KR20180060430A - Antibacterial active composition of strawberry leaf extract and method for controlling anthrax using the same - Google Patents
Antibacterial active composition of strawberry leaf extract and method for controlling anthrax using the same Download PDFInfo
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Abstract
Description
본 발명은 탄저병균에 항균 활성을 가지는 딸기잎 추출물 및 이를 포함하는 항균활성 조성물에 관한 것이다. 보다 구체적으로 본 발명은 탄저병균에 항균 활성을 가지는 딸기잎 추출물과 상기 추출물을 포함하는 탄저병균에 방제효과를 가지는 항균 조성물에 관한 것이다.The present invention relates to a strawberry leaf extract having antibacterial activity against anthrax bacteria and an antimicrobial active composition containing the same. More specifically, the present invention relates to a strawberry leaf extract having antibacterial activity against anthrax bacteria and an antimicrobial composition having an antifungal effect against anthrax bacteria containing the extract.
본 발명은 딸기잎에서 메탄올 추출물한 조성물로 탄저병 균에 대한 방제 효과가 있는 항균 조성물에 관한 것이다. The present invention relates to an antimicrobial composition having a controlling effect against anthrax bacteria by a methanol extract composition from strawberry leaves.
딸기는 쌍떡잎식물의 장미과에 속하는 딸기(Fragaria ananassa Duch )는 북반구의 온대지역이 원산지이나 남반구에서도 널리 심고 있다. 대부분의 재배종은 버지니아딸기와 칠레딸기의 변종들이다. The strawberry is a strawberry belonging to Rosaceae of the dicotyledonous plant ( Fragaria ananassa Duch) are extensively planted in the northern hemisphere in the country of origin or in the southern hemisphere. Most cultivars are varieties of Virginia strawberries and Chilean berries.
딸기는 수염뿌리가 나는데, 잎은 뿌리에서 나오며 잔잎이 3장으로 털이 많고 가장자리에는 톱니가 있다. 꽃은 거의 흰색이지만 붉은 꽃이 피기도 하며, 가느다란 꽃자루 위에 여러 개가 모여서 핀다. 꽃자루는 땅을 기는 줄기같이 생겼다. Strawberries have beard roots. Leaves come from roots. There are three leaves with many hairs. There are sawtooths on the edges. The flower is almost white, but the red flower also blooms, and several are gathered on the thin peduncle. The peduncle is like a stem that carries the earth.
꽃받침 속에는 흔히 씨라고 부르는 열매가 묻혀 있다. 비타민 C가 풍부하며, 철분과 다른 무기물도 들어 있다. In calyx, the fruit that is often called seed is buried. It is rich in vitamin C, and contains iron and other minerals.
현재 사용되고 있는 대부분의 항균제는 화학합성을 통해 제조된 것으로서 고비용이 소요되며 부작용을 유발하는 등의 많은 한계를 가지고 있다. 따라서, 최근에는 천연물로부터 새로운 항균물질을 분리하고자 하는 연구가 활발하게 이루어지고 있다. 천연물로부터 새로운 항균물질을 분리하기 위해서는 항균 스펙트럼이 광범위하고 장기간 투여하여도 부작용이 없이 안전하여야 한다는 점 등을 고려해야 한다.Most of the antimicrobial agents currently in use are manufactured through chemical synthesis, which is costly and has many limitations such as causing side effects. Therefore, recently, researches for separating new antimicrobial substances from natural products are actively conducted. In order to isolate new antimicrobial substances from natural products, it should be considered that the antimicrobial spectrum is wide and safe even without long-term administration without adverse effects.
탄저병(炭疽病)은 탄저병균의 감염에 의하여 생기는 식물의 병해로서 고추, 토마토 등의 가지과 및 벼, 콩과, 오이, 국화과 등의 작물과 감나무, 매화나무, 복숭아나무, 감귤나무, 밤나무, 사과나무 등의 과수와 호박, 바나나, 배추과 및 화훼류 등에서 각각 종류가 다른 탄저병균의 기생에 의해서 발생할 수 있다. Anthrax anthracnose is an anthrax disease caused by infection with anthrax. It is a kind of plant disease such as red pepper, tomato, etc., and crops such as rice, bean, cucumber and asteraceae, persimmon, plum tree, peach tree, It can be caused by the parasitism of different kinds of anthrax bacteria in fruit trees such as trees and amber, banana, cabbage and flower.
고추를 재배할 때 가장 피해가 큰 것이 탄저병에 의한 피해다. 그러나 고추 탄저병은 그 방제가 어렵고 피해가 크기 때문에 탄저병을 방제하기 위한 농약 등이 많이 개발되고 있으나, 효과적으로 방제하기 어려운 문제가 있다.The greatest damage to pepper is the damage caused by anthrax. However, since the anthracnose of pepper is difficult to control and the damage is large, a lot of pesticides and the like for controlling anthrax are developed, but it is difficult to effectively control.
고추 탄저병의 가장 효과적인 방제법은 비가림 시설로 빗물이 직접 과실에 튀는 것을 방지하는 것이다. 비가림에 의한 탄저병 방제효과는 95% 이상이라는 여러 연구결과들이 이미 보고된 바 있고, 무농약 고추재배시 비가림 시설이 무설치구에 비해 수량은 약 2배 증가하는 것으로 나타났다. 또한 한 개의 병든 과실에 탄저병균 전염원이 수천만 개 이상 형성되고 비바람에 의해 멀게는 수 km까지 날아가므로 병든 과실은 발견 즉시 제거하여 소각하는 것이 가장 중요하다.The most effective control of pepper anthracnose is to prevent rainwater from splashing directly into the fruit with a rainfed facility. Several studies have reported that the control of anthracnose by rain forests is more than 95%, and the yield of nonpolished red pepper plants is about twice as high as that of non - planted ones. In addition, it is most important to remove the sick fruids by discovery and incinerate as more than tens of millions of anthrax infectious agents are formed in one diseased fruit and they are blown away by several kilometers by wind and rain.
본 발명의 목적은 탄저병에 항균 활성을 가지는 딸기잎 메탄올 추출물을 제공하는 것이다. 또한 본 발명은 상기 딸기잎 메탄올 추출물을 포함하는 항균용 조성물을 제공하기 위한 것이다. It is an object of the present invention to provide a strawberry leaf methanol extract having antibacterial activity against anthrax. The present invention also provides an antimicrobial composition comprising the methanol extract of Strawberry Leaf.
본 발명은 작물의 탄저병 방제효과가 있는 딸기잎 메탄올 추출물을 제공하기 위한 것으로 딸기 잎을 분쇄기로 분쇄한 후 추출용매로 추출, 여과하는 단계를 거쳐 제조 한다. 상기에서 추출용매로는 물 또는 C1∼C5의 알코올이 사용되며, 추출물을 제조하기 위한 추출용매로는 메탄올(CH3OH)이 바람직하다. The present invention provides a methanol extract of strawberry leaf having an anthrax control effect of a crop, which is prepared by pulverizing strawberry leaves with a pulverizer, extracting with an extraction solvent, and filtering. Water or C1-C5 alcohol is used as an extraction solvent, and methanol (CH3OH) is preferable as an extraction solvent for preparing an extract.
본 발명의 딸기잎 추출물은 메탄올 1L에 딸기 잎 0.5~1.5Kg을 혼합한 후 28~32℃에서 10∼15시간 동안 방치하여 추출한 다음 여과한 후 농축함으로써 메탄올을 제거하여 추출물을 제조한다. 또한 증류수 10L에 딸기 잎 0.5~5Kg을 혼합한 후 25~30℃에서 12∼48시간 동안 방치하여 추출한 다음 여과한 후 농축함으로써 추출물을 제조할 수 있다.The strawberry leaf extract of the present invention is prepared by mixing 0.5-1.5 kg of strawberry leaves with 1 L of methanol and then allowing it to stand at 28-32 ° C for 10-15 hours to extract, then filter and concentrate to remove methanol to produce an extract. In addition, the extract can be prepared by mixing 0.5 to 5 kg of strawberry leaves in 10 L of distilled water, extracting the mixture at 25 to 30 ° C. for 12 to 48 hours, filtering, and concentrating.
상기 딸기 잎 메탄올 추출물의 탄저병 균사의 활성과 성장에 관한 기내 시험 결과를 보면 대조구, 메탄올, 0.31mg/ml 메탄올 추출물, 0.64mg/ml 메탄올 추출물, 2.50mg/ml 메탄올 추출물을 투입하였을 때 탄저병 균사의 성장이 억제되며, 메탄올 추출물을 투입량에 따라 탄저병 균사활성이 크게 저해된다.In vitro tests on the activity and growth of anthrax mycelia of the strawberry leaf methanol extract showed that when the control, methanol, 0.31 mg / ml methanol extract, 0.64 mg / ml methanol extract and 2.50 mg / ml methanol extract were added, Growth is inhibited, and anthrax mycelial activity is significantly inhibited by the amount of methanol extract.
딸기 잎 메탄올 추출물은 Colletotrichum coccodes(A), Fusarium oxyspourum(B), Glomerella cingulata(C), Rhizoctonia solani(D), Phytophthora cactorum(E), P. capsici(F) 균 활성의 억제에 효과가 있으며, 딸기의 잎(L), 꼭지(P), 덜익은 녹색딸기(G) 및 잘 익은 딸기(R)를 메탄올 추출하여 bioautogram으로 항균물질 2가지를 찾아냈다. Strawberry leaf methanol extract is colletotrichum coccodes (A) , Fusarium oxysporum (B) , Glomerella cingulata (C) , Rhizoctonia solani (D), Phytophthora cactorum (E), P. capsici (F) has an effect on the inhibition of bacterial activity, the strawberry leaves (L), the tap (P), the green unripe strawberry (G) and ripe strawberry ( R) was extracted with methanol and two kinds of antimicrobial substances were found by bioautogram.
상기 딸기 잎의 메탄올 추출물을 GC-MS chromatogram 방법으로 분석하여 항균물질이 tyrosol 및 sitosterol이라는 것을 알아냈다.The methanol extract of the strawberry leaf was analyzed by GC-MS chromatogram method, and it was found that the antibacterial substance was tyrosol and sitosterol.
가지과 탄저병(Colletotrichum coccodes)의 균사 성장에 관하여 tyrosol(A)과 β-sitosterol(B)에 의해 억제하는 것을 나타낸 것이다. 도 4에서 대조구, 메탄올 ㅁ및 0.01mg/ml tyrosol, 0.10mg/ml tyrosol, 1.00mg/ml tyrosol 을 각각 투입하였을 때 탄저병의 균 활성을 억제 정도를 나타낸 것으로 tyrosol의 투입량이 많을 수록 균 활성이 억제되었다. 또한 sitosterol의 항균 활성을 알아보기 위하여 탄저병 균 대조구와 메탄올, 0.01mg/ml β-sitosterol , 0.10mg/ml β-sitosterol , 1.00mg/ml β-sitosterol 을 투입하였으며, 대조구에 비하여 β-sitosterol 을 투입하였을 때 항균활성이 억제되었으며, 접종량에는 큰 차이는 없었다.(A) and β-sitosterol (B) on the mycelial growth of Colletotrichum coccodes . In FIG. 4, when the control, methanol, 0.01 mg / ml tyrosol, 0.10 mg / ml tyrosol and 1.00 mg / ml tyrosol were added respectively, the degree of inhibition of anthracnose activity was inhibited. . In order to investigate the antimicrobial activity of sitosterol, β-sitosterol, 0.01mg / ml of β-sitosterol, 0.10mg / ml of β-sitosterol and 1.00mg / ml of β-sitosterol were added to the control. Antimicrobial activity was inhibited and there was no significant difference in inoculation amount.
본 발명은 탄저병 방제효과가 있는 딸기잎 메탄올 추출물을 제공하기 위한 것이다. 상기 딸기잎 메탄올 추출물은 Colletotrichum coccodes(A), Fusarium oxyspourum(B), Glomerella cingulata(C), Rhizoctonia solani(D), Phytophthora cactorum(E), P. capsici(F) 균 활성을 억제하였다.The present invention is to provide a strawberry leaf methanol extract having an anthrax control effect. The strawberry leaf methanol extracts were colletotrichum coccodes (A) , Fusarium oxysporum (B) , Glomerella cingulata (C) , Rhizoctonia inhibited solani (D), Phytophthora cactorum ( E), P. capsici (F) bacteria activity.
딸기 잎 메탄올 추출물의 탄저병 균사의 활성의 억제 정도는 추출물의 함량이 증가할 수록 효과가 컸으며, 대조구에 딸기잎을 메탄올 추출물한 조성물로 처리한 경우에 그 효과가 증대되었다. 또한 메탄올 추출물은 그 함량이 증가할수록 효과가 증대되었으며, 0.31mg/ml 메탄올 추출물, 0.64mg/ml 메탄올 추출물,2.50mg/ml 메탄올 추출물을 각각 투입하였을 때 탄저병 균사의 성장을 억제하는 정도는 메탄올 함량이 증가할 수록 증대되었다.The inhibitory effect of methanol extract of strawberry leaves on the activity of anthrax mycelia was more effective as the content of extract increased, and the effect of methanol extract of strawberry leaves on the control was enhanced. The inhibitory effect of methanol extract, 0.64 mg / ml methanol extract and 2.50 mg / ml methanol extract on inhibition of mycelial growth of anthrax was investigated in terms of methanol content As the number of patients increased, the number of patients increased.
본발명의 딸기 잎 메탄올 추출물은 고추, 토마토 등의 가지과 작물은 물론 화훼류, 콩과작물, 배추과, 바나나 및 화박의 탄저병균에도 방제효과가 있는 것으로 나타났다. 딸기 잎의 메탄올 추출물을 분석한 결과 고추 탄저병 방제에 효과가 있는 항균물은은 tyrosol 및 sitosterol이라는 것을 알아냈다.The methanol extract of strawberry leaf of the present invention has an antifungal effect on not only branches and crops such as pepper, tomatoes but also anthracnose of flowers, soybean and crops, cabbage, banana, and firewood. Analysis of methanol extracts of strawberry leaves revealed that the antimicrobial agents effective against pepper anthracnose were silver tyrosol and sitosterol.
도 1은 딸기 잎 메탄올 추출물의 탄저병 균사의 활성과 성장에 관한 기내 시험 결과이다.
도 2는 탄저병원 균 등의 균 활성 억제 정도를 나타낸 그래프이다.
도 3은 딸기 잎의 메탄올 추출물을 분석하여 항균물질이 tyrosol 및 sitosterol이라는 것을 찾아낸 그래프이다.
도 4는 탄저병의 균사 성장에 관하여 tyrosol(A)과 β-sitosterol(B)에 의해 억제하는 것을 나타낸 것이다. Fig. 1 shows the results of in vitro tests on the activity and growth of anthrax mycelia of methanol extract of strawberry leaves.
2 is a graph showing the degree of inhibition of bacterial activity such as anthrax bacteria.
FIG. 3 is a graph showing that methanol extract of strawberry leaves is analyzed to find that the antimicrobial substance is tyrosol and sitosterol.
Figure 4 shows inhibition by tyrosol (A) and beta-sitosterol (B) on mycelial growth of anthrax.
본 발명에서 딸기 부위 중 딸기(Fragaria ananassa Duch)의 잎을 사용하며, 딸기 잎은 관부(crown)에서 잔잎이 3장으로 털이 많고 가장자리에는 톱니가 있다. 그 외에 산딸기( Rubus crataegifolius), 복분자 딸기(R. coreanus MIQ), 멍석딸기(R. parvifolius Linne var. tiphyllus), 청복분자딸기(R. coreanus for. concolor T.Lee), 곰딸기(R. phoenicolasius Maximowicz), 장딸기(R. hirsutus Thunberg), 수리딸기(R. corchorifoius L Fil), 섬딸기(R. ribesioideus Maisumira), 겨울딸기(R. buergeri Miquel) 및 줄딸기(R. oldhamil Maximowicz)의 잎을 사용할 수 있다.In the present invention, the leaves of the strawberry ( Fragaria ananassa Duch) are used, and strawberry leaves have three leaves with a long leaf in the crown and have sawtooth on the edge. Other than Rubus crataegifolius), bokbunja berries (R. coreanus MIQ), native raspberry (R. parvifolius Linne var. tiphyllus) , Zheng bokbunja strawberries (R. coreanus for. concolor T.Lee) , bear strawberry (R. phoenicolasius Maximowicz), jangttalgi ( R. hirsutus Thunberg, R. corchorifoius L Fil, R. ribesioideus Maisumira, R. buergeri Miquel and R. oldhamil Maximowicz can be used.
본 발명의 딸기잎 추출물의 제조에 사용되는 딸기 잎은 그 건체를 사용하는 것이 바람직하며, 추출 효율을 증대시키기 위해 분쇄기로 분쇄한 것을 사용할 수 있다. 상기 딸기 잎의 건조방법으로는 양건, 음건, 열풍건조, 동결건조 및 자연건조 방법을 모두 사용할 수 있다. 바람직하게는 동결건조 방법을 사용할 수 있다.The strawberry leaf used in the production of the strawberry leaf extract of the present invention is preferably used as the dried strawberry, and may be pulverized by a pulverizer to increase the extraction efficiency. As the drying method of the strawberry leaf, it is possible to use any of bion, shade, hot air drying, freeze drying and natural drying methods. Preferably, a freeze-drying method can be used.
또한, 본 발명의 딸기잎 추출물은 당 업계에 공지된 방법을 사용하여 제조할 수 있다. 즉, 본 발명의 추출물은 원료를 일정 크기로 절단하고 추출용매로 추출, 여과하는 단계를 거쳐 제조할 수 있다. 상기에서 추출용매로는 물 또는 C1∼C5의 알코올이 사용될 수 있다. 본 발명의 추출물을 제조하기 위한 추출용매로는 메탄올(CH3OH)이 바람직하다. In addition, the strawberry leaf extract of the present invention can be produced using a method known in the art. That is, the extract of the present invention can be prepared by cutting the raw material to a predetermined size, extracting it with an extraction solvent, and filtering it. As the extraction solvent, water or a C1 to C5 alcohol may be used. Methanol (CH3OH) is preferable as an extraction solvent for preparing the extract of the present invention.
본 발명의 딸기잎 추출물은 메탄올 1L에 딸기 잎 0.5~1.5Kg을 혼합한 후 28~32℃에서 10~15시간 동안 방치하여 추출한 다음 여과한 후 농축함으로써 메탄올을 제거하여 추출물을 제조한다. 상기와 같은 방법으로 여과 농축하게되면 딸기잎 메탄올 추출물은 최초 메탄올 용량(1L)의 1/100 수준으로 용량이 감소하게 된다. 이와 같이 얻은 딸기잎 메탄올 추출물을 물로 희석하여 고추 탄저병 예방에 사용하게 된다.The strawberry leaf extract of the present invention is prepared by mixing 0.5-1.5 kg of strawberry leaves with 1 L of methanol and then allowing to stand at 28 to 32 ° C for 10 to 15 hours for extraction, then filtering and concentrating to remove methanol to prepare an extract. When the filtrate is concentrated by the above method, the methanol extract of strawberry leaves is reduced to 1/100 of the original methanol capacity (1 L). The obtained methanol extract of strawberry leaf is diluted with water and used to prevent anthrax of pepper.
상기와 같이 추출한 딸기잎 메탄올 추출물에서 메탄올을 제거하고 농축한 것을 물에 희석하여 사용한다. 상기 여과 정제된 딸기잎 메탄올 추출물을 물에 희석할 때 상기 딸기잎 메탄올 추출물을 100 내지 300배로 희석하는 것이 바람직하며, 상기 딸기잎 메탄올 추출물 1중량부를 물 100중량부 내지 300중량부에 혼합 희석하여 사용한다.The methanol extract from the strawberry leaf extract thus extracted and concentrated is diluted with water and used. Preferably, the diluted methanol extract of strawberry leaves is diluted to 100 to 300 times when the methanol extract of filtered and purified strawberry leaves is diluted with water. 1 part by weight of the methanol extract of strawberry leaf is mixed and diluted with 100 parts by weight to 300 parts by weight of water use.
본 발명의 실시예에서는 80%~90%의 메탄올 1L에 딸기 잎 1Kg을 첨가하고 30℃에서 12시간 방치하여 추출한 후 여과한 다음 농축하여 딸기 잎 추출물을 제조하였다 In the example of the present invention, 1 kg of strawberry leaves was added to 1 L of methanol of 80% to 90%, and the mixture was allowed to stand at 30 ° C for 12 hours, filtered, and concentrated to produce a strawberry leaf extract
상기와 같은 방법으로 제조된 본 발명에 따른 딸기 잎 추출물을 다음의 실시예 1 및 실시예 2에서와 같이 시험장소를 달리하여 고추 탄저병에 대한 효과를 시험하였다.The strawberry leaf extract according to the present invention prepared as described above was tested for its effect on red pepper anthracnose by the different test sites as in Examples 1 and 2 described below.
그러나 본발명의 딸기 잎 메탄올 추출물은 고추, 토마토 등의 가지과 작물은 물론 화훼류, 콩과작물, 배추과, 바나나 및 화박의 탄저병균에도 방제효과가 있는 것으로 나타났다.However, the strawberry leaf methanol extract of the present invention has an antifungal effect on not only branches and crops such as pepper, tomatoes but also anthracnose of flowers, soybean, crops, cabbage, banana, and firewood.
도 1에서는 상기 딸기 잎 메탄올 추출물의 탄저병 균사의 활성과 성장에 관한 기내 시험 결과로서 대조구, 메탄올, 0.31mg/ml 메탄올 추출물, 0.64mg/ml 메탄올 추출물, 2.50mg/ml 메탄올 추출물을 투입하였을 때 탄저병 균사의 성장 크기를 보여 주는 것이다. 도 1를 보면 메탄올 추출물을 투입량에 따라 탄저병 균사활성이 크게 저해되고 있음을 알 수 있다.In FIG. 1, when the control, methanol, 0.31 mg / ml methanol extract, 0.64 mg / ml methanol extract and 2.50 mg / ml methanol extract were injected into the anthrax mycelium of the strawberry leaf methanol extract, It shows the growth size of mycelium. FIG. 1 shows that anthrax mycelial activity is significantly inhibited by the amount of the methanol extract.
도 2는 Colletotrichum coccodes(A), Fusarium oxyspourum(B), Glomerella cingulata(C), Rhizoctonia solani(D), Phytophthora cactorum(E), P. capsici(F) 균 활성의 억제 정도를 나타낸 것으로, 딸기의 잎(L), 꼭지(P), 덜익은 녹색딸기(G) 및 잘 익은 딸기(R)를 메탄올 추출하여 bioautogram으로 항균물질 2가지를 찾아냈다. 흰색 부분이 균 활성을 억제하는 것으로 딸기 잎(L)의 메탄올 추출물이 모든 균에서 항균활성을 나타내고 있다.Figure 2 is a graph coccodes (A) , Fusarium oxyspourum (B) , Glomerella cingulata (C) , Rhizoctonia solani (D) , Phytophthora shows an extent cactorum (E), P. capsici ( F) Inhibition of bacterial activity, extracts the leaf (L) of strawberry, tap (P), the green unripe strawberry (G) and ripe strawberries (R) methanol We found two antimicrobials in the bioautogram. The white part inhibits bacterial activity, and the methanol extract of strawberry leaf (L) shows antibacterial activity in all bacteria.
도 3은 상기 딸기 잎의 메탄올 추출물을 GC-MS chromatogram 방법으로 분석하여 항균물질이 tyrosol 및 sitosterol이라는 것을 알아냈다.FIG. 3 shows the results of GC-MS chromatogram analysis of the methanol extract of the strawberry leaf, and found that the antibacterial substance was tyrosol and sitosterol.
도 4는 탄저병(Colletotrichum coccodes)의 균사 성장에 관하여 tyrosol(A)과 β-sitosterol(B)에 의해 억제하는 것을 나타낸 것이다. 도 4에서 대조구, 메탄올 및 0.01mg/ml tyrosol, 0.10mg/ml tyrosol, 1.00mg/ml tyrosol 을 각각 투입하였을 때 탄저병의 균 활성을 억제 정도를 나타낸 것으로 tyrosol의 투입량이 많을 수록 균 활성이 억제되었다. 또한 sitosterol의 항균 활성을 알아보기 위하여 탄저병 균 대조구와 메탄올, 0.01mg/ml β-sitosterol , 0.10mg/ml β-sitosterol , 1.00mg/ml β-sitosterol 을 투입하였으며, 대조구에 비하여 β-sitosterol 을 투입하였을 때 항균활성이 억제되었으며, 접종량에는 큰 차이는 없었다.Figure 4 shows inhibition by tyrosol (A) and beta-sitosterol (B) on mycelial growth of anthracnose ( Colletotrichum coccodes ). In FIG. 4, when the control, methanol, 0.01 mg / ml tyrosol, 0.10 mg / ml tyrosol and 1.00 mg / ml tyrosol were added respectively, the degree of inhibition of anthracnose activity was inhibited. . In order to investigate the antimicrobial activity of sitosterol, β-sitosterol, 0.01mg / ml of β-sitosterol, 0.10mg / ml of β-sitosterol and 1.00mg / ml of β-sitosterol were added to the control. Antimicrobial activity was inhibited and there was no significant difference in inoculation amount.
<실시예 1>≪ Example 1 >
1. 시험개요1. Test Overview
실시예 1에서는 고추(단고추류 포함)의 탄저병에 대한 방제약제를 선발하기 위하여 상기 메탄올로 추출한 딸기 잎 추출물을 고추품종 '슈퍼마니따' 및 '파피레드' 품종에 시험하였다. 시험장소는 '경기도 화성시 남양읍 시리'이고, 시험기간은 2016년 8월 2일부터 8월 30일까지로 하였다.In Example 1, in order to select an anti-anthracnose agent for red pepper (including red pepper), the above-mentioned methanol-extracted strawberry leaf extract was tested in the pepper cultivar 'Super Maniata' and 'Papillot' varieties. The test site was 'Namyang-eup Siri' in Hwaseong City, Gyeonggi Province, and the test period was from August 2, 2016 to August 30, 2016.
2. 시험방법2. Test method
고추 품종(슈퍼마니따 및 파피레드)을 대상으로 탄저병(Colletotrichum spp.)을 대상으로 상기 딸기 잎 추출물을 하기와 같이 처리하였다. 약효 시험은 '슈퍼마니따'를 대상으로 하였고, 약해 시험은 '슈퍼마니따'와 '파피레드'를 대상으로 실시하였다.The strawberry leaf extract of Colletotrichum spp. Was subjected to the following treatments in the pepper cultivars (Super Manithe and Papi Red) as follows. The drug efficacy test was conducted on 'Super Manithe', and the weakness test was conducted on 'Super Manithe' and 'Pappy Red'.
시험약제
Test agent
함량
(중량%)chief ingredient
content
(weight%)
및 사용량Dilution factor
And usage
처리시기 및 방법
When and how to process
딸기 잎 메탄올
추출물
프로피네브
수화제(대조)
무처리
Strawberry leaf methanol
extract
Propyne
Wetting agent (contrast)
No treatment
0.5
(01+0.4)
70
-
0.5
(01 + 0.4)
70
-
300배
500
-
300 times
500
-
발병초 7일 간격
4회 경엽처리
(8/2, 8/9, 8/16, 8/23)
"
-
Every 7 days
4 foliage processing
(8/2, 8/9, 8/16, 8/23)
"
-
300배
(8/2)
-
-
300 times
(8/2)
-
-
150배
(8/2)
-
-
150 times
(8/2)
-
-
서울대학교
-
-
Seoul National University
-
-
상기 딸기 잎 추출물을 혼합한 희석액은 엔진동력살포기를 이용하여 10a당 150L를 살포하였다. 상기 표에서 딸기잎 메탄올 추출물 중에 주성분의 함량은 0.5중량% 함유된 것이며, 상기 0.5중량% 중에 tyrosol 이 0.1중량%, β-sitosterol이 0.4중량% 함유되어 있다. 즉, 딸기잎 메탄올 추출물에서 tyrosol과 β-sitosterol이 고추 탄저병의 균의 활성을 억제하는 주성분이 되며, 상기 주성분이 함유되어 있는 딸기잎 메탄올 추출물을 300배 희석하여 고추 탄저병의 약효시험 및 고추 약해시험을 실시하였다.The diluted solution containing the strawberry leaf extract was sprayed with 150 L / 10a using an engine power disperser. In the above table, the content of the main component in the strawberry leaf methanol extract is 0.5% by weight, and 0.1% by weight of tyrosol and 0.4% by weight of β-sitosterol are contained in 0.5% by weight of the extract. That is, in the methanol extract of strawberry leaves, tyrosol and? -Sitosterol are the main components for inhibiting the activity of the anthracnose bacterium, and the methanol extract of strawberry leaf containing the main ingredient is diluted 300 times, Respectively.
경작지에서 고추의 재배는 비닐 멀칭 및 노지재배를 하였으며, 정식일자는 2016. 5. 5., 재식거리는 150cm X 35cm로 하고, 시험기간 내 타 약제를 처리하거나 혼용하는 약제는 없었다.The cultivation of pepper in the cultivated area was carried out by vinyl mulching and cultivation. The official date was 5/5/2010, the planting distance was 150cm X 35cm, and no other medicines were treated or mixed during the test period.
시험구 배치와 재배 면적은 다음과 같이 난괴법 3반복으로 실시하였다.The placement of the test area and the cultivation area were carried out by the repeated three rounds as follows.
division
해about
year
약제 살포일의 기상상황은 수원 기상대의 측정치로 다음과 같았다.The meteorological situation at the spraying day was as follows.
3. 조사방법 및 판정기준3. Survey method and criteria
실시예 1에서의 조사방법 및 판정기준은 다음과 같이 실시하였다.The irradiation method and judgment criteria in Example 1 were carried out as follows.
4. 시험성적4. Test scores
가. 약효 시험에서 최종 약제 처리 후 7일 차에 고추 탄저병에 대한 약제방제 효과를 측정한 결과 다음과 같은 결과를 얻었다.end. In the efficacy test, the effect of controlling the anthracnose of pepper on the 7th day after the final treatment was measured and the following results were obtained.
시험약제
Test agent
(DMRT)Significant difference
(DMRT)
(%)The control
(%)
프로피네브 수화제 (대조)
무처리Strawberry leaf methanol extract
Propinab Wetting Agent (Contrast)
No treatment
5.9
30.69.4
5.9
30.6
6.4
33.011.8
6.4
33.0
5.4
28.96.0
5.4
28.9
5.9
30.89.1
5.9
30.8
c
ab
c
a
80.8
-70.5
80.8
-
상기 표에서 CV%(coefficient of variation %)는 8.1% 였다.The CV% (coefficient of variation%) in the above table was 8.1%.
Remarks
딸기 잎 메탄올 추출물
Strawberry leaf methanol extract
(슈퍼마니따, 파피레드)pepper
(Super Manithe, Papi Red)
0
0
0
0
약해 없음
No Weak
5. 실시예 1의 시험결과5. Test results of Example 1
본 시험을 수행하기 위하여 시험구 배치는 난괴법 3반복으로 시험을 수행하였으며, 시험의 약효시험 평가는 최종약제처리 7일 후 구당 200과 이상에 대한 이병과율을 조사하였으며, 약해시험 평가는 고추에 기준량, 배량 처리 후 약해 유무를 달관조사하였다. 약제간의 유의차 검정은 Duncan's multiple range test(DMRT)로 95% 수준에서 유의성을 검정하였다.In order to carry out this test, the experiment was performed by the repeated three rounds of the nodule method. In the test of drug efficacy test, the mortality rate of 200 or more per 7 days after the final drug treatment was examined. The amount of the standard dose and the weakness after the dose treatment were investigated. Significant differences between the drugs were tested by Duncan's multiple range test (DMRT) at a 95% significance level.
실시예 1에서 고추 탄저병의 무처리 평균 이병과율이 30.8로 약제를 평가하기에 충분한 조건이었으며, 공시자재(딸기잎 메탄올 추출물)의 방제효과는 최종약제 처리 7일 후 70.5%로 우수한 방제효과를 나타냈으며, 약제간의 통계적 유의성을 검정한 결과 통계적으로 유의성이 인정되었다. 또한 공시자재의 기준량과 배량을 경엽처리하여 약해 유무를 조사한 결과 약해가 없었다.In Example 1, the average rate of non-treatment of pepper anthracnose was 30.8, which was a sufficient condition to evaluate the drug. The control effect of the disclosed material (strawberry leaf methanol extract) was 70.5% after 7 days of the final treatment And statistical significance was confirmed by statistical significance between drugs. In addition, there was no weakness as a result of investigating whether or not the standard amount and the amount of the disclosure material were foliarized.
즉, 실시예 1에서 딸기잎 메탄올 추출물은 방제효과가 70.5%로 우수한 효과를 보았으며, 무처리구와 대비하여 통계적으로 유의성이 인정되었다. 약해 시험결과 각각의 기준량과 배량 모두에서 약해가 없어 딸기 잎 메탄올 추출물은 고추 탄저병에 대하여 실용성이 있다고 판단된다.That is, in Example 1, the methanol extract of Strawberry Leaf showed an excellent effect as a control effect of 70.5%, which was statistically significant as compared with control. As a result of the weak test, there was no weakness in both the standard amount and the dose, and the strawberry leaf methanol extract is considered to be practical for anthrax of pepper.
본발명의 딸기 잎 메탄올 추출물은 고추, 토마토 등의 가지과 작물은 물론 화훼류, 콩과작물, 배추과, 바나나 및 화박의 탄저병균에도 방제효과가 있는 것으로 나타났다. The methanol extract of strawberry leaf of the present invention has an antifungal effect on not only branches and crops such as pepper, tomatoes but also anthracnose of flowers, soybean and crops, cabbage, banana, and firewood.
<실시예 2>≪ Example 2 >
1. 시험개요1. Test Overview
실시예 2에서는 고추(단고추류 포함)의 탄저병에 대한 방제약제를 선발하기 위하여 상기 메탄올로 추출한 딸기 잎 추출물을 고추품종 '슈퍼마니따' 및 '파피레드' 품종에 시험하였으며, 시험장소는 '충북 청주시 청원구 북이면'이고, 시험기간은 2016년 8월 3일부터 8월 31일까지로 하였다.In Example 2, in order to select an antifungal agent for anthrax of pepper (including red pepper), the above-mentioned methanol-extracted strawberry leaf extract was tested in the pepper cultivar 'Super Manitta' and 'Pappy Red' Cheongju-si Cheongwon-gu, Seoul, and the test period was from August 3, 2016 to August 31, 2016.
2. 시험방법2. Test method
고추 품종(슈퍼마니따 및 파피레드)의 탄저병(Colletotrichum spp.)을 대상으로 상기 딸기 잎 추출물을 하기와 같이 처리하였다. 약효 시험은 '슈퍼마니따'를 대상으로 하였고, 약해 시험은 '슈퍼마니따'와 '파피레드'를 대상으로 실시하였다.Anthracnose of pepper varieties (Super Manithe and Papi Red) ( Colletotrichum spp.) were treated as follows. The drug efficacy test was conducted on 'Super Manithe', and the weakness test was conducted on 'Super Manithe' and 'Papill Red'.
시험약제
Test agent
함량
(%)chief ingredient
content
(%)
및 사용량Dilution factor
And usage
처리시기 및 방법
When and how to process
딸기 잎 메탄올
추출물
프로피네브
수화제(대조)
무처리
Strawberry leaf methanol
extract
Propyne
Wetting agent (contrast)
No treatment
0.5
(0.1+0.4)
70
-
0.5
(0.1 + 0.4)
70
-
300배
500
-
300 times
500
-
발병초 7일 간격
4회 경엽처리
(8/3, 8/10, 8/17, 8/24)
"
-
Every 7 days
4 foliage processing
(8/3, 8/10, 8/17, 8/24)
"
-
300배
(8/3)
-
-
300 times
(8/3)
-
-
150배
(8/3)
-
-
150 times
(8/3)
-
-
서울대학교
-
-
Seoul National University
-
-
상기 딸기 잎 추출물을 혼합한 희석액은 엔진동력살포기를 이용하여 10a당 150L를 살포하였다.The diluted solution containing the strawberry leaf extract was sprayed with 150 L / 10a using an engine power disperser.
상기 표에서 딸기잎 메탄올 추출물 중에 주성분의 함량이 0.5중량%이며, 상기 0.5중량% 주성분 중에 tyrosol 이 0.1중량%, β-sitosterol이 0.4중량% 함유되어 있다. 즉, 딸기잎 메탄올 추출물에서 tyrosol과 β-sitosterol이 고추 탄저병의 균의 활성을 억제하는 주성분이 되며, 상기 주성분이 함유되어 있는 딸기잎 메탄올 추출물을 300배 희석하여 고추 탄저병의 약효시험 및 고추 약해시험을 실시하였다. In the above table, the content of the main component in the strawberry leaf methanol extract is 0.5% by weight, and 0.1% by weight of tyrosol and 0.4% by weight of β-sitosterol are contained in the 0.5% by weight main component. That is, in the methanol extract of strawberry leaves, tyrosol and? -Sitosterol are the main components for inhibiting the activity of the anthracnose bacterium, and the methanol extract of strawberry leaf containing the main ingredient is diluted 300 times, Respectively.
경작지에서 고추의 재배는 비닐 멀칭 및 노지재배를 하였으며, 정식일자는 2016. 5. 17., 재식거리는 80cm X 35cm로 하고, 시험기간 내 타 약제를 처리하거나 혼용하는 약제는 없었다.The cultivation of pepper in the cultivated area was carried out by vinyl mulching and cultivation. The planting date was set on May 17, 2016, and the planting distance was 80 cm × 35 cm.
시험구 배치와 재배 면적은 다음과 같이 난괴법 3반복으로 실시하였다.The placement of the test area and the cultivation area were carried out by the repeated three rounds as follows.
division
해about
year
약제 살포일의 기상상황은 청주 기상대의 측정치로 다음과 같았다.The meteorological situation of the spraying day was as follows.
3. 조사방법 및 판정기준3. Survey method and criteria
실시예 1에서의 조사방법 및 판정기준은 다음과 같이 실시하였다.The irradiation method and judgment criteria in Example 1 were carried out as follows.
4. 시험성적4. Test scores
가. 약효 시험에서 최종 약제 처리 후 7일 차에 고추 탄저병에 대한 약제방제 효과를 측정한 결과 다음과 같은 결과를 얻었습니다.end. In the efficacy test, the efficacy of anti - anthracnose anti - anthracnose was measured on the 7th day after the final treatment, and the following results were obtained.
시험약제
Test agent
(DMRT)Significant difference
(DMRT)
(%)The control
(%)
프로피네브 수화제 (대조)
무처리Strawberry leaf methanol extract
Propinab Wetting Agent (Contrast)
No treatment
4.2
22.04.8
4.2
22.0
2.1
13.15.8
2.1
13.1
3.0
28.96.6
3.0
28.9
3.1
17.75.7
3.1
17.7
b
ab
b
a
82.5
-67.8
82.5
-
상기 표에서 CV%(coefficient of variation %)는 29.0% 였습니다.In the table above, CV% (coefficient of variation%) was 29.0%.
나. 약제 처리후 3일, 5일 및 7일 차의 약해 시험 결과는 다음과 같았습니다.I. The results of the weakness test on the 3rd, 5th and 7th day after the drug treatment were as follows.
Remarks
딸기 잎 메탄올 추출물
Strawberry leaf methanol extract
(슈퍼마니따, 파피레드)pepper
(Super Manithe, Papi Red)
0
0
0
0
약해 없음
No Weak
5. 실시예 2의 시험결과5. Test results of Example 2
본 시험을 수행하기 위하여 시험구 배치는 난괴법 3반복으로 시험을 수행하였으며, 시험의 약효시험 평가는 최종약제처리 7일 후 구당 200과 이상에 대한 이병과율을 조사하였으며, 약해시험 평가는 고추에 기준량, 배량 처리 후 약해 유무를 달관조사하였다. 약제간의 유의차 검정은 Duncan's multiple range test(DMRT)로 95% 수준에서 유의성을 검정하였다.In order to carry out this test, the experiment was performed by the repeated three rounds of the nodule method. In the test of drug efficacy test, the mortality rate of 200 or more per 7 days after the final drug treatment was examined. The amount of the standard dose and the weakness after the dose treatment were investigated. Significant differences between the drugs were tested by Duncan's multiple range test (DMRT) at a 95% significance level.
실시예 2에서 고추 탄저병의 무처리 평균 이병과율이 17.7%로 약제를 평가하기에 충분한 조건이었으며, 공시자재(딸기잎 메탄올 추출물)의 방제효과는 최종약제 처리 7일 후 67.8%로 대조약제와 비교하여 비등한 방제효과를 나타내었다. 또한 공시자재의 기준량과 배량을 경엽처리하여 약해 유무를 조사한 결과 약해가 없었다.In Example 2, the untreated average mortality of pepper anthracnose was 17.7%, which was sufficient to evaluate the drug. The control effect of the publicly available material (strawberry leaf methanol extract) was 67.8% after 7 days of the final treatment And showed a comparable control effect. In addition, there was no weakness as a result of investigating whether or not the standard amount and the amount of the disclosure material were foliarized.
즉, 실시예 1에서 딸기잎 메탄올 추출물은 방제효과가 67.8%로 우수한 효과를 보았으며, 무처리구와 대비하여 통계적으로 유의성이 인정되었다. 약해 시험결과 각각의 기준량과 배량 모두에서 약해가 없어 딸기 잎 메탄올 추출물은 고추 탄저병에 대하여 실용성이 있다고 판단된다.That is, in Example 1, the methanol extract of Strawberry Leaf showed an excellent effect as a control effect of 67.8%, which was statistically significant as compared with control. As a result of the weak test, there was no weakness in both the standard amount and the dose, and the strawberry leaf methanol extract is considered to be practical for anthrax of pepper.
본발명의 딸기 잎 메탄올 추출물은 고추, 토마토 등의 가지과 작물은 물론 화훼류, 콩과작물, 배추과, 바나나 및 화박의 탄저병균에도 방제효과가 있는 것으로 나타났다. The methanol extract of strawberry leaf of the present invention has an antifungal effect on not only branches and crops such as pepper, tomatoes but also anthracnose of flowers, soybean and crops, cabbage, banana, and firewood.
Claims (6)
상기 딸기잎 추출물에 포함되어 있는 탄저병에 항균활성을 가지는 물질은 tyrosol 및 sitosterol인 것을 특징으로 하는 탄저병 방제효과가 있는 항균활성 조성물
In a strawberry leaf extract having an antitumor effect against anthrax,
An antifungal active composition having an anthrax control effect characterized in that the substance having antibacterial activity against anthrax disease contained in the strawberry leaf extract is tyrosol and sitosterol
상기 딸기잎 추출물은 메탄올 1L에 딸기 잎 0.5~1.5Kg을 혼합한 후 28~32℃에서 10~15시간 동안 방치하여 추출하는 것을 특징으로 하는 탄저병 방제효과가 있는 항균활성 조성물.
The method according to claim 1,
Wherein the strawberry leaf extract is prepared by mixing 0.5-1.5 kg of strawberry leaves with 1 L of methanol and then allowing to stand for 10-15 hours at 28-32 ° C to extract an antifungal active composition.
상기 딸기잎 추출물은 증류수 10L에 딸기 잎 0.5~5Kg을 혼합한 후 25~30℃에서 12∼48시간 동안 방치하여 추출하는 것을 특징으로 하는 탄저병 방제효과가 있는 항균활성 조성물.
The method according to claim 1,
Wherein the strawberry leaf extract is prepared by mixing 0.5 to 5 kg of strawberry leaves with 10 L of distilled water and then allowing to stand for 12 to 48 hours at 25 to 30 캜 to extract an antifungal active composition.
상기 딸기잎 추출물에 사용되는 딸기잎은 딸기(Fragaria ananassa Duch) 및 산딸기(Rubus crataegifolius), 복분자 딸기(R. coreanus MIQ), 멍석딸기(R. parvifolius Linne var. tiphyllus), 청복분자딸기(R. coreanus for. concolor T.Lee), 곰딸기(R. phoenicolasius Maximowicz), 장딸기(R. hirsutus Thunberg), 수리딸기(R. corchorifoius L Fil), 섬딸기(R. ribesioideus Maisumira), 겨울딸기(R. buergeri Miquel) 및 줄딸기(R. oldhamil Maximowicz)의 잎을 C1∼C5의 알코올로 추출한 딸기잎 메탄올 추출물인 것을 특징으로 하는 탄저병 방제효과가 있는 항균활성 조성물.
4. The method according to any one of claims 1 to 3,
The strawberry leaves used for the strawberry leaf extract include Fragaria ananassa Duch and Rubus crataegifolius , R. coreanus MIQ, R. parvifolius Linne var. Tiphyllus, R. berry strawberry ( R. coreanus for concolor T.Lee, R. phoenicolasius Maximowicz, R. hirsutus Thunberg, R. corchorifoius L Fil, R. ribesioideus Maisumira, R. strawberry ( R. The present invention relates to an antimicrobial active composition having an anthrax control effect, which is characterized in that the leaves of buergeri Miquel and R. oldhamil Maximowicz are methanol extracts of strawberry leaves extracted with C1 to C5 alcohol.
상기 딸기잎 추출물이 탄저병 방제에 효과가 있는 작물은 고추, 토마토 등의 가지과 작물은 물론 화훼류, 콩과작물, 배추과, 바나나 및 호박으로 구성되는 농작물에 살포하였을 때 탄저병 방제에 효과가 있는 것을 특징으로 하는 항균활성 조성물.
4. The method according to any one of claims 1 to 3,
The strawberry leaf extract has an effect on anthrax control when it is sprayed on a crop comprising leaves, crops such as pepper and tomato, as well as crops such as flowers, soybean, crops, cabbage, banana and pumpkin Lt; / RTI >
상기 딸기잎 추출물의 제조방법은 메탄올 1L에 딸기 잎 0.5~1.5Kg을 혼합한 후 28~32℃에서 10~15시간 동안 방치하여 추출하되,
상기 딸기 잎에 혼합되는 메탄올은 80%~90%의 메탄올인 것을 특징으로 하는 항균활성을 가지는 딸기잎 추출물의 제조방법.
A method for producing a strawberry leaf extract having antibacterial activity,
The method of preparing the strawberry leaf extract comprises mixing 0.5-1.5 kg of strawberry leaves with 1 L of methanol and then allowing to stand for 10-15 hours at 28-32 ° C.,
Wherein the methanol mixed in the strawberry leaves is 80% to 90% methanol.
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