KR20200069765A - Composition for controlling cobweb disease comprising extract of Hosta longissima HONDA as an active ingradient and uses thereof - Google Patents
Composition for controlling cobweb disease comprising extract of Hosta longissima HONDA as an active ingradient and uses thereof Download PDFInfo
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Abstract
Description
본 발명은 산옥잠화(Hosta longissima HONDA) 추출물 또는 이의 분획물을 유효성분으로 포함하는 솜털곰팡이병 방제용 조성물 및 이의 용도에 관한 것으로, 상세하게는 산옥잠화(Hosta longissima HONDA) 추출물 또는 이의 분획물을 유효성분으로 포함하는 솜털곰팡이병 방제용 조성물, 이를 이용한 솜털곰팡이병 방제 방법 및 상기 방제용 조성물을 포함하는 솜털곰팡이병 방제제에 관한 것이다.The present invention relates to a composition for controlling fluffy fungal disease and its use, comprising an extract of Hosta longissima HONDA or a fraction thereof as an active ingredient, and in detail, an extract of Hosta longissima HONDA or a fraction thereof The present invention relates to a composition for controlling fluffy fungal disease, a method for controlling fluffy fungal disease using the same, and a fluffy fungal disease control agent comprising the composition for controlling.
솜털곰팡이병 또는 흰곰팡이병은 솜털곰팡이병원균(Cladobotryum mycophilum) 등에 의해 발병되며 우리나라에서는 새송이 버섯과 팽이 버섯에서 처음으로 발견되었고 양송이 버섯에서는 지난 2009년 경북지역의 양송이 재배농가에서 최초로 확인되었다.Downy mildew disease or white mold disease is caused by the downy mildew pathogen (Cladobotryum mycophilum), and was first discovered in oyster mushrooms and mushrooms in Korea, and was first confirmed in 2009 by a mushroom farm in Gyeongbuk in 2009.
솜털곰팡이병은 발병초기 복토 후 좁은 면적에서 하얗게 올라오는 곰팡이 균으로 시작되고 버섯균사와 색깔이 유사해 쉽게 구별하기 어려우나 시간이 지남에 따라 연분홍색이나 노란색으로 변한다. 특히 솜털곰팡이병은 버섯의 기부에서부터 솜털모양으로 잠식하다가 병이 진전되면서 버섯 전체에 퍼져서 부패하게 되며 공기 중에 포자를 비산시키는 성질이 있어 한번 발병하면 재배사 전체에 급격히 번질 수 있다. 따라서 솜털곰팡이병 발생이 확인된 경우 발병부위를 격리시켜 포자의 비산을 막는 것이 급선무다. 이와 함께 이 병원균은 높은 습도와 19~22℃의 온도조건에서 가장 많이 발생하는데 일단 병징이 발견된 후 이 병원균을 제거하는 것은 사실상 불가능하므로 예방이 최선이다.Fluffy fungal disease begins as a fungal fungus that rises white in a small area after the initial onset of the onset, and is similar in color to the fungal mycelia, making it difficult to distinguish easily, but changes over time to light pink or yellow. In particular, fluffy fungal disease is eroded from the base of the mushroom to the fluffy shape and spreads throughout the mushrooms as the disease progresses. It spreads spores in the air and spreads spores in the air. Therefore, when the occurrence of downy mildew is confirmed, it is urgent to isolate the affected area to prevent spores from scattering. Along with this, the pathogen is most likely to occur at high humidity and a temperature of 19~22℃. Once the disease is found, it is virtually impossible to remove the pathogen, so prevention is best.
한편, 산옥잠화(Hosta longissima HONDA)는 백합과의 여러해살이풀로 냇가의 바위틈에서 자란다. 잎은 모두 뿌리에서 나오고 타원형이거나 달걀모양으로 윤이나는 녹색이며 끝은 뾰족하고 가장자리가 밋밋하거나 우굴쭈굴하고 4~5쌍의 맥이 있다. 꽃은 7~8월에 자줏빛으로 피고 총상꽃차례에 한쪽으로 치우쳐서 달린다. 열매는 삭과(果)로서 긴 타원형이고 비스듬히 서며 3개로 갈라지며, 봄에 연한 잎을 나물로 먹는다. 효능은 이기(理氣), 보허, 화혈(和血), 지통(止痛)의 효능이 있고, 대하증, 변혈증, 복통, 어혈, 옹종(외옹), 종독, 진통, 치통, 타박상 몸과기를 보호하고 ,통증을 없애고, 염증을 삭히며 피를 멈추고 소변이 잘 나오게 하는 효능이 있다. Meanwhile, Hosta longissima HONDA is a perennial plant of the Liliaceae family that grows in the crevices of streams. The leaves are all from the root, oval or egg-shaped, shiny green, sharp at the end, flat or rugged, with 4 to 5 pairs of veins. Flowers bloom in purple in July-August and run sideways in the inflorescence. Fruits are oval, long oval, obliquely, split into three, and eat light leaves as herbs in spring. Efficacy (이), boheo, hwahyeol (和血), has the effect of pain (止痛), hypertrophy, septicemia, abdominal pain, blood, carcinoma (exorcism), poison, pain relief, toothache, bruises and protects the body, It has the effect of eliminating pain, stopping inflammation, stopping blood and making urine better.
산옥잠화와 관련하여, 등록특허 제10-1807954호에는 산옥잠화 추출물의 피하지방 감소 개선 효과가 개시되어 있으나, 산옥잠화의 항균력에 대한 연구는 부족한 실정이며 특히 식물 병해 방제를 위한 산옥잠화 연구는 전혀 이루어진 바가 없다.In connection with wild daylily, Patent No. 10-1807954 discloses an effect of improving the reduction of subcutaneous fat of the daylily, but research on the antibacterial activity of the daylily is insufficient, and in particular, the study of daylily for the control of plant diseases is completely absent. Nothing has been done.
이러한 배경 하에, 본 발명자들은 양송이 솜털곰팡이병의 생물학적 방제를 위하여 산옥잠화 추출물을 이용하여 화학농약 대체를 위한 친환경 방제법을 개발할 목적으로 수행하고, 이와 같은 효과를 확인하여 본 발명을 완성하였다.Under these backgrounds, the present inventors conducted the purpose of developing an eco-friendly control method for replacing chemical pesticides using wild oxidized extracts for biological control of the mushroom fungus disease, and completed the present invention by confirming such effects.
본 발명은 상기와 같은 요구에 의해 안출된 것으로, 화학농약을 대체할 수 있는 생물학적 방제를 위한 솜털곰팡이병 방제용 조성물을 제공하는데 목적이 있다.The present invention has been made by the above-mentioned needs, and has an object to provide a composition for controlling fluffy fungal diseases for biological control capable of replacing chemical pesticides.
본 발명의 다른 목적은 상기 방제용 조성물을 이용한 솜털곰팡이병 방제 방법을 제공하는 것이다.Another object of the present invention is to provide a method for controlling fluffy mold disease using the composition for controlling the above.
본 발명의 또 다른 목적은 상기 방제용 조성물을 포함하는 솜털곰팡이병 방제제를 제공하는 것이다.Another object of the present invention is to provide a fluffy fungal disease control agent comprising the composition for control.
상술한 과제를 해결하기 위하여, 본 발명의 솜털곰팡이병 방제용 조성물은 산옥잠화(Hosta longissima HONDA) 추출물 또는 이의 분획물을 유효성분으로 포함할 수 있다.In order to solve the above-mentioned problems, the composition for controlling fluffy fungi of the present invention may include an extract of Hosta longissima HONDA or a fraction thereof as an active ingredient.
본 발명의 바람직한 일 실시예에 있어서, 산옥잠화 추출물은 산옥잠화 뿌리로부터 추출한 것일 수 있다.In a preferred embodiment of the present invention, the wild daylily extract may be extracted from the root of wildfire daylily.
본 발명의 바람직한 일 실시예에 있어서, 산옥잠화 추출물은 C1 내지 C4 저급 알코올을 용매로 하여 추출한 것일 수 있다.In a preferred embodiment of the present invention, the acid daylily extract may be extracted with C 1 to C 4 lower alcohol as a solvent.
본 발명의 바람직한 일 실시예에 있어서, 산옥잠화 추출물은 메탄올을 용매로 하여 추출하는 것일 수 있다.In a preferred embodiment of the present invention, the acid daylily extract may be extracted using methanol as a solvent.
본 발명의 바람직한 일 실시예에 있어서, 산옥잠화 분획물은 산옥잠화 추출물을 n-헥산, 클로로포름, 에틸아세테이트 및 부탄올로 이루어진 군에서 선택된 어느 하나로 분획하여 제조된 것일 수 있다.In a preferred embodiment of the present invention, the oxidized fraction may be prepared by fractionating the oxidized extract from any one selected from the group consisting of n-hexane, chloroform, ethyl acetate, and butanol.
본 발명의 바람직한 일 실시예에 있어서, 산옥잠화 분획물은 산옥잠화 추출물을 n-헥산으로 분획하여 제조된 것일 수 있다.In a preferred embodiment of the present invention, the acid oxidized fraction may be prepared by fractionating the acid oxidized extract with n-hexane.
한편, 본 발명의 솜털곰팡이병 방제 방법은 앞서 언급한 솜털곰팡이병 방제용 조성물을 버섯에 처리하는 단계를 포함한다.On the other hand, the method for controlling fluffy mold disease of the present invention includes the step of treating the above-mentioned composition for controlling fluffy mold disease on mushrooms.
본 발명의 바람직한 일 실시예에 있어서, 상기 버섯은 양송이버섯, 새송이버섯, 팽이버섯, 느타리버섯 및 큰느타리버섯으로 이루어진 군에서 선택된 어느 1종일 수 있다.In a preferred embodiment of the present invention, the mushroom may be any one selected from the group consisting of mushrooms, oyster mushrooms, enoki mushrooms, oyster mushrooms and large oyster mushrooms.
나아가, 본 발명의 솜털곰팡이병 방제제는 앞서 언급한 솜털곰팡이병 방제용 조성물을 포함한다.Furthermore, the fluffy fungal disease control agent of the present invention includes the above-mentioned composition for controlling fluffy fungal disease.
본 발명의 바람직한 일 실시예에 있어서, 본 발명의 솜털곰팡이병 방제제는 수화제, 현탁제, 유제, 유탁제, 미탁제, 액제, 분산성액제, 입상수화제, 입제, 분제, 액상수화제, 입상수화제, 수면부상성입제 및 정제로 이루어진 군에서 선택된 어느 하나의 형태로 제제화된 것일 수 있다.In one preferred embodiment of the present invention, the fluffy fungal disease control agent of the present invention is a hydrating agent, a suspending agent, an emulsion, an emulsifying agent, an emulsifying agent, a liquid agent, a dispersing liquid agent, a granular hydrating agent, a granulating agent, a powdering agent, a liquid hydrating agent, a granulating hydrating agent, It may be formulated in any one form selected from the group consisting of hypnotic granules and tablets.
본 발명의 산옥잠화 추출물 또는 이의 분획물을 포함하는 솜털곰팡이병 방제용 조성물은 버섯에서 많은 문제가 되고 있는 솜털곰팡이병을 효과적으로 방제하여 버섯 생산성 향상에 유용하다.The composition for controlling fluffy fungal diseases comprising the extract of the wild daylily extract or a fraction thereof of the present invention is useful for improving mushroom productivity by effectively controlling fluffy fungal diseases that are a lot of problems in mushrooms.
또한, 본 발명의 산옥잠화 추출물 또는 이의 분획물을 포함하는 솜털곰팡이병 방제용 조성물을 이용한 방제 방법은 친환경적인 생물학적 방제방법이므로, 버섯의 친환경 재배를 가능하게 하여, 이를 통해 소비자의 신뢰 제고 및 버섯 소비 확대에 기여할 수 있을 것으로 예상된다.In addition, since the control method using the composition for controlling fluffy fungal diseases containing the extract of wildfire of the present invention or a fraction thereof is an eco-friendly biological control method, it enables eco-friendly cultivation of mushrooms, thereby enhancing consumer confidence and mushroom consumption It is expected to contribute to the expansion.
도 1은 산옥잠화 추출물 및 이로부터 분획물을 제조하는 과정을 모식도로 나타낸 것이다.
도 2는 산옥잠화 추출물의 솜털곰팡이병원균에 대한 항균 효과를 페이퍼 디스크법(paper disk)으로 확인한 결과를 나타낸 것이다.
도 3은 산옥잠화 분액층별 솜털곰팡이병원균에 대한 항균 효과를 페이퍼 디스크법으로 확인한 결과를 나타낸 것으로, Hexane는 n-헥산, CHCl3는 클로로포름, EA는 에틸아세테이트, BuOH는 부탄올을 나타낸다.
도 4는 산옥잠화 분액층별 함유물질의 TLC 분석을 나타낸 것으로, Hx는 n-헥산, CHCl3는 클로로포름, EA는 에틸아세테이트, BuOH는 부탄올을 나타낸다.
도 5는 솜털곰팡이병원균에 감염된 양송이버섯에 산옥잠화 n-핵산 분획물을 처리하여 방제 효과를 확인한 것이다.1 is a schematic diagram showing a process for preparing a mountain oxidized extract and a fraction therefrom.
Figure 2 shows the results confirmed by the paper disk method (paper disk) of the antibacterial effect on the fluffy fungi pathogens of the wild daylily extract.
Figure 3 shows the results of confirming the antimicrobial effect on the fluffy fungi pathogens of each layer of acid daylily by the paper disk method, Hexane is n-hexane, CHCl 3 is chloroform, EA is ethyl acetate, BuOH is butanol.
Figure 4 shows the TLC analysis of the inclusion material for each layer of acid oxidization, Hx is n-hexane, CHCl 3 is chloroform, EA is ethyl acetate, BuOH is butanol.
FIG. 5 shows the control effect by treating the wild-fermented fungus pathogen mushrooms with mushrooms with n-nucleic acid fractions.
이하, 본 발명을 보다 상세히 설명한다.Hereinafter, the present invention will be described in more detail.
상술한 바와 같이, 버섯에서 많은 문제가 되고 있는 솜털곰팡이병에 관한 연구 및 이를 효과적으로 방제할 수 있는 생물학적 방제제에 대한 연구는 미비한 실정이다.As described above, studies on fluffy fungal diseases, which are a lot of problems in mushrooms, and studies on biological control agents that can effectively control them are incomplete.
이에 본 발명은 산옥잠화 추출물 또는 이의 분획물을 유효성분으로 포함하는 솜털곰팡이병 방제용 조성물을 제공함으로써 상술한 문제의 해결방안을 모색하였다. 본 발명에 따른 솜털곰팡이병 방제용 조성물은 솜털곰팡이병을 효과적으로 방제하여 버섯 생산성 향상에 유용하다.Accordingly, the present invention sought to solve the above-mentioned problems by providing a composition for controlling fluffy fungal diseases, which includes an extract of daylily or a fraction thereof as an active ingredient. The composition for controlling fluffy mold disease according to the present invention is useful for improving mushroom productivity by effectively controlling fluffy mold disease.
또한, 본 발명의 산옥잠화 추출물 또는 이의 분획물을 포함하는 솜털곰팡이병 방제용 조성물을 이용한 방제 방법은 친환경적인 생물학적 방제방법이므로, 버섯의 친환경 재배를 가능하게 하여, 이를 통해 소비자의 신뢰 제고 및 버섯소비 확대에 기여할 수 있을 것으로 예상된다.In addition, since the control method using the composition for controlling fluffy fungal diseases containing the extract of the wild daylily extract or its fraction of the present invention is an eco-friendly biological control method, it enables eco-friendly cultivation of mushrooms, thereby increasing consumer confidence and mushroom consumption. It is expected to contribute to the expansion.
본 발명은 산옥잠화 추출물 또는 이의 분획물을 포함하는 솜털곰팡이병 방제용 조성물을 제공한다.The present invention provides a composition for controlling fluffy fungal disease comprising an extract of daylily or a fraction thereof.
상기 산옥잠화 추출물은 산옥잠화 전초, 꽃, 잎, 열매, 줄기, 종자 또는 뿌리로부터 추출된 것일 수 있으나, 바람직하게는 산옥잠화 뿌리로부터 추출된 것일 수 있다.The wild daylily extract may be extracted from a wild daylily, flower, leaf, fruit, stem, seed or root, but preferably may be extracted from the wild daylily root.
상기 산옥잠화 추출물 또는 이의 분획물은 하기의 단계들을 포함하는 제조방법에 의해 제조되는 것이 바람직하나 이에 한정되는 것은 아니다:It is preferred that the wild daylily extract or a fraction thereof is prepared by a manufacturing method including the following steps, but is not limited thereto:
1) 산옥잠화에 추출용매를 가하여 추출하는 단계;1) extracting by adding an extracting solvent to the wildfire;
2) 상기 1) 단계의 추출물을 여과하는 단계;2) filtering the extract of step 1);
3) 상기 2) 단계의 여과된 추출물을 감압농축한 후 건조하여 산옥잠화 추출물을 제조하는 단계; 및3) concentrating the filtered extract of step 2) under reduced pressure and drying it to prepare a mountain oxidized extract; And
4) 상기 3) 단계의 산옥잠화 추출물을 추가적으로 유기용매로 추출하여 분획물을 제조하는 단계.4) The step of preparing a fraction by extracting the acid oxidized extract of step 3) with an additional organic solvent.
상기 방법에 있어서, 1) 단계의 산옥잠화는 재배한 것 또는 시판되는 것 등을 제한없이 사용할 수 있다.In the above method, the oxidizing step 1) can be used without limitation, such as those grown or commercially available.
상기 추출용매는 알코올을 사용하는 것이 바람직하다. 상기 알코올로는 C1 내지 C4의 저급 알코올, 바람직하게는 C1 내지 C2의 저급 알코올을 사용할 수 있으나, 이로 한정되는 것은 아니다. 상기 C1 내지 C2의 저급 알코올로는 메탄올 또는 에탄올을 사용하는 것이 좋으며, 가장 바람직하게는 100% 메탄올을 사용하는 것이 가장 좋다.It is preferable to use alcohol as the extraction solvent. The alcohol may be C 1 to C 4 lower alcohol, preferably C 1 to C 2 lower alcohol, but is not limited thereto. As the lower alcohol of C 1 to C 2 , it is preferable to use methanol or ethanol, and most preferably, 100% methanol is used.
추출방법으로는 교반추출, 감압고온추출, 열탕추출, 환류추출, 열수추출, 냉침추출, 상온추출, 초음파 추출 또는 증기추출을 이용하는 것이 바람직하나 이에 한정되지 않는다. 상기 추출용매를 산옥잠화 식물 분량에 1 내지 10배 첨가하여 추출하는 것이 바람직하다. As an extraction method, it is preferable to use agitated extraction, reduced pressure high temperature extraction, hot water extraction, reflux extraction, hot water extraction, cold immersion extraction, room temperature extraction, ultrasonic extraction or steam extraction, but is not limited thereto. It is preferable to extract the extraction solvent by adding 1 to 10 times to the amount of the plant of daylily.
추출온도는 20℃ 내지 100℃인 것이 바람직하고, 20℃ 내지 40℃인 것이 더욱 바람직하며, 실온인 것이 가장 바람직하나 이에 한정되지 않는다. 추출시간은 2시간 내지 48시간인 것이 바람직하나 이에 한정되지 않는다. 아울러, 추출 회수는 2 내지 5회인 것이 바람직하고, 3회 내지 4회 반복 추출하는 것이 더욱 바람직하나 이에 한정되는 것은 아니다.The extraction temperature is preferably 20°C to 100°C, more preferably 20°C to 40°C, and most preferably room temperature, but is not limited thereto. The extraction time is preferably 2 hours to 48 hours, but is not limited thereto. In addition, the number of extractions is preferably 2 to 5 times, and it is more preferable to repeatedly extract 3 to 4 times, but is not limited thereto.
상기 방법에 있어서, 3) 단계의 감압농축은 진공감압농축기 또는 진공회전증발기를 이용하는 것이 바람직하나 이에 한정되지 않고, 건조는 감압건조, 진공건조, 비등건조, 분무건조 또는 동결건조하는 것이 바람직하나 이에 한정되지 않는다.In the above method, it is preferable to use a vacuum decompressor or a vacuum rotary evaporator in step 3) under reduced pressure, but the drying is preferably under reduced pressure drying, vacuum drying, boiling drying, spray drying or freeze drying. It is not limited.
상기 방법에 있어서, 4) 단계의 유기용매는 n-헥산, 클로로포름, 에틸아세테이트 또는 부탄올인 것이 바람직하고, n-헥산이 가장 바람직하나, 이에 한정되지 않는다. 상기 분획물은 산옥잠화 추출물을 물에 현탁시킨 후 n-헥산, 클로로포름, 에틸아세테이트 및 부탄올로 순차적으로 계통 분획하여 수득한 n-헥산 분획물, 클로로포름 분획물, 에틸아세테이트 분획물 또는 부탄올 분획물인 것이 바람직하나, 이에 한정되지 않는다. 상기 분획물은 상기 산옥잠화 추출물로부터 분획 과정을 1 내지 5회, 바람직하게는 3회 반복하여 수득할 수 있고, 분획 후 감압 농축하는 것이 바람직하나 이에 한정되지 않는다.In the above method, the organic solvent of step 4) is preferably n-hexane, chloroform, ethyl acetate or butanol, and n-hexane is most preferred, but is not limited thereto. The fraction is preferably an n-hexane fraction, a chloroform fraction, an ethyl acetate fraction or a butanol fraction obtained by sequential systemic fractionation with n-hexane, chloroform, ethyl acetate and butanol after suspending the acid daylily extract in water, It is not limited. The fraction can be obtained by repeating the fractionation process from 1 to 5 times, preferably 3 times, from the acid oxidized extract, and is preferably concentrated under reduced pressure after fractionation, but is not limited thereto.
본 발명의 구체적인 실시예에서, 본 발명자들은 산옥잠화 뿌리로부터 산옥잠화 추출물 및 이의 분획물을 제조하였으며, 제조 과정의 모식도는 도 1에 나타난 바와 같다.In a specific embodiment of the present invention, the present inventors prepared a wild oxidized extract and fractions thereof from the wild oxidized roots, and a schematic diagram of the manufacturing process is shown in FIG. 1.
본 발명의 산옥잠화 추출물 또는 이의 분획물의 솜털곰팡이병원균에 대한 항균 활성을 페이퍼 디스크법(paper disk)으로 측정한 결과, 추출물과 분획물 모두 솜털곰팡이병원균을 효과적으로 억제하였으며, 그 중에서도 n-헥산 분획물의 항균 활성이 가장 우수함을 확인하였다(도 2 및 3). 또한, 양송이버섯 자실체에 솜털곰팡이병원균을 접종한 후 산옥잠화 추출물과 분획물을 각각 처리하여 솜털곰팡이병해의 방제효과를 확인한 결과, 산옥잠화 n-헥산 분획물 처리구에서 87.9%의 우수한 방제 효과가 있음을 확인하였다(표 1 및 도 5).As a result of measuring the antibacterial activity against the fluffy fungal pathogen of the wild daylily extract or its fraction of the present invention by paper disk method, both the extract and the fraction effectively inhibited the fungal pathogen, and among them the antibacterial activity of the n-hexane fraction It was confirmed that the activity is the best (Fig. 2 and 3). In addition, after inoculating fluffy fungus pathogens on the mushroom fruiting body, the extract was treated with acid extracts and fractions respectively to confirm the control effect of fluffy fungal diseases. (Table 1 and Figure 5).
따라서, 본 발명의 산옥잠화 추출물 또는 이의 분획물은 솜털곰팡이병원균을 유의적으로 억제하므로, 솜털곰팡이병 방제용 조성물로 유용하게 사용될 수 있으며, 솜털곰팡이병 방제제, 종자 코팅제, 미생물 영양제, 토양 개량제 등의 전반적인 농약제로 사용될 수 있다.Therefore, since the acid daylily extract or fractions thereof of the present invention significantly inhibits the fungal pathogen, it can be usefully used as a composition for controlling fluffy fungal disease, fluffy fungal disease control agent, seed coating agent, microbial nutritional agent, soil improver, etc. It can be used as an overall pesticide.
본 발명은 또한, 전술한 솜털곰팡이병 방제용 조성물을 버섯에 처리하는 단계를 포함하는 솜털곰팡이병 방제 방법을 제공한다.The present invention also provides a method for controlling fluffy mold disease, comprising the step of treating the above-mentioned composition for controlling fluffy mold disease on mushrooms.
상기 솜털곰팡이병 방제용 조성물이 처리되는 버섯은 농가에서 재배되는 것이라면 특별히 제한하지 않으나, 바람직하게는 솜털곰팡이병원균에 의해 솜털곰팡이병이 발생하는 버섯 종일 수 있으며, 예를 들어, 양송이버섯, 새송이버섯, 팽이버섯, 느타리버섯 및 큰느타리버섯으로 이루어진 군에서 선택된 어느 1종일 수 있다.The mushrooms to which the composition for controlling fluffy fungus disease is processed are not particularly limited as long as they are cultivated in a farmhouse, and preferably, may be a mushroom species in which fluffy fungal diseases are caused by fluffy fungal pathogens, for example, mushroom mushrooms, king oyster mushrooms. , Enoki mushroom, oyster mushroom, and may be any one selected from the group consisting of oyster mushroom.
상기 방제 방법은 솜털곰팡이병 방제용 조성물은 분무살포를 포함하는 통상적인 방법으로 처리될 수 있으며, 솜털곰팡이병원균이 효과적으로 억제될 수 있는 처리 방법이라면 특별히 제한없이 사용될 수 있다.The control method, the composition for controlling fluffy mold disease can be treated by a conventional method including spray spraying, and can be used without particular limitation, as long as it is a treatment method that can effectively suppress fluffy mold pathogens.
본 발명은 또한, 전술한 방제용 조성물을 포함하는 솜털곰팡이병 방제제를 제공한다.The present invention also provides a fungal disease control agent comprising the above-mentioned control composition.
본 발명에 따른 솜털곰팡이병 방제제는 산옥잠화 추출물 또는 이의 분획물 이외에 당업계에서 통상적으로 사용되는 살충제 또는 살균제에 함유되는 물질을 추가로 포함할 수 있다.The downy mildew disease control agent according to the present invention may further include a material contained in an insecticide or fungicide commonly used in the art, in addition to the extract of daylily or a fraction thereof.
본 발명의 솜털곰팡이병 방제제는 수화제, 현탁제, 유제, 유탁제, 미탁제, 액제, 분산성 액제, 입상수화제, 입제, 분제, 액상수화제, 입상수화제, 수면부상성입제 및 정제로 이루어진 군에서 선택된 어느 하나의 형태로 제제화될 수 있다.Fluffy fungal disease control agent of the present invention is in the group consisting of hydration agent, suspension agent, emulsion, emulsion agent, mist agent, liquid agent, dispersible liquid agent, granular hydration agent, granule, powder, liquid hydrating agent, granular hydration agent, sleep injury granules and tablets It can be formulated in any one form selected.
이하, 본 발명을 실시예에 의해 상세히 설명한다. 단, 하기 실시예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예에 한정되는 것은 아니다.Hereinafter, the present invention will be described in detail by examples. However, the following examples are merely illustrative of the present invention, and the contents of the present invention are not limited to the following examples.
<실시예 1 : 병원균의 분리 및 병원성 조사><Example 1: Pathogen isolation and pathogenicity investigation>
양송이버섯 재배 농가에서 솜털곰팡이병 증상을 보이는 자실체의 이병부위를 표면살균한 후 1cm 크기로 절단하여 PDA(potato dextrose agar) 배지에 치상하여 25℃의 항온기에서 3∼4일 배양하면서 자라나온 균사 선단부위를 화염 멸균한 칼을 사용하여 5mm 크기로 잘라서 PDA에 다시 배양하여 병원균을 순수 분리하였다. 그리고 순수 분리한 균주를 유전자분석, 균학적특성, 병원성검정 등을 통하여 클라도보트리움 마이코필움(C. mycophilum) 균으로 동정하여 실험에 사용하였다.After disinfection of the diseased part of the fruiting body showing fluffy fungal disease in the mushroom cultivation farmhouse, it is cut into 1 cm size and densified in a PDA (potato dextrose agar) medium, cultured for 3-4 days in a thermostat at 25°C to grow hyphae. The site was cut to a size of 5 mm using a flame sterilized knife and cultured again on a PDA to isolate the pathogen purely. In addition, the purely isolated strain was identified as C. mycophilum bacteria through genetic analysis, mycological characteristics, and pathogenicity tests, and used in the experiment.
양송이버섯에서 분리된 클라도보트리움 마이코필움(C. mycophilum)균의 병원성 유무를 조사하기 위하여 멸균수에 병원균의 포자밀도가 55/ml이 되도록 희석하여 접종원으로 사용하였다. 양송이버섯의 갓 표면에 희석한 병원균 포자를 분무하여 25℃, 95%의 항온기에 넣어 2일 후 병원성유무를 확인하였다.To investigate the presence or absence of pathogenicity of C. mycophilum bacteria isolated from mushrooms, the mushroom was diluted with sterile water so that the spore density of the pathogen was 5 5 /ml and used as an inoculum. After spraying the pathogen spores diluted on the fresh surface of mushrooms with mushrooms, put them in a thermostat at 25℃ and 95%, and after 2 days, the presence or absence of pathogenicity was confirmed.
그 결과, 병원성은 균주에 따라 약간의 차이는 있었지만 양송이버섯 균주와 자실체에 전형적인 솜털곰팡이병 증상의 병원성을 보였다.As a result, although the pathogenicity was slightly different depending on the strain, it showed pathogenicity of fluffy fungal disease symptoms typical to the mushroom mushroom strain and fruiting body.
<실시예 2 : 산옥잠화 뿌리 추출물 및 이의 분획물 제조><Example 2: Preparation of root extracts of wild daylily and its fractions>
국립원예특작과학원 인삼특작부에서 보관 중인 식물 추출물 300점(메탄올 추출물 300)을 분양 받아 페이퍼 디스크법에 의해 1차 항균활성 추출물을 선발하였고, 여러 가지 식물 추출물 중 산옥잠화 추출물이 솜털곰팡이병원균인 클라도보트리움 마이코필움(C. mycophilum) 균에 대해 가장 높은 항균활성을 나타낸다는 것을 발견하였다.The primary antibacterial activity extract was selected by the paper disc method by pre-sale of 300 plant extracts (methanol extract 300) stored by the Ginseng Special Division of the National Academy of Horticultural Science, and the extract from wild plants of the various plant extracts is a fungal pathogen. It was found that it exhibits the highest antimicrobial activity against Bacillus mycophilum.
산옥잠화 추출물을 제조하기 위해, 산옥잠화 뿌리 건조분말 시료 5.0kg을 100% 메탄올 30L 넣고 상온에서 24시간 동안 교반 추출한 후 여과지(Advantec NO.2)로 여과하여 여액을 감압농축하여 추출물을 얻었다. 산옥잠화 시료의 추출은 3회 반복하여 수행하였다. 메탄올 추출물 100g을 취하여 3차 증류수에 현탁 및 용해시킨 후에 헥산(Hx), 클로로포름(CHCl3), 에틸아세테이트(EA), 부탄올(BuOH)로 순차적으로 분획하여 헥산층에서 16.5g의 분획물을 얻었다. 분리 물질별 TLC 확인(도 4)후 비슷한 band 끼리 모아 농축하였다. 100% 메탄올 추출물로부터의 분획물 제조 과정의 모식도는 도 1에 나타난 바와 같다.In order to prepare an extract of wild daylily, 5.0 kg of a dry root sample of wild daylily root was added with 30 L of 100% methanol, extracted with stirring at room temperature for 24 hours, filtered with filter paper (Advantec NO.2), and the filtrate was concentrated under reduced pressure to obtain an extract. Extraction of the sample of wild oxidization was repeated three times. After taking 100 g of methanol extract and suspending and dissolving it in tertiary distilled water, it was sequentially fractionated with hexane (Hx), chloroform (CHCl 3 ), ethyl acetate (EA), butanol (BuOH) to obtain a fraction of 16.5 g in the hexane layer. After confirming TLC for each separation material (FIG. 4), similar bands were collected and concentrated. A schematic diagram of a process for preparing a fraction from 100% methanol extract is shown in FIG. 1.
<실시예 3 : 산옥잠화 뿌리 추출물과 이의 분획물의 항균활성 측정><Example 3: Measurement of the antibacterial activity of the root extract of wild daylily and its fraction>
상기 실시예 2에서 제조된 산옥잠화 뿌리 추출물과 이의 분획물의 솜털곰팡이병원균에 대한 항균 활성을 페이퍼 디스크법(paper disk)으로 측정하였다.The antimicrobial activity against the fluffy fungal pathogen of the wildflower root extract and fractions thereof prepared in Example 2 was measured by a paper disk method.
상기 실시예 1에서 분리한 솜털곰팡이병원균은 PDA 배지에 5일간 배양 후 콕 보러(cork borer)를 이용하여 가장자리를 채취하여 검정용 PDA 배지의 양쪽 가장자리에 치상하고 정 가운데에 70㎕ 추출물과 분획물을 페이퍼 디스크(8mm)에 분주하여 생육저지환의 정도에 따라 항균 능력을 평가하였다.The fluffy fungal pathogen isolated in Example 1 was cultured in PDA medium for 5 days, and then the edges were collected by using a cork borer to dentate the teeth on both edges of the PDA medium for assay, and 70 μl extract and fraction in the middle. It was dispensed on a paper disk (8 mm) to evaluate the antibacterial ability according to the degree of growth inhibition.
그 결과, 도 2에 나타난 바와 같이 산옥잠화 뿌리 추출물은 솜털곰팡이병원균인 클라도보트리움 마이코필움(C. mycophilum)에 대해 높은 항균 활성을 보였다.As a result, as shown in FIG. 2, the root extract of San Daylily showed high antimicrobial activity against C. mycophilum, which is a fungal pathogen.
또한, 도 3에 나타난 바와 같이 n-헥산(Hexane), 클로로포름(CHCl3), 에틸아세테이트(EA) 및 부탄올(BuOH) 분획물 모두 솜털곰팡이병원균을 효과적으로 억제하였으며, 그 중 에서도 n-헥산 분획물이 가장 우수한 항균 활성을 나타냈다.In addition, as shown in Figure 3, n-hexane (Hexane), chloroform (CHCl 3 ), ethyl acetate (EA), and butanol (BuOH) fractions of all effectively suppressed the fungal pathogen, among which n-hexane fraction is the most It showed excellent antibacterial activity.
<실시예 4 : 솜털곰팡이병해의 방제효과><Example 4: Control effect of fluffy mold disease>
솜털곰팡이병해의 방제효과를 검정하기 위해 상자(30X20cm)에 재배된 양송이버섯을 실험에 사용하였다. 양송이버섯 자실체에 솜털곰팡이병원균 포자 현탁액을 분무살포하고 30분 후 상기 실시예 2에서 제조된 산옥잠화 뿌리 추출물과 이의 분획물을 분무살포하고 온도 16℃, 습도 90%의 생육실에서 재배하면서 병 발생율을 조사하였다.To test the control effect of fluffy fungal diseases, mushrooms grown in a box (30X20cm) were used in the experiment. After spraying the spore suspension of the fungus pathogen fungus on the fruiting body of the mushroom, 30 minutes later, spraying the spray of the wild rose extract and its fraction prepared in Example 2 above and cultivating it in a growing room at a temperature of 16°C and a humidity of 90% to increase the incidence of disease Was investigated.
그 결과, 표 1에 나타난 바와 같이 솜털곰팡이병원균 포자 현탁액을 분무살포한 후 아무것도 처리하지 않은 무처리구에서는 87.5%의 이병율을 보였지만 산옥잠화 n-헥산 분획물 처리구에서는 10.6%의 이병율을 보여 87.9%의 방제효과가 있음을 확인하였다. 무처리구와 산옥잠화 n-헥산 분획물 처리구에 대한 비교 사진은 도 5에 나타냈다.As a result, as shown in Table 1, after spray spraying the spore suspension of the fungal pathogen, 87.5% of the morbidity was observed in the untreated group but nothing was treated. Was confirmed. The comparative pictures of the untreated and acid-oxidized n-hexane fraction treatments are shown in FIG. 5.
버섯Mushroom
mushroom
이상에서 본 발명의 바람직한 실시예에 대하여 상세하게 설명하였지만, 본 발명은 상술한 특정의 실시예에 한정되지 아니하며, 청구범위에서 청구하는 본 발명의 요지를 벗어남이 없이 당해 발명이 속하는 기술분야에서 통상의 지식을 가진 자에 의해 다양한 변형 및 개량 실시가 가능한 것은 물론이고, 이러한 변형 및 개량 실시들은 본 발명의 기술적 사상이나 전망으로부터 개별적으로 이해되어져서는 안될 것이다.Although the preferred embodiments of the present invention have been described in detail above, the present invention is not limited to the specific embodiments described above, and is generally limited in the technical field to which the present invention pertains without departing from the gist of the present invention claimed in the claims. Of course, various modifications and improvements can be carried out by a person having knowledge of these, and these modifications and improvements should not be understood individually from the technical spirit or prospect of the present invention.
Claims (10)
Composition for controlling downy mildew disease, which includes an extract of Hosta longissima HONDA or a fraction thereof.
상기 추출물은 산옥잠화 뿌리로부터 추출한 것을 특징으로 하는 솜털곰팡이병 방제용 조성물.
According to claim 1,
The extract is a composition for controlling downy mildew disease, characterized in that it is extracted from the roots of daylily.
상기 추출물은 C1 내지 C4 저급 알코올을 용매로 하여 추출하는 것을 특징으로 하는 솜털곰팡이병 방제용 조성물.
According to claim 1,
The extract is a composition for controlling fluffy fungal diseases, characterized in that the extraction with C 1 to C 4 lower alcohol as a solvent.
상기 추출물은 메탄올을 용매로 하여 추출하는 것을 특징으로 하는 솜털곰팡이병 방제용 조성물.
According to claim 3,
The extract is a composition for controlling fluffy mold disease, characterized in that the extraction with methanol as a solvent.
상기 분획물은 산옥잠화 추출물을 n-헥산, 클로로포름, 에틸아세테이트 및 부탄올로 이루어진 군에서 선택된 어느 하나로 분획하여 제조된 것을 특징으로 하는 솜털곰팡이병 방제용 조성물.
According to claim 1,
The fraction is a composition for controlling fluffy fungal disease, characterized by being prepared by fractionating an acid daylily extract into one selected from the group consisting of n-hexane, chloroform, ethyl acetate, and butanol.
상기 분획물은 산옥잠화 추출물을 n-헥산으로 분획하여 제조된 것을 특징으로 하는 솜털곰팡이병 방제용 조성물.
The method of claim 5,
The fraction is a composition for controlling downy mildew disease, characterized in that it is prepared by fractionating the acid daylily extract with n-hexane.
A method for controlling fluffy fungal disease, comprising the step of treating the composition of any one of claims 1 to 6 with mushrooms.
상기 버섯은 양송이버섯, 새송이버섯, 팽이버섯, 느타리버섯 및 큰느타리버섯으로 이루어진 군에서 선택된 어느 1종인 것을 특징으로 하는 방제 방법.
The method of claim 7,
The mushroom control method, characterized in that any one selected from the group consisting of mushrooms mushrooms, king oyster mushrooms, enoki mushrooms, oyster mushrooms and large oyster mushrooms.
A fungal disease control agent comprising the composition of any one of claims 1 to 6.
상기 방제제는 수화제, 현탁제, 유제, 유탁제, 미탁제, 액제, 분산성액제, 입상수화제, 입제, 분제, 액상수화제, 입상수화제, 수면부상성입제 및 정제로 이루어진 군에서 선택된 어느 하나의 형태로 제제화된 것을 특징으로 하는 솜털곰팡이병 방제제.
The method of claim 9,
The control agent is any one form selected from the group consisting of a hydrating agent, a suspending agent, an emulsion, an emulsifying agent, an emulsifying agent, a liquid agent, a dispersing liquid agent, a granular hydrating agent, a granulating agent, a powder, a liquid hydrating agent, a granulating hydrating agent, a sleeping granulating agent and a tablet. Fluffy mold disease control agent characterized in that the formulation.
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