KR20150069594A - Streptomyces sp. KR-OO4 and use thereof - Google Patents

Abstract

The present invention relates to a herbicide composition for weed control, containing a Streptomyces sp. KR-OO4 strain or a culture medium thereof as an active ingredient. Specifically, the Streptomyces sp. KR-OO4 strain or the culture medium thereof exhibits excellent herbicidal activity against gramineous weeds, broad-leaved weeds and herbicide-resistant weeds as a result of the foliage treatment against the aforementioned weeds. In addition, the strain exhibits excellent weed control and herbicidal activity against different types of weeds under simple packaging conditions. Therefore, the invention can be effectively used as an herbicidal composition for weed control, wherein Streptomyces sp. KR-OO4 strains or the culture medium thereof are contained as an active ingredient.

Description

Streptomyces sp. KR-OO4 strain and its use {Streptomyces sp. KR-OO4 and use thereof}

The present invention relates to a novel strain of Streptomyces sp. KR-OO4 and herbicidal use of the strain.

Weeds or weeds are plants that are not absolute and are not appropriate for the time and place that has arisen as human beings began their agricultural life in relative terms. Weeds are the main control target to reduce the productivity and merchantability of crops, compete with light, nutrients, water, and deodorize them, or to make medicines and agriculture work of pests difficult. Weeds occur throughout the year, regardless of location, season, or environment, and require much effort and cost to control. In addition, because weeds are many kinds and have various weeds, they are classified into weeds, weeds, weeds, weeds and weeds according to plant taxonomic classification and morphological classification. Weeds are divided into annual weeds, perennial weeds and perennial weeds. Therefore, in general, weeds are classified into weeds, perennial weeds, perennial weeds, perennial weeds, perennial weeds, and perennial weeds and perennial weeds and perennial weeds, and on the contrary, they are classified into weeds, weeds, weeds, weeds or perennial weeds, weeds, .

So far, various organic synthetic herbicides have been used for controlling weeds, but the emergence of resistant weeds, environmental hazards and residual toxicity have been pointed out as social problems. In addition, in modern agriculture, organic synthetic herbicides are widely used worldwide due to their low cost and high herbicidal power, but most synthetic herbicides strictly limit their use in many countries even though their impact on environment and ecosystem is not large. In addition, the emergence of resistant weeds due to long-term use of herbicides has been recognized as a serious socio-economic problem (Riches et al. 1996). However, herbicides with a new point of action that can solve the problem of resistance have not been developed for more than 10 years after the development of triketones herbicides (Prisbylla et al. 1993; Schultz et al. 1993). Therefore, the development of synthetic herbicides with new points of new structure is required. However, due to the increase of interest in environmentally friendly agriculture and the emphasis on quality of life and quality of life, there is a strong regulation of each country to reduce the use of chemical pesticides It is a fact that the condition of the development of organic synthetic herbicide is not recorded.

As a solution to the problems of organic synthetic herbicides, interest in natural products that have been useful for mankind for a long time is increasing. It is used as a basic framework of indicator substance or synthetic substance having a new action mechanism by exploiting a variety of natural substances directly or by searching for a substance having a new chemical structure. The validity of such a natural substance-derived substance as a crop protection agent is actively studied (Copping and Duke 2007). Environmentally friendly herbicide, the genus Streptomyces (Streptomyces sp.) or an active substance derived from natural materials originating in plants have been used. In a case study of a foreign country for natural herbicides derived from soil microorganisms and plants, Collet storage compartment global EO Spokane Rio Edith (Colletorichum gloeosporioides), pie Saratov look up Sora palmitate (Phytophthora palmivora), geulradi Pseudomonas (Pseudomonas gladioli ) as a herbicidal herbicide for Zygomycetes japonica, Ivy and ivy, and for the control of herbicides. In the case of research on herbicidal active substances in plants, anthraquinone, naphtha, Recent studies have reported that natural compounds based on naphthoquinone or benzoquinone inhibit hydroxyphenylpyruvate dioxygenase (HPPD).

However, successful commercialization of natural herbicidal active substances derived from plants is extremely limited. The main reason for this is that the herbicidal activity is relatively low compared with the organic synthetic herbicide. Therefore, . In the case of 'Maize gluten meal' using by-products from the grinding process of corn kernels, WeedBan ^™ , Corn Weed Blocker ^™ , But require at least 2 tonnes of throughput per hectare to achieve a satisfactory effect (Quarles 1999). In addition, pelargonic acid, a C9 fatty acid derived from plant extracts, has been developed as a herbicide for foliar treatment and is marketed under the trade name Scythe ^{TM. It} has a rapid-acting and non-selective control of a wide range of weeds (Fukuda et al. 2004) Is so high that it has not been commercially successful. As described above, when a natural herbicidal active material derived from a plant is used as a crop protection agent, a relatively high throughput is required as compared with a synthetic herbicide, which is a fundamental disadvantage of a herbicide derived from a natural material and causes undesirable effects on the environment Which can be an obstacle. Therefore, even if a plant-derived natural substance whose herbicidal activity has been confirmed through the evaluation at the greenhouse level, the applicability of the natural substance in the actual packaging condition must be promptly determined, so that it can be determined whether or not to enter the development stage.

Accordingly, the present inventors have sought to find an environmentally friendly herbicide-derived herbicide, and discovered a novel strain of Streptomyces sp. KR-OO4, and found that the strain or a culture thereof is in a different kind (weed, light and heat) The KR-OO4 strain or its culture was inoculated with a natural herbicide-controlled herbicide (KR-OO4), and the herbicide-resistant KR-OO4 strain The present invention has been accomplished on the basis of these findings.

It is an object of the present invention to provide a novel strain of KR-OO4 belonging to the genus Streptomyces.

It is still another object of the present invention to provide a herbicide using the strain and a culture solution thereof.

It is still another object of the present invention to provide a herbicidal method using the strain and a culture solution thereof.

In order to achieve the above object,

The present invention provides a Streptomyces sp. Strain KR-OO4 deposited with Accession No. KCTC 12412BP.

The present invention also provides a herbicide containing as an active ingredient at least one selected from the group consisting of Streptomyces sp. KR-OO4, a culture thereof, a concentrate of the culture, and a dried product of the culture.

In addition, the present invention relates to a herbicidal herbicidal composition comprising a herbicide or a herbivorous herb comprising one or more selected from the group consisting of Streptomyces sp. KR-OO4, a culture thereof, a concentrate of said culture, and a dried product of said culture .

The KR-OO4 strain of the genus Streptomyces or the culture thereof of the present invention exhibited excellent fungicidal activity on weeds of different weeds, as a result of foliar application to weeds, weeds, light weeds and heating weeds. The strain exhibits excellent control and herbicidal effect on different kinds of weeds, and thus can be usefully used as a herbicidal herbicidal composition for controlling weeds containing the strain Streptomyces sp. KR-OO4 or a culture thereof as an active ingredient.

FIG. 1 shows that strain KR-OO4 of the genus Streptomyces is 99% homologous to Streptomyces agroblogelles NBRC 14001 and is a novel strain.
FIG. 2 is a graph showing that the culture medium of Streptomyces sp. KR-OO4 effectively inhibits KAPAS.
FIG. 3 is a graph showing the viability of KR-OO4 strain treated with Streptomyces sp. KR-OO4.
FIG. 4 is a graph showing the viability of KR-OO4 culture broth of Streptomyces sp.
FIG. 5 is a graph showing the viability of KR-OO4 cultured strain of Streptomyces genus by foliar application to 6 main crops including three weeds, three weeds and three weeds.
FIG. 6 is a graph showing the effect of controlling the effect of the culture of Streptomyces sp. KR-OO4 on the tortoises.
FIG. 7 is a graph showing the effect of controlling the effect of the culture of Streptomyces sp. KR-OO4 on violets.
Fig. 8 is a graph showing the effect of controlling the effect of treating the culture medium of Streptomyces sp. KR-OO4 in a greenhouse under visible light.
FIG. 9 is a view showing the effect of the control by treating the cultured strain of Streptomyces sp. KR-OO4 in a packaging condition under visible light.
10 is a view showing the effect of controlling the culture medium of Streptomyces sp. KR-OO4 on the light leaf weeds such as red ginseng, Lepidoptera, Lepidoptera, Lepidoptera, and Lepidoptera under packaging conditions.
Fig. 11 is a view showing the effect of controlling the effect of the culture medium of Streptomyces sp. KR-OO4 on the light leaf weeds such as carnivorous, rape, rape, and morning glory under the packaging conditions.
Fig. 12 is a view showing the effect of controlling the effect of the culture medium of Streptomyces sp. KR-OO4 on the weed weevil clover, mugwort and hornblende in the packaging condition.
Fig. 13 is a view showing the effect of controlling the effect of treating the culture medium of Streptomyces sp. KR-OO4 strain with the heating weed seedlings under the packaging conditions.

Hereinafter, the present invention will be described in detail.

The present invention provides a Streptomyces sp. Strain KR-OO4 deposited with Accession No. KCTC 12412BP.

The strain is preferably isolated from the soil, but is not limited thereto.

Preferably, the strain is composed of the base sequence of SEQ ID NO: 1, but is not limited thereto.

In a specific embodiment of the present invention, the novel strains of KR-OO4 belonging to the genus Streptomyces according to the present invention are isolated from the forest soil around Daecheong Lake, Cheongwon-gun, Chungbuk province (Korea Research Institute of Bioscience and Biotechnology, KRIBB) The strain, which was preserved by freeze drying in a deep freezer at -70 ° C, was purchased and used. In order to identify the isolated strain, 16S rRNA gene sequencing analysis was carried out. As a result, it was confirmed that the isolated strain was a novel strain showing 99% homology with Streptomyces agroblogelles NBRC 14001 (Fig. 1 reference), Streptomyces strain in the KR-004 (Streptomyces sp. KR-004), and deposited on June 03, 2013 as the deposit number KCTC 12412BP in the Life Resource Center, Korea Research Institute of Bioscience and Biotechnology. As a result of measuring the inhibitory activity of KAPAS (7-keto-8-aminopelargonate synthase) at the new herbicide action point of Bacterium of KR-004 strain of Streptomyces sp., The culture broth of Streptomyces sp. KR-004 effectively inhibited KAPAS see Fig. 2), the crude extract was confirmed to exhibit a concentration dependent response in the state, and 50% inhibitory concentrations (I ₅₀ value) was 70.8 μg mL ^- was ^1. As a result of measuring the herbicidal activity against Bacillus cereus of a strain of Streptomyces sp. KR-004, it was found that Bacillus treated with Streptomyces sp. Strain KR-004 was completely inhibited (100%) as compared with Bacillus sp. And the major symptom was leaf burn-down, and necrosis symptoms were observed in some individuals (see FIG. 3). As a result of evaluating the concentration of KR-004 strain of Streptomyces sp. Cultures, 100, 100, 95 and 40%, respectively, were obtained at 1, 1/2, 1/4 and 1/8 concentrations See Fig. 4 and Table 3). As a result of measuring the foliar treatment effect of the culture broth of Streptomyces sp. KR-004, symptoms of Streptomyces sp. KR-004 were observed after 1 day, It was confirmed that the culture broth of the strain exhibited remarkably strong caustic force (see Fig. 5 and Table 4). At the 1 concentration of the strain culture solution (dilution drainage), it was confirmed that 100% of all the test samples were completely controlled. At 1/2 concentration, 95% It was controlled. At the concentration of 1/4, 95, 80 and 95%, respectively, were observed in the case of Acanthopanax senticosus, Tortoise and Tortoise, and the other species were completely controlled. In addition, symptoms of tobacco growth were observed from 1 day after treatment, but continued to increase in vigor, and major symptom was defolination after leaf burn-down, accompanied by necrosis Lt; / RTI > When the culture broth of Streptomyces sp. KR-004 was evaluated, the culture broth of the strain was treated at 1, 1/2, and 1/4 concentration, 80%, and the efficacy was continuously increased with time after 100, 100 and 95% after 15 days of the culture, respectively. Burn-down was noted as the main symptom and necrotizing symptoms were observed in some leaves. In addition, even when the treatment time elapsed in the culture medium concentration range of the strain, there was no occurrence of new leaf, and no growth was observed in the leaves treated with the strain. Therefore, after 7 days of treatment, (See Fig. 6 and Table 5). ≪ tb >< TABLE > When the effect of controlling the violet control of the strains of Streptomyces sp. KR-004 was evaluated, after 7 days from 1, 1/2 and 1/4 concentration of the culture, the spikelets were 100, 100 and 100% , And it was confirmed that even after the treatment time elapsed in the above concentration range, no regeneration or generation of new leaves occurred and that 100% was maintained even after 15 days from the treatment of the culture broth. In addition, burn-down was noted as the main symptom and necrosis symptoms were observed in some leaves (see FIG. 7 and Table 6). When the effect of controlling the visibility of the culture broth of Streptomyces sp. KR-004 strain was evaluated under greenhouse conditions, when the culture broth of the strain was treated with 1, 1/2, and 1/4 concentration, And the control of the culture broth of the strain showed a control effect of 95, 40, and 20%, respectively, in the broth treated with the 1/8, 1/16, and 1/32 concentrations, Was very effective for the control of visibility. In addition, burn-down was observed as a main symptom, and it was not confirmed that the culture broth of the strain was treated with a concentration of 1/4 or more, and thus the growth point was completely destroyed (See FIG. 8 and Table 7). When the culture broth of Streptomyces sp. KR-004 strain was tested under the packaging conditions, the culture broth of the strain was treated with 1 and 1/2 concentration of the culture broth. As a result, And the control effect was 60% and 40%, respectively. It was confirmed that the control effect was maintained even after 2 weeks of treatment with the culture broth of the strain, and the efficacy persistence was also excellent (see Fig. 9 and Table 8). As a result of evaluating the effect of controlling the light leaf weeds (Anopheles mellonus, Lycopersicon esculentum, Lycopersicon esculentum, Lycopersicon esculentum, Lycopersicon esculentum, and Dogus mugwort) in the culture medium of Streptomyces sp. Strain KR-004 under the packaging conditions, 100, 90, 100, and 95%, respectively, in the cultivars treated with the culture solution at the concentration of 1 and 12 days after the treatment of the culture broth, the light leaf weeds 100, 100, 95, 100 and 100%, respectively, and it was confirmed that the efficacy was maintained even after the treatment period and most of the stem was completely damaged. 100%, 95%, 100% and 100%, respectively, after 5 days of treatment with the culture solution of the strain, and that the viable cell counts were identical even after 12 days (see FIG. 10 and Table 9) ). As a result of confirming the control effect on legumes, rape seedlings, turnip, and morning glory belonging to the light leaf weed which is naturally grown under the packaging conditions of Streptomyces sp. Strain KR-004 strain, 1 day after the strain treatment, , 100%, 100%, 90% and 70% after 4 days of treatment, respectively, and the treatment time was longer The effect is sustained, and most of the stem (See Fig. 11 and Table 10). As a result of confirming the control effect against the herbaceous weeds and weeds which are naturally produced under the packing conditions of Streptomyces sp. Strain KR-004, the culture broth of the strain was treated at 1 and 1/2 concentration, and after 5 days, And 100% and 40%, respectively, in the barnacles. On the other hand, when the culture broth of the strain was treated with 1 and 1/2 concentration, the spreading power was 70 and 40%, respectively (see Table 11). As a result of confirming the control effect against the weeds such as clover, mugwort, and hanam weevil, which are difficult to control even with the conventional herbicide, the culture broth of Streptomyces sp. KR-004 was treated with clover, mugwort, The killing power of the clover, mugwort, and hanwax cv. Treated with 1 strain of the culture medium was 95, 95 and 95%, respectively. The cultivation of the mugwort and the hornblende Were 60 and 50%, respectively. In the heating weeds and the like treated with the culture broth of the strain at a concentration of 1, it was not possible to identify the individuals to be maintained and maintained after 2 weeks of the treatment with the drug. In addition, leaf burn-down was observed as a major symptom, and it was confirmed that necrosis symptoms were expressed in some individuals treated with the culture medium at a concentration of 1/2 (see FIG. 12 and Table 12 Reference). As a result of checking the control effect on the weeds, which are heating weeds of the strains of KR-004 strain of Streptomyces sp. KR-004, the seedling harvesting ability of the seedlings at 1, 1/2, 1/4, 70, 50, 30 and 20%, respectively, after 10 days of treatment, and the effective dose was increased to 98, 70, 50 and 30% after 10 days of treatment, (See Fig. 13 and Table 13).

Thus, genus Streptomyces of the present invention (Streptomyces sp.) KR-OO4 strain or its culture showed significant herbicidal activity against weeds, weeds, weeds and heating weeds, and even when weed conditions were simple, weeds, weeds, The herbicidal composition of the present invention can be effectively used as an effective ingredient of a herbicidal herbicidal composition containing the Streptomyces genus KR-OO4 or a culture thereof as an active ingredient.

The present invention also provides a herbicide containing as an active ingredient at least one selected from the group consisting of Streptomyces sp. KR-OO4, a culture thereof, a concentrate of the culture, and a dried product of the culture.

 The herbicide is preferably used for controlling at least one herbicide selected from the group consisting of herbaceous weeds, weed weevils and weed weeds, but is not limited thereto.

The herbaceous weeds and weeds are preferably, but not limited to, any one or more weeds selected from the group consisting of barnyardgrass, sorghum, and spinach.

It is preferable that the light leaf and the weed are for controlling against any one or more weeds selected from the group consisting of amphipoda, olivine, molluscum, molluscum, cedarwood, carrot, parsley, morning glory and morning glory.

Preferably, the heating weeds are those for controlling at least one weed selected from the group consisting of horseshoe, clover, mugwort, and hornblende, but the present invention is not limited thereto.

The herbicide is preferably used for controlling at least one herbicide selected from the group consisting of spiny mackerel, violaceous mackerel, and rodent mackerel. However, the herbicide is not limited thereto.

The herbicide is preferably formulated for foliar treatment or soil treatment, but is not limited thereto.

The herbicide is preferably, but not limited to, the strain, the culture solution thereof, the concentrated solution of the culture solution, and the dried product of the culture solution in a concentration of 1 to 1/8 (v).

Genus Streptomyces of the present invention (Streptomyces sp.) KR-OO4 strain or its culture showed significant herbicidal activity against weeds, weeds, weeds and heating weeds, and even when weed conditions were simple, weeds, weeds, The herbicidal composition of the present invention can be effectively used as an effective ingredient of a herbicidal herbicidal composition containing the Streptomyces genus KR-OO4 or a culture thereof as an active ingredient.

The present invention also relates to a herbicide or herbicide comprising one or more selected from the group consisting of Streptomyces sp. KR-OO4, a culture thereof, a concentrate of the culture, and a dried product of the culture, ≪ / RTI >

The weeds are composed of weevils and weeds such as weevil, sorghum and mulberry, broad-leaved weevil, mulberry leafhopper, mulberry leafhopper, mulberry leafhopper, , But it is not limited thereto.

Preferably, the weeds are one or more selected from the group consisting of spiny mackerel, violet mackerel, and rodent mackerel. However, the present invention is not limited thereto.

The treatment is preferably foliar treatment or cross treatment, but is not limited thereto.

The treatment is preferably carried out under greenhouse conditions or packaging conditions, but is not limited thereto.

The treatment is preferably, but not limited to, the strain, the culture solution thereof, the concentrated solution of the culture solution, and the dried product of the culture solution in a concentration of 1 to 1/8 (v).

The KR-OO4 strain of the genus Streptomyces or the culture thereof showed significant herbicidal activity against weeds, weeds, weeds, and weeds of warming weeds, and the weeds of weeds, weeds, The herbicidal herbicidal composition containing the Streptomyces sp. Strain KR-OO4 or a culture thereof as an active ingredient can be effectively used as a herbicidal herbicidal herbicide containing the herbicidal herbicidal composition for controlling weeds .

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

However, the following examples illustrate the present invention in detail, and the present invention is not limited by the following examples.

< Example  1> Isolation of strain

Humic acid-vitamin medium was used for isolation from forest soil around Daecheong Lake, Chungwon-do, Chungcheongbuk-do, and cyclohexamide or nalidixic acid was added at an appropriate concentration to inhibit the growth of bacteria and fungi. Respectively. Each 1 g of soil sample was added to sterilized physiological saline solution (10 ml), mixed thoroughly with vortex, and 100 μl of each sample was plated on a separate culture medium and cultured at 28 ° C for a certain period of time. The culture conditions were observed periodically during the culture period, and the cultured colonies were inoculated on a fresh medium to isolate single colonies. The colonies were selected by observing with a microscope. The culture broth and culture conditions were as follows (Table 1).

The strains isolated from each strain were cultured on an agar medium. Cells were collected, suspended in 20% sterilized glycerol, and stored in a -70 ° C ultra-low temperature refrigerator. The isolated strains were cultured in a solid medium, suspended in sterilized 10% skim milk, lyophilized and stored at 4 ° C or below.

Actinomycetes isolated culture medium and culture conditions microbe Separation medium and composition pH Incubation temperature Incubation period


Actinomycetes HV agar (g / l)
Humic acid 1
Na ₂ HPO ₄ 0.5
KCl 1.71
MgSO ₄ 7H ₂ O 0.05
FeSO ₄ 7H ₂ O 0.01
CaCO ₃ 0.02
Agar 18


pH 7.2


28 ℃


7-30 days

< Example  2> Streptomyces  genus( Streptomyces sp .) KR -004 Identification of strain

For the identification of the strains isolated in Example 1, 16S rRNA gene sequencing was performed.

Specifically, to obtain the nucleotide sequence of the 16S rRNA gene of the strain, genomic DNA was isolated using Qiagen DNeasy Blood & Tissue kit and PCR was performed with 27f and 1492r primers as a template. The amplified PCR product was purified and sequenced by Genotech Corp. (SEQ ID NO: 1). The nucleotide sequence was compared with the nucleotide sequence of GenBank database using NCBI BLAST. Sequence groups with high similarity were sorted using Clustal X and a tree was created using the neighborhood-joining algorithm using the PHYDIT program version 3.0 (Fig. 1).

As a result, as shown in Fig. 1, it was confirmed that this strain was a novel strain showing 99% or more homology with Streptomyces agroblogelles NBRC 14001 (Fig. 1). Strain used in the present invention are Streptomyces in KR-004 (Streptomyces sp. KR-004), and deposited on June 03, 2013 as the deposit number KCTC 12412BP in the Life Resource Center, Korea Research Institute of Bioscience and Biotechnology.

< Example  3> Streptomyces  genus( Streptomyces sp .) KR -004 Inhibitory effect of KAPAS on the culture medium

In order to measure the inhibitory effect of KAPAS (7-keto-8-aminopelargonate synthase) on the new herbicide action point of the fermentation broth of Streptomyces sp. Strain KR-004 in Example 2, the following experiment was conducted.

Specifically, the enzyme activity inhibition was determined by measuring the absorbance at the A ₃₄₀ wavelength of NADH generated by the reaction of KAPAS. Reaction solutions were prepared by adding 10 mM α-ketoglutarate, 25 mM thiamin pyrophosphate, 10 mM NAD ⁺ , 30 mM MgCl ₂ and 20 mM potassium phosphate buffer to each plate well. 50 μl of 5 mM pyridoxal 5-phosphate, 50 μl of pimeloyl-CoA, and 10 μl of culture medium were added to the reaction mixture. (3 μg / μl) and 45 μl of DW to prepare a final amount of 250 μl. At this time, the culture solution for enzyme activity inhibition assay was prepared by diluting the culture solution to a final concentration of 1250 to 5 μg mL ^-1 by dissolving the crude extractant in distilled water. Reaction temperature conditions were maintained at 37 ° C and absorbance measurements were taken using a microplate spectrophotometer for 60 minutes at 1 minute intervals.

As a result, as shown in Fig. 2, it was confirmed that the culture medium of Streptomyces sp. KR-004 effectively inhibited KAPAS (Fig. 2) and showed a concentration-dependent response in the crude extract condition. 50% inhibitory concentrations (I ₅₀ value) was 70.8 μg mL ^- was ^1.

< Example  4> Streptomyces  genus( Streptomyces sp .) KR -004 for the culture of the strain culture Live

In order to measure the herbicidal activity of the fermentation broth of KR-004 strain of Streptomyces sp. Cultured in Example 2, the following experiment was carried out.

Specifically, 500 ml of GSS liquid medium was added to a 500 ml baffled flask, and 1 ml of the culture obtained by pre-incubation was inoculated and cultured at 27 DEG C and 180 rpm for 7 days in a culture medium of Streptomyces sp. KR-004 take the supernatant surface area 38.5 cm ² (Tween-20, containing 0.1%) was treated with 5 ml per pot for 7 days after seeding on polystyrene cups in a greenhouse condition (30 ± 5 ° C, light / dark = 14/10 hours) The herbaceous powers were assessed by questionnaire survey using the drug effectiveness / weakness criteria. The composition of the GSS medium is shown in Table 2 below.

As a result, as shown in Fig. 3, it was confirmed that the barley treated with culture medium of Streptomyces sp. Strain KR-004 was completely inhibited (100%) compared to the barley treated with the strain culture solution, (leaf burn-down), and necrosis symptoms were also observed in some individuals (FIG. 3).

Actinomycetes culture medium composition component Soluble starch
Glucose
Soybean meal
Beef extract
Yeast extract
NaCl
K ₂ HPO ₄
CaCO ₃
pH 10g
20g
25g
1g
4g
2g
0.25 g
2g
7.2

< Example  5> Streptomyces  genus( Streptomyces sp .) KR -004 Assessment of the concentration response of culture broth

The same procedure as in Example 4 was conducted to evaluate the concentration response of the strain of Streptomyces sp. KR-004 in Example 2, and the culture of Streptomyces sp. KR-004 was diluted 1, 2, 1/4, and 1/8 times, and evaluated 5 days after the treatment.

As a result, as shown in Fig. 4 and Table 3, the barnyard weights at the 1, 1/2, 1/4 and 1/8 fold concentrations of Streptomyces sp. KR-004 culture medium were 100, 100, 95, 40% (Fig. 4 and Table 3).

Concentration response of KR-004 strain of Streptomyces sp. Treatment concentration
(Dilution factor) Fertility (%) One 100 1/2 100 1/4 95 1/8 40

< Example  6> Streptomyces  genus( Streptomyces sp .) KR -004 Effect of foliar treatment on culture broth

In order to measure the foliar treatment effect of the culture medium of Streptomyces sp. Strain KR-004 of Example 2, the following method was performed.

Specifically, three types of weeds and weeds (Syringae, Dipipoda, Bungli) and three kinds of weedy leaf weevils (Zygosaurus, Lepidoptera, Diptera) were sown in greenhouse condition (30 ± 5 ℃, light / (Tween-20, 0.1%) containing 1, 1/2, and 1/4 times the culture medium of Streptomyces sp. Strain KR-004 was treated with 14 mL per pot. After 6 days, Symptoms and efficacy were investigated according to the weak criteria.

As a result, as shown in Fig. 5 and Table 4, the symptoms of the culture of the strain Streptomyces sp. KR-004 were observed from the 1st day after the treatment of all the six test samples. (Fig. 5 and Table 4). At the 1 concentration of the strain culture solution (dilution drainage), it was confirmed that all of the six test species were 100% controlled. At 1/2 concentration, only 95% of the tested strains were controlled to 100% . In addition, 95%, 80% and 95% of kernels were observed at the concentration of 1/4, respectively, and the other species were completely controlled. The symptoms of the herbicidal activity were expressed from the 1st day after treatment, but the sustained vigorousness was increased. The main symptom was defolination after leaf burn-down, and some necrosis accompanied by some necrosis (Fig. 5 and Table 4).

Effect of foliar treatment of Streptomyces sp. Treatment concentration
(Dilution factor) Fertility (%) Sorghum Peck wire grass Japonica The A toddler One 100 100 100 100 100 100 1/2 100 100 100 100 95 100 1/4 95 100 100 100 80 95

< Example  7> Streptomyces  genus( Streptomyces sp .) KR -004 Effect of culture media on the control of rats

The following method was carried out to measure the effect of the culture of Streptomyces sp. KR-004 strain of Example 2 on the control of Rutaceae.

Specifically, a horticultural soil was filled in a 38.5 cm ² polystyrene cup, and 10 to 15 livers were inoculated in a low-temperature dry state and stored in a low-temperature dry state. The seedlings were seeded in greenhouse conditions (30 ± 5 ° C., light / Hour) for 13 days. The culture broth was diluted to a concentration of 1, 1/2 and 1/4 (Tween-20, 0.1%) and treated with 5 mL per pot. After 7 days and 15 days, (0; no effect; 100; complete control).

As a result, as shown in Fig. 6 and Table 5, the test results of the culture broth of the strain at 1, 1/2, and 1/4 concentration after 7 days of treatment were 100, 95 and 80% And 100%, 100% and 95% after 15 days of the culture, respectively. As a result, it was confirmed that the efficacy was continuously increased with time. The main symptom was burn-down, and necrotic symptoms were observed in some leaves. In addition, even when the treatment time elapsed in the culture medium concentration range of the strain, there was no occurrence of new leaf, and no growth was observed in the leaves treated with the strain. Therefore, after 7 days of treatment, (Fig. 6 and Table 5). &Lt; tb &gt;&lt; TABLE &gt;

Effect of culture medium of Streptomyces sp. After processing Fertility (%) One 1/2 1/4 7 100 95 80 15 100 100 95

< Example  8> Streptomyces  genus( Streptomyces sp .) KR -004 Effect of culture media on the control of violets

In order to measure the violet controlling effect of the culture medium of Streptomyces sp. Strain KR-004 of Example 2, the following method was performed.

Specifically, a polystyrene cup having a surface area of 38.5 cm ² was filled with horticultural soil, and 10-15 lobes of the violet seedlings kept in a low-temperature dry state were inoculated and seeded in a greenhouse (30 ± 5 ° C, light / dark = 14/10 hours) Lt; / RTI &gt; for 13 days. The culture was diluted to a concentration of 1, 1/2, and 1/4 (Tween-20, 0.1%) and treated with 5 mL per pot. After 7 days and 15 days, (0; no effect; 100; complete control).

As a result, as shown in FIG. 7 and Table 6, after 7 days of treatment of the culture broth with 1, 1/2, and 1/4 concentration of violet, the violet treated in the concentration range was 100, 100 And 100%, and it was confirmed that even when the treatment time elapsed in the concentration range, regeneration or no generation of new leaf was observed, and that 100% was maintained even after 15 days from the treatment of the culture broth. The main symptom was burn-down, and necrosis symptoms were also observed in some of the leaves (Fig. 7 and Table 6).

Effect of Streptomyces sp. Culture solution on violet control in greenhouse condition After processing Fertility (%) One 1/2 1/4 7 100 100 100 15 100 100 100

< Example  9> In a greenhouse condition Streptomyces  genus( Streptomyces sp .) KR -004 of the culture broth Thorny night  Control effect

The following method was performed to determine the effect of the culture broth of Streptomyces sp. KR-004 of Example 2 on the greenhouse condition.

Specifically, the seeds were stored at 40 ° C in a dry condition under low temperature. The seeds were sown in a greenhouse (30 ± 5 ° C, light / dark = 14/10 hours) And transplanted into a 38.5 cm ² polystyrene cup. After the transplantation, the cultured spikelets grown in the same greenhouse conditions to 3.2 leaf stage were diluted to a concentration of 1, 1/2, 1/4, 1/8, 1/16, and 1/32 (Tween -20, 0.1%) treated with 12 mL per pot and examined 5 days later.

As a result, as shown in FIG. 8 and Table 7, it was confirmed that when the culture broth of the strain was treated with visible bacteria at a concentration of 1, 1/2, and 1/4, When the culture broth was treated at 1/8, 1/16 and 1/32 concentrations, the inhibition effect was 95, 40 and 20%, respectively. I confirmed that it is very excellent. The main symptom was burn-down, and it was confirmed that the strain was not regenerated even after a period of time of treating the strain with a concentration of 1/4 or more, and the growth point was completely eliminated (Fig. 8 and Table 7).

Effect of the culture of Streptomyces sp. KR-004 in the greenhouse condition Treatment concentration
(Dilution factor) Fertility (%) One 100 1/2 100 1/4 100 1/8 95 1/16 40 1/32 20

< Example  10> Under packaging conditions Streptomyces  genus( Streptomyces sp .) KR -004 of the culture broth Thorny night  Control effect

The following methods were carried out to determine the effect of the culture broth of Streptomyces sp. KR-004 in the above <Example 2> on packaging conditions.

Specifically, as shown in <Example 9>, it was confirmed that the culture broth had a strong controlling effect on the green leaf in the greenhouse condition, and the application evaluation was carried out under actual packaging conditions. The test site was a natural wilderness area of Kashim Park located in Byeong-yong, Yanghwa-myeon, Chungcheongnam-do. The testing period was from August 14, 2019 to August 27, 2013. During the treatment of the strain, the growth conditions of spikelet were 7 to 10 leaflets, and the length was 3 m and the treatment area was 1 m ² . The culture broth was treated with 1, 1/2, 1/4, and 1/8 concentrations of visible bacteria and treated with CO ₂ spray at 200 L ha ^{-1 for} each concentration. The culture broth of the strain was treated with visible light and irradiated with naked eyes 1, 2, 3, 5, 7 and 14 days later.

As a result, as shown in FIG. 9 and Table 8, when the culture broth of the strain was treated with 1 and 1/2 concentration of visiblast, naturally occurring visible bark was completely controlled at this concentration, 8 concentrations were 60 and 40%, respectively. It was confirmed that the control effect was maintained even after 2 weeks of treatment with the culture broth of the strain, and the efficacy persistence was also excellent (FIG. 9 and Table 8).

14 days after the treatment under the packaging conditions, the effect of the control of the visceral effect of the culture medium of Streptomyces sp. Treatment concentration
(Dilution factor) Fertility (%) One 100 1/2 100 1/4 60 1/8 40

< Example  11> Under packaging conditions Streptomyces  genus( Streptomyces sp .) KR -004 of the culture broth Light weed (Angora, Lepidoptera, Lepidoptera, Gag  And dog mugwort) Control effect

The following methods were carried out in order to measure the effect of controlling the light leaf weeds (Anopheles mellonus, Lycopersicon esculentum, Lycopersicon esculentum, Corynebacterium glutinosa) in the culture medium of Streptomyces sp. Strain KR-004 of Example 2 under the packaging conditions.

Specifically, in the packaging of Korean Chemical Research Institute, Changdeok, Yuseong - gu, Daejeon, korean ginseng (4 ~ 5 leaves), mulberry leaves (4 ~ 5 leaves), mulberry leaves (15 leaves) ) Were selected and 50 × 50 cm was added to the treatment area. The chemical treatment was June 5, 2013. The concentration of the culture broth of the strain was diluted to 1 and 1/2 (Tween-20, 0.1%), treated with foliar in an amount of 60 mL per each treatment. After 5 days and 12 days after the culture, Respectively.

As a result, as shown in Fig. 10 and Table 9, after 5 days from the cultivation of the culture broth of the strain to the light leaf weeds, the culture broth of the strain was treated with concentration 1, 100, 95, 100, and 100%, respectively. After 12 days, the weed weights of the weed weevils were increased to 100, 100, 95, 100 and 100%, respectively. It was confirmed that it was dead. 100%, 95%, 100%, and 100% of the weedy weeds were observed at 5 days after the treatment, and that the viable counts were similar even after 12 days (FIGS. 10 and 11) Table 9).

Effect of light leaf weeds (Amaranthus sinensis L., Lycopersicon esculentum, Lycopodium spec. Treatment concentration
(Dilution factor) After processing Fertility (%) Anoptera Japonica Turned Gag Shag One 5 90 100 90 100 95 12 100 100 95 100 100 1/2 5 40 100 95 100 100 12 40 100 95 100 100

< Example  12> Under packaging conditions Streptomyces  genus( Streptomyces sp .) KR -004 of the culture broth Light weed (Purlins, rape seeds, garlands, and morning glory)

The following methods were carried out in order to confirm the control effect against the leaves of the leguminous weeds, the extracts of the rape seedlings, the seedlings and the morning glory belonging to the wild-weed seedlings grown under the packaging conditions of the strain of Streptomyces sp. KR-004 of Example 2 above .

Specifically, we selected four branches of weed species, including four broad-leaf weed species (10 cm), leggings (8 cm), rape herb (5 cm) and morning glory (15 cm) in the packaging of the Dongbang Agro Technology Research Institute in Yonghwa-myeon, 100 × 100 cm, and the treatment was October 1, 2013. The treatment concentration was adjusted to 1, 1/2, 1/4, and 1/8 (Tween-20, 0.1%) and treated with 200 mL per treatment. After 1 and 4 days of treatment, Respectively.

As a result, as shown in Fig. 11 and Table 10, after 1 day of the above-mentioned strain liquor treatment, the extractive powders for carrot, rape seed oil, turnip and morning glory were found to be 60, 50, 85 and 35% And after 4 days of treatment, 100, 100, 90, and 70%, respectively, increase the herbicidal power, and the efficacy continues even after the treatment period. (Fig. 11 and Table 10).

< Example  13> Under packaging conditions Streptomyces  genus( Streptomyces sp .) KR -004 of the culture broth Weed and weeds  Check control effect

The following procedure was carried out to confirm the control effect on the weeds and weeds which are naturally grown under the packaging conditions of the culture medium of Streptomyces sp. KR-004 of Example 2 above.

Specifically, the sites in which the barnacles (3.5 to 4.5 leaves) and the peridotites (4 to 5 leaf beds) were naturally grown in the packaging of the Korea Research Institute of Chemical Technology, Yuseong-gu, Daejeon Metropolitan City were selected to provide treatment at 50 x 50 cm, It was June 5th. The treatments were diluted to 1 and 1/2 (Tween-20, 0.1%) and treated with 60 mL per treatment. After 5 and 12 days of treatment,

As a result, as shown in Table 11, the culture broth of the strain was treated at 1 and 1/2 concentration, and after 5 days, the broth treated at the above concentration showed 100 and 40% It was confirmed that the grazing power was maintained even after that. On the other hand, when the culture broth of the strain was treated with 1 and 1/2 concentration, the spreading power was 70 and 40%, respectively (Table 11).

Effect of weed control on Streptomyces sp. Treatment concentration
(Dilution factor) After processing Fertility (%) wire grass Peck One 5 100 70 12 100 70 1/2 5 40 40 12 40 40

< Example  14> Under packaging conditions Streptomyces  genus( Streptomyces sp .) KR -004 of the culture broth Heating agent  weed( Clover , Mugwort, and Hwangsam)

The culture broth of Streptomyces sp. Strain KR-004 of Example 2 was tested by the following method to check the control effect against weeds such as clover, mugwort, and hornblende which were difficult to control even by applying herbicide .

Specifically, the area where the clover, mugwort, and hornblende are grown in the vicinity of the pavement of the Korea Research Institute of Chemical Technology, Yuseong - gu, Daejeon, was selected and the treatment plot was constructed at 50 x 50 cm. The culture broth was diluted to 1 and 1/2 times (Tween-20, 0.1%) and treated with 70 mL of each treatment. The broth was treated with only 1-fold concentration on the clover. After 6 days of treatment, the degree of immersion was visually examined. The test period was from June 3, 2013 to September 10, 2013.

As a result, as shown in FIG. 12 and Table 12, the cultivars of clover, wormwood, and hornbuckle vine, which had been treated with the strain culture solution 1 at 6 days after treatment of the culture broth with clover, mugwort, 95 and 95%, respectively. It was confirmed that the mortality of mugwort and hornblende treated with the concentration of the strain was 60 and 50%, respectively. In the heating weeds and the like treated with the culture broth of the strain at a concentration of 1, it was not possible to identify the individuals to be maintained and maintained after 2 weeks of the treatment with the drug. Leaf burn-down was observed as a main symptom, and necrosis symptoms were expressed in some individuals treated with the culture medium at a concentration of 1/2 (FIG. 12 and Table 12).

Effect of Clover, Mugwort, and Panax ginseng on Streptomyces sp. Treatment concentration
(Dilution factor) Fertility (%) Clover Mugwort Horseshoe One 95 95 95 1/2 - 60 50

< Example  15> Under packaging conditions Streptomyces  genus( Streptomyces sp .) KR -004 of the culture broth Heating agent  Weed control effect

In order to confirm the control effect of the herbicide of the Streptomyces sp. KR-004 strain of Example 2, which is difficult to control even by applying the conventional herbicide, to the controlling effect of the weed seeds of the weeds and the weeds of the heating weeds, the following method was performed.

Specifically, the locality where the rhizomes were naturally cultivated in the non-agricultural land pavements in the Kyuam-myeon area, Chungcheongnam-do, was selected as 100 x 100 cm and the treatment was October 1, 2013. The treatment concentration was adjusted to 1, 1/2, 1/4 and 1/8 (Tween-20, 0.1%) and treated with 200 mL per treatment lot. After 3 and 10 days of treatment, Respectively.

As a result, as shown in FIG. 13 and Table 13, after 3 days from the treatment, 70, 50, and 30 days after the treatment of the seedling at the concentration of 1, 1/2, And 20%, respectively. After 10 days of treatment, the viability increased to 98, 70, 50 and 30%, respectively, and the efficacy was maintained even after the treatment period. (Fig. 13 and Table 13).

Effect of the Streptomyces sp. Treatment concentration
(Dilution factor) After processing Fertility (%) Pecker One 3 70 10 98 1/2 3 50 10 70 1/4 3 30 10 50 1/8 3 20 10 30

Korea Biotechnology Research Institute KCTC12412BP 20130603

<110> DONGBANGAGRO          Korea Research Institute of Bioscience and Biotechnology          Korea Research Institute of Chemical Technology <120> Streptomyces sp. KR-OO4 and use thereof <130> 13P-09-47 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 1328 <212> DNA <213> Streptomyces sp. <400> 1 gggcaatctg ccctgcactc tgggacaagc cctggaaacg gggtctaata ccggatacga 60 gcctccaccg catggtgggg gttggaaagc tccggcggtg caggatgagc ccgcggccta 120 tcagcttgtt ggtgaggtaa cggctcacca aggcgacgac gggtagccgg cctgagaggg 180 cgaccggcca cactgggact gagacacggc ccagactcct acgggaggca gcagtgggga 240 atattgcaca atgggcgaaa gcctgatgca gcgacgccgc gtgagggatg acggccttcg 300 ggttgtaaac ctctttcagc agggaagaag cgaaagtgac ggtacctgca gaagaagcgc 360 cggctaacta cgtgccagca gccgcggtaa tacgtagggc gcaagcgttg tccggaatta 420 ttgggcgtaa agagctcgta ggcggcttgt cacgtcggtt gtgaaagccc ggggcttaac 480 cccgggtctg cagtcgatac gggcaggcta gagttcggta ggggagatcg gaattcctgg 540 tgtagcggtg aaatgcgcag atatcaggag gaacaccggt ggcgaaggcg gatctctggg 600 ccgatactga cgctgaggag cgaaagcgtg gggagcgaac aggattagat accctggtag 660 tccacgccgt aaacggtggg cactaggtgt gggcaacatt ccacgttgtc cgtgccgcag 720 ctaacgcatt aagtgccccg cctggggagt acggccgcaa ggctaaaact caaaggaatt 780 gacgggggcc cgcacaagcg gcggagcatg tggcttaatt cgacgcaacg cgaagaacct 840 taccaaggct tgacatacac cggaaagcat tagagatagt gccccccttg tggtcggtgt 900 acaggtggtg catggctgtc gtcagctcgt gtcgtgagat gttgggttaa gtcccgcaac 960 ggcgcaacc cttgtcccgt gttgccagca ggcccttgtg gtgctgggga ctcacgggag 1020 accgccgggg tcaactcgga ggaaggtggg gacgacgtca agtcatcatg ccccttatgt 1080 cttgggctgc acacgtgcta caatggccgg tacaaagagc tgcgataccg tgaggtggag 1140 cgaatctcaa aaagccggtc tcagttcgga ttggggtctg caactcgacc ccatgaagtc 1200 ggagtcgcta gtaatcgcag atcagcattg ctgcggtgaa tacgttcccg ggccttgtac 1260 acaccgcccg tcacgtcacg aaagtcggta acacccgaag ccggtggccc aaccccttgt 1320 gggaggga 1328

Claims (17)

Streptomyces sp. KR-OO4 deposited with accession number KCTC 12412BP.
The strain according to claim 1, wherein the strain is isolated from the soil.
2. The strain of claim 1, wherein the strain comprises a base sequence of SEQ ID NO: 1.
A herbicide containing as an active ingredient at least one selected from the group consisting of Streptomyces genus KR-OO4 strain of claim 1, a culture thereof, a concentrate of said culture, and a dried product of said culture.
5. The herbicide according to claim 4, wherein the herbicide is a herbicide for at least one weed selected from the group consisting of herbaceous weeds, weed weeds, and weed weeds.
6. The herbicide according to claim 5, wherein the herbicide and the weed are for controlling against at least one weed selected from the group consisting of barnyardgrass, sorghum, and coriander.
[Claim 7] The herbicide according to claim 5, wherein the light leaf and the weed are suitable for controlling at least one weed selected from the group consisting of amphipathic larvae, red mats, lobsters, pine needles, pine needles,
6. The herbicide according to claim 5, wherein the heating weeds are for controlling against at least one weed selected from the group consisting of horseshoes, clover, mugwort and hornblende.
[Claim 6] The herbicide according to claim 5, wherein the herbicide is a herbicide for at least one weed selected from the group consisting of rhizomes, violets and rats.
5. The herbicide according to claim 4, wherein the herbicide is formulated for foliar treatment or soil treatment.
5. The herbicide according to claim 4, wherein the herbicide comprises the strain, the culture solution thereof, the concentrated solution of the culture solution, and the dried product of the culture solution at a concentration of 1 to 1/8 (v).
A herbicidal herbicidal herb comprising the step of treating the herb or its habitat with one or more selected from the group consisting of Streptomyces genus KR-OO4 strain of claim 1, a culture thereof, a concentrate of said culture, and a dried product of said culture.
13. The method according to claim 12, wherein the weeds are selected from the group consisting of weevil, sorghum and perennial weeds, weeds, migratory birds, wolves, weevils, hornblends, Wherein the weed or its habitat comprises one or more selected from the group consisting of weeds or herbicide weeds.
13. The herbicidal method according to claim 12, wherein the weeds include one or two or more selected from the group consisting of rhizomes, violets, and rodents.
13. The method according to claim 12, wherein the treatment is foliar treatment or cross treatment.
13. The method according to claim 12, wherein the treatment is carried out under greenhouse conditions or packaging conditions.
13. The herbicidal method according to claim 12, wherein the strain, the culture solution thereof, the concentrated solution of the culture solution, and the dried product of the culture solution are contained at a concentration of 1 to 1/8 (v).

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