KR20060019865A - A novel streptomyces sp. and biopesticide containing the strain for extermination of anthracnose - Google Patents
A novel streptomyces sp. and biopesticide containing the strain for extermination of anthracnose Download PDFInfo
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Abstract
본 발명은 방선균속(Streptomyces sp.) 신규균주 및 상기 균주를 함유하는 미생물농약제제에 관한 것으로, 더욱 상세하게는 고추탄저병 방제에 유용한 항균성물질을 생성하는 방선균 속 신규균주, 상기 균주를 함유하는 고추탄저병 방제용 미생물농약제제 및 상기 미생물농약제제를 이용하는 것을 특징으로 하는 고추탄저병의 방제방법에 관한 것이다. The present invention relates to a novel strain of Streptomyces sp. And a microbial pesticide preparation containing the strain, and more particularly, a novel strain of the actinomycetes which produces an antimicrobial substance useful for controlling pepper anthrax, and pepper containing the strain. It relates to a microbial pesticide preparation for anthrax control and pepper anthrax control method characterized in that using the microbial pesticide preparation.
본 발명에 따른 방선균속(Streptomyces sp.) KACC 91028 균주는, 기존의 화학농약에서 나타날 수 있는 과다사용으로 인한 생태계의 파괴 없이도, 기존 화학농약보다 우수한 고추탄저병 방제효과를 가지므로 생물농약으로서 유용하다. Streptomyces sp. KACC 91028 strain according to the present invention is useful as a biopesticide because it has a superior pepper anthrax control effect than conventional chemical pesticides, without destroying the ecosystem due to overuse that may occur in conventional chemical pesticides. .
방선균, 고추탄저병, 방제, 미생물제제Actinomycetes, Pepper Anthrax, Control, Microbial
Description
도 1은 신규한 방선균속(Streptomyces sp.) KACC 91028 균주를 Bennet 배지에서 배양하여 주사현미경으로 관찰한 결과이다.FIG. 1 shows the results of observing the novel Streptomyces sp. KACC 91028 strain in Bennet medium with an injection microscope.
도 2는 신규한 방선균속(Streptomyces sp.) KACC 91028 균의 16S rRNA 유전자 분석결과이다.Figure 2 is a 16S rRNA gene analysis of the novel Streptomyces sp. KACC 91028.
도 3은 고추탄저병이 발병된 고추에 방선균속 KACC 91028 균의 배양액을 처리한 결과를 나타내는 사진으로, 오른쪽은 고추탄저병이 발병된 고추에 방선균속 KACC 91028 균의 배양액을 처리한 결과이고, 왼쪽은 처리하지 않은 결과이다.Figure 3 is a photograph showing the result of processing the culture medium of actinomycetes KACC 91028 bacteria in the pepper with the onset of pepper anthrax, the right side is the result of processing the culture medium of actinomycetes KACC 91028 bacteria in the pepper with the onset of pepper anthrax, The result is not processed.
본 발명은 방선균속(Streptomyces sp.) 신규균주 및 상기 균주를 함유하는 미생물농약제제에 관한 것으로, 더욱 상세하게는 고추탄저병 방제에 유용한 항균성 물질을 생성하는 방선균 속 신규균주, 상기 균주를 함유하는 고추탄저병 방제용 미생물농약제제 및 상기 미생물농약제제를 이용하는 것을 특징으로 하는 고추탄저병의 방제방법에 관한 것이다. The present invention relates to a novel strain of Streptomyces sp. And a microbial pesticide preparation containing the strain, and more particularly, a novel strain of the actinomycetes which produces an antimicrobial substance useful for controlling pepper anthrax, and pepper containing the strain. It relates to a microbial pesticide preparation for anthrax control and pepper anthrax control method characterized in that using the microbial pesticide preparation.
고추탄저병(anthracnose)은 진균계의 자낭균문에 속하는 콜레토트리쿰 코코드(Colletotrichum coccodes)에 의해 발병되며, 주로 과실에서 발생한다. 이 병원균은 자낭포자와 분생포자를 형성하고 자낭포자는 자낭 내에서 8개씩 생성되며 방추형으로 되어 있다. 분생포자는 타원형으로 되어 있다.Anthracnose is caused by Colletotrichum coccodes , which belongs to the fungal bacterium , and mainly occurs in fruit. These pathogens form follicular spores and conidia, and eight of them are fusiform in the capsule. Conidia are oval-shaped.
콜레토트리쿰 코코드의 생육온도는 5~35℃ 사이이나 26~28℃ 사이가 적정온도이다. 이 병의 증상은 초기에는 과실에 수침상으로 원형반점이 생기고 병이 진전됨에 따라 원형으로 확대된다. 병이 심하게 진전되면 담황색의 포자덩어리가 형성되어 과실이 말라버리게 된다. 콜레토트리쿰 코코드는 자낭각과 균사로 겨울을 지내고, 병의 전염은 주로 분생포자로 이루어진다. 병의 발생은 7월 초순부터 노지재배된 풋고추에서 시작되어 수확기까지 진전된다. 특히 고온 다습하면 좀 더 심하게 발병한다. The growth temperature of choletotricum cocord is between 5 ~ 35 ℃ but between 26 ~ 28 ℃. Symptoms of the disease initially appear as a circular spot on the fruit with a water soak, and expand as the disease progresses. When the disease progresses severely, a pale yellow spore mass forms, causing the fruit to dry. Colletotricum cocode spends winter with asymptomatic and hyphae, and the transmission of disease mainly consists of conidia. The outbreak begins in early July and grows in green-grown green peppers until the harvest season. Especially when the temperature and humidity are more severe.
고추탄저병의 약제방제는 델란, 다코닐, 포룸만, 톱신엠, 크린힛 등이 사용되고 있는데 이들은 모두 화학농약이다. 화학농약이 근래에는 소비자들로부터 안전성 문제로 외면 받고 있는 현실을 감안할 때 새로운 방제방법의 개발이 필요한 실정이다.Drug control of pepper anthrax is used as delan, daconyl, formum, sawcinm, cleanse, etc., all of which are chemical pesticides. Considering the fact that chemical pesticides have recently been ignored by consumers as a safety issue, it is necessary to develop new control methods.
이에 본 발명자들은 고추탄저병 방제에 효율적인 미생물제제를 개발하고자 예의 노력한 결과, 방선균속에 속하는 신규균주가 고추탄저병 방제활성을 가진다는 것을 확인하고 본 발명을 완성하게 되었다. Accordingly, the present inventors have made intensive efforts to develop an effective microbial agent for controlling pepper anthrax. As a result, new strains belonging to the actinomycetes have confirmed that pepper anthrax control activity has been completed.
결국 본 발명의 주된 목적은 고추탄저병 방제효과를 나타내는 방선균 속 (Streptomyces sp.) 신규균주를 제공한다.After all, the main object of the present invention is to provide a novel Streptomyces sp . Strain showing the effect of controlling pepper anthrax.
본 발명의 다른 목적은 상기 균주를 함유하는 고추탄저병 방제용 미생물농약제제 및 상기 미생물농약제제를 이용하는 것을 특징으로 하는 고추탄저병의 방제방법을 제공하는데 있다.
Another object of the present invention is to provide a method for controlling pepper anthrax, which comprises using the microbial pesticide preparation for controlling pepper anthrax and the microbial pesticide preparation containing the strain.
상기의 목적을 달성하기 위하여, 본 발명은 고추탄저병에 대하여 병원성을 나타내는 방선균 속(Streptomyces sp.) 균주를 제공한다. 본 발명에 있어서, 상기의 균주는 방선균 속(Streptomyces sp.) KACC 91028인 것을 특징으로 할 수 있고, 서열번호 1의 16S rRNA 유전자 염기서열을 가지는 것을 특징으로 할 수 있다.In order to achieve the above object, the present invention provides a Streptomyces sp . Strain showing pathogenicity against pepper anthrax. In the present invention, the strain may be characterized in that the Streptomyces sp . KACC 91028, characterized in that it has a 16S rRNA gene sequence of SEQ ID NO: 1.
본 발명은 또한, 상기 균주 또는 그 배양액을 유효성분으로 함유하는 고추탄저병 방제용 미생물농약제제 및 상기 미생물농약제제를 이용하는 것을 특징으로 하는 고추탄저병의 방제방법을 제공한다.The present invention also provides a control method for pepper anthrax, characterized in that the microbial pesticide preparation for pepper anthrax control containing the strain or its culture as an active ingredient and the microbial pesticide preparation.
본 발명은 또한, 상기 균주를 배양하는 것을 특징으로 하는 고추탄저병 방제용 미생물농약제제의 제조방법을 제공한다. 본 발명에 있어서, 배양은 베넷(Bennet)배지를 사용하여 25~30℃에서 약 10일간 수행하는 것을 특징으로 할 수 있 다.The present invention also provides a method for producing a microbial pesticide preparation for pepper anthrax, characterized in that the culture of the strain. In the present invention, the culture may be characterized by performing about 10 days at 25 ~ 30 ℃ using Bennett (Bennet) medium.
본 발명의 균주는 방선균 속(Streptomyces sp.) KACC 91028은 2003년 2월 24일 대한민국 농업생명공학원구원(경기도 수원시 권선구 서둔동 225 (우)441-707)에 방선균 속(Streptomyces sp.) lim 8이라는 균주명과 기탁번호 KACC 91028로 기탁하였다.The strain of the present invention is Streptomyces sp . KACC 91028 is a Streptomyces sp . Lim 8 in the Korea Agricultural Biotechnology Park (February 22, 2003 225 (R) 441-707, Seodun-dong, Gwonseon-gu, Suwon-si, Gyeonggi-do) The strain name and accession number KACC 91028 were deposited.
본 발명의 미생물제제는 본 발명의 균주 또는 균주의 배양액 또는 배양물을 포함하는 다양한 제제의 형태, 즉 액제, 수화제, 분제, 입제, 유제 등으로 제조할 수 있다. 그 제조방법은 당업계의 일반적인 제조방법에 따른다.The microbial agent of the present invention can be prepared in the form of a variety of preparations, including cultures or cultures of the strains or strains of the invention, i.e. solutions, hydrates, powders, granules, emulsions and the like. The manufacturing method is according to the general manufacturing method in the art.
이하 실시예를 통하여 본 발명을 보다 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 구체적으로 설명하기 위한 것으로 본 발명의 요지에 따라 본 발명의 범위가 실시예에 의하여 구체적으로 제한되지 않는다는 것은, 당 업계에서 통상적인 지식을 가진 자에게 자명할 것이다. Through the following examples will be described the present invention in more detail. It will be apparent to those skilled in the art that these examples are only for explaining the present invention in detail, and the scope of the present invention is not specifically limited by the examples according to the gist of the present invention.
실시예 1: 신규한 방선균속(Streptomyces sp.) KACC91028 균의 동정 Example 1 Identification of Novel Streptomyces sp.
균주 KACC 91028의 현탁배양을 위하여 방선균배양에 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 strain KACC 91028. 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 KACC 91028 균주의 주사현미경 (Scanning Electron Microscope) 관찰결과, 방선균속(Streptomyces sp.)과 매우 유사하였다 (도 1).Scanning Electron Microscope observations of the cultured KACC 91028 strain were very similar to Streptomyces sp. (FIG. 1).
신규균 방선균속(Streptomyces sp.) KACC 91028 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 분리한 다음, Genomic DNA Extraction Kit (Intron)를 이용하여 DNA를 추출하였다.Genetic analysis of Streptomyces sp. KACC 91028 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, the cells were centrifuged to separate the cells, and the DNA was extracted using Genomic DNA Extraction Kit (Intron).
PCR은 반응용액(10mM Tris, pH 8.3; 50mM KCl; 1.5mM MgCl2; 0.2mM dNTPs; 1 unit Taq DNA polymerase)에 30ng genomic DNA, 0.25μM 16S rRNA 유전자 primer 1쌍(fD1과 rP2)을 이용하여 증폭하였다. 증폭된 절편은 약 1500bp의 크기를 보였다. PCR(초기 94℃ 5분 1회; 94℃ 1분, 58℃ 1분, 72℃ 2분, 35cycle; 72℃ 10분 1회)을 실시한 후, 증폭된 16S rRNA 유전자 조각을 pGEM-T Easy vector(Promega)에 ligation한 후, 대장균 DH5α에 형질전환하였다.PCR was performed using 30ng genomic DNA, 0.25μM 16S rRNA gene primer (fD1 and rP2) in reaction solution (10mM Tris, pH 8.3; 50mM KCl; 1.5mM MgCl 2 ; 0.2mM dNTPs; 1 unit Taq DNA polymerase). Amplified. The amplified fragment was about 1500bp in size. After PCR (initial 94 ℃ 5 minutes once; 94
형질전환된 대장균은 선발배지(1.5% agar, 50mg/L ampicillin, 100μM IPTG, 50mg/L X-gal)에 도발하여 37℃에서 12시간 배양하였다. 증폭된 16S rRNA 유전자조각이 들어 있는 vector의 삽입은 blue/white colony가 나타난 배지에서 white colony만을 LB배지에 접종하여 배양하였다. 배양액은 plasmid분리를 위하여 원심분리한 후 Alkaline lysis방법을 이용하였다. 분리된 plasmid는 제한효소 EcoRI으로 절단한 후 전기영동을 통하여 증폭된 16S rRNA 유전자조각의 삽입을 확인하였다 (도 2).Transformed Escherichia coli was provoked in selection medium (1.5% agar, 50mg / L ampicillin, 100μM IPTG, 50mg / L X-gal) and incubated at 37 ° C for 12 hours. Insertion of the vector containing the amplified 16S rRNA gene fragment was cultured by inoculating only LB medium with white colony in a medium showing blue / white colony. The culture was centrifuged for plasmid separation and Alkaline lysis was used. The isolated plasmid was digested with restriction enzyme EcoRI and confirmed insertion of amplified 16S rRNA fragment through electrophoresis (FIG. 2).
증폭된 16S rRNA 유전자 염기서열(서열번호 1)은 제작된 4개의 sequencing primer (표 1)와 Big dye terminator v.3.0 (Applied Biosystems)를 이용하여 PCR 단편을 direct sequencing reaction을 수행한 후 염기서열분석장치(ABI3100)를 이용하여 분석하였으며, NCBI(National Center for Biotechnology Information)의 BLAST를 이용하여 homology를 확인하였다. 그 결과 방선균 스트렙토마이세스 프라보그리세우스 (Streptomyces flavogriseus)와 99%의 상동성을 보였다.Amplified 16S rRNA gene sequence (SEQ ID NO: 1) was sequenced after direct sequencing reaction of PCR fragments using four prepared sequencing primers (Table 1) and Big dye terminator v.3.0 (Applied Biosystems). The device was analyzed using ABI3100, and homology was confirmed using BLAST of the National Center for Biotechnology Information (NCBI). The results showed 99% homology with Streptomyces flavogriseus .
실시예 2: 방선균속 KACC 91028 균 배양액의 고추탄저병 방제 효과 Example 2: Control effect of pepper anthrax on the actinomycetes KACC 91028 culture
시험 대상 고추 품종은 향촌[동부한농화학(주)]을 사용하였으며, 종자를 원예용 상토(부농사)에 파종하고 25±5℃의 온실에서 본엽 4-5엽기 고추가 되도록 재배한 후에 실험에 사용하였다. 고추 탄저병균은 Colletotrichum coccodes 2-25 균주를 사용하였으며, C. coccodes 2-25의 균사조각을 감자즙액배지(potato dextrose broth)에 접종하여 25℃, 암상태에서 7일 동안 진탕배양하였다. 이를 Waring blender로 마쇄하고 오트밀 배지(oatmeal agar)에 접종하여 25℃ 항온기에서 배양하였다. 멸균한 붓으로 오트밀 배지에 자란 C. coccodes의 균총을 긁고 광처리하여 포자를 형성하였으며, 형성된 포자는 증류수로 수확하고 3겹의 가제로 거른 후에 광학현미경하에서 혈구계(hematocytometer)를 사용하여 포자 농도를 측정하고, 포자의 농도를 ml 당 3x105가 되도록 조정하여 접종원으로 사용하였다. The pepper varieties tested were Hyangchon [Eastern Hanwha Chemical Co., Ltd.]. The seeds were sown in horticultural soils (vice farms) and grown in a greenhouse at 25 ± 5 ℃ to 4-4 leaves of red pepper. Used. Pepper anthrax bacterium was used as the Colletotrichum coccodes 2-25 strain, the mycelia of C. coccodes 2-25 were inoculated in potato juice broth (potato dextrose broth) and cultured for 7 days in a dark condition at 25 ℃. This was ground with a Waring blender and inoculated in oatmeal agar and incubated in a 25 ° C thermostat. Sterile brush was scraped and light- treated to obtain spores of C. coccodes grown on oatmeal medium. The spores were harvested with distilled water and filtered with three layers of gauze, and then spore concentration was measured using a hematocytometer under an optical microscope. The spores were measured and used as inoculum by adjusting the concentration of spores to 3 × 10 5 per ml.
배양한 미생물은 전착제로 Tween 20을 첨가하여 최종농도가 250㎍/ml이 되도록 준비한 후, 4-5엽기의 고추 유묘에 미생물 배양액을 흘러내릴 정도로 살포하였다. 처리한 고추 유묘는 온실에서 1일 동안 재배한 후에 고추 탄저병균의 포자현탁액을 분무접종 하였다. 이때 무처리구는 Tween 20 250㎍/ml 용액을 살포하고 처리구와 동일하게 접종하였다. 접종한 고추 유묘는 25℃ 습실상에서 48시간 동안 습실처리한 후에 항온항습실(25℃, 상대습도 70%)에서 1-2일 동안 발병을 유도하고 고추 유묘의 병반면적율을 조사하였다. 다음과 같은 수학식 1에 따라 방제가를 계산하였다. The cultured microorganisms were prepared by adding Tween 20 as an electrodeposition agent to a final concentration of 250 µg / ml, and then spraying the microbial culture solution to 4-5 leaves of pepper seedlings. The treated pepper seedlings were grown in a greenhouse for 1 day, and then sprayed with a spore suspension of pepper anthrax. At this time, the untreated group was sprayed with Tween 20 250㎍ / ml solution and inoculated in the same manner as the treated group. The inoculated pepper seedlings were incubated for 48 hours in a 25 ° C. humidity chamber, and then induced for 1-2 days in a constant temperature and humidity room (25 ° C., 70% relative humidity). The control value was calculated according to
<수학식 1> <
방제가(%) = [(1- 처리구의 병반면적율)/무처리구의 병반면적율] × 100Control Value (%) = [(Large Area Ratio of 1-Treatment) / Large Area Ratio of No-Treatment] × 100
실시예 1의 방법으로 배양한 방선균속(Streptomyces sp.) KACC 91028 균의 배양액(미생물 세포내 물질이 포함된)을 처리한 결과와 무처리구를 비교하였다 (도 3). 일반적으로 Colletotrichum coccodes에 의해 고추 탄저병이 발생하면 잎에 둥근 병반이 생긴다. KACC 91028 균의 배양액 처리구(오른쪽)와 무처리구(왼쪽) 모두 Colletotrichum coccodes을 접종하였으나, 도 3에 나타난 바와 같이, 무처리구에서는 잎에 탄저병 병반이 관찰된 반면, 처리구에서는 방제효과로 인해 탄저병이 발생 되지 않았으며, 이때, 방제가는 90%였다. 이 결과로부터 본 발명에 따른 방선균속(Streptomyces sp.) KACC 91028 균주 또는 그 배양액은 고추 등 농작물의 탄저병 방제에 효율적이라는 것을 확인할 수 있었다.The results of treatment of the culture solution (containing microbial intracellular material) of Streptomyces sp. KACC 91028 cultured by the method of Example 1 were compared with the untreated group (FIG. 3). Generally, pepper anthrax caused by Colletotrichum coccodes results in round lesions on leaves. In the culture solution treatment group (right) and untreated group (left) of KACC 91028, both were inoculated with Colletotrichum coccodes . However, as shown in FIG. 3, anthrax lesions were observed in the leaves, whereas in the treatment group, anthrax did not occur due to the control effect. At this time, the control value was 90%. From these results, it was confirmed that the Streptomyces sp. KACC 91028 strain or the culture medium according to the present invention is effective for controlling anthrax of crops such as pepper.
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 고추탄저병 방제에 유용한 방선균 속 신규균주, 상기 균주를 함유하는 고추탄저병 방제용 미생물농약제제 및 상기 미생물농약제제를 이용하는 것을 특징으로 하는 고추탄저병의 방제방법을 제공하는 효과가 있다. 본 발명에 따른 방선균속(Streptomyces sp.) KACC 91028 균은 고추탄저병 방제용 농용미생물제제로 유용하므로, 이는 기존의 화학농약에서 나타날 수 있는 과다사용으로 인한 생태계의 파괴를 방지할 수 있다.As described in detail above, the present invention provides a novel strain of actinomycetes useful for controlling pepper anthrax, a microbial pesticide preparation for controlling pepper anthrax containing the strain, and a method for controlling pepper anthrax, comprising the microbial pesticide preparation. It is effective. Actinomycetes ( Streptomyces sp.) KACC 91028 according to the present invention is useful as agricultural microorganisms for the control of pepper anthrax, which can prevent the destruction of the ecosystem due to overuse that may appear in conventional chemical pesticides.
<110> Konkuk University Industrial Cooperation Corp <120> A Novel Streptomyces sp. and Biopesticide Containing the Strain for Extermination of Anthracnose <130> P04-353 <160> 7 <170> KopatentIn 1.71 <210> 1 <211> 1489 <212> DNA <213> Streptomyces sp. <400> 1 agagtttgat cctggctcag gacgaacgct ggcggcgtgc ttaacacatg caagtcgaac 60 gatgaagccc ttcggggtgg attagtggcg aacgggtgag taacacgtgg gcaatctgcc 120 cttcactctg ggacaagccc tggaaacggg gtctaatacc ggataacact ctgtgccgca 180 tgggacgggg ttaaaagctc cggcggtgaa ggatgagccc gcggcctatc agcttgttgg 240 tggggtgatg gcctaccaag gcgacgacgg gtagccggcc tgagagggcg accggccaca 300 ctgggactga gacacggccc agactcctac gggaggcagc agtggggaat attacacaat 360 gggcgaaagc ctgatgcagc gacgccgcgt gagggatgac ggccttcggg ttgtaaacct 420 ctttcagcag ggaagaagcg aaagtgacgg tacctgcaga agaagcgccg gctaactacg 480 tgccagcagc cgcggtaata cgtagggcgc aagcgttgtc cggaattatt gggcgtaaag 540 agctcgtagg cggcttgtca cgtcggatgt gaaagcccgg ggcttaaccc cgggtctgca 600 ttcgatacgg gctagctaga gtgtggtagg ggagatcgga attcctggtg tagcggtgaa 660 atgcgcagat atcaggagga acaccggtgg cgaaggcgga tctctgggcc attactgacg 720 ctgaggagcg aaagcgtggg gagcgaacag gattagatac cctggtagtc cacgccgtaa 780 acgttgggaa ctaggtgttg gcgacattcc acgtcgtcgg tgccgcagct aacgcattaa 840 gttccccgcc tggggagtac ggccgcaagg ctaaaactca aaggaattga cgggggcccg 900 cacaagcagc ggagcatgtg gcttaattcg acgcaacgcg aagaacctta ccaaggcttg 960 acatataccg gaaagcatca gagatggtgc ccccttgtgg tcggtataca ggtggtgcat 1020 ggctgtcgtc agctcgtgtc gtgagatgtt gggttaagtc ccgcaacgag cgcaaccctt 1080 gttctgtgtt gccagcatgc ccttcggggt gatggggact cacaggagac tgccggggtc 1140 aactcggagg aaggtgggga cgacgtcaag tcatcatgcc ccttatgtct tgggctgcac 1200 acgtgctaca atggccggta caaagagctg cgatgccgcg aggcggagcg aatctcaaaa 1260 agccggtctc agttcggatt ggggtctgca actcgacccc atgaagtcgg agttgctagt 1320 aatcgcagat cagcattgct gcggtgaata cgttcccggg ccttgtacac accgcccgtc 1380 acgtcacgaa agtcggtaac acccgaagcc ggtggcccaa ccccttgtgg gagggagctg 1440 tcgaaggtgg gactggcgat tgggacgaag tcgtaacaag gtagccgta 1489 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> PCR primer <400> 2 agagtttgat cctggctcag 20 <210> 3 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> PCR primer <400> 3 acggctacct tgttacgact t 21 <210> 4 <211> 17 <212> DNA <213> Artificial Sequence <220> <223> PCR primer <400> 4 tattaccgcg gctgctg 17 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> PCR primer <400> 5 ggcagcagtg gggaatattg 20 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> PCR primer <400> 6 tagataccct ggtagtccac 20 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> PCR primer <400> 7 tcgtcagctc gtgtcgtgag 20 <110> Konkuk University Industrial Cooperation Corp <120> A Novel Streptomyces sp. and Biopesticide Containing the Strain for Extermination of Anthracnose <130> P04-353 <160> 7 <170> KopatentIn 1.71 <210> 1 <211> 1489 <212> DNA <213> Streptomyces sp. <400> 1 agagtttgat cctggctcag gacgaacgct ggcggcgtgc ttaacacatg caagtcgaac 60 gatgaagccc ttcggggtgg attagtggcg aacgggtgag taacacgtgg gcaatctgcc 120 cttcactctg ggacaagccc tggaaacggg gtctaatacc ggataacact ctgtgccgca 180 tgggacgggg ttaaaagctc cggcggtgaa ggatgagccc gcggcctatc agcttgttgg 240 tggggtgatg gcctaccaag gcgacgacgg gtagccggcc tgagagggcg accggccaca 300 ctgggactga gacacggccc agactcctac gggaggcagc agtggggaat attacacaat 360 gggcgaaagc ctgatgcagc gacgccgcgt gagggatgac ggccttcggg ttgtaaacct 420 ctttcagcag ggaagaagcg aaagtgacgg tacctgcaga agaagcgccg gctaactacg 480 tgccagcagc cgcggtaata cgtagggcgc aagcgttgtc cggaattatt gggcgtaaag 540 agctcgtagg cggcttgtca cgtcggatgt gaaagcccgg ggcttaaccc cgggtctgca 600 ttcgatacgg gctagctaga gtgtggtagg ggagatcgga attcctggtg tagcggtgaa 660 atgcgcagat atcaggagga acaccggtgg cgaaggcgga tctctgggcc attactgacg 720 ctgaggagcg aaagcgtggg gagcgaacag gattagatac cctggtagtc cacgccgtaa 780 acgttgggaa ctaggtgttg gcgacattcc acgtcgtcgg tgccgcagct aacgcattaa 840 gttccccgcc tggggagtac ggccgcaagg ctaaaactca aaggaattga cgggggcccg 900 cacaagcagc ggagcatgtg gcttaattcg acgcaacgcg aagaacctta ccaaggcttg 960 acatataccg gaaagcatca gagatggtgc ccccttgtgg tcggtataca ggtggtgcat 1020 ggctgtcgtc agctcgtgtc gtgagatgtt gggttaagtc ccgcaacgag cgcaaccctt 1080 gttctgtgtt gccagcatgc ccttcggggt gatggggact cacaggagac tgccggggtc 1140 aactcggagg aaggtgggga cgacgtcaag tcatcatgcc ccttatgtct tgggctgcac 1200 acgtgctaca atggccggta caaagagctg cgatgccgcg aggcggagcg aatctcaaaa 1260 agccggtctc agttcggatt ggggtctgca actcgacccc atgaagtcgg agttgctagt 1320 aatcgcagat cagcattgct gcggtgaata cgttcccggg ccttgtacac accgcccgtc 1380 acgtcacgaa agtcggtaac acccgaagcc ggtggcccaa ccccttgtgg gagggagctg 1440 tcgaaggtgg gactggcgat tgggacgaag tcgtaacaag gtagccgta 1489 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> PCR primers <400> 2 agagtttgat cctggctcag 20 <210> 3 <211> 21 <212> DNA <213> Artificial Sequence <220> PCR primers <400> 3 acggctacct tgttacgact t 21 <210> 4 <211> 17 <212> DNA <213> Artificial Sequence <220> PCR primers <400> 4 tattaccgcg gctgctg 17 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> PCR primers <400> 5 ggcagcagtg gggaatattg 20 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> PCR primers <400> 6 tagataccct ggtagtccac 20 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> PCR primers <400> 7 tcgtcagctc gtgtcgtgag 20
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