KR20160000659A - The composition of biocontroller against soil nematode using plant extracts and microorganism - Google Patents
The composition of biocontroller against soil nematode using plant extracts and microorganism Download PDFInfo
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Abstract
Description
본 발명은 토양에서 서식하는 토양 선충을 방제할 수 있는 조성물에 관한 것으로서, 더욱 정확하게는 항선충 활성을 가진 천연식물추출물과 단백질분해효소를 분비하는 미생물을 혼합하여 액상 제형한 조성물을 특징으로 하는 것이다.
The present invention relates to a composition capable of controlling soil nematides in soil, more precisely, a composition comprising a natural plant extract having anti-nematic activity and a microorganism that secretes protease .
농작물, 채소류 및 과수류의 연작재배로 인해 토양 내에는 보이지 않는 기생선충이 확산하여 고품질 안전 농산물 생산에 큰 문제를 제기하고 있다. 선충에 피해를 받게 되면 지상부의 생육은 육안상으로 큰 차이가 없어 보이나 열매 및 종자의 크기나 품질이 떨어져 20~30%의 수량 감소가 일어난다. 식물병원성 기생선충이 감염된 식물은 방어체계가 약화되기 때문에 여러 식물병원균의 감염을 촉진시켜 복합적인 병을 발병시켜 농작물을 조기에 고사시킨다. 세계적으로 식물기생선충으로 인한 농작물 피해는 점차 늘어가고 있는 추세이며, 식량작물 11% 및 경제작물 14%의 수량적인 손실이 발생하고 있다(미국선충학회, 1987년). Because of the cultivation of crops, vegetables and edible crops, invisible parasitic nematodes spread to produce high quality safe agricultural products. When the nematode is damaged, the growth of the ground part does not seem to be visually significant, but the size and quality of the fruit and seed are reduced, resulting in a reduction in yield of 20 to 30%. Plant pathogens Parasitic nematode plants are infected with a weak defense system, which can accelerate the infestation of several plant pathogens, leading to the development of complex diseases and early destruction of crops. Globally, damage to crops due to plant parasitic nematodes is increasing and there is a numerical loss of 11% of food crops and 14% of economic crops (US Nematode Society, 1987).
전세계적으로 식물기생선충 중 뿌리혹선충(Meloidogyne spp.)에 의한 피해가 가장 심각하며 세계적으로는 78종이 분포하고 있다. 국내에서 큰 문제가 되는 뿌리혹선충은 M.
arenaria(땅콩뿌리혹선충), M.
hapla(당근뿌리혹선충), M. incognita(고구마뿌리혹선충) 등 3종이다. 뿌리혹선충에 의한 피해는 전세계적으로 분포되어 있는데, 한대지방보다는 온도가 높아 선충의 번식횟수가 많은 열대 혹은 아열대지방에서 특히 더 심각하다. 최근 국내 시설재배지의 면적이 계속 증가하고 시설재배지에는 겨울 동안에도 시설난방을 이용하여 채소류를 연중 재배하기 때문에 아열대지방에 적응하는 뿌리혹선충의 발생이 많은 편이다. 우리나라에서는 주요 시설원예작물인 참외, 메론, 오이, 고추, 토마토, 가지, 딸기 등 시설재배지의 과채류 경제작물에서 많은 피해를 주는데, 뿌리혹선충에 심하게 감염된 식물은 조기에 고사하거나, 수확량이 감소하는 등 그 피해가 심각하다. 특히 참외, 고추, 딸기 등과 같이 10년 이상 동안 동일 포장에서 매년 같은 작물을 연작하는 재배지와 오이, 멜론 증 1년 2작기 이상 봄 재배에 이어 가을재배를 하는 포장은 년중 식물기생선충의 증식에 좋은 환경 조건을 조장함으로써 뿌리혹선충의 밀도가 높고 피해가 증가하고 있다.
Globally, the damage caused by the root-knot nematode ( Meloidogyne spp.) Among plant parasitic nematodes is the most serious and 78 species are distributed worldwide. The three most common root-knot nematodes in Korea are M. arenaria (peanut root nematode), M. hapla (carrot root nematode), and M. incognita (sweet potato root nematode). Damage caused by root-knot nematodes is distributed throughout the world, especially in tropical or subtropical regions where the temperature is higher than in one region and the number of nematode breeding is high. In recent years, the area of domestic plant growing area has been continuously increasing, and planting facilities have been growing vegetables throughout the year by using facility heating during the winter, so that the root - knot nematodes adapted to the subtropical regions are generated. In Korea, major crops such as melon, melon, cucumber, pepper, tomato, eggplant, strawberry, etc., are damaged in major crops such as melon, horticultural crops, and plants that are severely infested by root-knot nematodes. The damage is serious. In particular, the cultivated crops such as melon, red pepper, strawberry, etc., which produce the same crop every year in the same package for more than 10 years, followed by cucumber, By promoting environmental conditions, the density of root-knot nematodes is high and damage is increasing.
국내에서는 선충방제를 위해서 대부분 유기합성 살선충제를 사용하는데 이러한 화학적 방제는 수질 및 토양의 환경오염을 야기한다. 국외에서는 선충방제에 토양훈증제를 많이 사용하고 있으나 이것도 환경문제로 인해 사용이 제한되고 있어 새로운 친환경적 살선충제의 개발이 필요한 실정이다. In Korea, organic nematocides are mostly used for nematode control, which causes environmental pollution of water quality and soil. There are many uses of soil fumigants in nematodes control outside the country, but their use is restricted due to environmental problems. Therefore, it is necessary to develop a new environmentally friendly nematocide.
국내의 경우 뿌리혹선충 억제를 위해서 토양개량, 침수, 태양열소독 등의 물리적 방제, 윤작이나 저항성 품종 등 작부체계에 따른 경종적 방제, 천적인 곰팡이나 세균을 이용한 생물학적 방제, 그리고 살선충제를 이용한 화학적 방제 등 다양한 방법이 사용되고 있다. 국내의 선충연구에 관한 역사는 짧지만, 최근 미생물 및 식물추출물을 이용한 선충 생물학적 방제 연구가 이뤄지고 있으나 방제효율이 화학적 방제에 비해 다소 낮은 편이다. In Korea, physical control such as soil improvement, inundation, solar thermal disinfection, anti-segregation control based on cropping system, biological control using natural fungi and bacteria, chemical treatment using nematicide And a variety of methods are being used. Although the history of nematode research in Korea is short, recent studies on nematode biosynthesis using microorganisms and plant extracts have been carried out, but the efficiency of the control has been somewhat lower than that of chemical control.
천연식물추출물의 장점은 생분해가 잘 되고 저농도에서도 활성을 나타낼 수 있으며 속효성이 있기 때문에 이에 대한 연구자들의 관심이 점차 커지고 있다. 또한 천연화합물들은 보다 안전한 유기합성농약의 개발을 위한 신규성 작용기작을 제공하는 장점이 있다. The advantages of natural plant extracts are biodegradation, activity at low concentrations, and rapidity of interest. Natural compounds also have the advantage of providing a novel mechanism for the development of safer organic synthetic pesticides.
선충의 천적으로는 톡톡이, 응애, 세균, 포식성곰팡이, 기생성 곰팡이 등 여러 종류가 있는데, 그 중에서 가장 연구 역사가 깊은 천적은 포식성 곰팡이이며 포식성곰팡이는 세계적으로 거의 모든 토양으로부터 발견되는 공통종이다. 선충 천적의 장점은 효능의 지속성이나, 속효성이 없고 토양환경의 영향을 크게 받는다는 것이 단점이다. There are many kinds of nematode, mites, bacteria, predatory fungi, and fungi. Among them, predator is the predominant fungus, and predatory fungus is the common species found in almost all the world. The advantage of the nematode is that it has persistence of efficacy, but it has no short-lived effect and is greatly affected by the soil environment.
본 발명에서는 살선충 활성을 가진 천연식물추출물과 세균의 장점을 혼합하여 선충방제에 효과적인 제형을 확립하고 이에 대한 선충방제활성을 확인하는 것으로 한다.
In the present invention, an effective formulation for nematode control is established by mixing the advantages of a natural plant extract having a nematode activity and a bacterium, and the nematode controlling activity thereof is confirmed.
친환경농법을 위해서는 유기화학농약 사용을 최소화하고, 생물학적 그리고 생화학적 소재를 활용하여 병충해를 방제해야한다. 본 발명에서는 선충 방제의 속효성 및 토양환경 조건에 구애를 받지 않는 식물유래 추출물의 장점과 토양에 서식하면서 단백질 분해효소 등의 대사산물을 생산하면서 지속적인 효과를 발휘할 수 있는 미생물의 장점을 선택하여 친환경 선충방제제를 조성하기 위한 유효성분으로 선택하였다. 또한 이들의 장점을 최적화할 수 있도록 유화제, 분산제 및 산화방지제를 부가적으로 첨가하여 선충 방제에 적합한 조성물로 제형화하는 것을 특징으로 할 수 있다.
For eco-friendly farming, the use of organic chemical pesticides should be minimized and biological and biochemical materials should be used to control pest infestation. In the present invention, the advantages of the plant-derived extract which does not depend on the quick-acting nature of the nematode control and the soil environmental conditions, and the advantages of the microorganisms which can produce a sustained effect while producing the metabolites such as proteolytic enzymes in the soil, And was selected as an active ingredient for the formulation of a controlling agent. In order to optimize the advantages of the present invention, an emulsifier, a dispersant, and an antioxidant may be additionally added to form a composition suitable for controlling nematodes.
본 발명은 식물추출물과 미생물을 제형화하여 선충방제를 위한 조성물을 제공하는 효과가 있다. 본 발명은 식물유래 추출물인 님오일과 고삼추출물 그리고 단백질 분해 효소 생산 미생물인 Bacillus
sp. K4 와 Brevibacillus sp. K9을 혼합한다. 혼합 제형 제조시 유화제, 분산제 및 산화방지제를 부가적으로 첨가하여 액상 제형을 한다. 이 제형은 선충을 방제하는 효과가 확인되었다. 따라서 본 발명에서 제시한 조성으로 선충을 방제할 수 있는 새로운 선충방제제가 될 수 있을 것으로 기대된다.
The present invention is effective in providing a composition for nematode control by formulating a plant extract and a microorganism. The present invention relates to plant-derived extracts such as nymph oil and gossam extract, and protease-producing microorganisms such as Bacillus sp. K4 and Brevibacillus sp. Mix K9. An emulsifier, a dispersant and an antioxidant are additionally added in the preparation of the mixed formulation to form a liquid formulation. This formulation was found to be effective in controlling nematodes. Therefore, it is expected that the composition of the present invention can be a new nematide controlling agent capable of controlling nematodes.
도 1은 토양에서 분리한 선충의 모습
도 2는 NGM 배지에서의 선충 성장 곡선
도 3은 님오일의 농도에 따른 선충 억제 효과
도 4는 고삼추출물의 농도에 따른 선충 억제 효과
도 5는 단백질 분해 미생물의 배양 농축액의 선충 억제 효과
도 6은 제형화된 제제의 농도에 따른 선충 억제 효과Figure 1 shows a nematode isolated from soil
Fig. 2 shows the nematode growth curve in the NGM medium
Fig. 3 shows the effect of nematode inhibition
Fig. 4 shows the effect of inhibiting nematodes
FIG. 5 is a graph showing the nematode inhibitory effect of a culture concentrate of a proteolytic microorganism
Figure 6 shows the effect of the nematocide inhibitory effect
1. 토양 선충 분리1. Isolation of soil nematode
실험에 사용한 선충은 포장에 재배중인 오이의 근권 토양으로부터 분리하여 이용하였다. 선충의 채취 방법은 토양 표면 5~10 Cm 을 걷어내고, 오이 뿌리를 채취하여 뿌리혹선충이 감염되어 있는지 유무를 확인하 후, 삽을 이용하여 약 5kg 정도의 토양을 채취하였다. 채취한 토양을 잘게 부순 뒤 물을 첨가하여 토양 현탁액을 만들고 2 mm 체로 걸러 돌을 걸러냈다. 토양 현탁액을 표준망체(60, 325, 400 mesh)를 사용하여 통과시키고 최종적으로 400 mesh를 통과한 토양 현탁액을 깔대기법으로 선충을 분리하였다. 깔대기 아래에 모여 있는 선충을 현미경으로 관찰하였으며, 도 1과 같다. The nematodes used in the experiment were separated from the rhizosphere soil of the cultivated cucumber. 5 ~ 10 Cm of the surface of the nematode was removed and cucumber roots were collected and it was confirmed whether or not the root nematode was infected. Then, about 5 kg of soil was collected using a shovel. The soil was crushed and water was added to make a soil suspension, which was filtered with a 2 mm sieve. The soil suspension was passed through a standard netting (60, 325, 400 mesh) and the soil suspension, which had finally passed through 400 mesh, was separated by funnel method. The nematodes collected under the funnel were observed under a microscope, as shown in Fig.
2. 토양 선충 기내 배양 및 증식2. Cultivation and proliferation in soil nematode
깔대기법으로 분리된 선충을 항생제가 함유된 멸균 증류수에서 약 30분 동안 세척을 한 후 다시 항생제가 함유되지 않은 멸균 증류수에서 약 30분 동안 세척을 하였다. 이러한 세척을 2~3회 반복 수행하였다. 세척한 선충을 NGM agar plate(Agar 20g/L, Peptone 4g/L, NaCl 1.5g/L, Cholesterol 0.0008g/L, Potassium phosphate monobasic 3g/L, Potassium phosphate dibasic 0.5g/L, pH 5.5 ~ 6.5)에 접종하였다. Plate당 약 100~150 마리 정도를 접종하고 25℃, 암조건에서 5일간 배양하였다. 배양된 plate를 멸균된 칼로 약 가로 세로 약 1cm 로 자른 후 60mm plate로 에 넣고 2mL NGM broth(Peptone 4g/L, NaCl 1.5g/L, Cholesterol 0.0008g/L, Potassium phosphate monobasic 3g/L, Potassium phosphate dibasic 0.5g/L, pH 5.5 ~ 6.5)를 첨가하고 25℃, 암조건에서 3일간 배양하였다. Agar block 속에서 자란 선충이 액상 배지로 이동하여 성장하였다. 액상 배지에서 성장한 선충을 NGM borth에서 25℃, 암조건에서 7일간 계대배양하였다. 선충의 성장속도를 확인하기 위해서 50마리씩 3개 plate에 분리하여 10일 동안 살아 있는 선충 개체수를 조사하였다. 이 실험은 3 반복 수행되었고, 그 결과 도 2와 같다. 1일차에서는 평균 62마리, 2일차에서는 평균 1052마리로 폭발적 증식을 하였고, 4일차에는 평균 1313마리, 6일차에는 평균 1225마리, 8 일차에는 평균 1139 마리, 10일차에는 평균 1066마리로 확인되었다. 6일차부터 생존한 선충의 개체수가 감소하는 것으로 확인되었다.
The nematodes separated by the funnel method were washed in sterile distilled water containing antibiotics for about 30 minutes and then for 30 minutes in sterile distilled water containing no antibiotics. This washing was repeated 2-3 times. The washed nematode was treated with NGM agar plate (Agar 20g / L, Peptone 4g / L, NaCl 1.5g / L, Cholesterol 0.0008g / L, Potassium phosphate monobasic 3g / L, Potassium phosphate dibasic 0.5g / Lt; / RTI > About 100 ~ 150 per plate were inoculated and cultured at 25 ℃ for 5 days under dark condition. The cultured plates were cut into approximately 1 cm length with a sterilized knife and placed in a 60 mm plate. 2 mL of NGM broth (Peptone 4 g / L, NaCl 1.5 g / L, Cholesterol 0.0008 g / L, Potassium phosphate monobasic 3 g / dibasic 0.5 g / L, pH 5.5 ~ 6.5) was added and incubated at 25 ℃ for 3 days under dark conditions. Nematodes grown in the agar block migrated to the liquid medium and grew. Nematodes grown in liquid medium were subcultured in NGM borth at 25 ℃ for 7 days under dark conditions. In order to confirm the growth rate of nematodes, 50 nematodes were separated into 3 plates for 10 days. This experiment was repeated three times, and the result is shown in FIG. On the first day, the average number was 62 and on the second day, the average was 1052. The average number of 1313 on the 4th day, 1225 on the 6th day, 1139 on the 8th day and 1066 on the 10th day. It was confirmed that the number of surviving nematodes decreased from day 6.
1. 항선충 식물추출물 검토1. Review of nematode plant extracts
님 오일, 고삼 추출물(냉수 추출) 및 목초액을 구매하여 선충방제를 위한 식물추출물을 검토하였다. 대조군 및 처리군에 각각 100마리씩의 선충을 2mL NGM broth에 접종하였고, 식물추출물은 1/100, 1/500 및 1/1000 의 배율로 선충이 접종된 처리군에 희석하여 25 ℃, 암조건에서 배양하였다. 식물추출물을 처리한 1일차와 4일차에 현미경으로 선충의 치사율을 검토하였다. 이 실험은 3 반복 수행되었고, 그 결과는 도 3과 도 4와 같다. 도 3은 님오일에 대한 선충의 치사율을 검토한 것으로서 대조군에서는 선충의 증식이 관찰되었다. 님오일 1/100 처리군에서는 1일차에 65%, 4일차에 61%의 치사율을 보였으며, 님오일 1/500 처리군에서는 1일차에 54%, 4일차에 47%의 치사율을 보인 반면에, 님오일 1/1000 처리군에서는 1일차에 17%의 치사율을 보였지만 4일차에는 선충이 증식한 것으로 확인되었다. 도 4는 고삼추출물에 대한 선충의 치사율을 검토한 것으로서 대조군에서는 선충의 증식이 관찰되었다. 고삼추출물 1/100 처리군에서는 1일차에 46%, 4일차에 43%의 치사율을 보였으며, 고삼추출물 1/500 처리군에서는 1일차에 27%, 4일차에 25%의 치사율을 보인 반면에, 고삼추출물 1/1000 처리군에서는 1일차에 1%의 치사율을 보였지만 4일차에는 선충이 증식한 것으로 확인되었다. 마지막으로 목초액은 선충 억제 효과가 없는 것으로 확인되었다. 이상의 결과로 선충 방제에는 님오일이 고삼추출물보다 우수한 것으로 확인되었다.
Nematode, ginseng extract (cold water extract) and wood vinegar were purchased to examine plant extracts for nematode control. The control and treatment groups were each inoculated with 2 mL of NGM broth at a rate of 1/100, 1/500, and 1/1000, respectively, and diluted in the treatment group inoculated with nematode at 25 ° C. under dark conditions Lt; / RTI > On the 1st and 4th day after plant extract treatment, nematode mortality was examined under a microscope. This experiment was repeated three times, and the results are shown in FIG. 3 and FIG. FIG. 3 shows the nematode mortality rate of nematocysts. In the control group, nematode proliferation was observed. In the nim oil 1/100 treatment group, the mortality rate was 65% in the first day and 61% in the fourth day treatment group. In the nim oil treatment group, the mortality rate was 54% in the first day and 47% in the fourth day , The nematode treated group showed a mortality rate of 17% on day 1, but the nematode was proliferated on day 4. FIG. 4 is a graph showing the mortality rate of nematodes against the extract of Gosam. In the control group, proliferation of nematodes was observed. In the 1/100 treatment group, the mortality rate was 46% in the 1st day and 43% in the 4th day treatment. In the 1/500 treatment group, the mortality rate was 27% in the first day and 25% in the fourth day , And 1/1000 treatment group, respectively, but it was confirmed that the nematodes were proliferated on the 4th day. Finally, it was confirmed that the wood vinegar had no nematode inhibitory effect. As a result, nematicide was found to be superior to goose extract in nematode control.
1. 단백질 분해 미생물 검토 1. Protein-degrading Microorganisms Review
선충 방제법 중에 미생물이 분비하는 분해효소나 항생물질과 같은 2차 대사산물 등에 의한 것으로 방제할 수 있다는 연구보고가 있다. Bacillus cereus가 생산하는 단백질 분해효소에 의해 M. javanica의 표피가 분해되었으며, B. ambifaria의 배양액내의 단백질분해효소와 키틴분해효소에 의해 알 부화와 유충이 억제되었다는 연구보고가 있다. It has been reported that it can be controlled by secondary metabolites such as decomposition enzymes and antibiotics secreted by microorganisms during nematocarcinoma control. Bacillus There was a report that M. javanica epidermis was degraded by the protease produced by S. cereus , and the protease and chitinolytic enzyme in the culture medium of B. ambifaria inhibited egg hatching and larvae.
단백질 분해 미생물을 분리하기 위해서 토양 시료를 nutrient skim milk agar(NSKA: nutrient broth 0.8%, agar 1.5%, skim milk 1%, Difco) 배지 상에서 10-3~10-5의 농도로 희석평판하여 30℃에서 24 시간동안 배양하여 colony 주변에 투명환 형성균을 1차 선발하였다. 1차 선발된 미생물중에 형태를 분석한 결과 그람양성, 간균, 내생포자형성, 운동성이 있는 후보 미생물 2종을 분리하였다. 이 2종을 동정한 결과 Bacillus sp. 균주와 Brevibacillus sp.로 확인되었으며 이를 Bacillus sp. K4 와 Brevibacillus sp. K9으로 명명하였다. 이 2종의 균주를 1% skim milk가 함유된 nutrient broth 접종하고 30℃에서 48 시간 동안 배양하였다. 배양 후 원심분리하여 배양상등액을 취하고 10 kDa membrane filter로 배양상등액을 약 1/10으로 농축하였다. 이 농축된 배양상등액을 1:1 비율로 선충이 배양된 배지에 첨가하여 선충 방제활성을 검토하였다. 각 샘플당 선충은 100 마리로 하였으며, 25℃에서 배양하였고, 48 시간 후에 선충의 치사율을 확인하였다. 이 실험은 3반복 수행되었고, 그 결과는 도 5와 같다. 농축 배양상등액에 대한 선충의 치사율을 검토한 것으로서 대조군에서는 선충의 증식이 관찰되었다. K4 배양액 처리군에서는 81%, K9 배양액 처리군에서는 86%, K4와 K9 혼합배양액 처리군에서는 89%의 치사율을 보였다. 미생물은 생화학적 성분에 비해 속효성이 떨어지지만 지속적인 효과면에서 우수하기 때문에 높은 농도에서 선충방제효과를 검토하였다. 각각 단독 처리군 보다 K4와 K9 혼합배양액 처리군에서 다소 높은 선충 치사율이 확인되었다.
To isolate proteolytic microorganisms, soil samples were diluted to a concentration of 10 -3 to 10 -5 on nutrient skim milk agar (NSKA: nutrient broth 0.8%, agar 1.5%, skim milk 1%, Difco) For 24 hrs, and a transparent ring - forming bacterium was firstly selected around the colony. As a result of the morphological analysis of the selected microorganisms, two candidate microorganisms having Gram - positive, Bacillus, endogenous spore formation and motility were isolated. As a result of identification of these two species, Bacillus sp. And Brevibacillus sp., Which were identified as Bacillus sp. K4 and Brevibacillus sp. K9. These two strains were inoculated with nutrient broth containing 1% skim milk and cultured at 30 ℃ for 48 hours. After culturing, the culture supernatant was centrifuged and the culture supernatant was concentrated to about 1/10 with a 10 kDa membrane filter. This concentrated culture supernatant was added to the culture medium of the nematodes at a ratio of 1: 1 to examine the nematocidal activity. The number of nematodes per each sample was 100. After incubation at 25 ° C, the mortality of the nematodes was confirmed after 48 hours. This experiment was repeated three times, and the results are shown in FIG. The nematode lethality of the concentrated culture supernatant was examined. In the control group, nematode proliferation was observed. 81% in the K4 medium, 86% in the K9 medium, and 89% in the K4 and K9 medium. Since microorganisms have less rapid effect than biochemical components but are superior in terms of sustained effects, the effect of controlling nematodes at high concentrations was examined. The higher nematode mortality was observed in the K4 and K9 mixed cultures treated group than the single treated group, respectively.
1. 제형화1. Formulation
선충방제제를 제형화 하기 위해서 유효성분으로 님오일, 고삼추출물 및 K4 / K9 균주 배양액을 선택하였다. 제형화를 위해서 산화방지제, 유화제 및 분산제를 검토하였다. 님오일 경우 식물성 오일이기 때문에 공기 중 노출 시 물리화학적 안정성 및 지속적인 살충효과유지를 위해서는 산화방지제를 이용한 안정성이 확보되어야 한다. 식품용 및 사료용으로 널리 사용중인 Ethoxyquin을 0.04% 처리 시 20일 후 수면전재 양상이 가장 양호한 것으로 확인되었다. 유화성분이 유화되도록 하기 위해서 완전한 유화가 되어야 한다. 유화제는 액상제형에 적합한 농업영 계면활성제를 대상으로 실험한 결과 Polyoxyethylene(1,1,3,3-tetramethylbutyl) phenyl ether, Polyethylene glycol, Polyoxyethylene nonylphenol 10%에서 물성 및 유화정도가 우수한 것으로 확인되었다. 최종적으로 취급성과 단가 등을 판단하여 Polyethylene glycol을 선택하였다. 효력증진을 위한 분산제로는 Polyoxyethylene sorbitan monoleate 10%에서 유효성분의 뭉침 정도와 분산정도가 양호한 것으로 나타났다. 최종적인 구성은 표 1과 같다.In order to formulate nematocides, nematode, ginseng extract and K4 / K9 culture medium were selected as effective ingredients. Antioxidants, emulsifiers and dispersants were studied for formulation. Since nem oil is a vegetable oil, stability with the use of antioxidants should be ensured in order to maintain physico-chemical stability and continuous insecticidal effect in the air. Ethoxyquin, which is widely used for foods and feeds, was found to have the best sleeping pattern after 20 days of 0.04% treatment. In order to emulsify the emulsifying component, it must be completely emulsified. The emulsifier was found to be excellent in physical properties and emulsification in 10% of polyoxyethylene (1,1,3,3-tetramethylbutyl) phenyl ether, polyethylene glycol, and polyoxyethylene nonylphenol. Polyethylene glycol was selected by judging final handling and unit cost. As a dispersing agent for improving efficacy, 10% of polyoxyethylene sorbitan monoleate showed good degree of aggregation and dispersion of active ingredients. The final configuration is shown in Table 1.
1. 선충 방제 테스트1. Nematode Control Test
표 1의 구성으로 제형화된 제제로 선충 방제 효과를 검토하기 위해서 2일령된 선충 100마리씩을 2mL NGM borth/plate 에 옮기고 12시간 동안 선충을 안정화 시킨 후에 제형된 제제를 1/100, 1/500 및 1/1,000 비율로 투입했다. 실험 조건은 암조건에 25℃, 4일 동안 관찰하였고, 선충의 치사율을 검토하였다. 이 실험은 3반복 수행되었고, 그 결과는 도 6과 같다. 제제를 처리하지 대조군에서는 선충이 4일차에 선충의 개체수가 1,000 마리 가량 증식한 것으로 확인되었다. 제제 1/100 처리군에서는 1일차에 97%, 2일차에 98%, 3일차에 98%, 4일차에 100%의 치사율을 보였다. 제제 1/500 처리군에서는 1일차에 88.3%, 2일차에 91%, 3일차에 97.3%, 4일차에 99%의 치사율을 보였다. 제제 1/1000 처리군에서는 1일차에 92%, 2일차에 90.6%, 3일차에 95.3%, 4일차에 97.6% 의 치사율을 보였다. In order to examine the nematocidal effect of the formulation formulated in Table 1, 100 nematodes of two days old were transferred to a 2 mL NGM borth / plate and the nematode was stabilized for 12 hours. And 1 / 1,000 ratio. Experimental conditions were observed for 4 days at 25 ℃ in dark condition, and the mortality of nematodes was examined. This experiment was repeated three times, and the results are shown in FIG. In the control group treated with the preparation, it was confirmed that the number of nematodes increased by about 1,000 on the 4th day of the nematode. In the 1/100 treatment group, the mortality rate was 97% on day 1, 98% on day 2, 98% on
이상의 결과로 식물성분인 님오일과 고삼추출물 그리고 단백질 분해효소를 분비하는 미생물 배양액을 제형한 제제가 약 90% 이상의 선충 방제 효과를 확인하였다.
As a result, it was confirmed that the formulation containing the nimule oil, the ginseng extract and the proteolytic enzyme - secreting microorganism was more than 90% in controlling the nematode.
Claims (3)
In nematode control, a composition consisting of nimoo oil, gossam extract, microorganism producing protease, emulsifier, dispersant and antioxidant
The method according to claim 1, wherein the microorganism producing the protease is Bacillus sp. K4, Brevibacillus sp. K9, a nematocidal composition comprising polyethylene glycol as the emulsifier, polyoxyethylene sorbitan monoleate as the dispersing agent, and Ethoxyquin as the antioxidant
Characterized in that it comprises 30 to 50%, 25 to 35%, 25 to 35%, and 5 to 15% of an emulsifier, 5 to 15%, and 0.1 to 0.01% of an antioxidant Of nematode
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KR20120111568A (en) | 2011-04-01 | 2012-10-10 | 한양대학교 에리카산학협력단 | Contamination measuring device, robot system and methode for maintaining outer wall of building |
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2014
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Patent Citations (3)
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KR100860726B1 (en) | 2002-01-24 | 2008-09-29 | 주식회사 프로바이오닉 | Bacillus thuringiensis Probio-35 strain for controlling parasitic nematode and method of treating nematode infections in plants |
KR101089318B1 (en) | 2009-04-28 | 2011-12-05 | (주) 에코윈 | Formulation for plant parasitic nematodes and method for preparing the same |
KR20120111568A (en) | 2011-04-01 | 2012-10-10 | 한양대학교 에리카산학협력단 | Contamination measuring device, robot system and methode for maintaining outer wall of building |
Non-Patent Citations (2)
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[문헌 1] Sela S. et al. 1998. Purification and characterization of Bacillus cereus collagenolytic/proteolytic enzyme and its effect on Meloidogyne javanica cuticular proteins. Eur. J. Plant. Pathol. 104. pp. 59-67 |
[문헌 2] Li W. et al. 2002. Broad spectrum anti-biotic activity and disease suppression by the potential biocontrol agent Burkholderia ambifaria BC-F. Crop Protection. 21. pp. 129-135 |
Cited By (3)
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WO2019088319A1 (en) * | 2017-11-02 | 2019-05-09 | 구경본 | Composition containing geraniol for control of plant-parasitic nematodes |
CN111295095A (en) * | 2017-11-02 | 2020-06-16 | 具炅本 | Composition containing geraniol for controlling plant parasitic nematodes |
US11278024B2 (en) | 2017-11-02 | 2022-03-22 | Kyung Bon KOO | Composition containing geraniol for control of plant-parasitic nematodes |
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