KR20160049911A - Composition comprising N-benzoyltryptamine for controlling plant disease and use thereof - Google Patents

Abstract

The present invention is N - relates to a control method of plant disease pathogens using the agricultural chemical formulation, fertilizer and the composition comprising the composition for controlling plant diseases, the composition comprising a benzoyl tryptamine (N -benzoyltryptamine).

Description

N-benzoyltryptamine, and a use thereof. ≪ Desc / Clms Page number 2 >

The present invention is N - relates to a control method of plant disease pathogens using the agricultural chemical formulation, fertilizer and the composition comprising the composition for controlling plant diseases, the composition comprising a benzoyl tryptamine (N -benzoyltryptamine).

Rice is an important food resource for the world's population and is one of the most used grains. Therefore, bacterial diseases caused by pathogenic bacteria infection can be linked to large-scale deaths, and in severe cases, the cultivating farmers cause a decrease in income due to a sharp decrease in yield, while the supply of goods to the consumers is rapidly increased due to imbalance of demand and supply Resulting in enormous economic confusion.

There are three major strains of bacteria reported in Korea: bacterial grain rot, bacterial brown streak, and bacterial leaf blight. It has been reported that it occurs in Asia, North and South America and Europe. The bacterial disease each Burkholderia glue sheets (Burkholderia glumae , < / RTI > Acidovorax avanae or Xanthomonas oryzae ), and the seeds infected by the pathogens may become a primary source of infection and cause a major outbreak. In addition, the pathogens are spread through water, for example, irrigation water. Therefore, it is required to develop a more effective disinfectant for efficiently blocking disease caused by such infection and securing a healthy seed

In addition, although fungicides have a controlling effect, they are potentially dangerous to human life and ecosystem, and studies are being conducted to develop a fungicide that is environmentally friendly and low in side effects. In this regard, Korean Patent Registration No. 10-0713857 discloses an eco-friendly bactericide containing a fenugreek extract and veratramine isolated therefrom. In Korean Patent No. 10-1230367, a floretine compound and a The present invention discloses a composition for controlling plant diseases that contains an apple extract. However, no antibacterial activity of a compound extracted from rice treated with ultraviolet rays has been reported.

Under these circumstances, the inventors of the present invention have made extensive efforts to develop a fungicide that can exhibit an environmentally friendly and excellent controlling effect, and as a result, it has been confirmed that rapidly accumulated compounds in rice treated with ultraviolet light exhibit high antimicrobial activity to complete the present invention .

An object of the present invention are N - to provide a composition for controlling plant diseases comprising a benzoyl tryptamine (N -benzoyltryptamine).

Another object of the present invention is to provide an agricultural chemical preparation comprising the composition.

It is another object of the present invention to provide a fertilizer comprising the composition.

Another object of the present invention is to provide a method for controlling plant pathogenic pathogens using the composition.

In one embodiment of the present invention for achieving the abovementioned objects it is N - provides for the plant disease controlling composition containing benzoyl tryptamine (N -benzoyltryptamine).

In the present invention, "N - benzoyl tryptamine (N -benzoyltryptamine)" is the following, the compound represented by the formula (I) is a substance which is involved in depression of the central nervous system as a kind of flavonoids. N - benzoyltryptamine is one of the phytoalexins identified in the leaves of rice infected with Magnaporthe grisea , the causative agent of rice blast.

In one embodiment of the present invention, compounds 1 to 5 rapidly accumulating in rice treated with ultraviolet light were identified and separated, and NMR analysis revealed that compound 3 was N- benzoyltryptamine (Table 3).

In addition, in one embodiment of the present invention, it was observed that N- benzoyltryptamine of Chemical Formula 1 exhibited antibacterial activity against bacterial pathogens (Table 6), and it was confirmed that this compound could be used as a broad-spectrum bactericide.

In the present invention, " plant disease " means a disease that occurs in a plant, and refers to an abnormality in its original form and physiological function. The cause of the disease may be caused by a pathogen-caused disease, lack of nutrients in the soil, meteorological condition, toxic gas, and the like. Specifically, the plant may be rice, and the plant disease may be a disease or a disease occurring in rice. More specifically, it may be caused by infection from fungal pathogens or bacterial pathogens.

Specifically, the plant disease, but may be being caused by the Xanthomonas Ducks re hospital-type ducks again (Xanthomonas oryzae pv. Oryzae) or Xanthomonas Ducks re hospital-type origination Cola (Xanthomonas oryzae pv. Oryzicola), this But is not limited to.

Specifically, the plant disease may be a bacterial leaf blight of rice or a stripe disease of rice, but is not limited thereto.

In the present invention, the term " bacterial leaf blight " refers to Xanthomonas oryzae pv . oryzae ). It is a disease that occurs extensively throughout the world and is a major problem in rice growing areas in Southeast Asia, including Korea, Japan, Taiwan, the Philippines, India and Indonesia.

The pathogen has a size of (1 to 2 μm) × (0.8 μm), rounded end, and has a single flagella. Gram-negative bacteria do not have spores. The surface of the microbial cells is encapsulated with a polysaccharide viscous liquid (capsule), and when the microbial cells are combined, the microbial cells can be stably present in a severe environmental change. It forms a yellow, round and smooth colonies on nutrient media and is normally able to grow at 5-10 ° C, but the optimum temperature is 26-30 ° C. The death temperature is 53 ° C for 10 minutes, while the dry type bacteria are slightly longer at 56 ° C for 10 minutes.

The disease is a ductal disease mainly occurring in the leaves. The damage caused by the disease is a decrease in rice yield and a deterioration in the quality due to a decrease in the ripening rate due to loss of photosynthesis function in the leaves. It is transmitted through water, infected by infestation of pathogens through pores, hand or wounds of leaves, and propagation in water pipes and / or body tubes. Therefore, inundation and typhoons promote disease development. In case of high temperature and high humidity, The damage is big because it is strong.

In one embodiment of the present invention, it was confirmed that Compounds 1 to 4 isolated from rice treated with ultraviolet light showed an inhibitory effect on Xanthomonas oryzae pv. Oryzae ( Xanthomonas oryzae pv. Oryzae ) (Fig. 3 and Table 6 ), And the compound 3 of the above four compounds was found to be excellent in the inhibitory activity of N -benzoyltryptamine.

In the present invention, the term " stripe disease of rice " refers to symptoms of a gangrene symptom of brown streaks on leaves, stems, and the like, and Xanthomonas oryzae pv . oryzicola ) is known to be caused by infection. Especially when the drainage management is bad, it occurs frequently, and it is transmitted through seeds and surrounding weeds. Therefore, it is necessary to use healthy seeds and care about weed management.

In one embodiment of the present invention, the compounds 1 to 5 isolated from ultraviolet-treated rice were all treated with Xanthomonas < RTI ID = 0.0 > oryzae pv. it was found to exhibit an inhibitory effect against oryzicola) (Figure 3 and Table 6), in particular the compound N 3 - was found to have excellent inhibitory activity of benzoyl tryptamine.

In the present invention, the term " controlling " means preventing a plant such as a crop from damaging a pest, removing a pathogen, or preventing the spread of a pathogen. Methods for creating an inappropriate environment for pathogens by using cultivation and spinning of resistant plants as well as chemical control using fungicides or insecticides which are toxic to pests, methods for removing pathogens by heat-treating plants or treating soil , A method of attracting and capturing pests using an attractant or sex pheromone, and the like.

In one example of the present invention, it was confirmed that compounds 1 to 5, which were rapidly accumulated in rice treated with ultraviolet rays, exhibited excellent antimicrobial activity (FIGS. 3 and 6), and the composition containing the compound was applied to plants, And can be used to inhibit the growth of fungal or bacterial pathogens. Specifically, damage from the pathogen can be prevented by inhibiting infection from fungal or bacterial pathogens in rice.

The composition for controlling the present invention may further contain a substance contained in a commonly used insecticide or bactericide. In addition to the active ingredient, a pharmaceutically acceptable solid carrier, a liquid carrier, a liquid diluent, a liquefied gaseous diluent, A solid diluent or other adjuvant such as an emulsifier, a dispersing agent or a surfactant such as a foaming agent. The controlling composition of the present invention may be formulated into an agricultural composition by mixing the active ingredient with the above excipient. The composition may be formulated into an agricultural composition, and any methods conventionally used in the art may be applied.

In addition, the composition for prevention of the present invention may be provided in the form of a wettable powder, a granule, a powder, an oil, a spray, a film, a capsule or a gel, and is provided with a contact agent of a formulation such as a donut type .

Another embodiment of the present invention provides an agricultural chemical formulation comprising the composition.

In the present invention, the term " pesticide " means a medicament used for protection and growth of crops. Specifically, the present invention relates to a growth regulator for use in enhancing or suppressing the physiological function of a crop, as well as an agent for controlling diseases, pests, weeds, germs, nematodes and mites occurring during cultivation and storage of the crop, May be included.

In one embodiment of the present invention, it was observed that rapidly accumulated compounds 1 to 5 in rice treated with ultraviolet light showed excellent antimicrobial activity and could be used for controlling pathogens to plants (FIGS. 3 and 6) Can be used.

Another embodiment of the present invention provides a fertilizer comprising the composition.

In the present invention, the term " fertilizer ", also referred to as fertilizer, means nutrients that fertilize land and promote the growth of vegetation. The fertilizer containing the composition of the present invention can be easily used by mixing with the soil before sowing, and can be fertilized when spraying pesticides and the like.

In particular, the compound of the present invention is a natural compound extracted from rice and is an environmentally friendly composition, which is harmless to human beings and livestock and soil, so that it is more safe than chemical fertilizer and can prevent disease. The crops in which the fertilizer of the present invention can be used include, but are not limited to, rice.

In another embodiment of the present invention, the composition for prevention is applied to Xanthomonas oryzae pv. oryzae or Xanthomonas oryzae pv. oryzicola , which comprises administering to a subject infected with Xanthomonas oryzae pv. oryzicola .

In addition, by treating the above-described composition for controlling a plant or an area requiring control, bacterial leaf blight of rice or stripe disease of rice can be controlled. Specifically, the controlling composition for plant diseases for a pathogen, i.e., Xanthomonas duck material hospital type duck material (Xanthomonas oryzae pv. oryzae ) or Xanthomonas oryzae pv. oryzicola ), and it is possible to use a method of admixing the soil or directly spraying the crop.

The compound of the present invention exhibits excellent antimicrobial activity and antifungal activity and can be used not only as a composition for controlling plant pathogens but also as a natural compound extracted from rice treated with ultraviolet rays as a natural disinfectant and a fertilizer containing it .

Fig. 1 shows the results of detection of compounds 1 to 5, which are rapidly increased by UV treatment.
FIG. 2 shows the results of measurement of accumulated amounts of compounds 1 to 5 in rice leaves according to time after UV treatment, showing rapid accumulation of compounds 1 to 5 after UV treatment (FIG. 2A, 2, B in FIG. 2, C in FIG. 2, D in FIG. 2, D in FIG. 2, E in FIG.
FIG. 3 is a graph showing the inhibitory activity against the pathogen of each of the compounds 1 to 5 (FIG. 3 A: Compound 1, B in FIG. 3, Compound 2, ; Compound 4, E in Fig. 3, Compound 5, Xoo (), Xoc () and B. glumae (O).

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited by the following examples.

Example  1. Isolation of compounds accumulated by UV treatment

1-1. UV treatment

Rice ( Oryza sativa L. spp japonica cv. Dongjin ) were grown in the greenhouse for 8 weeks under the conditions of 28 캜 during the day and 20 캜 at night. For the next 2 hours, 8 weeks old rice was treated with ultraviolet rays in a growth chamber equipped with five 20 W sterilizing lamps (maximum emission at 254 nm, 7.5 W UV output, Sankyo Denshika, Japan). UV treated rice was transferred to the greenhouse and leaves were collected at 1, 24, 48, 72 and 96 hours after UV treatment for phytochemical analysis.

1-2. By UV treatment Increased  Isolation and Identification of Compounds

The present inventors have confirmed that there are compounds that are rapidly increased by ultraviolet ray treatment of the above-mentioned 1-1 (Fig. 1). In addition, it was confirmed that the compounds rapidly accumulated in rice leaves after UV treatment (FIG. 2). These compounds were referred to as compounds 1 to 5, and the compounds 1 to 5 were isolated and confirmed.

The ultraviolet treated rice leaves were soaked in liquid nitrogen and extracted with 70% methanol (MeOH) - water for 1 hour. After centrifugation, an aqueous methanol extract was obtained and dried under vacuum. The residue was then fractionated by dissolving the residue in a mixture of ethyl acetate (EtOAc) -water (1: 1) to enrich the phenolic compound, and the ethyl acetate phase was dried in vacuo. The residue was dissolved in methanol and analyzed by Park, HL et. al., J. Korean Soc . Appl . Biol . Chem . 2013, 56, 237-241 using a Sunfire C ₁₈ column (Waters, Milford, MA) equipped with a reversed phase HPLC according to the "property of ultraviolet-induced rice phytoalexins Cytotoxic human colon carcinoma HCT-116 cells to" elution conditions described in Respectively.

The silica gel column was applied to isolate ultraviolet-derived compounds from the sample from the rice leaves collected on day 1 after UV treatment. The sample was eluted with hexane-benzene (1: 1), hexane-benzene (1: 2), hexane-benzene (1: 3), benzene-chloroform (1: 1), chloroform- Acetone. The eluted fractions were analyzed by reverse phase HPLC on a Sunfire C ₁₈ column (Waters, Milford, MA) according to the elution conditions. The fractions containing the ultraviolet-induced compound were then collected separately and dried in vacuum. The fraction of each ultraviolet-derived compound was purified on a preparative Sunfire C ₁₈ column (Waters, Milford, MA) with 25-60% acetonitrile linear gradient elution in 3% acetic acid-water for 25 min (flow rate 3 ml / min) ). ≪ / RTI >

The separated compounds were identified by NMR spectroscopy and positive electron spray ionization mass spectrometry (ESI MS). NMR spectra of the separated compounds were recorded on a CD ₃ OD with a Bruker Avance 600 NMR analyzer (Germany) using TMS as an internal standard.

NMR analysis of the chemical shift (chemical shift) of ¹ H of the isolated compounds 1 to 5 (600MHz) and ¹³ C (150MHz) are shown in the following Table 1 to Table 5, respectively.

From the NMR analysis results of Table 1, it was confirmed that the compound 1 was Sakuranetine.

From the NMR analysis results of Table 2, compound 2 is N - was found to be - (trans -cinnamoyltyramine N) - trans-cinnamoyl tyramine.

Table 3 Analysis of the NMR results from the compound 3 is N - could be confirmed that the benzoyl-tryptamine (N -benzoyltryptamine).

The compound from the NMR results in Table 44 are N - it was identified cinnamoyl tryptamine (N-trans-cinnamoyltryptamine) - trans.

In addition, in the heteronuclear multiple bond correlation (HMBC) spectrum of the heteronuclear core, the methine proton signal of δ 3.60 (H-11) was found in the carnauba moiety of δ 168.8 (C-9 ' Nil carbon signal, indicating that the tripmit tamil moiety is linked to the cinnamoyl moiety via an amino bond. Further, it was confirmed that the compound 4 was N-trans-cinnamoyltryptamine through detection of a pseudomolecular ion peak at m / z 291.3 [M + H] ⁺ in positive ESI MS.

Table 5 for compound 5 from the NMR analysis was found to be Np- Koh mellow one serotonin (Np-coumaroylserotonin).

In identifying compound 5, it was found that compound 5 had a structure similar to that of compound 4 when the ¹ H and ¹ H- ¹ H correlation spectra were observed. It was found that two proton signals for H-6 and H-4 are not present in compound 5. Carbon signals for C-6 and C-4 in compound 5 were observed at δ 151.3 and δ 160.7, respectively, indicating that hydroxylation occurred at this position. Thus, compound 5 was found to be composed of serotonin and p -coumaroyl moieties, and the methine proton signal at δ 3.56 (H-11) serotonin moiety in the HMBC spectrum and δ 169.4 (C-9 ' ) p of the - Koh mellow one serotonin moiety through a carbonyl carbon signal in the T and p-Kou mellow one moiety that could be seen that the connection via an amino bond.

From this, it was confirmed that the compound 5 was Np -coumaroyl serotonin.

Example  2. Identification of antimicrobial activity of compounds isolated from ultraviolet-treated rice

From the above Example 1, it was found that both of the phenylamide compound as well as the saccharin were increased in the ultraviolet treated rice, and the present inventors measured the antibacterial activity of the compounds.

Specifically, three kinds of phenolic compounds isolated from ultraviolet light treated rice leaves, Magnaporthe , a pathogenic fungus of rice, grisea PO6-6), Rhizoctonia solani AG-1) and non- polar oryzae ( Bipolaris oryzae ). Each compound was dissolved in DMSO and potato dextrose was added to PDA medium (Beckton, Dixon & Co., Sparks, Maryland, USA) at a final concentration of 1, 5, 10 and 20 ug / mL. The final DMSO concentration in the medium was 1% (v / v). Mycerial discs (1 mm in diameter) were cut from the edge of each pathogen cultured on a PDA dish for two days, and then Mycelial discs were placed in a PDA dish containing various compounds at various concentrations. Pathogenic fungi were left at 28 ° C for 3 days and the Mycelial radius of each fungus was measured to confirm growth inhibition by UV-induced compounds. The experiment was repeated three times.

In addition, the pathogenic bacterium Xanthomonas Ducks re hospital-type ducks again (Xanthomonas oryzae pv. Oryzae (Xoo )), Xanthomonas Ducks re hospital-type origination Cola (Xanthomonas oryzae pv. Oryzicola (Xoc )) and Burkholderia glue The antibacterial activity against Burkholderia glumae was investigated. Each of the above strains was cultured at 28 ° C in NB medium (Becton, Dickson Mann Company, Spark, Maryland, USA). The cultured bacteria were inoculated into NB medium containing 1, 5, 10 and 20 ug / ml of compound, respectively. The final DMSO concentration in the medium was 1% (v / v). In order to confirm the inhibitory activity of each compound, bacterial growth was measured by using a UV / Vis spectrophotometer at OD ₆₀₀ during the growth period of each bacterium. The experiment was repeated three times.

As a result, as shown in Fig. 3, the concentrations of the compounds 1 to 5 of each of the three bacterial pathogens, Xanthomonas oryzae hospital duck, Xanthomonas oryzae hospital type oryzicola and burgunderia glume The compounds showed different degrees of antimicrobial activity. Compounds 1 and 4 showed an inhibitory effect on all bacterial pathogens (A and D in FIG. 3), whereas compounds 2 and 3 exhibited inhibitory effects on Xanthomonas oryzae hospital duck and Xanthomonas oryzae hospital type origin And showed antibacterial activity against cola (B and C in Fig. 3). In addition, it was found that Compound 5 inhibited the growth of Orchitococcus from Xanthomonas oryzae (Fig. 3E).

In addition, IC ₅₀ (ug / ml) indicating the inhibitory activity against fungal or bacterial pathogens of the compounds 1 to 5 isolated from rice leaves treated with ultraviolet rays was measured.

As shown in Table 6 below, Compound 1 and Compound 4 showed an inhibitory effect against both fungal pathogens and bacterial pathogens. Particularly, in the case of Compound 1, fungal pathogens such as Magnaporthe grisea, Lyocalonia solani and nonpolaris dories, bacterial pathogens such as xanthomonas oryzae hospital type duck, xanthomonas oryzae hospital type arijikola, It was found that the antibacterial activity against all of the allergic glume was exhibited.

The IC ₅₀ of Compound 1, which exhibited 50% inhibition of mycelial growth of Magnaporthe grisea, was 6.44 ug / ml, and IC ₅₀ values of 54.04 and 19.05 ug / Of the antifungal activity.

In addition, although most of the phenylamide compounds did not exhibit an inhibitory effect on fungal pathogens, Compound 4 exhibited antifungal activity with an IC ₅₀ value of 26.92 ug / ml for non-polar dorsiflexors.

Furthermore, with respect to the antimicrobial activity against bacterial pathogens, Compound 1 and Compound 4 showed the strongest inhibition of growth of Xanthomonas oryzae aspirin by the IC ₅₀ of 2.36 and 2.45 ug / ml, respectively.

Compound 4 similarly inhibited the growth of xanthomonas oryzae duck material compared to Compound 1 and exhibited relatively weak antibacterial activity against Burkholderia glume (Fig. 3 D and Table 6). Compounds 2 and 3 strongly inhibited the growth of Xanthomonas oryzae asiatica colonies with an IC ₅₀ of 3.18 and 3.72 ug / ml, respectively, despite somewhat weaker activity compared to Compound 1. In the case of Compound 5, the IC ₅₀ of 54.54 ug / ml showed antibacterial activity only against Xanthomonas oryzae hospital type oryzae cola (Table 6).

From the above results, it was found that the rapidly accumulated compounds 1 to 5 in rice treated with ultraviolet rays showed excellent antimicrobial activity and could be used for the control of pathogens to plants. Especially, natural compounds extracted from rice, It can be used for the application.

From the above description, it will be understood by those skilled in the art that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. In this regard, it should be understood that the above-described embodiments are to be considered in all respects as illustrative and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention without departing from the scope of the present invention as defined by the appended claims.

Claims (7)

N - benzoyltryptamine ( N - benzoyltryptamine).
2. The method of claim 1, wherein the plant disease is Xanthomonas < RTI ID = 0.0 > oryzae pv. oryzae ) or Xanthomonas oryzae pv. oryzicola . < / RTI >
The composition according to claim 1, wherein the plant bottle is a bacterial leaf blight of rice or a stripe disease of rice.
The composition of claim 1, wherein the N -benzoyltryptamine is extracted from rice treated with ultraviolet light.
An agricultural chemical formulation comprising the composition of claim 1.
A fertilizer comprising the composition of claim 1.
The composition of claim 1 is applied to Xanthomonas < RTI ID = 0.0 > oryzae pv. oryzae ) or Xanthomonas oryzae pv. oryzicola ) to an individual infected with the plant pathogenic fungi.

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