Non-patent document 22: Mukherjee, A. K., and Das, K. (2005). Correlation between diverse cyclic lipopeptides production and regulation of growth and substrate utilization by Bacillus subtilis strains in a particular habitat. FEMS Microbiol Ecol 54 (3): 479-89.

Accordingly, it has been confirmed that the present invention has resistance inducing effect and plant growth promoting effect by treating the endophytic bacterium Bacillus oryzicola YC7011 on the rhizosphere or the seed of the host plant and separating and purifying the substances produced by the strain, It is an object of the present invention to provide a structure of a novel substance capable of inducing resistance to plant diseases and insects and promoting plant growth.

The present invention to solve the above-mentioned problem is novel microorganism Bacillus origination coke (Bacillus oryzicola) YC7011 or the Bacillus origination coke (Bacillus oryzicola) Bacillus origination coke (Bacillus having the same pattern YC7011 and genetic analysis (BOX-PCR) Results oryzicola .

In addition, the Bacillus originality Cola (Bacillus oryzicola ) YC7011 provides Bacillus oryzicola , which comprises 16S rRNA having the nucleotide sequence shown in SEQ ID NO: 1.

In addition, the Bacillus originality Cola (Bacillus oryzicola YC7011 culture medium.

In addition, the microbial preparation comprises an active ingredient having a structure in which a C8-C20 beta amino acid chain is bonded to a cyclic structure of a basilopeptin-based structure represented by the following formula (1).

 (Formula 1)

And the active ingredient has a structure of any one of the following formulas (2) to (10).

 (2)

(Formula 3)

(Formula 4)

(Formula 5)

(Formula 6)

(Formula 7)

(Formula 8)

(Formula 9)

(Formula 10)

Also, the microbial agent provides a microbial agent having an aphid control, a control of a brownfly, a growth inhibition of a fungal pathogen, a host resistance induction of a plant rhizome, or a plant growth promoting effect.

In addition, the above-mentioned hospital fungus is caused by Fusarium fujikuroi or Candida causing human inflammation albicans . < / RTI >

According to the present invention, the Bacillus origination coke (Bacillus oryzicola ) YC7011 and the materials isolated from the culture broth of this strain were found to induce the resistance of the host plants to control plant pests and promote growth. By using these multifunctional microorganisms and substances to promote the growth of plants, inhibit plant pathogens and induce host pest resistance, it is possible to produce excellent microbial agents capable of simultaneously acting as multifunctional natural plant protection agents, plant strengthening agents and microbial fertilizers .

1 shows the nucleotide sequence of a gene (rDNA) encoding 16S rRNA of Bacillus orienticola YC7011 strain,
Fig. 2 shows the results of a 16S rRNA gene sequence analysis of Bacillus orienticola YC7011 strain,
Fig. 3 shows the result of gene analysis (BOX-PCR) of Bacillus oryzae Cola YC7011 strain and a similar strain,
Fig. 4 is a schematic diagram of a process for separating and purifying a substance from a culture broth of Bacillus oryzicola YC7011,
FIG. 5 is a schematic view of the separation and purification process of F2D2 from fraction F,
FIG. 6 is a schematic view of the separation and purification process of G4B and G4C from fraction G in the first separated material,
FIG. 7 is a schematic diagram of the separation and purification process of Bacillopeptin A (H4E) from the primary separation material H,
FIG. 8 shows a method for separating and purifying I4B, Bacillopeptin C (I4D), Bacillopeptin B (I4C), I4E and I4F from fraction I,
9 shows the structural formula of F2D2,
10 is a 1 H-NMR analysis graph for the solvent CD3OD for the spectroscopic structure of F2D2,
11 is a 1 H-NMR analysis graph in solvent DMSO-d6 for the spectroscopic structure of F2D2,
12 is a 13C-NMR analysis graph in solvent DMSO-d6 for the spectroscopic structure of F2D2,
13 is a graph obtained using HR-ESI-TOF-MS for mass analysis of F2D2,
14 shows the structural formula of G4B,
15 is a 1 H-NMR analysis graph for the solvent CD3OD for the spectroscopic structure of G4B,
16 is a 1 H-NMR analysis graph in solvent DMSO-d6 for the spectroscopic structure of G4B,
17 is a 13C-NMR analysis graph in solvent DMSO-d6 for the spectroscopic structure of G4B,
18 is a COSY-NMR analysis graph for identifying the spectroscopic structure of G4B,
19 shows HSQC-NMR analysis graphs for the spectroscopic structure of G4B,
FIG. 20 shows the HMBC-NMR analysis graph for the spectroscopic structure of G4B,
21 is a TOCSY-NMR analysis graph for identifying the spectroscopic structure of G4B,
22 shows a ROESY-NMR analysis graph for identifying the spectroscopic structure of G4B,
23 is a graph obtained using HR-ESI-TOF-MS for mass analysis of G4B,
Figure 24 shows the structural formula of Bacillopeptin A (H4E)
25 is a 1 H-NMR analysis graph for the solvent CD3OD for the spectroscopic structure of bacillopeptin A (H4E)
26 is a 1 H-NMR analysis graph in solvent DMSO-d6 for the spectroscopic structure of bacillopeptin A (H4E)
27 is a 13C-NMR analysis graph for the solvent DMSO-d6 for the spectroscopic structure of bacillopeptin A (H4E)
28 is a graph obtained using HR-ESI-TOF-MS for mass spectrometry analysis of bacillopeptin A (H4E)
29 shows a structural formula of I4E,
30 is a graph of < 1 > H-NMR analysis on solvent CD3OD for the spectroscopic structure of I4E,
31 is a graph of < 1 > H-NMR analysis in solvent DMSO-d6 for the spectroscopic structure of I4E,
32 is a 13C-NMR analysis graph in solvent DMSO-d6 for the spectroscopic structure of I4E,
33 is a graph obtained using HR-ESI-TOF-MS for mass spectrometry of I4E,
34 shows the structural formula of bacillopeptin B (I4C)
35 is a 1 H-NMR analysis graph for solvent CD3OD for the spectroscopic structure of bacillopeptin B (I4C)
FIG. 36 is a 13C-NMR analysis graph for the solvent CD3OD for the spectroscopic structure of bacillopeptin B (I4C)
FIG. 37 is a graph showing the 13C-NMR analysis in solvent DMSO-d6 for the spectroscopic structure of bacillopeptin B (I4C)
FIG. 38 shows a COSY-NMR analysis graph for the spectroscopic structure of bacillopeptin B (I4C)
FIG. 39 shows the HSQC-NMR analysis graph for the spectroscopic structure of bacillopeptin B (I4C)
FIG. 40 shows the ROSEY-NMR analysis graph for the spectroscopic structure of bacillopeptin B (I4C)
41 is a TOCSY-NMR analysis graph for identifying the spectroscopic structure of bacillopeptin B (I4C)
Figure 42 is a graph obtained using HR-ESI-TOF-MS for mass spectrometry analysis of bacillopeptin B (I4C)
Figure 43 shows the structural formula of Bacillopeptin C (I4D)
Figure 44 is a 1 H-NMR analysis graph for solvent CD3OD for the spectroscopic characterization of bacillopeptin C (I4D)
45 is a 1 H-NMR analysis graph in solvent DMSO-d6 for the spectroscopic structure of bacillopeptin C (I4D)
46 is a 13C-NMR analysis graph in solvent DMSO-d6 for the spectroscopic structure of bacillopeptin C (I4D)
47 is a graph obtained using HR-ESI-TOF-MS for mass analysis of bacillopeptin C (I4D)
48 shows the structural formula of G4C,
49 is a 1 H-NMR analysis graph in solvent DMSO-d6 for the spectroscopic structure of G4C,
50 is a 13C-NMR analysis graph in solvent DMSO-d6 for the spectroscopic structure of G4C,
51 is a graph obtained using HR-ESI-TOF-MS for mass analysis of G4C,
52 shows a structural formula of I4B,
53 is a 1 H-NMR analysis graph for solvent CD3OD for the spectroscopic structure of I4B,
54 is a 1 H-NMR analysis graph in solvent DMSO-d6 for the spectroscopic structure of I4B,
55 is a 13C-NMR analysis graph in solvent DMSO-d6 for the spectroscopic structure of I4B,
56 is a COSY-NMR analysis graph for identifying the spectroscopic structure of I4B,
57 is a graph showing the HSQC-NMR analysis graph for the spectroscopic structure of I4B,
58 is an HMBC-NMR analysis graph for identifying the spectroscopic structure of I4B,
59 is a graph obtained using HR-ESI-TOF-MS for mass analysis of I4B,
Figure 60 shows the structure of I4F,
61 is a 1 H-NMR analysis graph for solvent CD3OD for the spectroscopic structure of I4F,
62 is a 1 H-NMR analysis graph in solvent DMSO-d6 for the spectroscopic structure of I4F,
63 is a 13C-NMR analysis graph in solvent DMSO-d6 for the spectroscopic structure of I4F,
64 is a graph obtained using HR-ESI-TOF-MS for mass spectrometry of I4F,
FIG. 65 is a photograph showing the effect of controlling the resistance of rice bacilli arising from the resistance of the substances Bacillopeptin A, B and the novel substance I4E,
FIG. 66 is a graph showing a control effect result of a control aphid on a YC7011 culture medium,
67 is a graph showing a control effect of the YC7011 culture medium on the brown planthopper.

Hereinafter, the present invention will be described in detail with reference to examples. Prior to this, terms and words used in the present specification and claims should not be construed as limited to ordinary or dictionary terms, and the inventor should appropriately interpret the concepts of the terms appropriately The present invention should be construed in accordance with the meaning and concept consistent with the technical idea of the present invention. Accordingly, it is to be understood that the constituent features of the embodiments described herein are merely the most preferred embodiments of the present invention, and are not intended to represent all of the inventive concepts of the present invention, so that various equivalents, And the like.

The present inventors have found that a multifunctional endophytic bacterium having both a disease resistance inducing effect and a plant growth promoting effect can be isolated and mass-cultured by broadly inhibiting the growth of a plant pathogenic fungus and a bacterium, , We found a new strain that could be used as a new form of pesticide with microbial fertilizer efficacy.

As a result of searching for the bacterium isolated from the pot experiment in Jeil Green Industrial Co., Ltd., it was confirmed that the isolated strain had all the above effects, and the 16S rRNA gene was sequenced and analyzed, and gene analysis (BOX-PCR) As a result of conducting experiments and the like, it was identified as a microorganism belonging to the genus Bacillus ( Bacillus sp.) And named as " Bacillus oryzicola YC7011".

The isolated microorganism Bacillus oryzae coli according to the present invention can be obtained from Dunlap CA et al. (Dunlap CA, Kim SJ, Kwon SW, and Rooney AP (2015) Bacillus velezensis is not a lie heteotypic synonym of Bacillus amyloliquefaciens ; Bacillus methylotrophicus , Bacillus amyloliquefaciens subsp. plantarum and? Bacillus oryzicola ? are later heterotypic synonyms of Bacillus velezensis based on phylogenomics. Int J Syst Evol Microbiol 66: 1212-1217) has become a taxonomic dispute among closely related species. Marques AS et al., Marchaison A., Garden L. et al., ≪ RTI ID = 0.0 > M. < / RTI & and Samson R. (2008) BOX- PCR-based indentification of bacterial species belonging to Pseudomonas syringae -P viridiflava group Genetics and Molecular Biology 31 (1):... 106-115) or a paper such as Cherif (Cherif A., Brusetti L., Borin S., Rizzi A., Boudabous A., Khyami-Horani H., and Daffonchio D. (2003) Genetic relationship in the Bacillus cereus group? by rep-PCR fingerprinting and sequencing of Bacillus anthracis -specific rep-PCR fragment. J Micro Biol 94: 1108-1119), it is possible to clarify the classification of closely related species through a gene-analysis (BOX-PCR) method. Therefore, as a result of gene analysis (BOX-PCR), the same pattern of microorganisms can be treated as the same microorganism as Bacillus oryzae cola of the present invention.

According to a preferred embodiment of the present invention, the Bacillus oryzicola may include 16S rRNA having the nucleotide sequence shown in SEQ ID NO: 1.

In addition, the present inventors isolated the substances having a controlling effect against plant pests caused by resistance induction from the culture medium of the novel Bacillus oryzae Cola YC7011 and having a growth promoting effect, and found out the structure thereof, Respectively.

In the present invention, substances obtained from the culture of Bacillus oryzae coli YC7011 can be obtained from Kajimura et al. (Non-Patent Document 17: Kajimura Y., et al, 1995 Bacillopeptins, New cyclic lipopetide Antibiotics from Bacillus Bacillopeptin-based cyclic lipopeptides including Bacillopeptin A, B, and C, which were published in the subtilis FR-2, and the novel substances in the isolated bacillopeptins induce resistance from the host plants, thus exhibiting a control effect against rice blight and brownflies At the same time, growth promotion effect was also confirmed. In addition, the plant pathogenic fungus Fusarium causing rice seedling disease In addition to fujikuroi , Candida and the growth inhibition effect on albicans was confirmed.

Hereinafter, the present invention will be described in detail with reference to examples.

Isolation and Identification of YC7011 Strain

(1) Isolation of YC7011 strain

The YC7011 strain was isolated from the root tissue of rice plant rhizosphere during pot experiment in Jeil Green Industrial Technology Development Co., First, in order to isolate endophytic bacteria, the surface of root cut pieces was immersed in 1% hypochlorite (NaOCl) solution for 10 minutes to sterilize the surface. These fragments were placed on a 1/10 TSA medium (3 g of Tryptic Soy Broth, 16 g of agar / 1 L of distilled water), and bacterial growth was observed during culturing for about 2 to 3 days to confirm sterilization of the surface. Was taken for surface sterilization is a root piece 1.0g confirmation piece 10-fold diluted with sterile distilled water 9.0㎖ (10 ^-3) and further diluted ^(10-4 and ^10-5) are sequentially tenfold, the 1/10 dilution TSA medium and spread evenly. The cultured medium was incubated at 28 ° C for 2-3 days, and then a single colony was isolated purely, which was named as YC7011 strain.

(2) Identification of YC7011 strain

The isolated YC7011 strain was Gram-positive, rod-like aerobic bacteria and had no mobility. YC7011 strain grows at a temperature of 13 to 60 占 폚 at a pH of 4 to 12.0 and is obtained by hydrolysis of gelatin, degradation of carboxymethylcellulose, abutine, starch, dilactose, diglycoside, glycogen, enmeaseyl-glucosamine, Naphthol-AS-bi-phosphohydrolase, and n-acetyl-beta-glucosaminidase enzymes.

As a result of homology searching using Genbank and EzTaxon-e database server, 100% of Bacillus oryzicola YC7010 and 100% of Bacillus ( Bacillus sp. methylotrophicus KACC13105 ^T. Fig. 2 shows the phylogeny of 16S rRNA gene sequence analysis of YC7011 strain. In FIG. 2, the intersection number represents the bootstrap value from 1000 repetitions.

In addition, Versalovic et al. (Versalovic J., Schneider M., Bruijn FJ and Lupski JR (1994) Genomic fingerprinting of bacteria using repetitive sequence-based polymerase chain reaction for more detailed comparison with YC7011 strain and similar closely related species. Methods Mol Cell Biol 5: 25-40) was performed to perform gene analysis (BOX-PCR). From the DNA extracted from each strain, GoTaq Green Master Mix (Promega) was used for gene analysis and BOXAR1 (5? -CATCGGCAAGGCGACGCTGACG-3?) Was used as a primer. The denaturation, annealing, and extension of the initial denaturation at 95 ° C for 7 minutes and 35 times were 30 seconds at 90 ° C, 1 minute at 40 ° C, 72 ° C for 3 minutes, and final extension for 72 minutes for 10 minutes. The PCR product was electrophoresed on 1% LE agarose gel (Seakem). As shown in Fig. 2, the strain YC7011 exhibited a pattern similar to that of strain YC7007 and strain YC7010, and that of Bacillus velezensis KCTC13102 and Bacillus methylotrophicus KACC13105 ^T.

Therefore, the microorganism of the present invention was named as Bacillus oryzicola YC7011 strain based on the above-mentioned genetic analysis result, and received the accession number KCTC13085BP from KCTC (Korean Collection for Type Cultures) on August 24,

Example  One: Bacillus Orizicola  Separation and purification of material from the culture medium of YC7011 strain

Substance separation and purification can be divided into the cultivation step of Bacillus oryzae Cola YC7011 and the step of purifying the substance from the culture medium after culturing. The culture conditions of Bacillus oryzae Cola YC7011 were determined under the optimal conditions of 1/10 TSB medium (Tryptic Soy Broth 3 g / casein decomposition 1.7 g, soybean degradation product 0.3 g, glucose 0.25 g, sodium chloride 0.5 g, diphosphate potassium 0.25 g / distilled water 1 L) 28 [deg.] C, an optimal pH of 7.0, and a culture time of 48 hours at 120 rpm. After culturing, 20 liters of the resulting culture was concentrated with methanol, and the resulting brown solid was dissolved in water and chromatographed on a C18 flash column. Each fraction was developed by mixing water and methanol at a certain ratio. Among them, the active fractions were collected, and 9 kinds of substances were separated and purified using HPLC and Silica MPLC (FIG. 4-8).

The purified sample was dissolved in methanol and analyzed by HR-ESI-TOF-MS mass spectrometry to determine the molecular weight and molecular formula of the separated active substance. Also, 1D-NMR or 2D-NMR (1H, 13C, COZY, HSQC, HMBC, TOCSY, ROESY) was used to analyze the structure of the material. The nine separated materials were found to be cyclic Bacillopeptin family structures containing Bacillopeptin A, B, and C (Table 1). All materials are separated sequence of the ring structure _{Bacillopeptin (L -Asn1- D -Tyr- D -Asn2-} L -Ser1- L -Glu- D -Ser2- L -Thr-β-AA) and other beta-amino acids for each substance Chain structure (C _{10 -} C ₁₇ ), and the NMR results for analyzing the structure and structure of the identified substance are shown in FIG. 4 to FIG. 64. Among these substances, the novel substances identified in the present invention were designated as Bacillopeptin D, F, G, H, I, X (Table 1).

Separation material Molecular Weight Molecular formula Structure type Chemical structure Reference diagram F2D2 964.4502 C ₄₂ H ₆₄ N ₁₀ O ₁₆ C ₁₀ -Normal Bacillopeptin F ^* 6 G4B 992.4815 C ₄₄ H ₆₈ N ₁₀ O ₁₆ C ₁₂ -Normal Bacillopeptin G ^* 11 H4E 1020.5128 C ₄₆ H ₇₂ N ₁₀ O ₁₆ C ₁₄ -Normal Bacillopeptin A ¹⁾ 21 I4E 1048.5441 C ₄₈ H ₇₆ N ₁₀ O ₁₆ C ₁₆ -Normal Bacillopeptin X ^* 26 I4C 1034.5284 C ₄₇ H ₇₄ N ₁₀ O ₁₆ C ₁₅ -iso Bacillopeptin B ¹⁾ 31 I4D 1048.5441 C ₄₈ H ₇₆ N ₁₀ O ₁₆ C ₁₆ -iso Bacillopeptin C ¹⁾ 40 G4C 1006.4971 C ₄₅ H ₇₀ N ₁₀ O ₁₆ C ₁₃ -Anteiso Bacillopeptin D ^* 45 I4B 1034.5284 C ₄₇ H ₇₄ N ₁₀ O ₁₆ C ₁₅ - Anteiso Bacillopeptin I ^* 49 I4F 1085.5495 C ₄₉ H ₇₈ N ₁₀ O ₁₆ C ₁₇ -Anteiso Bacillopeptin H ^* 57

* New substances

1) Non-Patent Document 17

Example  2: Bacillus Orizicola  Of YC7011 culture separation material Host  Test for effectiveness of controlling resistance to brown planthopper caused by resistance induction

The control effect of Bacillus oryzae Cola YC7011 isolate was investigated as follows.

Rice seeds (Shindongjin) were immersed in tap water and only seeds that were sunk were selected. Seeded seeds were immersed in 2% sodium hypochlorite (NaOCl) for 5 minutes, sterilized on the first surface, and then put in 70% alcohol again for 5 minutes to sterilize the second surface and rinse with sterile distilled water several times. The surface sterilized seeds were immersed in sterilized distilled water and subjected to shaking at 30 ° C for 3 days. Sprouted seeds were seeded on Murashige & Skoog medium (MS 2.2 g / L, agar 8 g / L, pH 6.0) and cultivated for 12 days in plant growth medium (28-30 ℃, relative humidity 80% or more). 100 μg / ml of nine kinds of pure substances isolated from the seedlings grown for 12 days were treated with 300 μl / week. After the material was treated, the plants were placed on a growing plant and transferred to a pot containing 100 g of sterilized soil. Two days after the transfer, the plants were irradiated with brown planthopper (15 generations / second) . The damage level of the rice leaves was confirmed 7 days after the radiation of the brown planthopper.

Table 2 shows the results of the investigation of the effectiveness of controlling the resistance of each substance to the brown planthopper.

matter Degree of damage
(Index: 0-9) ^* Control (%) ^** No treatment 8.50 ± 0.50 b - F2D2 8.50 ± 0.50 b - G4B 8.00 ± 1.00 b 5.88 H4E 2.50 ± 1.66 a 70.59 I4E 1.00 ± 0.00 a 88.24 I4C 1.25 + 0.63 a 85.29 I4D 6.00 ± 1.29 b 29.41 G4C 8.50 ± 0.50 b - I4B 5.00 ± 0.82 b 41.18 I4F 5.25 ± 1.93 b 38.24

* 0: sound, 1-3: minor damage, 4-6: moderate damage, 7-8: severe damage, 9:

** Control (%) = (1-treatment damage / non-treatment damage) x 100

Nine species of the isolates were treated in rice seedlings and then irradiated with the brown planthopper. All seven treatments except F2D2 and G4C treatments showed resistance to rice plants by resistance induction. Among them, the treatments with 70% or more control were H4E (Bacillopeptin A), I4C (Bacillopeptin B) and I4E new material treatment, respectively, and showed 70.59%, 85.29% and 88.24% control, respectively.

As a result, the treatment of the isolate from the YC7011 strain resulted in the control of the resistance to the rice plants and the resistance effect of the novel substance I4E was the highest.

Example  3: Bacillus Orizicola  Of YC7011 culture separation material Host  Resistance inducing rice Egg wilt  Control efficacy test

Bacillus oryzae cola YC7011 culture isolate was investigated as follows to determine the control efficacy of rice blight-induced resistance induction by host resistance.

The seeds were soaked in constant water, and the seeds were soaked in 2% sodium hypochlorite for 5 minutes to sterilize the surface first, then sterilize the surface with 70% alcohol for 5 minutes. The surface sterilized seeds were rinsed several times with sterile distilled water. The surface sterilized seeds were immersed in sterilized distilled water and subjected to shaking at 30 ° C for 3 days. Sprouted seeds were seeded on MS medium (MS 2.2 g / L, agar 8 g / L, pH 6.0) and grown for 5 days in plant growth medium (28-30 ° C, relative humidity 80% or more). 100 μg / ㎖ of 9 kinds of pure substances isolated from the seedlings grown for 5 days were treated with 300 ㎕ / week and grown for 5 days in plant growth.

Five days after the treatment of the nine substances, rice blight ( Bukholderia glumae suspension (5x10 ⁷ cfu / ml, OD ₆₀₀ = 0.1) was inoculated on three leaves in three replicates on each leaf. The pathogenic bacteria suspension was cultured in R2A medium for 24 hours, centrifuged, and suspended in 10 mM MgSO ₄ solution. The lesion necrosis was examined after 10 days of inoculation. The disease severity was classified into 0 to 3 (0: no symptoms, 1: small necrotic spots, 2: several necrotic spots were merged into large gall bladder, 3: total necrosis) Respectively.

Table 3 shows the results of controlling rice blight resistance by inducing rice resistance of the isolate. A comparative photograph of the substances Bacillopeptin A, B and the novel substance I4E, which shows a high control effect against rice blight, is shown in FIG.

matter The incidence (score: 0-3) Control (%) No treatment 1.93 ± 0.05 b - F2D2 1.00 ± 0.09 ab 48.19 G4B 1.53 + - 0.20 b 20.73 H4E 0.13 0.05 a 93.26 I4E 0.33 + 0.27 a 82.90 I4C 0.53 + 0.11 a 72.54 I4D 1.00 ± 0.09 ab 48.19 G4C 1.07 ± 0.14 ab 44.56 I4B 0.67 ± 0.11 ab 65.28 I4F 0.73 ± 0.30 ab 62.18

As shown in Table 3, the effect of controlling the rice seed blight by inoculating pathogens 5 days after treatment with 9 kinds of substances was found to be 93.26%, which is the highest among the other treatments of H4E (Bacillopeptin A) And 82.4% of fresh substance I4E and 72.54% of I4C (bacillopeptin B), respectively. It was confirmed that the control effect against rice blight disease was shown by the resistance of rice by the treatment of the separation material, although the control value was slightly different according to the treatment of the separated material.

Example  4: Bacillus Orizicola  YC7011 Growth Promotion Effectiveness of Rice by Treatment of Culture Substance

Bacillus oryzae cola YC7011 culture medium In order to test the growth promoting effect of rice by the treatment of the separation material, the following plant growth promoting effect was examined.

Only seeds soaked in water were used for rice seeds. Seeded seeds were soaked in 2% sodium hypochlorite for 5 minutes and then sterilized on the surface for 1 minute. Then, the surface was sterilized on a second surface for 5 minutes with 70% alcohol and rinsed several times with sterilized distilled water. The surface sterilized seeds were immersed in sterilized distilled water and subjected to shaking at 30 ° C for 3 days. Sprouted seeds were seeded on MS medium (MS 2.2 g / L, agar 8 g / L, pH 6.0) and grown for 5 days in plant growth medium (28-30 ° C, relative humidity 80% or more). The length of stem and the weight of stem were measured after treating for 5 days in each plant.

Table 4 shows the results of promoting rice growth after treatment with 9 kinds of substances.

matter Stem length (cm) Fresh weight (mg / plant) Biomass Growth Rate (%) No treatment 11.4 ± 0.5 j 48.2 ± 2.33 f - F2D2 14.8 ± 0.2 ef 60.4 ± 1.33 cd 25.3 G4B 15.6 ± 0.24 63.2 ± 2.06 cd 31.1 H4E 17.2 + - 0.2 c 75.6 ± 1.69 b 56.8 I4E 21.4 ± 0.24 a 85.0 ± 2.24 a 76.3 I4C 19.0 ± 3.74 b 78.0 ± 3.74 b 61.8 I4D 13.8 ± 0.20 g 59.2 ± 1.77 cda 22.8 G4C 14.4 ± 0.24 fgh 58.8 ± 2.22 cde 22.0 I4B 16.4 ± 0.24 cd 65.0 ± 2.23 c 34.9 I4F 15.6 ± 0.24 65.2 + 2.03 c 35.3

As shown in Table 4, after 5 days from the treatment with 9 kinds of substances isolated from YC7011, the stem length and fresh weight of the rice were measured. As a result, compared with the control and BTH treatments, . In the case of the new substance I4E, the effect of promoting growth of rice was 76.3% higher than that of the other substances.

Example  5: Bacillus Orizicola  YC7011 Plant and Human Hospital of culture-separated material Inhibitory effect on fungi  Research

In order to investigate the inhibitory effect of nine pure substances isolated from Bacillus oryzicola YC7011 on plants and human pathogenic fungi, antibacterial activity was investigated as follows.

The antimicrobial activity of 9 species is important for the fungus Fusarium 1 fujikuroi (Rice Kidari germ), a fungal species that causes inflammation in humans Candida albicans (Candida) by the confrontation bioassay method.

The phytopathogenic fungi and the nine species of substances were inhibited by growth on 1/5 PDA and R2A in a growth medium (16 g agar / distilled water 1 L) containing 5 g of Potato agar sugar medium (PDB, Difco) using a paper disc method The effect was investigated. A paper disc (5 mm in diameter) was placed at the same distance of 1 cm from the edge of the incubator. After 1,000 μg / ml of the 9-type substance was treated with 200 μl of each paper disk and sufficiently impregnated, Mycelial disks (6 mm) were placed and incubated at 28 ° C for 3-5 days.

Inhibition of human pathogens was performed by incubating Candida albicans were suspended in sterilized distilled water (OD ₆₀₀ = 0.1), 0.1 ml of the suspension was applied to the medium, and then a paper disk impregnated with 9 kinds of substances (1,000 μg / ml, 200 μl) The degree of inhibition of growth was measured.

Table 5 below shows the results of the antimicrobial activity test for nine plant seed and plant pathogenic fungi.

matter Plant pathogenic fungi
Fusarium fujikuroi Human hospital fungus
Candida albicans F2D2 - - G4B - - H4E + - I4E ++ + I4C + - I4D - - G4C - - I4B + - I4F + +

[Note] Suppression distance: -, 0 mm; +, ≪ 3 mm; ++, 4-5 mm

As shown in Table 5, the antimicrobial activities of 9 kinds of substances isolated from YC7011 culture broth were found in H4E, I4E, I4C, I4B and I4F, and the antimicrobial activity against human pathogenic fungi was observed in the growth of I4E and I4F substances Inhibitory effect. I4E, a novel substance, showed inhibitory activity against plant and human pathogenic fungi.

Example  6: Bacillus Orizicola  Of the YC7011 culture Host  Effectiveness of aphid control by resistance induction

In order to test the effect of host resistance induction of Bacillus oryzicola YC7011 culture medium on the control of aphid control, YC7011 strain and Bacillus oryzae cola YC7007, Bacillus velezensis KCTC13012), Bacillus methylation in trophy Syracuse (Bacillus methylotrophicus KACC13105), Bacillus amyl Lori rake Pacifico Enschede Francisco tarum (Bacillus amyloliquefaciens subsp. plantarum KACC13105) were also investigated as follows.

Plant saengyuksang (28-30 ℃, relative humidity 80%) was transferred in to a port containing the grown Arabidopsis seedlings sterile bed soil (100g) for 4 weeks, then transferred simeunji 7 days suspension of the above strains for each treatment (2x10 ⁷ cfu / L) was dispensed into the supernatant. The suspension preparation was carried out by shaking culture in 1 / 10TSB medium at 28 ° C overnight, centrifuging the suspension, suspending the bacterial cells in 10 mM MgSO ₄ solution and adjusting the concentration appropriately. After 7 days of treatment, aphids (5 rats / week) were irradiated to the treatments for 8 weeks per treatment, and the number of aphids per each port was confirmed 7 days after aphid irradiation.

FIG. 66 shows the results of controlling aphid resistance by inducing host resistance after the culture of each strain.

As shown in FIG. 66, the number of aphids in B. oryzicola YC7007 and YC7011 treated groups was significantly higher than that of B. velezensis KCTC13012, B. methylotrophicus KACC13105, B. amyloliquefaciens subsp. plantarum KACC13105 was significantly lower than the number of aphids. Therefore, it was confirmed that host resistance was induced by treatment of B. oryzicola YC7007 and YC7011 cultures on Arabidopsis thaliana.

Example  7: Bacillus Orizicola  Of the YC7011 culture Host  Test for effectiveness of controlling resistance to brown planthopper caused by resistance induction

In order to test the effectiveness of controlling Bacillus oryzae Cola YC7011 host resistance by inducing host resistance, YC7011 strain and Bacillus oryzae cola YC7007, Bacillus velezensis KCTC13012), Bacillus methylation in trophy Syracuse (Bacillus methylotrophicus KACC13105), Bacillus amyl Lori rake Pacifico Enschede Francisco tarum (Bacillus amyloliquefaciens subsp. plantarum KACC13105) were also investigated as follows.

The rice seedlings raised for 10 days were transferred to a pot containing 100 g of sterilized soil (at 28-30 ° C., relative humidity of 80% or higher), and after 2 days of transplanting, a suspension of the above strains (2 × 10 ⁷ cfu / L) was dispensed into the supernatant. The suspension preparation was carried out by shaking culture in 1 / 10TSB medium at 28 ° C overnight, centrifuging the suspension, suspending the bacterial cells in 10 mM MgSO ₄ solution and adjusting the concentration appropriately. Five days after the treatment, the brown rice hatchlings (15 grains / week) were sprinkled into the treatments for 5 weeks, and the degree of damage of the rice leaves per each port was confirmed 7 days after the irradiation of the brown planthopper.

FIG. 67 shows the results of controlling the resistance to host disease after treatment with the culture solution of each strain.

As shown in FIG. 67, the degree of damage by B. oryzicola YC7007 and YC7011 treated plants was not affected by the treatment of B. velezensis KCTC13012, B. methylotrophicus KACC13105, B. cerevisiae, and B. licheniformis after 5 days from the treatment of each strain . amyloliquefaciens subsp. plantarum KACC13105 treatment. Therefore, treatment of B. oryzicola YC7007 and YC7011 cultures with rice rootstock showed that host resistance was induced and the effect of controlling.

While the preferred embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that various modifications, additions and substitutions are possible, Such modifications and changes are to be considered as falling within the scope of the following claims.

Institution name: Korea Biotechnology Research Institute

Accession number: KCTC13085BP

Checked on: 20160824

<110> Jgreen Inc.          INDUSTRY-ACADEMIC COOPERATION FOUNDATION GYEONGSANG NATIONAL UNIVERSITY <120> Novel compounds produced by Bacillus oryzicola YC7011 with          activities of induced resistance against plant pathogens and          insect and plant growth promotion <130> NP16-06075 <160> 1 <170> KoPatentin 3.0 <210> 1 <211> 1512 <212> RNA <213> Unknown <220> <223> Bacillus oryzicola <400> 1 cagatcgagc ggacagatgg gagcttgctc cctgatgtta gcggcggacg ggtgagtaac acgtgggtaa 120 cctgcctgta agactgggat aactccggga aaccggggct aatactggat ggttgtttga 180 accgcatggt tcagacataa aaggtggctt cggctaccac ttacagatgg acccgcggcg 240 cattagctag ttggtgaggt aacggctcac caaggcgacg atgcgtagcc gacctgagag 300 ggtgatcggc cacactggga ctgagacacg gcccagactc ctacgggagg cagcagtagg 360 gaatcttccg caatggacga aagtctgacg gagcaacgcc gcgtgagtga tgaaggtttt 420 cggatcgtaa agctctgttg ttagggaaga acaagtgccg ttcaaatagg gcggcacctt 480 gacggtacct aaccagaaag ccacggctaa ctacgtgcca gcagccgcgg taatacgtag 540 gtggcaagcg ttgtccggaa ttattgggcg taaagggctc gcaggcggtt tcttaagtct 600 gatgtgaaag cccccggctc aaccggggag ggtcattgga aactggggaa cttgagtgca 660 gaagaggaga gtggaattcc acgtgtagcg gtgaaatgcg tagagatgtg gaggaacacc 720 agtggcgaag gcgactctct ggtctgtaac tgacgctgag gagcgaaagc gtggggagcg 780 aacaggatta gataccctgg tagtccacgc cgtaaacgat gagtgctaag tgttaggggg 840 tttccgcccc ttagtgctgc agctaacgca ttaagcactc cgcctgggga gtacggtcgc 900 aagactaaac tcaaaggaat tgacgggggc ccgcacaagc ggtggagcat gtggtttaat 960 tcgaagcaac gcgaagaacc ttaccaggtc ttgacatcct ctgacggtcc tagagatagg 1020 acgtcccctt cgggggcaga gtgacaggtg gtgcatggtt gtcgtcagct cgtgtcgtga 1080 gatgttgggt taagtcccgc aacgagcgca acccttgatc ttagttgcca gcattcagtt 1140 gggcactcta aggtgactgc cggtgacaaa ccggaggaag gtggggatga cgtcaaatca 1200 tcatgcccct tatgacctgg gctacacacg tgctacaatg gacagaacaa agggcagcga 1260 aaccgcgagg ttaagccaat cccacaaatc tgttctcagt tcggatcgca gtctgcaact 1320 cgactgcgtg aagctggaat cgctagtaat cgcggatcag catgccgcgg tgaatacgtt 1380 cccgggcctt gtacacaccg cccgtcacac cacgagagtt tgtaacaccc gaagtcggtg 1440 aggtaacctt ttaggagcca gccgccgaag gtgggacaga tgattggggt gaagtcgtaa 1500 caaggtgccg ta 1512

Claims (7)

New microorganism Bacillus oryzicola YC7011 (accession number: KCTC13085BP). The method according to claim 1,
The Bacillus originality Cola (Bacillus oryzicola ) Bacillus oryzicola characterized in that YC7011 comprises 16S rRNA having the nucleotide sequence shown in SEQ ID NO: 1. Bacillus &lt; RTI ID = 0.0 &gt; ( Bacillus &lt; oryzicola ) YC7011. The method of claim 3,
Wherein the microbial agent comprises an active ingredient having a structure in which a beta amino acid chain of (C8 to C20) is bonded to a cyclic structure of a basilopeptin-based structure represented by the following formula (1).
(Formula 1)

5. The method of claim 4,
Wherein the active ingredient has a structure of any one of the following formulas (2) to (10).
(2)

(Formula 3)

(Formula 4)

(Formula 5)

(Formula 6)

(Formula 7)

(Formula 8)

(Formula 9)

(Formula 10)

The method of claim 3,
Wherein the microorganism preparation has the effect of inhibiting the growth of Candida albicans causing Fusarium fujikuroi or human inflammation and promoting plant growth, which cause aphid control, a control of the brown planthopper , a control of rice blight, rice hind legs disease. delete

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