KR100597319B1 - A Novel Streptomyces sp. and Biopesticide Containing the Strain for Extermination of Rice Blast Disease - Google Patents

A Novel Streptomyces sp. and Biopesticide Containing the Strain for Extermination of Rice Blast Disease Download PDF

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KR100597319B1
KR100597319B1 KR1020040068543A KR20040068543A KR100597319B1 KR 100597319 B1 KR100597319 B1 KR 100597319B1 KR 1020040068543 A KR1020040068543 A KR 1020040068543A KR 20040068543 A KR20040068543 A KR 20040068543A KR 100597319 B1 KR100597319 B1 KR 100597319B1
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rice blast
rice
streptomyces
microbial
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KR20060019866A (en
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임융호
김범준
이용섭
조광연
최경자
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건국대학교 산학협력단
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20Bacteria; Culture media therefor
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/20Bacteria; Substances produced thereby or obtained therefrom
    • A01N63/28Streptomyces

Abstract

본 발명은 벼 도열병에 대한 방제효과를 가지는 방선균속 신규균주, 상기 균주를 함유하는 미생물농약제제 및 상기 미생물농약제제를 이용한 벼 도열병의 방제방법에 관한 것이다. The present invention relates to a new strain of actinomycetes having a control effect against rice blast, a microbial pesticide formulation containing the strain, and a method for controlling rice blast using the microbial pesticide formulation.

본 발명에 따른 균주는, 기존의 화학농약에서 나타날 수 있는 과다사용으로 인한 생태계의 파괴 없이도, 기존 화학농약보다 우수한 벼 도열병 방제효과를 가지므로 생물농약으로서 유용하다.The strain according to the present invention is useful as a biopesticide because it has a superior rice pest control effect than conventional chemical pesticides without destroying the ecosystem due to overuse that may occur in conventional chemical pesticides.

미생물농약, 벼도열병, 방선균, 스트렙토마이세스, 항균제Microbial Pesticides, Rice Fever, Actinomycetes, Streptomyces, Antibacterials

Description

방선균속 신규균주 및 상기 균주를 함유하는 벼도열병 방제용 미생물농약제제 {A Novel Streptomyces sp. and Biopesticide Containing the Strain for Extermination of Rice Blast Disease} New strains of actinomycetes and microbial pesticides for control of rice fever containing the strains {A Novel Streptomyces sp. and Biopesticide Containing the Strain for Extermination of Rice Blast Disease}             

도 1은 신규 방선균속(Streptomyces sp.) KACC 91011 균주의 주사현미경 사진이다.1 is a scanning micrograph of the novel Streptomyces sp. KACC 91011 strain.

도 2는 신규 방선균속(Streptomyces sp.) KACC 91011 균의 16S rRNA 유전자분석 결과이다.Figure 2 is a 16S rRNA gene analysis of the novel Streptomyces sp. KACC 91011.

도 3은 도열병이 심하게 발병된 무처리벼(왼쪽), 방선균속(Streptomyces sp.) KACC 91011 균의 배양액을 처리하여 도열병이 방제된 벼(중앙) 및 기존에 벼도열병 방제를 위해 사용되는 화학농약 트리사이클라졸(tricyclazole)을 처리하여 도열병이 방제된 벼(오른쪽)를 나타낸 것이다.Figure 3 is treated with a culture solution of untreated rice (left), Streptomyces sp. KACC 91011 bacteria severely affected by blasting rice (central) and chemical pesticides used for conventional rice blast control Treated tricyclazole (tricyclazole) shows the rice (right) control of the blast disease.

본 발명은 벼 도열병에 대한 방제효과를 가지는 방선균속 신규균주, 상기 균 주를 함유하는 미생물농약제제 및 상기 미생물농약제제를 이용한 벼 도열병의 방제방법에 관한 것이다.The present invention relates to a new strain of actinomycetes having a control effect against rice blast, a microbial pesticide formulation containing the strain and a method for controlling rice blast using the microbial pesticide formulation.

우리나라에서 벼에 발생하는 병들 중 가장 무서운 병이 벼도열병인데 이는 마그나포르테 그리세아(Magnaporthe grisea)에 의해 발병하고 이 균은 진균계의 자낭균문에 속한다. 이 병원균의 균사는 주로 27~28℃에서 발육되며 벼로 침입하는 시간은 품종과 온도에 따라 정도의 차이는 있으나 일반적으로 6-8 시간 정도 소요된다. 비가 자주 오고 일사량이 부족하면 병이 발생하는데, 발병부위에 따라 목도열병, 잎도열병, 벼알도열병, 마디도열병 등으로 불리우나 목도열병과 잎도열병의 피해가 가장 심하다. 병원균은 분생포자 형태로 월동한 후 바람을 타고 이동하여 기주가 되는 벼에 도착하면 병을 일으킨다. 벼 외의 밀, 피, 강아지풀 등도 기주식물이 된다.The most frightening disease of rice disease in Korea is rice fever, which is caused by Magnaporthe grisea , which belongs to the fungal system of Aspergillus. Mycelia of the pathogen are mainly developed at 27 ~ 28 ℃ and the time to invade the rice, depending on the variety and temperature, but usually takes about 6-8 hours. Rain often occurs and the amount of insolation causes illness. Depending on the area of occurrence, it is called as fever, leaf fever, rice fever, and fever, but the most severe damage is caused by fever and fever. Pathogens overwinter in the form of conidia spores, then move by wind to reach host rice, which causes disease. Other than rice, wheat, blood, and foxtail are also host plants.

이와 같은 벼도열병을 방제하기 위해 현재 사용되고 있는 방법으로는 질소질 비료의 과용을 피한다거나, 균형시비, 또는 채식밀도 조절 등을 사용하고 있고, tricyclazole과 blasticidin-S 등과 같은 화학농약을 사용한다.Current methods used to control rice fever are to avoid overdose of nitrogenous fertilizers, balanced fertilization, or vegetarian density control, and chemical pesticides such as tricyclazole and blasticidin-S.

그러나 화학농약에 의한 토양오염과 소비자들의 거부감이 점차 커짐에 따라 새로운 방법의 방제 방법이 필요하게 되었고 이의 하나로 미생물을 이용한 방제 방법이 대두되었다. 이와 같은 방법을 이용하여 방제한 예로는 1880년에 독일에서 곰팡이로 딱정벌레목해충을 방제하기 위한 시도가 최초로 1880년에 보고 되었고 그 이후로 현재까지 약 70여종의 미생물을 이용한 방제 방법이 실용화 되었다. However, as soil pollution caused by chemical pesticides and consumer's rejection gradually increased, a new method of control was needed, and one of them was the control method using microorganisms. For example, in 1880, an attempt to control beetle pests with a fungus in Germany was first reported in 1880, and since then, control methods using about 70 microorganisms have been put to practical use.

미생물로 방제하는 방법은 여러 가지가 있으나 대표적인 방제방법 중 하나는 미생물 배양액을 직접 방제에 사용하는 경우이다. 미생물 배양액은 미생물이 살아가기 위한 일차대사물 외에 특수한 목적에 사용되는 이차대사물을 포함하고 있는데 이와 같은 미생물 배양액이 항균효과를 보이는 경우가 종종 있고 대표적인 예로서 방선균의 배양액으로부터 발견된 스트렙토마이신을 들 수 있다. 미생물 배양액에 함유된 항생물질을 단일물질로 사용하여 방제에 사용하면 화학농약이나 별반 다를 바가 없기 때문에 앞서 언급한 바와 같이 토양오염에 대한 우려를 떨쳐내기 어렵다. 그러나 배양액을 사용하는 경우는 토양에 미생물 배양액을 가하여 병원균을 방제한다고 하여도 배양액 대부분이 물이므로 토양을 오염시킬 우려를 제거할 수 있다는 장점을 갖는다. 그러나 미생물이 살아 있는 형태로 사용되는 것이 아니고 미생물의 배양액을 사용하는 것이므로 미생물제제이기 보다는 생화학제제로 분류되는 것이 타당하다.There are many methods for controlling microorganisms, but one of the representative methods for controlling microorganisms is to use microbial cultures for direct control. Microbial cultures contain secondary metabolites that are used for special purposes in addition to the primary metabolites for microorganisms. Such microbial cultures often have antimicrobial effects. Can be. When antibiotics contained in microbial cultures are used as a single substance for control, there is no difference between chemical pesticides and the soil contamination, as mentioned above. However, in the case of using a culture solution, even if the microorganism culture solution is added to the soil to control pathogens, most of the culture solution is water, which has the advantage of eliminating the risk of contaminating the soil. However, because the microorganism is not used in a live form, but using a culture solution of the microorganism, it is reasonable to be classified as a biochemical agent rather than a microbial agent.

따라서 미생물, 특히 토양으로부터 얻은 방선균 배양액이 병원균에 대한 방제 효과를 갖는다면 새로운 생화학농약제제로 사용이 가능하다. Therefore, microbial, especially if the actinomycetes cultured from the soil has a control effect against the pathogen can be used as a new biochemical pesticide formulation.

이에 본 발명자들은 벼 도열병 방제에 뛰어난 효과를 나타내면서, 환경친화적인 생화학농약제제를 개발하고자 예의 노력한 결과, 토양에서 분리한 방선균(Streptomyce sp.)이 벼 도열병에 탁월한 방제효과를 나타내는 것을 확인하고 본 발명을 완성하였다.Therefore, the present inventors have shown an excellent effect on the control of rice blast, and as a result of intensive efforts to develop an eco-friendly biochemical pesticide formulation, it was confirmed that the Streptomyce sp. Isolated from the soil shows excellent control effect on rice blast Was completed.

결국, 본 발명의 주된 목적은 벼 도열병에 대한 방제효과를 가지는 방선균속 신규균주를 제공하는데 있다. After all, the main object of the present invention is to provide a new strain of actinomycetes having a control effect against rice blast.

본 발명의 다른 목적은 상기 균주를 함유하는 미생물농약제제 및 상기 미생물농약제제를 이용하는 것을 특징으로 하는 벼 도열병의 방제방법을 제공하는데 있다.
Another object of the present invention is to provide a microbial pesticide formulation containing the strain and a method for controlling rice blast, characterized in that using the microbial pesticide formulation.

상기의 목적을 달성하기 위하여, 본 발명은 벼 도열병에 대하여 방제효과를 가지는 방선균 속(Streptomyces sp.) 균주를 제공한다. 본 발명에 있어서, 상기의 균주는 방선균 속(Streptomyces sp.) BG2-17(KACC 91011)인 것을 특징으로 할 수 있고, 상기 균주는 서열번호 1의 16S rRNA 염기서열을 가지는 것을 특징으로 할 수 있다.In order to achieve the above object, the present invention provides a Streptomyces sp . Strain having a control effect against rice blast. In the present invention, the strain may be characterized in that the Streptomyces sp . BG2-17 (KACC 91011), the strain may be characterized by having a 16S rRNA sequence of SEQ ID NO: 1. .

본 발명은 또한, 상기 균주 또는 그 배양액을 유효성분으로 함유하는 벼 도열병 방제용 미생물농약제제 및 상기 미생물농약제제를 이용하는 것을 특징으로 하는 벼 도열병의 방제방법을 제공한다.The present invention also provides a microbial pesticide preparation for rice blast control, which contains the strain or its culture as an active ingredient, and a microbial pesticide control method comprising using the microbial pesticide preparation.

본 발명은 또한, 상기 균주를 배양하는 것을 특징으로 하는 벼 도열병 방제용 미생물농약제제의 제조방법을 제공한다. 본 발명에 있어서, 배양은 베넷(Bennet)배지를 사용하여 25~30℃에서 약 10일간 수행하는 것을 특징으로 할 수 있다.The present invention also provides a method for preparing a microbial pesticide for controlling rice blast, characterized in that the strain is cultured. In the present invention, the culture may be characterized by performing about 10 days at 25 ~ 30 ℃ using Bennett (Bennet) medium.

본 발명의 신규한 방선균속(Streptomyces sp.) KACC 91011 균은 항균성을 가진 세균으로서, 서열번호 1의 16S rRNA의 유전자 염기서열을 가지며, 16S rRNA의 전체 염기서열에 있어서 방선균 스트렙토마이세스 라벤둘라(Streptomyces lavendulae)와 96%의 상동성을 가지고, 벼 도열병에 있어서 탁월한 방제효과를 가진다. 본 발명의 방선균 속(Streptomyces sp.) 균주는, 2003년 10월 10일 대한민국 농업생명공학원구원(경기도 수원시 권선구 서둔동 225 (우)441-707)에 방선균 속(Streptomyces sp.) BG2-17이라는 균주명과 기탁번호 KACC 91011로 기탁하였다.The novel Streptomyces sp. KACC 91011 of the present invention is an antimicrobial bacterium, which has a gene sequence of 16S rRNA of SEQ ID NO: 1, and acts as Streptomyces ravendula in the entire sequence of 16S rRNA. Streptomyces lavendulae ) and 96% homology, and excellent control effect in rice blast. Streptomyces sp. Strain of the present invention, a strain called Streptomyces sp. BG2-17 in the Korea Institute of Agricultural Biotechnology Parkwon (225 (W) 441-707, Seodun-dong, Suwon-si, Suwon-si, Gyeonggi-do) on October 10, 2003 It was deposited with the name and accession number KACC 91011.

본 발명의 미생물농약제제는 본 발명의 균주의 배양액 또는 배양물을 포함하는 다양한 제제의 형태 즉 액제, 수화제, 분제, 입제, 유제 등으로 제조하여 사용할 수 있다. Microbial pesticides of the present invention can be prepared and used in the form of a variety of preparations including the culture or culture of the strain of the present invention, that is, liquids, hydrates, powders, granules, emulsions and the like.

이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것으로서, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지는 않는 것은 당업계에서 통상의 지식을 가진 자에게 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention, it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as being limited by these examples.

실시예 1. 신규 방선균속(Example 1 New Actinomycetes Streptomyces Streptomyces sp.) KACC 91011 균주의 분리 및 동정sp.) Isolation and Identification of KACC 91011 Strains

선발된 균주 KACC 91011의 현탁배양을 위하여 방선균 배양에 이용되는 Bennet 배지를 이용하였다. 배지조성은 beef extract 1g/L, yeast extract 1g/L, tryptone 2g/L, glycerol 10g/L이며, 배양온도 28℃와 pH 7.2에서 진탕시키며 10일간 배양하였다.Bennet medium used for actinomycetes culture was used for suspension culture of the selected strain KACC 91011. Medium composition consisted of beef extract 1g / L, yeast extract 1g / L, tryptone 2g / L, glycerol 10g / L, and incubated for 10 days with shaking at 28 ℃ and pH 7.2.

상기 배양된 BG2-17(KACC 91011) 균주의 주사현미경 (Scanning Electron Microscope) 관찰결과, 방선균속(Streptomyces sp.)과 매우 유사하였다 (도 1).Scanning Electron Microscope observation of the cultured BG2-17 (KACC 91011) strain was very similar to the Streptomyces sp. (Fig. 1).

신규균 방선균속(Streptomyces sp.) BG2-17(KACC 91011) 균의 유전적 분석을 실시하고, 이를 동정하기 위하여 16S rRNA 유전자의 염기서열을 다음과 같이 분석하였다. DNA 추출을 위하여 Bennet 배지에 7일간 현탁배양한 후 배양액을 원심분리 하여 cell을 분리하였다. DNA 추출은 Genomic DNA Extraction Kit (Intron)를 이용하여 추출하였다. Genetic analysis of the novel Streptomyces sp. BG2-17 (KACC 91011) was performed, and the nucleotide sequence of the 16S rRNA gene was analyzed as follows. The suspension was cultured in Bennet medium for 7 days for DNA extraction, and then the cells were separated by centrifugation. DNA extraction was performed using Genomic DNA Extraction Kit (Intron).

PCR은 반응용액(10mM Tris, pH 8.3; 50mM KCl; 1.5mM MgCl2; 0.2mM dNTPs; 1unit Taq DNA polymerase)에 0.2㎍ genomic DNA , 0.25μM 16S rRNA 유전자 프라이머 4 쌍이 표 1과 같이 조합하여 증폭하였다. 증폭된 DNA 절편의 크기가 A조합은 900bp, B조합은 480bp, C조합은 1200bp, D조합은 800bp가 되도록 하였다. PCR(초기 94℃ 5분 1회; 94℃ 1분, 55℃ 1분, 72℃ 1분, 35cycle; 72℃ 5분 1회)을 수행한 후, 증폭된 16S rRNA 유전자 일부분을 pGEM-T Easy vector (Promega co.)에 ligation한 후, 대장균 DH5α에 형질전환하였다.PCR was amplified by combining four pairs of 0.2 μg genomic DNA and 0.25 μM 16S rRNA gene primers in a reaction solution (10 mM Tris, pH 8.3; 50 mM KCl; 1.5 mM MgCl 2 ; 0.2 mM dNTPs; 1 unit Taq DNA polymerase) as shown in Table 1. . The size of the amplified DNA fragment was 900bp for A combination, 480bp for B combination, 1200bp for C combination, and 800bp for D combination. After PCR (initial 94 ° C. 5 min once; 94 ° C. 1 min, 55 ° C. 1 min, 72 ° C. 1 min, 35 cycles; 72 ° C. 5 min once), a portion of the amplified 16S rRNA gene was transferred to pGEM-T Easy. After ligation into the vector (Promega co.), E. coli DH5α was transformed.

형질전환된 대장균을 선발배지(1.5% agar, 50mg/L ampicillin, 100μM IPTG, 50mg/L X-gal)에 도발하여 37℃에서 12 시간 배양하였다. 증폭된 16S rRNA 유전자조각의 vector로의 삽입은 blue/white colony로 1차 선발하고, 이중 white colony만을 LB배지에 접종하여 배양하였다. 배양액은 플라스미드분리를 위하여 원심분리한 후 Alkaline lysis방법을 이용하였다. 분리된 플라스미드는 제한효소 EcoRI으로 절단 후 전기영동을 통하여 증폭된 16S rRNA 유전자조각의 삽입을 확인하였다.Transformed Escherichia coli was provoked in selection medium (1.5% agar, 50 mg / L ampicillin, 100 μM IPTG, 50 mg / L X-gal) and incubated at 37 ° C. for 12 hours. Insertion of the amplified 16S rRNA fragment into the vector was first selected as a blue / white colony, and only white colony was inoculated into LB medium and cultured. The culture was centrifuged for plasmid separation and Alkaline lysis was used. The isolated plasmid was digested with restriction enzyme Eco RI and confirmed insertion of amplified 16S rRNA fragment through electrophoresis.

증폭된 16S rRNA 유전자를 염기서열분석장치(ABI3100)를 통하여 분석하였다. 이를 NCBI(National Center for Biotechnology Information)의 BLAST를 이용하여 homology를 확인하였고, 그 결과 방선균 스트렙토마이세스 라벤둘라 (Streptomyces lavendulae)와 96%의 상동성을 보였다 (도 2).The amplified 16S rRNA gene was analyzed through a sequencing device (ABI3100). The homology was confirmed using BLAST of the National Center for Biotechnology Information (NCBI), and as a result, it showed 96% homology with Streptomyces lavendulae (FIG. 2).

16S rRNA 유전자의 PCR 및 염기서열분석에 사용된 프라이머Primers used for PCR and sequencing of 16S rRNA genes 프라이머 쌍Primer pair 염 기 서 열Base standing 서열번호SEQ ID NO: AA 5'-CAAGCGTTGTCCGGAATTAT-3'5'-CAAGCGTTGTCCGGAATTAT-3 ' 22 5'-TTCGGGTGTTACCGACTTTC-3'5'-TTCGGGTGTTACCGACTTTC-3 ' 33 BB 5'-CAAGCGTTGTCCGGAATTAT-3'5'-CAAGCGTTGTCCGGAATTAT-3 ' 44 5'-ATCTCTGGATGTTTCCGGT-3'5'-ATCTCTGGATGTTTCCGGT-3 ' 55 CC 5'-GATACGACTCAGGACCGCAT-3'5'-GATACGACTCAGGACCGCAT-3 ' 66 5'-TTCGGGTGTTACCGACTTTC-3'5'-TTCGGGTGTTACCGACTTTC-3 ' 77 DD 5'-GATACGACTCAGGACCGCAT-3'5'-GATACGACTCAGGACCGCAT-3 ' 88 5'-ATCTCTGGATGTTTCCGGT-3'5'-ATCTCTGGATGTTTCCGGT-3 ' 99

실시예 2: 방선균속 BG2-17(KACC 91011) 균 배양액의 벼 도열병 방제효과Example 2: Effect of Actinomycetes BG2-17 (KACC 91011) Culture on Rice Blast Control

시험 대상 벼 품종으로 Oryza sativa cv Nakdong을 사용하였고, 벼 도열병균으로 Magnaporthe grisea을 사용하였으며, 벼풍종 낙동에 대하여 감수성인 KJ201 레이스를 사용하였다. 벼 종자를 최아시켜 수도용상토(부농사)에 파종하고 25± 5℃의 온실에서 약 4주 동안 재배하여 준비한 약 3~4엽기의 벼를 사용하여 방제효과를 조사하였다. Oryza sativa cv Nakdong was used as a rice cultivar to be tested, Magnaporthe grisea was used as rice blast bacterium, and KJ201 lace, which was susceptible to rice cultivation, was used. Rice seeds were seeded and sown on aquatic soils (sub-farms), and the control effect was investigated using about 3-4 leaf rice plants prepared by cultivation for about 4 weeks in a greenhouse of 25 ± 5 ℃.

벼 도열병균을 감자즙액배지(potato dextrose broth, PDB)에 접종하고 25℃에서 7일 동안 진탕배양 하였다. 액체배지에서 자란 균사체를 Waring blender를 사용하여 자른 후에 멸균하여 식힌 쌀겨배지(쌀겨 20g, dextrose 10g, agar 2g, 증류 수 1L)에 넣고 잘 흔들어주고 Petri-dish에 부어 굳힌 후에 25℃ 항온기에서 7일 동안 배양하였다. 배양한 균의 공중균사를 제거하고 형광등 하에 두어 형성된 포자를 수확하고 포자농도를 ml 당 5 x 105 개로 조정하고 이를 접종원으로 사용하였다. Rice blast bacteria were inoculated in potato juice medium (potato dextrose broth, PDB) and shaken at 25 ° C. for 7 days. Mycelium grown in liquid medium was cut using a Waring blender, sterilized and put in a cooled rice bran medium (rice bran 20g, dextrose 10g, agar 2g, distilled water 1L), shaken well, poured into Petri-dish and hardened. Incubated for Spores formed by removing the aerial hyphae of the cultured bacteria and placed under a fluorescent lamp were harvested, and the concentration of spores was adjusted to 5 x 10 5 per ml and used as an inoculum.

미생물 배양액의 방제효과를 조사하기 위하여 미생물 배양액에 250mg/L 농도가 되도록 Tween 20을 첨가하고 벼 폿트에 경엽처리 한 후에 24시간 동안 풍건하였다. 미생물 배양액을 처리한 벼 잎에 위에서 준비한 벼 도열병균 포자현탁액을 spray하여 접종하였다. 접종한 벼는 25℃ 습실상에 넣고 1일 동안 습실처리한 후에 25℃, 상대습도 80%의 항온항습실에서 4일 동안 발병을 유도하며 재배하고 병반면적율을 조사하였다. In order to investigate the control effect of the microbial culture medium, Tween 20 was added to the concentration of 250 mg / L in the microbial culture medium, and the leaves were air-dried in rice pots for 24 hours. The rice leaves treated with the microbial culture solution were inoculated by spraying the spore suspension of rice blast fungus prepared above. Inoculated rice was incubated for 1 day after incubation at 25 ° C. incubation for 4 days in a constant temperature and humidity room at 25 ° C. and a relative humidity of 80%.

실시예 1과 같은 방법으로 배양한 방선균속(Streptomyces sp.) BG2-17(KACC 91011) 균의 배양액(미생물 세포내 물질이 포함된)을 처리한 결과와 무처리구를 비교하였다 (도 3). 일반적으로 Magnaporthe grisea에 의해 벼 도열병이 심하게 발생하면 잎이 죽게 된다. 모두 Magnaporthe grisea를 접종하여 발병시킨 결과, 도 3에 나타난 바와 같이, 무처리구(왼쪽)에서는 잎이 모두 죽은 것으로 관찰되었고, 방선균속(Streptomyces sp.) BG2-17(KACC 91011) 균의 배양액 처리구 (중앙)와 기존에 도열병 방제를 위해 사용되고 있는 화학농약 트리사이클라졸(tricyclazole) 처리구(오른쪽)에서는 방제효과로 인해 살아남은 것으로 관찰되었다.The results of treatment of the culture solution (containing microbial intracellular material) of Streptomyces sp. BG2-17 (KACC 91011) cultured in the same manner as in Example 1 were compared with the untreated group (FIG. 3). In general, a severe rice infestation caused by Magnaporthe grisea causes the leaves to die. As a result of inoculation with Magnaporthe grisea , as shown in FIG. 3, all leaves were observed to be dead in the non-treated group (left), and cultured treatment group of Streptomyces sp. BG2-17 (KACC 91011) ) And the chemical pesticide tricyclazole treatment group (right), which has been used for the control of blasting disease (right), has been observed to survive the control effect.

병의 발생을 억제하는 방제가는 다음의 식으로 계산하였다.The control value which suppresses disease occurrence was calculated by the following formula.

방제가(%)={1-(처리구의 병반면적율/무처리구의 병반면적율)}x100Control value (%) = {1- (Legn area ratio of treated area / Long area ratio of untreated area)} x100

방선균속(Streptomyces sp.) KACC 91011 균의 배양액을 처리한 경우, 벼 도열병 방제가가 90%로 나타났다. 이 결과로부터, 본 발명의 KACC 91011 균은 우수한 벼 도열병 방제효과를 가지는 것을 확인할 수 있었다.In case of treatment with the culture solution of Streptomyces sp. KACC 91011, rice blast control value was 90%. From these results, it was confirmed that the KACC 91011 bacterium of the present invention has excellent rice blast control effect.

이상으로 본 발명 내용의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적 기술은 단지 바람직한 실시양태일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의하여 정의된다고 할 것이다.Having described the specific part of the present invention in detail, it is obvious to those skilled in the art that such a specific description is only a preferred embodiment, thereby not limiting the scope of the present invention. something to do. Thus, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

이상 상세히 기술한 바와 같이, 본 발명은 벼 도열병 방제효과를 가진 신규 방선균 속(Streptomyces sp.) BG2-17(KACC 91011) 균주, 상기 균주를 함유하는 미생물농약제제 및 상기 미생물농약제제를 이용한 벼 도열병의 방제방법을 제공하는 효과가 있다. 본 발명에 따른 균주는, 기존의 화학농약에서 나타날 수 있는 과다사용으로 인한 생태계의 파괴 없이도, 기존 화학농약보다 높은 벼 도열병 방제효과를 나타내므로 생물농약으로 유용하다.As described in detail above, the present invention provides a novel Streptomyces sp. BG2-17 (KACC 91011) strain having a rice blast control effect, a microbial pesticide formulation containing the strain, and a rice blast bottle using the microbial pesticide formulation. It is effective to provide a control method. The strain according to the present invention is useful as a biopesticide because it exhibits a higher rice blast control effect than conventional chemical pesticides without destroying the ecosystem due to overuse that may occur in conventional chemical pesticides.

서열목록 전자파일 첨부 Attach sequence list electronic file  

Claims (8)

벼 도열병에 대하여 방제효과를 가지는 것을 특징으로 하는 방선균 속(Streptomyces sp.) BG2-17 (KACC 91011) 균주. Streptomyces sp . BG2-17 (KACC 91011) strain, characterized in that it has a control effect against rice blast. 삭제delete 제1항에 있어서, 상기 균주는 서열번호 1의 16S rRNA 염기서열을 가지는 것을 특징으로 하는 균주.The strain of claim 1, wherein the strain has a 16S rRNA nucleotide sequence of SEQ ID NO: 1. 제1항 또는 제3항에 있어서, 상기 균주 또는 그 배양액을 유효성분으로 함유하는 미생물농약제제.The microbial pesticide preparation according to claim 1 or 3, wherein said microorganism pesticide formulation contains said strain or its culture solution as an active ingredient. 제4항에 있어서, 벼 도열병 방제용인 것을 특징으로 하는 미생물농약제제.The microbial pesticide preparation according to claim 4, which is for controlling rice blast. 제1항 또는 제3항에 있어서, 상기 균주를 배양하는 것을 특징으로 하는 벼 도열병 방제용 미생물농약제제의 제조방법.The method for producing a microbial pesticide preparation for rice blast control according to claim 1 or 3, wherein the strain is cultured. 제6항에 있어서, 배양은 베넷(Bennet)배지를 사용하여 25~30℃에서 10일간 수행하는 것을 특징으로 하는 방법.The method of claim 6, wherein the culturing is carried out for 10 days at 25 ~ 30 ℃ using Bennett (Bennet) medium. 제4항의 미생물농약제제를 이용하는 것을 특징으로 하는 벼 도열병의 방제방법.A method for controlling rice blast, characterized by using the microorganism pesticide preparation of claim 4.
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Citations (3)

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KR950026979A (en) * 1994-03-17 1995-10-16 신준식 New Antibiotic Antifungal Antagonist Strains Streptomyces Violaseoniger
KR20020017557A (en) * 2000-08-31 2002-03-07 서형원 Antifungal Biocontrol Agents and Use thereof
KR20020050672A (en) * 2000-12-21 2002-06-27 복성해 A new Streptomyces sp. AG-P(KCTC 8965P), and an agent for control of plant diseases using this strain

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Publication number Priority date Publication date Assignee Title
KR950026979A (en) * 1994-03-17 1995-10-16 신준식 New Antibiotic Antifungal Antagonist Strains Streptomyces Violaseoniger
KR20020017557A (en) * 2000-08-31 2002-03-07 서형원 Antifungal Biocontrol Agents and Use thereof
KR20020050672A (en) * 2000-12-21 2002-06-27 복성해 A new Streptomyces sp. AG-P(KCTC 8965P), and an agent for control of plant diseases using this strain

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