KR20220048662A - Composition for relieving brown spot disease of crops - Google Patents
Composition for relieving brown spot disease of crops Download PDFInfo
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Abstract
Description
본 발명은 친환경적인 작물의 깨씨무늬병 완화를 위한 조성물에 관한 것이다.The present invention relates to a composition for alleviating sesame seed disease in environmentally friendly crops.
깨씨무늬병은 옥수수 등의 잎에서 갈색을 띤 작은 반점으로 시작되어 방추형 혹은 타원형 병반으로 확대되며 병반은 암갈색으로 변한다. 주로 생육과정에서 온도가 높고 강우량이 많을 때 우리나라 전역에서 병 발생이 심하다고 알려졌다. 또한, 작물의 영양 불균형 및 결핍된 조건에서 쉽게 감염되는 병으로도 알려져 있다. Sesame seed disease starts as small brownish spots on the leaves of corn, etc., then expands to spindle-shaped or oval lesions, and the lesions turn dark brown. It is known that disease outbreaks are severe throughout Korea when the temperature is high and there is a lot of rainfall during the growth process. In addition, it is also known as a disease that is easily infected in nutrient imbalance and deficient conditions of crops.
감염원으로써는 진균인 코츨리오볼로우Cochliobolus heterostrophus(Derechs.) Drechs.)라는 자낭균에 속하는 곰팡이로써 불완전세대는 바이오폴라리스 메디어스(Bipolaris maydis), 바이폴라리스 제이콜라(Bipolaris zeicola) 및 바이폴라리스 오리지에 (Biopolaris oryzae)라는 곰팡이가 깨씨무늬병의 원인균이다.As an infectious agent, the fungus Cochliobolus heterostrophus (Derechs.) Drechs. (Biopolaris oryzae) is the causative agent of sesame seed blotch.
곰팡이는 포장에 있는 이병 잔재물에서 포자나 균사로 월동하며 포자는 바람이나 빗물에 의해 식물체로 전이되어 일차 감염을 일으키고, 병반에 형성된 포자가 다시 1, 2차 감염원이 된다. 특히, 벼와 옥수수는 국내에서 생산량이 가장 많은 중요한 식량작물로써 경제적 가치가 매우 높다. 그리하여 병이 발생하면 생산량의 감소에 직접 영향을 줄 수 있으며, 이에 따른 경제적 피해도 크다고 할 수 있다.The fungus overwinters as spores or mycelium in the remnants of disease in the field, and the spores are transferred to the plant by wind or rainwater to cause primary infection, and the spores formed on the lesion become primary and secondary infection sources again. In particular, rice and corn are the most important food crops in Korea and have very high economic value. Therefore, when a disease occurs, it can directly affect the decrease in production, and the economic damage can be said to be large.
깨씨무늬병의 일반적인 제어법은 병원균을 박멸하거나 그 밀도를 줄이는 방법, 주위환경을 병원균의 생장 및 활동조건에 맞지 않게 조절하는 방법, 그리고, 병에 견디는 품종을 재배하는 방법이 있다. 이 중, 병원균의 제어를 위하여 유기합성 농약의 사용은 효과적이나, 최근의 빈번하고 광범위한 살포로 인하여 환경오염, 독성, 약제내성 저항성 돌연변이 병충해를 유발하는 등 많은 문제가 초래되고 있다.General control methods for sesame seed blotch include methods of eradicating pathogens or reducing their density, adjusting the surrounding environment to be unsuitable for growth and activity conditions of pathogens, and cultivating disease-resistant varieties. Among these, the use of organic synthetic pesticides for the control of pathogens is effective, but due to recent frequent and extensive spraying, many problems such as environmental pollution, toxicity, and drug-resistant mutant pests are caused.
그리하여, 유기합성 농약을 대체할 친환경적인 방법으로 생물학적 방법들이 연구되고 있는 추세이다.Therefore, biological methods are being studied as an eco-friendly way to replace organic synthetic pesticides.
본 발명은 친환경적인 작물의 깨씨무늬병 완화를 위한 조성물을 제공하는 것을 목적으로 한다.An object of the present invention is to provide a composition for alleviating sesame seed disease in environmentally friendly crops.
본 발명의 목적은 유산균의 상등액을 포함하는 작물의 깨씨무늬병 완화용 조성물에 의해 달성된다. 상기 작물은 옥수수를 포함할 수 있다. 상기 유산균은 피도박테리움 락티스 IDCC 4301 및 락토바실러스 람노서스 IDCC 3201 중 적어도 어느 하나를 포함할 수 있다.The object of the present invention is achieved by a composition for alleviating sesame seed disease of crops comprising a supernatant of lactic acid bacteria. The crop may include corn. The lactic acid bacteria may include at least one of Pidobacterium lactis IDCC 4301 and Lactobacillus rhamnosus IDCC 3201.
본 발명에 따르면 친환경적인 작물의 깨씨무늬병 완화를 위한 조성물이 제공된다.According to the present invention, there is provided a composition for alleviating sesame seed disease in environmentally friendly crops.
도 1은 본 발명의 일실시예에서 유산균 상등액을 얻는 공정을 나타낸 것이고,
도 2는 본 발명의 실험예에서 각 유산균 상등액의 사용에 따른 깨씨무늬병의 완화정도를 나타낸 표이다.1 shows the process of obtaining a lactic acid bacteria supernatant in an embodiment of the present invention,
Figure 2 is a table showing the degree of relief of the sesame seed pattern disease according to the use of each lactic acid bacteria supernatant in the experimental example of the present invention.
본 명세서 및 청구범위에 사용된 용어나 단어는 통상적이거나 사전적인 의미로 한정해서 해석되어서는 안 되며, 발명자는 그 자신의 발명을 가장 최선의 방법으로 설명하기 위해 용어의 개념을 적절하게 정의할 수 있다는 원칙에 입각하여 본 발명의 기술적 사상에 부합하는 의미와 개념으로 해석되어야만 한다.The terms or words used in this specification and claims should not be construed as being limited to their ordinary or dictionary meanings, and the inventor may properly define the concept of the term in order to best describe his invention. Based on the principle that there is, it should be interpreted as meaning and concept consistent with the technical idea of the present invention.
따라서 본 명세서에 기재된 실시예와 도면에 도시된 구성은 본 발명의 가장 바람직한 실시예에 불과할 뿐이고 본 발명의 기술적 사상을 모두 대변하는 것은 아니므로, 본 출원시점에 있어서 이들을 대체할 수 있는 다양한 균등물과 변형예들이 있을 수 있음을 이해하여야 한다.Accordingly, the embodiments described in this specification and the configurations shown in the drawings are only the most preferred embodiments of the present invention and do not represent all the technical spirit of the present invention, so various equivalents that can be substituted for them at the time of the present application It should be understood that there may be variations and variations.
이하의 설명에서는 옥수수의 깨씨무늬병을 예시하여 설명하지만 본 발명은 옥수수에 한정되지 않는다. 본 발명의 조성물은 작물의 깨씨무늬병에 사용될 수 있으며, 구체적으로는 벼, 배추, 옥수수 등의 깨시무늬 병에 사용될 수 있다.In the following description, sesame seed disease of corn will be exemplified, but the present invention is not limited to corn. The composition of the present invention may be used for sesame seed pattern disease of crops, and specifically may be used for sesame seed pattern disease such as rice, Chinese cabbage, and corn.
본 발명에서의 '완화'는 항균효과, 방제효과, 질병의 치료, 예방 및 완화를 모두 포함하는 개념이다.In the present invention, 'alleviation' is a concept that includes all of the antibacterial effect, the control effect, and the treatment, prevention and alleviation of disease.
본 발명에 따른 조성물은 유산균 상등액을 유효 성분으로 포함한다.The composition according to the present invention includes a lactic acid bacteria supernatant as an active ingredient.
본 발명에서의 '상등액'이란 유산균 제조에 있어 발효 후 원심분리과정에서 얻어지는 액체로 상청액, 상층액, 상증액 또는 상징액으로도 불린다.In the present invention, 'supernatant' is a liquid obtained in the centrifugation process after fermentation in the production of lactic acid bacteria, and is also called supernatant, supernatant, supernatant or supernatant.
본 발명에 따른 조성물은 액제, 입제, 수화제 등의 형태일 수 있다. 또한 본 발명에 따른 조성물은 분말, 펠렛 또는 과립, 마이크로캡슐 등으로 제형화할 수 있다. 조성물은 표면 활성화제, 비활성 담체, 보존제, 습윤제, 공급 촉진제, 유인제, 캡슐화제, 결합제, 유화제, 염료, U.V. 보호제, 완충제 등 목적 병원균 및 작물에 따라 적용하기 용이한 다른 성분을 부가할 수 있다.The composition according to the present invention may be in the form of a solution, granules, wettable powder, and the like. In addition, the composition according to the present invention can be formulated as a powder, pellets or granules, microcapsules, and the like. The compositions may contain surface active agents, inert carriers, preservatives, wetting agents, supply promoters, attractants, encapsulating agents, binders, emulsifiers, dyes, U.V. Other ingredients that are easy to apply, such as protective agents and buffers, may be added depending on the target pathogen and crop.
본 발명의 조성물은 적용하기 전에 적당한 양의 물 또는 다른 희석제로 희석을 하여 직접 적용하기에 적합한 형태 또는 농축액 또는 1차 조성물일 수 있다. 농도는 특히 농축 또는 직접 사용하거나 하는 것은 특정 제제의 성질, 적용 작물의 종류, 재배지역의 토양 및 기후 등에 따라 다양할 수 있음은 당업계에 명백히 이해될 것이다.The composition of the present invention may be in a form or concentrate or primary composition suitable for direct application by dilution with an appropriate amount of water or other diluent prior to application. It will be clearly understood by those skilled in the art that the concentration, particularly concentration or direct use, may vary depending on the nature of the particular formulation, the type of crop applied, the soil and climate of the cultivation area, and the like.
본 발명의 조성물에 포함되는 미생물의 양에 관해서는, 본 발명의 효과를 손상하지 않는 한 특별히 제한은 없지만, 전체 조성물에 대하여 106 ~ 1010 CFU (colony forming unit)/mL가 되도록 하여 사용하는 것이 바람직하고, 106 ~ 109 CFU/mL 하는 것이 더욱 바람직하고, 106 ~ 109 CFU/mL 하는 것이 보다 바람직하다.Regarding the amount of microorganisms contained in the composition of the present invention, there is no particular limitation as long as the effect of the present invention is not impaired, but 10 6 to 10 10 CFU (colony forming unit) / mL for the entire composition. Preferably, it is more preferably 10 6 to 10 9 CFU/mL, more preferably 10 6 to 10 9 CFU/mL.
본 발명의 미생물 농약 조성물의 제형으로서는 특별히 제한은 없고, 입상, 분상, 액상 등이라도 좋지만, 액상이 바람직하다.There is no restriction|limiting in particular as a dosage form of the microbial pesticide composition of this invention, Although granular, powdery form, liquid etc. may be sufficient, a liquid is preferable.
본 발명의 미생물 농약 조성물의 사용 방법에 관해서는 특별히 제한은 없지만, 예컨대 500배, 1000배, 2000배, 4000배 등, 농약등록시에 정해진 소정의 희석 배율이되도록 물로 희석하여 살포액을 제작하고, 그 살포액을 동력 분무기 등을 사용하여 작물 전체에 안개 형상으로 분무 처리하는 등으로 이용할 수 있다.Although there is no particular limitation on the method of using the microbial pesticide composition of the present invention, for example, 500 times, 1000 times, 2000 times, 4000 times, etc., diluted with water to a predetermined dilution ratio determined at the time of registration of pesticides to prepare a spray solution, The spraying liquid can be used by spraying the whole crop in the form of mist using a power sprayer or the like.
본 발명에서는 영양소가 풍부하며 인체에도 무해한 8종의 유산균 상등액을 깨씨무늬병에 유도된 옥수수에 적용하였고, 그 결과, 비피도박테리움 락티스 IDCC 4301와 락토바실러스 람노서스 IDCC 3201의 상등액이 깨씨무늬병 제어에 매우 높은 효과가 있음을 확인하였다.In the present invention, supernatant of 8 kinds of lactobacilli, which is rich in nutrients and harmless to the human body, was applied to corn induced in sesame seed pattern disease. As a result, the supernatant of Bifidobacterium lactis IDCC 4301 and Lactobacillus rhamnosus IDCC 3201 It was confirmed that there was a very high effect on disease control.
이하 실험예를 통해 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail through experimental examples.
유산균 상등액의 제조Preparation of lactic acid bacteria supernatant
유산균 배양을 위한 액체배지는 각각의 유산균 특성을 고려하여 구성성분을 달리하였으며 [탄소원(포도당, 말토오스, 설탕, 트레할로스) 1.0~3.0%(w/v), 질소원(대두펩톤, 카제인펩톤, 효모추출물) 0.5~3.0%(w/v), 미량원소(구연산나트륨, 초산나트륨, 염화칼슘, 인산일수소칼륨, 인산이수소칼륨, 황산마그네슘, 황산망간) 0.001~3.0%(w/v), 탄산칼슘 0.1~0.5%(w/v)] 121℃, 30분 이상 멸균하여 준비하였다.The liquid medium for culturing lactic acid bacteria has different components in consideration of the characteristics of each lactic acid bacteria. ) 0.5 to 3.0% (w/v), trace elements (sodium citrate, sodium acetate, calcium chloride, potassium monohydrogen phosphate, potassium dihydrogen phosphate, magnesium sulfate, manganese sulfate) 0.001 to 3.0% (w/v), calcium carbonate 0.1~0.5% (w/v)] It was prepared by sterilization at 121°C for 30 minutes or more.
유산균은 다음의 8가지를 사용하였다.The following 8 types of lactic acid bacteria were used.
BBL, Bifidobacterium lactis IDCC 4301; BC, Bacillus coagulans IDCC 1201; BS, Bacillus subtilis IDCC 1101; CB, Clostridium butyricum IDCC 9207; ECF, Enterococcus faecium IDCC 2102; LBP, Lactobacillus plantarum IDCC 3507; LBR, Lactobacillus rhamnosus IDCC 3201; LCL, Lactococcus lactis IDCC 2301; BBL , Bifidobacterium lactis IDCC 4301; BC , Bacillus coagulans IDCC 1201; BS , Bacillus subtilis IDCC 1101; CB , Clostridium butyricum IDCC 9207; ECF , Enterococcus faecium IDCC 2102; LBP , Lactobacillus plantarum IDCC 3507; LBR , Lactobacillus rhamnosus IDCC 3201; LCL , Lactococcus lactis IDCC 2301;
각 유산균은 보리의 맥아, BS, CB, 유아의 변, 김치, 가정식 치즈 등에서 분리하였다.Each lactic acid bacteria was isolated from barley malt, BS, CB, infant feces, kimchi, and homemade cheese.
멸균된 액체배지에 미리 키워둔 유산균을 1.0~5.0%(v/v) 접종하여 30~45℃온도에서 16시간이상 유산균 수가 >1ㅧ107 CFU (colony forming unit)/mL가 될 때까지 배양하였다. 일부 유산균은 배양종료 후에 25~30% 수산화나트륨 용액을 첨가하여 pH 6.0~7.0 범위로 중화하였고, 배양종료 후에 연속식 원심분리기를 이용하여 7000~7500 rpm으로 유산균체와 상등액을 분리하여 각각의 목적에 따라 회수하였다(도 1 참조).Inoculate 1.0~5.0% (v/v) of previously grown lactic acid bacteria in a sterilized liquid medium and incubate at a temperature of 30~45℃ for more than 16 hours until the number of lactic acid bacteria becomes >1ㅧ10 7 CFU (colony forming unit)/mL did Some lactic acid bacteria were neutralized to a pH of 6.0 to 7.0 by adding 25-30% sodium hydroxide solution after the end of the culture. was recovered according to (see FIG. 1).
유산균 상등액 8종의 옥수수 깨씨무늬병에 적용Lactobacillus supernatant applied to 8 types of corn sesame seed bottle
미생물 배양액의 옥수수 깨씨무늬병에 대한 저항성을 유도하는지 확인하기 위하여 미생물 배양액을 3∼4엽기의 옥수수가 심겨져 있는 폿트에 옥수수 개체마다 5mL씩 관주한 후 옥수수를 온실 및 비닐하우스에서 7일간 배양했다.In order to check whether the microbial culture induces resistance to corn sesame disease, 5 mL of the microbial culture was irrigated into pots in which corn of the 3-4 leaf stage was planted, and then the corn was cultured in greenhouses and plastic greenhouses for 7 days.
옥수수 깨씨무늬병균 접종을 위하여 국내 옥수수에서 발생한 깨씨무늬병 균주를 분리한 균주를 사용하였다. 깨씨무늬병 균주를 PDA(Potato dextrose agar; Dextrose 20g, potato infusion 4g, potato stach 4g, potato extract 4g, Agar 15g, 증류수 1L)배지에 접종 후 25℃에 7일 이상 배양하였다.For inoculation with corn sesame seed bacterium, a strain isolated from domestic corn sesame seed streak strain was used. Sesame seed disease strain was inoculated in PDA (Potato dextrose agar; Dextrose 20g, potato infusion 4g, potato stach 4g, potato extract 4g, Agar 15g, distilled water 1L) medium and then cultured at 25℃ for more than 7 days.
배양된 병원균의 균총(직경 5mm)을 쌀겨 평판배지(쌀겨 20g, 설탕 20g, 한천 20g)에서 8~10일간 암상태로 배양했다. 배지 전면에 자란 균총을 고무 브러쉬를 이용하여 기중균사를 제거한 후 40W 형광등을 24시간 조명하면서 3일간 배양 페트리디쉬 뚜겅을 열고 포자 형성을 유도했다. 포자가 형성되면 0.03% Tween 20 용액을 포자가 형성된 배지 표면에 부어 고무 브러쉬를 이용하여 포자를 수거했다. 소독관 가제로 여과하여 현탁액을 제조했다. 이때 포자 농도는 105∼106cell/mL 정도로 조정하여 사용하며 포자현탁액을 접종장치나 스프레이를 사용하여 골고루 분무했다.The cultured pathogen colonies (5 mm in diameter) were cultured in a dark state for 8 to 10 days in rice bran plate medium (rice bran 20 g, sugar 20 g, agar 20 g). After removing the aerial mycelium using a rubber brush for the flora grown on the entire surface of the medium, spore formation was induced by opening the lid of the Petri dish cultured for 3 days while illuminating a 40W fluorescent lamp for 24 hours. When spores were formed, 0.03% Tween 20 solution was poured onto the surface of the medium on which the spores were formed, and the spores were collected using a rubber brush. A suspension was prepared by filtration with sterile tube gauze. At this time, the spore concentration was adjusted to about 105~106 cells/mL and the spore suspension was sprayed evenly using an inoculation device or a sprayer.
병을 접종한 옥수수는 24시간동안 암조건에서 포화습도(90%이상)조건을 유지시켰다. 암조건 습실처리가 끝나면 시료를 온실 및 비닐하우스등에서 생육시키면서 병 발생을 관찰하였는데, 필요에 따라 병 발생을 조장하기 위하여 습도를 유지시켰다.The inoculated corn was maintained at saturated humidity (90% or more) under dark conditions for 24 hours. After the dark conditions and wet room treatment was completed, the samples were grown in greenhouses and plastic houses, and disease occurrence was observed. If necessary, humidity was maintained to promote disease occurrence.
위와 같이 처리된 옥수수를 온실로 옮긴 후 습도를 보완시켜 7일간 정치하여 발병시켰다. 옥수수 깨씨무늬병은 유산균 상등액의 처리유무에 따라 일주일 동안 관찰하였고, 저항성 검정법에 따라 병심각도(severity)를 0∼5까지 단계를 나누어 심각도를 측정하였다. 심각도는 0 (미발생)부터 5 (고사)까지이다.After the corn treated as above was transferred to the greenhouse, the humidity was supplemented, and the disease was developed by standing still for 7 days. Corn sesame seed blotch was observed for a week depending on whether or not the lactobacillus supernatant was treated, and the severity was measured by dividing the severity from 0 to 5 according to the resistance test method. Severity ranges from 0 (non-occurring) to 5 (fatal).
그 결과, 도 2와 같이 무처리 군(CON)에서는 심각도 평균 4, 즉 고사상태와 근접한 결과가 나왔으며, BC, BS, CB, LBP, ECF 및 LCL의 상등액 처리군에서도 무처리군과 유사하게 심각도가 3∼4.5의 범위로 옥수수 깨시무늬병의 호전상태를 관찰할 수 없었다. 그러나, BBL(Bifidobacterium lactis) 및 LBR(Lactobacillus rhamnosus)의 상등액을 처리한 군에서는 심각도가 1.3∼1.8의 범위로 옥수수 깨시무늬병의 제어효과가 뛰어남을 유의적으로 관찰할 수 있었다.As a result, as shown in FIG. 2, in the untreated group (CON), the average severity of 4, that is, close to the dead state, was obtained, and in the supernatant treated group of BC, BS, CB, LBP, ECF and LCL, similarly to the untreated group. In the range of 3 to 4.5 in severity, no improvement was observed in the cornstarch disease. However, in the group treated with the supernatant of Bifidobacterium lactis (BBL) and Lactobacillus rhamnosus (LBR), the severity ranged from 1.3 to 1.8, and it was significantly observed that the control effect of cornstarch disease was excellent.
Claims (3)
상기 작물은 옥수수를 포함하는 조성물.In claim 1,
The crop composition comprising corn.
상기 유산균은 피도박테리움 락티스 IDCC 4301 및 락토바실러스 람노서스 IDCC 3201 중 적어도 어느 하나를 포함하는 조성물.In any one of claims 1 and 2,
The lactic acid bacteria is a composition comprising at least one of Pidobacterium lactis IDCC 4301 and Lactobacillus rhamnosus IDCC 3201.
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