KR100986653B1 - Biological control of plant diseases using pseudomonas sp. pf-1 - Google Patents

Biological control of plant diseases using pseudomonas sp. pf-1 Download PDF

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KR100986653B1
KR100986653B1 KR1020100028677A KR20100028677A KR100986653B1 KR 100986653 B1 KR100986653 B1 KR 100986653B1 KR 1020100028677 A KR1020100028677 A KR 1020100028677A KR 20100028677 A KR20100028677 A KR 20100028677A KR 100986653 B1 KR100986653 B1 KR 100986653B1
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Abstract

PURPOSE: An agent for preventing plant diseases using Pseudomonas sp. PF-1 without problems such as remaining toxicity and environmental harmfulness. CONSTITUTION: A Pseudomonas sp. PF-1(deposit number: KACC91536P) prevents plant pathogen of Sclerotium cepivorum, Pythium myriotylum, or Sphaerotheca macular in garlic, strawberry or watermelon. A microorganism agent for preventing plant pathogen contains Pseudomonas sp. PF-1(deposit number KACC91536P) or culture medium thereof as an active ingredient.

Description

슈도모나스 에스피 피에프-1 균주를 이용한 식물병 방제제{Biological control of plant diseases using Pseudomonas sp. PF-1}Biological control of plant diseases using Pseudomonas sp. PF-1}

본 발명은 토양에서 분리한 슈도모나스 에스피 피에프-1(Pseudomonas sp . PF-1, 미생물기탁번호 KACC91536P)균주와 이를 배양한 배양물을 이용한 식물병 방제제에 대한 것이다.The present invention is Pseudomonas Pseudomonas sp. sp . PF-1, Microbial Accession No. KACC91536P) strain and a plant disease control agent using the culture culture thereof.

식물의 병해충에 대한 방제방법은 크게 유기합성 방제제 등을 이용한 화학적 방제법과 천적 등을 이용한 생물학적 방제법으로 나눌 수 있다. 1900년대 초부터 사용하기 시작한 유기합성 방제제는 근래까지 작물보호의 수단으로서 매우 유효하게 사용되어 식량 생산의 증대, 노동력 절감 등에 크게 기여하였다. 그러나 최근들어, 선진국을 중심으로 농식품의 안전성, 생태계의 환경 악화 등이 크게 인식되면서 친환경적 작물 생산에 관한 관심이 높아짐에 따라 이러한 합성 방제제에 대한 사회적 환경은 매우 엄격해지고 있다. Plant pest control methods can be largely divided into chemical control methods using organic synthetic control agents and biological control methods using natural enemies. Organic synthetic control agents, which have been in use since the early 1900s, have been very effective as a means of crop protection until recently, contributing to the increase of food production and labor reduction. Recently, however, with the growing awareness of agri-food safety and deterioration of the environment of ecosystems, especially in developed countries, the social environment for these synthetic control agents has become very strict.

따라서, 최근에는 식물병의 생물학적 방제에 대한 관심이 증대되고 있다. 생물학적 방제는 소비자의 안전한 농산물 요구증대에 부응하여 환경에 유해한 화학적 농약을 사용하는 대신에 미생물 또는 식물 추출물을 이용하여 환경, 다른 식물 및 인축에는 영향을 미치지 않으면서 대상 병을 선택적으로 방제한다. 이로 인해, 미생물 살균제를 이용한 친환경 농업을 추구하는 재배농가들이 급속히 증가하는 추세이고 효율적이며 보다 안전한 농산물 생산을 위한 방제기술 개발이 요구되고 있다(Dennis와 Davis, 1979; Hunter 등, 1987; Katan, 1980; Kim 등, 1995).Therefore, in recent years, interest in biological control of plant diseases has increased. Instead of using chemically harmful pesticides that are harmful to the environment in response to increasing consumer demand for safe agricultural products, biological control selectively uses microorganisms or plant extracts to selectively control the disease without affecting the environment, other plants, and killing. Accordingly, the growing number of farmers pursuing eco-friendly agriculture using microbial fungicides is rapidly increasing, and development of control techniques for producing more efficient and safer agricultural products is required (Dennis and Davis, 1979; Hunter et al., 1987; Katan, 1980; Kim et al., 1995).

국내에 보고된 딸기, 상추, 수박 등의 식물병은 방제가 까다로우면서도 그 효과가 그다지 높지 않다. 또한 방제비용도 비싸고 반복적으로 사용할 때는 약제에 대한 저항성도 발생할 수 있다. 또한 화학농약 자체가 사람과 동물에게 유해하기 때문에 농약 잔류 문제 등의 문제점을 가지고 있다(Asari, et al., 1994; Erikson et al., 1998). 특히, 상추의 경우는 쌈 등으로 이용되고 있기 때문에 소비자로부터 농약에 대한 안전성 문제가 계속 대두되고 있고 이로 인해 화학농약의 사용도 회피되고 있어 농가에서 많은 어려움을 겪고 있다. Plant diseases such as strawberries, lettuce, and watermelon reported in Korea are difficult to control, but the effect is not so high. In addition, control costs are high and resistance to drugs can occur when used repeatedly. In addition, because chemical pesticides are harmful to humans and animals, they have problems such as pesticide residues (Asari, et al., 1994; Erikson et al., 1998). In particular, since the lettuce is used as a wrapper, safety problems for pesticides continue to emerge from consumers, and thus, the use of chemical pesticides is being avoided, which causes many difficulties in farmers.

이를 위해 본 발명에서는 국내 부존자원인 유용 미생물을 분리, 선발, 이용하여 식물병원균의 발생을 억제함으로써 안전하고 품질이 좋은 딸기, 상추, 수박을 비롯한 작물들을 생산하고자 한다. 본 발명의 신규한 슈도모나스 에스피 피에프-1 (Pseudomonas sp . PF-1) 균주는 여러 식물 병원균의 생장 억제효과가 우수하며 화학농약을 대체할 수 있는 미생물로서, 잔류농약으로 인한 농업 생태계 문제점을 방지하고 친환경적 방제에 효과적으로 이용할 수 있다.To this end, the present invention is to produce crops, including strawberries, lettuce and watermelon, which are safe and of good quality by inhibiting the generation of phytopathogens by separating, selecting and using useful microorganisms, which are domestic resources. Pseudomonas novel Pseudomonas sp. F-1 of the present invention sp . PF-1) strain is a microorganism that has excellent growth inhibitory effect of various plant pathogens and can replace chemical pesticides, and can effectively prevent agricultural ecosystem problems caused by residual pesticides and effectively use it for environmentally friendly control.

본 발명은 식물 병원균에 대해 항균활성을 갖는 슈도모나스 에스피 피에프-1(Pseudomonas sp. PF-1, 미생물기탁번호 KACC91536P) 균주를 이용하여 환경 친화적으로 여러 가지 식물 병원균을 방제하는 미생물 제제를 제공하는 데에 있다. The present invention provides a microbial agent for controlling various plant pathogens in an environmentally friendly manner using Pseudomonas sp. PF-1 (Microorganism Accession No. KACC91536P) strain having antibacterial activity against plant pathogens . Is in.

본 발명은 슈도모나스 에스피 피에프-1을 이용한 식물병 방제제와 방제 방법을 그 구성으로 한다.This invention makes the composition the plant disease control agent and the control method using Pseudomonas sp.

먼저 토양에서 균주를 분리하여, 생물검정을 통하여 식물병 진전 억제 효과가 우수한 균주를 선발하고 이 후에 형태적, 생리 생화학적 특성 및 DNA 프로파일(profiles) 비교 등을 통하여 균주를 동정하였고 상기 균주는 슈도모나스 에스피 피에프-1(Pseudomonas sp . PF-1, 미생물기탁번호 KACC91536P)으로 동정되었다. First, isolates were isolated from soil, and biopsies were used to select strains with excellent inhibitory effects on plant disease. Afterwards, strains were identified through morphological, physiological, biochemical properties, and DNA profiles. Pseudomonas- 1 sp . PF-1, microbial accession number KACC91536P).

본 발명은 슈도모나스 에스피 피에프-1 균주의 배양조건을 제공한다. 산업적 사용을 위해서는 상기 균주를 단시간 내에 대량으로 배양할 수 있는 방법이 필수적이다. 본 발명의 슈도모나스 에스피 피에프-1 균주는 배양 배지로서 NA(Nutrient agar) 배지 상에서 잘 자라며, 15~37℃ 온도범위에서 생육이 가능하고, 생육 pH 범위는 5.0~7.5이고, 배양시간은 72시간에서 잘 자란다. The present invention provides a culture condition of Pseudomonas sp. PI-1 strain. For industrial use, a method of culturing the strain in large quantities in a short time is essential. Pseudomonas S.P.P-1 strain of the present invention grows well on the culture medium (Nutrient agar) as a culture medium, it is possible to grow in the temperature range of 15 ~ 37 ℃, the growth pH range is 5.0 ~ 7.5, the culture time is 72 hours Grows well in

본 발명은 슈도모나스 에스피 피에프-1 균주를 이용한 식물 병원균 방제제 및 이를 이용한 식물 병원균의 방제방법을 제공한다.The present invention provides a plant pathogen control agent using Pseudomonas S.P.P. strain 1 and a method for controlling plant pathogens using the same.

본 발명의 슈도모나스 에스피 피에프-1 균주를 이용하여 방제할 수 있는 처리 대상균은 스크레로티움 세피보룸(Sclerotium cepivorum), 피시움 미리오틸륨(Pythium myriotylum), 콜레토트리쿰 글로에스포리오이데스(Colletotrichum gloeosporioides), 파이토프토라 캡시시(Phytophthora capsici), 보트리티스 시네리아(Botrytis cinerea), 스크레로티니아 스크레로티오룸(Sclerotinia sclerotiorum), 스파에로테카 마쿠라(Sphaerotheca macular), 후사리움 옥시스포룸 프라가리에(Fusarium oxysporum f. sp. fragariae) 등이 해당된다. 상기 균주들은 딸기, 상추, 수박, 오이, 마늘, 배추, 고추를 비롯한 식물의 잿빛곰팡이병, 흰가루병, 시들음병, 탄저병, 역병, 균핵병, 흑색썩음균핵병, 뿌리썩음병을 비롯한 다양한 식물병을 유발한다. Treatment target bacteria that can be controlled using the Pseudomonas S.P.P. strain 1 of the present invention are Sclerotium cepivorum , Pythium myriotylum , and choletotricum gloespo Colletotrichum gloeosporioides , Phytophthora capsici , Botrytis cinerea , Sclerotinia sclerotiorum , Sphaerotheca macular ), Fusarium oxysporum f . Sp.fragariae And the like. These strains cause various plant diseases including strawberries, lettuce, watermelon, cucumber, garlic, cabbage, pepper, asymptomatic disease, powdery mildew, wilting disease, anthrax, late blight, fungal disease, black rot fungal disease, root rot disease.

본 발명의 슈도모나스 에스피 피에프-1 균주는 상기 식물 병원균의 생육을 저지하여 방제할 수 있다. Pseudomonas sp. PI-1 strain of the present invention can be controlled by inhibiting the growth of the plant pathogen.

이상에서 상세히 설명한 바와 같이 본 발명에 따른 신규 슈도모나스 에스피 피에프-1(Pseudomonas sp . PF-1, 미생물기탁번호 KACC91536P) 균주를 이용한 미생물 제제는 딸기, 상추, 수박을 비롯한 여러 작물의 식물 병원균들을 효과적으로 방제할 수 있었다.As described in detail above, novel Pseudomonas sp. F-1 according to the present invention ( Pseudomonas sp . PF-1, microbial accession number KACC91536P) strain was able to effectively control plant pathogens of various crops, including strawberries, lettuce, watermelon.

또한 안전한 농산물을 생산가능하게 함으로써 농가의 소득증대 및 소비자의 농약에 대한 문제 제기를 불식시키고 화학농약의 사용을 절감하거나 대체할 수 있기 때문에 환경오염을 줄일 수 있다.In addition, by enabling the production of safe agricultural products, it is possible to reduce the environmental pollution by increasing the income of farmers and raising the issue of pesticides of consumers and reducing or replacing the use of chemical pesticides.

따라서 본 발명은 재배 중에 발생하는 각종 채소류의 균핵병 등에 대한 생물농약으로서 기능할 수 있어 농약 잔류로 인한 문제점을 방지할 수 있고, 농업 생태계 보전과 병해 종합방제를 위한 획기적인 생물학적 방제방법을 제공하는 효과가 있다. Therefore, the present invention can function as a biopesticide against fungal nucleus diseases of various vegetables occurring during the cultivation to prevent problems caused by pesticide residues, and to provide an innovative biological control method for the preservation of agricultural ecosystems and comprehensive control of the disease. have.

도 1은 슈도모나스 에스피 피에프-1 균주를 NA(Nutrient Agar)배지에 배양하였을 때의 콜로니 사진이다.
도 2는 슈도모나스 에스피 피에프-1 균주의 16S rDNA gene의 염기서열이다.
도 3은 슈도모나스 에스피 피에프-1 균주의 16S rDNA gene의 계통학적 위치를 나타내는 서식도이다.1 is a photograph of colonies when Pseudomonas sp. PI-1 strain was cultured in NA (Nutrient Agar) medium.
Figure 2 is the nucleotide sequence of 16S rDNA gene of Pseudomonas S.P.P. strain.
Figure 3 is a format diagram showing the phylogenetic location of the 16S rDNA gene of Pseudomonas S.P.P-1 strain.

이하에서는 본 발명에 따른 슈도모나스 에스피 피에프-1(Pseudomonas sp. PF-1, 미생물기탁번호 KACC91536P)를 이용한 미생물 제제 및 이용방법의 바람직한 실시예를 참고하여 상세히 설명하기로 한다. 하기 실시예는 단지 본 발명을 구체적으로 설명하고자 하는 것으로, 본 발명의 보호범위가 하기 실시예에 한정되는 것은 아니다.Hereinafter, with reference to the preferred embodiment of the microorganism preparation and the method using Pseudomonas sp. PF-1 (Microbial Accession Number KACC91536P) according to the present invention will be described in detail. The following examples are only intended to illustrate the present invention in detail, and the protection scope of the present invention is not limited to the following examples.

<실시예 1 : 길항미생물의 분리 및 항균 활성 측정>Example 1 Isolation and Antibacterial Activity of Antagonists

본 발명의 길항미생물은 토양에서 분리하였다. 미생물의 분리를 위해 토양 시료를 잘 혼합한 후 멸균수에 희석하여 NA(Nutrient agar) 배지에 희석평판법으로 미생물을 분리하여 27℃에서 5일간 배양하였다. 이곳에서 분리된 세균을 NA 배지로 옮겨 순수분리하여 보관 후, 한천확산법(Paper disc method)을 이용하여 주요 식물병원균과 대치배양하여 균사생장을 억제하는 길항 미생물을 선별하였고 그 중 가장 우세한 길항미생물의 길항 결과를 표 1에 나타내었다. Antagonist microorganisms of the present invention were isolated from the soil. Soil samples were mixed well for separation of microorganisms, diluted in sterile water, and microorganisms were separated by dilution plate method in NA (Nutrient agar) medium and incubated at 27 ° C. for 5 days. After transferring the isolated bacteria to NA medium for pure separation and storing, the antagonistic microorganisms inhibiting mycelial growth were selected by replacing them with major phytopathogens using the paper disc method. Antagonistic results are shown in Table 1.

Figure 112010020312632-pat00001
Figure 112010020312632-pat00001

표 1은 슈도모나스 에스피 피에프-1의 길항력을 측정한 결과를 나타낸다. 표 1의 결과를 확인하면, 본 발명의 길항미생물은 흑색썩음균핵병, 뿌리썩음병, 탄저병, 역병, 잿빛곰팡이병, 균핵병, 흰가루병, 시들음병에 우수한 항균활성 효과가 있는 것으로 조사되었다. Table 1 shows the results of measuring the antagonistic force of Pseudomonas sp. Confirming the results of Table 1, the antagonistic microorganisms of the present invention was found to have an excellent antimicrobial activity against black rot fungal nucleus disease, root rot disease, anthrax disease, late blight, gray mold fungus, fungal nucleus disease, powdery mildew, wilt disease.

균주의 장기 보관을 위해 상기 길항미생물 슈도모나스 에스피 피에프-1은 NB(Nutrient broth)에서 약 3일간 배양하여 배양액을 멸균된 20% 글리세롤(glycerol) 튜브에 넣고 잘 혼합하여 보존균주를 만들어 -70℃ 냉동고에 저장하였다. For long-term storage of the strain, the antagonist Pseudomonas S.P.P-1 was incubated for about 3 days in NB (Nutrient broth), and the culture medium was added to a sterile 20% glycerol tube and mixed well to make a conserved strain. Store in freezer.

<실시예 2 : 길항미생물 슈도모나스 에스피 피에프-1의 16S ribosomal DNA (rDNA) gene 염기서열 분석> <Example 2: 16S ribosomal DNA (rDNA) gene sequencing analysis of the antagonist Pseudomonas S.P.P-1>

분자생물학적 동정은 DNA를 분리(Benzyl chloride법을 변형한 방법 이용)하여 16S rDNA의 염기서열을 밝혀 계통분류학적 위치를 확인하였다. 길항미생물 슈도모나스 에스피 피에프-1의 16S rDNA를 증폭하기 위해 27F(5′-AGA GTT TGA TCC TGG CTC AG-3′)와 1492r(5'-GGT TAC CTT GTT ACG ACT T-3') 프라이머를 사용하였다. 16S rDNA의 PCR 증폭산물은 PCR Product Purification Kit (Qiagen)를 사용하여 정제하였고, 16S rDNA 유전자의 염기서열은 Genetic analyzer 310A(Applied Biosystems)를 사용하여 분석하였다. 염기서열은 NCBI/Genebank와 Ribosomal Database Project Ⅱ(RDP Ⅱ)의 데이터베이스에서 상동성 검색을 수행하고, CLUSTAL X 프로그램(Thompson et al., 1994) 및 PHYLIP 프로그램(Felsenstein, 1993)을 이용하여 계통학적 위치를 확인하였다. 16S rDNA의 염기서열(도 2 참조)에 기초한 분자계통학적 분석 결과, 슈도모나스(Pseudomonas)속의 종을 포함하는 계통학적 그룹에 속하는 균주로서 슈도모나스 프라기 ATCC4973(Pseudomonas fragi ATCC4973, AF094733)와 99.6%, 슈도모나스 싸이크로필라(Pseudomonas psychrophila, AB041885)와 99.4%의 유연관계를 나타내어 슈도모나스 에스피(Pseudomonas sp.)임이 확인되었다.(도 3 참조).Molecular biological identification was confirmed by identifying the nucleotide sequence of 16S rDNA by separating DNA (using modified method of Benzyl chloride method). Primers 27F (5′-AGA GTT TGA TCC TGG CTC AG-3 ′) and 1492r (5′-GGT TAC CTT GTT ACG ACT T-3 ′) primers were used to amplify 16S rDNA of antagonist Pseudomonas sp. Used. PCR amplification products of 16S rDNA were purified using PCR Product Purification Kit (Qiagen), and the base sequence of 16S rDNA gene was analyzed using Genetic analyzer 310A (Applied Biosystems). The sequences were subjected to homology searches in the NCBI / Genebank and Ribosomal Database Project II (RDP II) databases, and systematically located using the CLUSTAL X program (Thompson et al., 1994) and the PHYLIP program (Felsenstein, 1993). It was confirmed. Molecular systematic analysis based on the nucleotide sequence of 16S rDNA (see FIG. 2), Pseudomonas ( Pseudomonas fragi ATCC4973, 99.6% Pseudomonas fragi ATCC4973, Pseudomonas psychrophila, AB041885) and 99.4% Pseudomonas strain Pseudomonas sp. Was confirmed (see FIG. 3).

상기 결과에 따라, 본 발명의 길항미생물은 슈도모나스 에스피(Pseudomonas sp.)로 동정되었고 슈도모나스 에스피 피에프-1(Pseudomonas sp. PF-1)으로 명명하여 농촌진흥청 한국농업미생물자원센터에 미생물기탁번호 KACC91536P로 기탁하였다.Depending on the result, the antagonistic microorganism of the present invention has been identified as Pseudomonas sp (Pseudomonas sp.) Pseudomonas sp blood F -1 (Pseudomonas sp. PF-1 ) to the deposit of micro-organisms named in the Korea National Institute of Agricultural Microbiology Resource Center number KACC91536P Was deposited.

<실시예 3 : 슈도모나스 에스피 피에프-1 균주의 딸기 흰가루병 방제효과> <Example 3: Strawberry powdery mildew control effect of Pseudomonas sp. PI-1 strain>

길항미생물 슈도모나스 에스피 피에프-1의 딸기 흰가루병 방제효과를 딸기에서 직접 확인하기 위해 330㎡의 비닐하우스에 흰가루병 균주 스파에로테카 마쿠라(Sphaerotheca macular)를 밀곡립배지에서 25일간 배양한 병원균 접종원을 1㎡ 당 4.5g씩 접종하고, 24시간 후에 슈도모나스 에스피 피에프-1을 NB 배지에서 3일간 진탕배양한 후 배양을 종료하였다. 상기 배양물은 분무건조기(spray dryer) 또는 동결건조기(freeze dryer)를 이용하여 배양액을 분말화하였고, 분말화한 배양물 300g, 계면활성제(습윤제) 10g, 증량제 670g, 보존제 10g, 무기염 10g을 혼합 후 에어밀링(air milling)을 통해 수화제(WP) 제형을 제조하였다. In order to confirm the control of strawberry powdery mildew against the antagonist Pseudomonas sp.PI-1 in strawberry, a pathogen inoculum cultivated with powdery mildew strain Sphaerotheca macular in wheat grain medium for 25 days in 330㎡ plastic house. 4.5 g per 1 m 2 was inoculated, and after 24 hours, Pseudomonas sp. PI-1 was shaken in NB medium for 3 days, and then culture was terminated. The cultured powder was cultured using a spray dryer or freeze dryer, 300g powdered culture, 10g surfactant (wetting agent), 670g extender, 10g preservative, 10g inorganic salt Hydrating (WP) formulations were prepared via air milling after mixing.

상기 제형제는 물에 500배로 희석하여 동력분무기로 7일 간격으로 흰가루병이 주로 발생하는 기간에 딸기에 3회 경엽처리하였다. 최종 약제처리 7일 후 딸기흰가루병의 발생정도를 시험구 전체에 있는 딸기과실수 대비 딸기흰가루병이 발생한 과실수인 이병과율로 조사하여 방제효과를 평가하였고 각 실험은 세 번 반복되었다. 대조약제로 화학약제인 Triflumizole 30% WP(일반명: 리프졸 수화제)를 사용하였다. 표 2의 결과는, 본 발명의 수화제가 딸기흰가루병에 91.4%의 방제효과를 보여 70.3%의 방제효과를 나타내는 대조약제인 화학농약 Triflumizole 30% 수화제보다 월등한 것을 나타낸다. The formulation was diluted 500 times in water, and three leaves were treated with strawberries three times in the period of powdery mildew disease at 7 days intervals with a power sprayer. Seven days after the final treatment, the incidence of strawberry powdery mildew was evaluated by the fruiting rate, which is the number of fruits of strawberry powdery mildew compared to that of the whole fruit, and each control was repeated three times. As a control drug, Triflumizole 30% WP (general name: Repazole hydrous) was used. The results of Table 2 show that the hydrating agent of the present invention was superior to the chemical pesticide Triflumizole 30% hydrating agent, a control agent exhibiting a 91.4% control effect on strawberry powdery mildew disease, showing a 70.3% control effect.

Figure 112010020312632-pat00002
Figure 112010020312632-pat00002

상추 균핵병을 방제할 수 있는 미생물은 파종 전에 토양에 혼화처리해야 하기 때문에 토양 내 정착을 위해 당분야에서 통상적으로 사용하는 탈지강(85중량%)+피마자박(10중량%)+제올라이트(5중량%)의 혼합조건의 담체를 사용하였다. Microorganisms capable of controlling lettuce fungal diseases must be mixed with the soil prior to sowing, so that the degreasing steel (85 wt%) + castor foil (10 wt%) + zeolite (5 wt. %) Mixed carrier was used.

길항미생물 슈도모나스 에스피 피에프-1의 균핵병 방제효과를 상추에서 직접 확인하기 위해 330㎡의 비닐하우스에 균핵병균(Sclerotinia sclerotiorum)을 밀곡립배지에서 25일간 배양한 병원균 접종원을 1㎡ 당 4.5g씩 접종하고, 24시간 후에 슈도모나스 에스피 피에프-1을 NB 배지에서 3일간 진탕배양한 배양액(108cfu/㎖)을 원심분리하였다. 이후 균체만 수확하여 멸균수에 108cfu/㎖ 농도로 희석하여 담체(탈자강 85중량%, 피마자박 10중량%, 제올라이트 5중량%)에 처리하여 토양에 혼화처리 한 후 방제효과를 정식기부터 수확 직전까지 10일 간격으로 조사하여 표 3에 그 결과를 나타내었고 각 실험은 세 번 반복되었다. In order to confirm the control effect of the antagonist Pseudomonas sp.PI-1 on lettuce, the bacterium Sclerotinia sclerotiorum ) was inoculated with 4.5g / m2 of the pathogen inoculum incubated for 25 days in wheat grain medium, and cultured with Pseudomonas sp.PI-1 in shaken culture for 3 days in NB medium (10 8 cfu / ml). Centrifuged. After that, only the cells were harvested and diluted to a concentration of 10 8 cfu / ml in sterile water, treated with a carrier (85% demagnetized steel, 10% by weight castor foil, 5% by weight zeolite), mixed with soil, and the control effect from the planting season. The results were shown in Table 3 at 10-day intervals immediately before harvest, and each experiment was repeated three times.

Figure 112010020312632-pat00003
Figure 112010020312632-pat00003

표 3의 결과를 확인하면 길항미생물 슈도모나스 에스피 피에프-1 균주는 상추 정식전 토양혼화 처리하였을 때 발병주율이 현저하게 줄어들었으며 90.7%의 우수한 방제효과를 나타냈다. 생육기간 중 발병초기에 슈도모나스 에스피 피에프-1 균주 배양액을 1회 토양관주하면 방제효과는 더 높아질 것으로 판단된다. 또한 표 1의 결과를 참고로 슈도모나스 에스피 피에프-1를 이용하는 것이 상용화되면 균핵병 이외에도 각각의 식물 병해를 광범위하게 방제할 수 있으면서도 효과가 좋은 미생물 방제제로 농가에서 실용화될 것으로 판단된다.As shown in the results of Table 3, the antagonist Pseudomonas S.P.P-1 strain significantly reduced the incidence rate when the soil was mixed with lettuce before treatment and showed excellent control effect of 90.7%. In the early stages of the growth period, once the soil culture of Pseudomonas S.P.P.-1 strain culture is done, the control effect will be higher. In addition, if the use of Pseudomonas S.P.P-1 is commercialized with reference to the results shown in Table 1, it is expected that it will be practically used in farms as an effective microbial control agent capable of controlling a wide range of plant diseases in addition to fungal diseases.

국립농업과학원 National Academy of Agricultural Science KACC91536PKACC91536P 2010030220100302

서열목록 전자파일 첨부Attach an electronic file to a sequence list

Claims (7)

마늘, 딸기 및 수박으로 이루어진 군에서 선택된 1종 이상의 식물에 대한 스크레로티움 세피보룸(Sclerotium cepivorum), 피시움 미리오틸륨(Pythium myriotylum) 및 스파에로테카 마쿠라(Sphaerotheca macular)로 이루어진 군에서 선택된 1종 이상의 식물 병원균을 방제할 수 있는 슈도모나스 에스피 피에프-1(Pseudomonas sp. PF-1, 미생물기탁번호 KACC91536P).Consisting of Sclerotium cepivorum , Pythium myriotylum and Sphaerotheca macular for one or more plants selected from the group consisting of garlic, strawberries and watermelon Pseudomonas sp. PF-1 (Microbial Accession No. KACC91536P) capable of controlling one or more plant pathogens selected from the group. 슈도모나스 에스피 피에프-1(Pseudomonas sp. PF-1, 미생물기탁번호 KACC91536P) 균주의 균체 또는 그 배양액을 유효성분으로 함유하는 마늘, 딸기 및 수박으로 이루어진 군에서 선택된 1종 이상의 식물에 대한 스크레로티움 세피보룸(Sclerotium cepivorum), 피시움 미리오틸륨(Pythium myriotylum) 및 스파에로테카 마쿠라(Sphaerotheca macular)로 이루어진 군에서 선택된 1종 이상의 식물 병원균의 방제를 위한 미생물 제제.Sclerotis of one or more plants selected from the group consisting of bacteria, Pseudomonas sp. PF-1 (Microorganism Accession No. KACC91536P) strains, or garlic, strawberries and watermelons containing the culture medium as an active ingredient. A microbial agent for the control of at least one plant pathogen selected from the group consisting of Sclerotium cepivorum , Pythium myriotylum and Sphaerotheca macular . 삭제delete 삭제delete 제 1 항의 균주 또는 제 2 항의 미생물 제제를 마늘, 딸기 및 수박으로 이루어진 군에서 선택된 1종 이상의 식물에 처리하여 스크레로티움 세피보룸(Sclerotium cepivorum), 피시움 미리오틸륨(Pythium myriotylum) 및 스파에로테카 마쿠라(Sphaerotheca macular)로 이루어진 군에서 선택된 1종 이상의 식물 병원균을 방제하는 방법.The microbial agent of claim 1 or the microbial agent of claim 2 is treated with at least one plant selected from the group consisting of garlic, strawberries, and watermelon, Sclerotium cepivorum , Pythium myriotylum and A method of controlling one or more plant pathogens selected from the group consisting of Sphaerotheca macular . 삭제delete 삭제delete
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KR100997677B1 (en) 2009-04-21 2010-12-02 한국화학연구원 Pseudomonas geniculata mh102 strain and method for the biological control of plant diseases using same
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