Enzymatic properties of a bee venom serine protease from the bumblebee Bombus ignitus

추영무,진병래,Yuling Qiu,윤형주,이경용,손흥대 한국응용곤충학회 2011 Journal of Asia-Pacific Entomology Vol.14 No.3

We previously reported that bumblebee (Bombus ignitus) venom serine protease (Bi-VSP) acts as a prophenoloxidase-activating factor in arthropods and a fibrin(ogen)olytic enzyme in mammals. In the present study, wecharacterized the enzymatic properties of Bi-VSP purified fromB. ignitus venom. The 34-kDa active form of Bi-VSP was purified from the venomof B. ignitusworker bees.Glycoprotein staining showed that approximately 20%of the total molecularmass of Bi-VSP is due to carbohydratemoieties. Bi-VSP had an optimal pH and temperature of pH 9.0 and 40 °C, respectively, and was stable at 50 °C for at least 10 min. Bi-VSP activity decreased abruptly below pH 6.0, indicating that Bi-VSP activity is almost completely inhibited at pH 5.4 of B. ignitus venom. The protease activity of Bi-VSP was strongly inhibited by typical serine protease inhibitors such as phenylmethanesulfonyl fluoride, leupeptin, and soybean trypsin inhibitor.

거짓쌀도둑거저리 (Tribolium castaneum Herbst)의 생활사 및 난황단백질에 관한 연구

김성렬,추영무,임태승,진병래,손흥대 한국생명과학회 2000 한국생명과학회 학술발표회 Vol.29 No.-

거짓쌀도둑거저리 ( Tribolium castaneum Herbst ) 난황단백질의 특성

김성렬,추영무,임태승,진병래,손흥대 한국잠사학회 2000 한국잠사학회 잠사과학 100주년 국제심포지엄 및 추계학술대회 Vol.- No.-

곤충병원성선충, Steinernema arenarium의 수확시기와 유충 체장이 병원성과 발육 및 증식에 미치는 영향

한건영,이동운,추영무,추호렬,Han, Gun-Yeong,Lee, Dong-Woon,Choo, Young-Moo,Choo, Ho-Yul 한국응용곤충학회 2012 한국응용곤충학회지 Vol.51 No.1

곤충병원성선충, $Steinernema$ $arenarium$ 유충의 체장은 724 to 1408 ${\mu}m$로 변이가 크다. 따라서 $S.$ $arenarium$의 수확시기와 침입태 유충의 체장이 병원성과 발육 및 증식에 미치는 영향을 꿀벌부채명나방($Galleria$ $mellonella$) 노숙유충을 대상으로 조사하였다. $S.$ $arenarium$ 침입태 유충의 수확시기는 병원성에 영향을 미쳐 10일째에 수확한 것이 3일째 수확한 것에 비하여 병원성이 높았다. 10일째에 수확한 선충은 선충의 체장에 관계없이 처리 48시간 후에 꿀벌부채명나방 유충을 100% 치사시켰으나 3일째 수확한 체장이 작은 집단 접종 시에는 40%의 치사율을 보였으며 체장이 큰 집단은 18%의 치사율을 보였다. 기주 체내에 정착한 선충의 수도 침입태 유충의 체장에 따라 차이가 있었다. 3일째 수확한 선충들 중 체장이 작은 집단과 큰 집단, 혼합 집단 접종 시 기주에 정착한 선충 수는 각각 1.8, 3.3, 3.2마리였으며 10일째 수확한 선충들은 각각 5.3, 7.4, 7.6마리가 정착하였다. 3일째 수확한 유충의 체장이 작은 집단과 큰 집단을 구분하여 처리하였을 때 기주 체내의 암컷 체장은 각각 7,070.5 ${\mu}m$와 7,893.9 ${\mu}m$였고, 수컷 체장은 각각 1,460.5 ${\mu}m$와 1,688.2 ${\mu}m$였다. 10일째 수확한 체장이 작은 집단과 큰 집단 접종 시에는 기주 체내에서 성숙한 암컷의 체장은 각각 7,573.6 ${\mu}m$와 8,305.4 ${\mu}m$였고, 수컷의 체장은 1,733.4 ${\mu}m$와 1,794.4 ${\mu}m$였다. 수확시기와 접종 선충의 체장은 침입태 유충의 체장과 증식수에 영향을 미치지 않았다. The size of infective $Steinernema$ $arenarium$ juveniles is variable and ranges from 724 to 1408 ${\mu}m$. Effects of harvest time and infective juvenile size on pathogenicity, development, and reproduction were examined in the last instar of the great wax moth, $Galleria$ $mellonella$. Harvest time of infective juveniles (IJs) of $S.$ $arenarium$ affected pathogenicity. IJs harvested at the 10th day from trapping were more pathogenic than those harvested the 3rd day from trapping. Mortality of $G$. mellonella also depending on harvest time, $i.e$, 100% died within 48h when IJs were harvested at the 10th day, without relation to size. However, mortality was 40% in the small size group (SSG) compared with 18% in the large size group (LSG) within 48h when IJs were harvested at the 3rd day. Establishment of $S.$ $arenarium$ within the host was different depending on IJ size. The number of established IJs was 1.8 in the SSG, 3.3 in the LSG, and 3.2 in the mixed size group (MSG) when IJs were harvested at the 3rd day, and 5.3 in the SSG, 7.4 in the LSG, and 7.6 in the MSG when IJs were harvested at the 10th day. The length of the female adult was 7,070.5 ${\mu}m$ in the SSG and 7,893.9 ${\mu}m$ in the LSG and that of the male was 1,460.5 ${\mu}m$ in the SSG and 1,688.2 ${\mu}m$ in the LSG when IJs were harvested at the 3rd day. The length of the female adult was 7,573.6 ${\mu}m$ in the SSG and 8,305.4 ${\mu}m$ in the LSG and that of the male adult was 1,733.4 ${\mu}m$ in the SSG and 1,794.4 ${\mu}m$ in the LSG when IJs were harvested at the 10th day. Harvest time and size of IJs did not influence numbers of progeny or size of IJS.

Molecular cloning and characterization of two peptide toxins from the spider Araneus ventricosus

Hu Wan,이광식,추영무,제연호,Jianhong Li,진병래 한국응용곤충학회 2013 Journal of Asia-Pacific Entomology Vol.16 No.1

Spider toxins have great potential in the development of biopesticides. Here, we report the molecular cloning and characterization of two peptide toxins from the spider Araneus ventricosus. Two cDNAs encoding peptide toxins were cloned from A. ventricosus. Analysis of the cDNA sequence shows that the mature peptides of AvT-39 and AvT-48 consist of 39-amino acid residues and 48-amino acid residues, respectively. Both of the mature peptides include six conserved cysteine residues and a principal structural motif typical of spider toxins. The AvT-39 and AvT-48 cDNAs, which encode the mature peptide, were expressed in baculovirusinfected insect cells. AvT-39 and AvT-48 expression in insect cells significantly decreased cell viability. Additionally,the median lethal time (LT50) of Spodoptera exigua larvae inoculated with recombinant AcNPV expressing AvT-48 was approximately 1 day shorter than that of larvae expressing wild-type AcNPV, demonstrating that the recombinant virus reduced LT50 by approximately 25%. Taken together, our findings describe the molecular characterization of two peptide toxins from A. ventricosus and demonstrate the potential for these toxins to be used as biopesticides.

Dermal and Eye Irritation Studies on Bactonematicide, Photorhabdus temperata Isolated from Heterorhabditis megidis (Nematoda: Heterorhabditidae) in Rabbit

김진호,박순환,정남준,추영무,김영준 한국유기농업학회 2022 韓國有機農業學會誌 Vol.30 No.3

The acute dermal and eye irritation tests of Photorhabdus temperata, a symbiotic bacterium of Heterorhabditis megidis Gwangju strain, were carried out in New Zealand white rabbit (Oryctolagus cuniculus), following the guidelines of OECD and Rural Development Administration (RDA) of Korea. In both tests, neither dermal nor eye responses were found from all the P. temperata treated rabbits and the results were classified as non-irritating. That is, erythema, eschar, edema, and any other dermal critical signs were not observed from all the experimental rabbits in the dermal irritation test. In the acute eye irritation test, no clinical signs of cornea, iritis, conjunctiva (redness, edema, lacrima, and chemosis) were observed from all the experimental rabbits. Individual ocular irritation, mean ocular irritation, and acute ocular irritation were calculated as 0.0. The results of dermal and eye irritation studies on P. temperata indicated that this bacterium could be a safe and effective alternative bionematicide against the most serious plant-parasitic root-knot nematodes in the genus Meloidogyne.

사체의 유기장소별 부패진행에 따른 발생 파리류 비교

임채석,조태호,이동운,추영무,추호렬,Lim, Chae-Seak,Jo, Tae-Ho,Lee, Dong-Woon,Choo, Young-Moo,Choo, Ho-Yul 한국응용곤충학회 2012 한국응용곤충학회지 Vol.51 No.3

사체의 부패단계에 따라 발생하는 곤충은 차이가 있다. 본 결과는 사체의 유기조건에 따라 발생하는 절족동물의 종류를 법의학적으로 활용하고자 유기 가능성이 높은 여러 장소에서 수행한 일련의 연구 중 부패단계를 용이하게 구분할 수 있었던 몇몇 장소에서 사체의 부패단계에 따라 발생하였던 파리류를 비교 분석한 것이다. 부패단계에 따라 발생한 파리류는 유기장소(야산, 들판, 하천, 온실, 옥상)와 유기계절(봄, 여름, 가을, 겨울) 및 사체의 매장 유무에 따라 부패의 진행에 차이가 있음에 따라 차이가 있었다. 사체의 부패는 여름에 가장 빨리 진행되었으며 겨울철 온실에서 유기한 사체는 다른 장소에 비하여 빨리 진행되었다. 그리고 매장한 사체에 비하여 노출되었던 사체의 부패가 빨리 진행되었다. 모든 유기장소에서 노출처리된 사체의 부패초기 단계인 신선기(fresh stage)에 발생한 파리류는 구리금파리(Phaenicia sericata)와 연두금파리(Lucilia illustris), Lucilia sp., 큰검정빰금파리(Chrysomyia pinguis), 검정빰금파리(C. megacephala) 등 검정파리과(Calliphoridae)로 계절에 따라 차이가 있기는 하였지만 대체로 2일 이내에 성충이 사체를 찾아 들었다. 반면 매장처리에서는 신선기에 사체를 찾아드는 파리가 없었다. 쉬파리과(Sarcophagidae) 파리들은 유기 장소와 시기에 관계없이 사체에 유인되었으나 매장처리에 비하여 노출처리에 빨리 유인되었다. 또한 쉬파리과 파리는 온실에 유기한 사체에 더 빨리 유인되었으며 발생하는 시간도 짧았다. 온실에서 봄과 여름에는 신선기(fresh)에서 부패활성기(active decay) 사이에 유인되었으나 가을에는 신선기에서 부패진전기(advanced decay) 사이에 유인되었고, 겨울에는 부패진전기(advanced decay)와 유지기(remains) 사이에 유인되어 계절에 따른 부패단계에서도 발생하는 시기에 차이가 있었다. 큰검정파리와 두꼬리검정파리, 털검정파리는 봄과 겨울에 유기한 사체에 발생하는 우점종으로 fresh~active decay 단계에 주로 발생하여 활동시기가 다른 종과는 차이를 보였다. 매장된 사체에서는 검정파리과 유충보다는 쉬파리과 유충들이 먼저 출현하였다. Because insect visitors was different depending on decomposition stage, dipterans occurred on chicken carcasses depending on postmortem intervals were compared by abandoned sites and decomposition from a serial study on arthropod occurrence from carcasses as forensic indicator. Species occurrence and decomposition of carcasses were different depending on abandoned sites (forest hill, open field, stream, greenhouse and roof), seasons (spring, summer, fall and winter), and situation (burying and non-burying). Development of carcass decomposition was faster at all sites in summer, carcass in greenhouse in winter, and unburied carcasses. Although visiting time of blow flies (Calliphoridae) (Phaenicia sericata, Lucilia illustris, Lucilia sp., Chrysomyia pinguis, and Chrysomyia megacephala) was different depending on abandoned season, their adults were generally collected from carcasses within 2 days. However, there were no visited flies at fresh stage of buried carcasses. The flesh flies (Sarcophagidae) were collected from all sites and seasons, and much faster from unburied carcasses than buried carcasses. Those were collected earlier from carcasses in greenhouse than other sites and occurrence was also shorter. In greenhouse, occurrence time of flesh flies were different depending on season; spring and summer - from fresh to active decay stage, fall - fresh to active decay stage, and winter - advanced decay to remains stage. Calliphora lata, Tricerotopyga calliphoroides, and Aldrichana grahami were dominant species and occurrences were different from other flies. These flies were active mainly from fresh to active stage. Larvae of sarcophagid flies were occurred earlier than those of calliphorid flies on buried carcasses.

곤충병원성 선충을 이용한 목화바둑명나방(Palpita indica Saunder)의 생물적방제

김형환,추호렬,박정규,이상명,추영무 한국응용곤충학회 2001 한국응용곤충학회지 Vol.40 No.3

목화바둑명나방에 대한 곤춘병원성 선충의 병원성은 선충의 종과 농도에 따라 차이가 있었으며, 치사율은 령기에 따라 달랐다. 실험에 사용한 S. carpocapsae 포천 계통, S. glaseri 동래 계통, S. longicaudum 논산 계통, H. bacteriophora 함양 계통과 Heterorhabditis sp. 경산 isolate 중 S. carpocapsae의 효과가 가장 좋았다. 목화바둑명나방 유충 한 마리당 S. carpocapsae를 20마리 이상 처리하였을 때 72시간 후 5령충과 전용을 제외하고는 모든 령기에서 거의 100%의 치사율을 나타내었다. 또한 어린 유충일수록 감수성이 높았다. 곤충병원성 선충의 목화바둑명나방에 대한 반수치사농도는 선충의 종에 따라 차이가 있었는데, S. carpocapsae의 경우 령기에 따라 4.9~8.2마리였다. 각 령기에서 유충 한 마리당 선충을 20마리 접종하였을 때 가장 짧은 반수치사시간은 S. carpocapsae의 경우 령기에 따라 3.4~9.2시간이었다. 목화바둑명나방의 S. carpocapsae에 대한 치사율은 실험장소, 처리농도, 오이 잎의 위치에 따라 차이가 있었다. S. carpocapsae의 ha당$ 1$\times$10^{9}$ /마리 처리구보다 $3$\times$10^{9}$ 마리구에서, 노지보다 시설하우스에서, 상엽과 중엽보다 하엽에서 치사율이 높았다. Entomopathogenic nematodes were effective in the control of the cotton caterpillar, Palpita indica Saunder. P. indica mortality was significantly different depending on nematode species, treatment concentration, and instar. S. carpocapsae Pocheon strain was more effective against P. indica than S. glaseri Dongrae strain, S. longicaudum Nonsan strain, Heterorhabditis sp. Gyeongsan isolate, and H. bacteriophora Hamyang strain. When S. carpocapsae was treated with the rate of > 20 infective juveniles (ijs)/larva, mortality was ca. 100% at the 1st-4th instars of P. indica in 72h. $LC_{50}$ of entomopathogenic nematodes were significantly different depending on nematode species. The lowest $LC_{ 50}$ value was obtained by S. carpocapsae with 4.9-8.2 ijs in the 1st-prepupa while the highest $LC_{50}$ by Heterorhabditis sp. with 5.5-21.9 ijs, the shortest LT$_{50}$ value of P. indica was induced by S. carpocapsae when 20 ijs/larva were inoculated. The $LT_{50}$ s were 3.4-9.2h against the 1st-prepupa. The mortality of P. indica in fields was significantly different depending on field condition, nematode concentration, and leaf location. Mortality of P. indica by S. carpocapsae was higher at greenhouse,$ 3$\times$10^{9}$ ijs/ha and upper leaves than at field, 1$\times$10$^{9}$ ijs/ha and lower and middle leaves, respectively.

Newly Recorded Noctuid Pest, Leucapamea askoldis (Lepidoptera: Noctuidae) from Amur Silver Grass, Miscanthus sacchariflorus

정영학,유은주,안종웅,박정준,추영무,추호열,이동운 한국잔디학회 2017 Weed & Turfgrass Science Vol.6 No.4

Amur silver grass, Miscanthus sacchariflorus is one of the promising biofuel weeds. A damage of noctuid pest, Leucapamea askoldis was firstly observed from Amur silver grass in Hwasun silver grass plantation during the survey of insect pests of Amur silver grass in Iksan, Hwasun, and Sancheong plantation areas in Korea. The host of L. askoldis was not known yet in Korea. The L. askoldis damage was observed as larval feeding on newly grown shoots and roots of M. sacchariflorus close to soil surface from early May in 2013. Investigated larval density was 1.6±1.1 per m2 on April 4 and damage rate of shoots was 0.8±0.4% on May 4, 2013. The larvae bore into shoots of M. sacchariflorus and feed on the inside of plant. The damaged shoots are easily pulled out and distinguished by the boring hole on the shoots. L. askoldis was potential insect pest in M. sacchariflorus plantation areas.

곤충병원성 선충을 이용한 느타리버섯해충, 긴수염버섯파리 (Lycoriella mali)의 생물적방제

김형환,추호렬,이흥수,박정규,이동운,진병래,추영무 한국응용곤충학회 2001 한국응용곤충학회지 Vol.40 No.1

긴수염버섯파리 (Lycoriella mali)에 대한 곤충병원성 선충 Steinernema carpocapsae 포천계통과 Heterorhabditis bacteriophora함양 계통의 병원성을 실내와 포장에서 실험하였다. 곤충병 원성 선충에 의한 긴수염버섯파리의 치사율은 선충의 종, 농도, 온도 및 긴수염버섯파리의 발육단 계에 따라 차이가 있었다. S. carpocapsae가 H. bacteriophora보다, 긴수염버섯파리 2령충보다는 3령충과 4령충에서 효과가 높았다. 그리고 온도에 따른 긴수염버섯파리의 치사율은 $20^{\circ}C$보다는 $25^{\circ}C$에서 높았다. 반수치사농도가 가장 낮았던 값은 $25^{\circ}C$에서 S. carpocapsae의 긴수염버섯파리 2령 충에서 20.0마리, 3령충에서 27.5마리였다. S. carpocapsae에 의한 긴수염버섯파리의 치사율은 알과 1령충을 제외한 전 발육단계에서 병원성이 높았으며, 가장 치사율이 높았던 것은 암컷 성충으로 2$0^{\circ}C$에서 74.0%, $25^{\circ}C$에서 80.0%였다. S. carpocapsae는 긴수염버섯파리 암컷의 산란수를 감소시 켰고, 긴수염버섯파리의 성충에 의하여 감염이 된 후 전파가 되었다. 포장실험에서 S. carpocapsae를 $1.5\;\textrm{m}^2$ 당$2.25{\times}10^{5}마리와\;4.5{\times}10^{5}$ 마리로 처리한 14일째 유충감소율은 42.2%와 81.6%였으며 ,버섯재배사에서 곤충병원성 선충은 14일까지 지속되고 있었다. 곤충병원성 선충은 느타리버섯의 생장에는 아무런 영향을 미치지 않았다 The potential of two entomopathogenic nematodes, Sreinernema carpocapsae Pocheon strain and Heterorhabditis bacteriophora Hamyang strain as biological control agents was evaluated against mushroom ny, Lycoriella mali in laboratory and field. Mortality of L. mali was significantly different according to nematode species, concentration, temperature, and developmental stage of fly S. carpocapsae was more effective than H. bacteriophora. Mortality of L. mali was higher at $25^{\circ}C$ than at $20^{\circ}C$. In addition, the 3rd instal and the 4th instar of L. mali were more susceptible than the 2nd instar. The lowest $LC^{50}$ value was represented by S. carpocapsae, 20.0 infective juveniles (Ijs) in the 3rd instar, 27.5 Ijs in the 4th instar at $25^{\circ}C$. S. carpocapsae infected all the developmental stages of L. mali except egg stage and the 1st instar of larva. The highest mortality was shown in adult female representing 74.0% at$20^{\circ}C$ and 80.0% at $25^{\circ}C$.L. mali female adult was influenced by S. carpocapsae in oviposition. The number of eggs by L. mali female infected by nematodes was much lower than uninfected females. S. carpocapsae was dispersed by infected L. mali adult with higher numbers by females than males. When S. carpocapsae was applied at the rate of $2.25{\times}10^{5}\;and\;4.5{\times}10^{5}\;Ijs/1.5\;\textrm{m}^2$ in the mushroom house, mortalities were 42.2% and 81.6%, respectively. The infective juveniles of nematodes survived for 14 days in the mushroom medium. However, nematodes did not affect mushroom growth.