간행물
한국응용곤충학회 학술대회논문집
- 발행기관 한국응용곤충학회
- 자료유형 학술대회
- 간기 반년간
- 수록기간 2000 ~ 2024
- 주제분류 농수해양 > 농학 농수해양 분류의 다른 간행물
- 십진분류KDC 495DDC 595
권호리스트/논문검색
2009년도 한국응용곤충학회 춘계학술발표회 (2009년 5월) 209건
181.
2009.05
구독 인증기관·개인회원 무료
This study was performed to investigate the bacterial diversity isolated from the twospotted spider mite and to interpret their correlation between insect bacteria and acaricide resistance. Twospotted spider mite, Tetranychus urticae was used the resistance strains, which developed over eight years to the six acaricides such as abamectin, acequinocyl, bifenozate, etoxazole, fenpropathrin, and pyridaben, respectively. After cultivating the bacteria from body maceration, bacterial colony was selected and identified through 16S rRNA gene sequences. We are identified six genus from Pyridaben resistant strain, five genus from acequinocyl, three genus from abamectin, bifenozate, etoxazole, and two genus from fenpropathrin. However, we could not found correlation between bacterial density and diversity (phylotypes) among these resistant strains. By analyzing the diversity of population microorganisms, fenpropathrin was showed 40% of Cs value (Similarity coefficient) with susceptible strain, however, abamectin and pyridaben were perfectly different (0%) with susceptible strain. It remains to be learned about how microorganisms co-evolutionary developed with their host insect correlating to the resistance and how microorganisms play role in acaricide resistant mite.
182.
2009.05
구독 인증기관·개인회원 무료
Insensitive acetylcholinesterase (AChE) was determined to be basically involved in a EPN-resistant (ER) strain of diamondback moth (DBM, Plutella xylostella L.), as estimated by the AChE inhibition assay using DDVP and thiodicarb as inhibitors in nondenaturing electrophoresis gel. AChEs were clearly separated into four different bands (a major band and three minor bands) in susceptible strain (CS) and all bands inhibited by used inhibitors almost same level, however, only two bands (a major band and a minor band) showed in that of ER strain and major band showed higher insensitivity. Moreover, ER strain showed cross resistance against used inhibitors and DDVP highly inhibited esterase in both strains. About 2kb of ace1,2 cloned and point mutations were detected in ER strain.
183.
2009.05
구독 인증기관·개인회원 무료
과원 문제해충의 하나인 꼬마배나무이의 월동성충 방제를 위한 기계유 유제 살포적기는 2월 1일부터 일일 최고기온이 6℃ 이상인 날의 수가 16-21일 에 도달하는 시기로 농가에서 활용하고 있다. 이 시기에 꼬마배나무이 성충이 낳은 알의 부화율을 2008-09년에 포장에서 채취한 배 단과지를 현미경하에서 조사하는 한편, 부화율 모델을 고안하고자 월동성충 방제적기 모델과 유사하 게 2월 1일부터 일일 최고기온 6℃ 이상의 온도를 누적하였다. 2008년에 50% 알 부화일은 4월 11일이었으며 누적적산온도는 429.7일도였고, '09년에는 4월 6일이었으며 417.6일도였다. 한편 농가에서 기계유유제로 월동성충을 방제할 때 살충제를 혼용하는 경우가 많은데 기계유유제와 살충제혼용, 살충제 단용 살포, 기계유유제 단용살포에 대한 꼬마배나무이 살충율을 조사하였다. 기계 유유제와 살충제 혼용살포는 혼합약종에 따라 다소의 차이가 있었으며 처리 7일후 살충율이 약 75-90%를 보였고, 살충제만 처리하였을 경우는 약 30~65%, 기계유유제만 살포한 경우는 살충제와 혼용하여 살포하는 것과 유사 한 결과를 보였다. 따라서 꼬마배나무이 월동성충을 방제할 때 살충제를 혼용 또는 단용 살포하는 것은 경제성이 없는 것으로 판단되었다.
184.
2009.05
구독 인증기관·개인회원 무료
Hyung Keun Oh, Chang Hwan Bae, Man Il Kim, Xinlong Wan, Byong Yong Park, Jae Kook Lee, Seong Han Oh, Yeon Soo Han, Iksoo Kim
Root knot nematode species, such as Meloidogyne hapla, M. incognita, M. arenaria and M. javanica are economically most notorious nematode pests, causing serious damage to the various crops throughout world. In this study, DNA sequence analyses of the D1-D3 expansion segments of the 28S gene in the ribosomal DNA were conducted to characterize genetic variation of the four Meloidogyne species obtained from Korea and United States. PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism), SCAR (Sequence Characterized Amplified Region) marker and RAPD (Random Amplification of Polymorphic DNA) also were used to develop the methods for exact and rapid species identification. In the sequence analysis of the D1-D3 expansion segments, only a few nucleotide sequence variation were detected among M. incognita, M. arenaria, and M. javanica, except for M. hapla. The PCR-RFLP analysis that involves amplification of the mitochondrial COII and lrRNA region yielded one distinct amplicon for M. hapla at 500 bp, enabling us to distinguish M. hapla from M. incognita, M. arenaria, M. javanica reproduced by obligate mitotic parthenogenesis. SCAR markers successfully identified the four root knot nematode species tested. We are under development of RAPD primers specific to the three root knot nematodes found in Korea.
185.
2009.05
구독 인증기관·개인회원 무료
The pinewood nematode (PWN, Bursaphelenchus xylophilus) is known as a virulent factor of the pine wilt disease, transmitted to pinewoods by the pine sawyer beetle, Monochamus alternatus. It is very hard to discriminate B. xylophilus from B. mucronatus because these Bursaphelenchus species are genetically and biochemically very close. Therefore, it has been necessary to detect PWN-infected trees for the prevention of pine wilt disease transmission in a short time. We developed polyclonal antibodies against B. xylophilus in BalbC mice and primarily screened with ELISA. Positive clones releasing polyclonal antisera revealed B. xylophilus-specific immuno-reactivity, which were at least two times higher than that of B. mucronatus. Two clones, D9-F10 and 1F3, were finally selected and exhibited specific immuno-reactivity for B. xylophilus, not for B. mucronatus in Western blot analysis. D9-F10 clone was reactive with a 43-kDa whereas 1F3 clone with two proteins, 40- and 45-kDa. Their isotypes against mouse Ig family were identical, kappa-light chain. These results suggest that these monoclonal antibodies can be useful for the development of diagnostic kit for the pine wilt disease.
186.
2009.05
구독 인증기관·개인회원 무료
This experiment was conducted to prevent the contamination of parasite egg on vegetables for supporting safety production of leafy vegetables. After the chinese cabbages and soil samples were collected at 5 cities of Gyeonggido during 2007-2008, the existence of parasite egg was surveyed. All collected samples had no parasite egg, so the chinese cabbages produced in Gyeonggi area were assumed parasite egg-free. To examine the characteristics of parasite eggs, the pig roundworms, Ascaris suum, were collected from the intestine of infected pig and the parasite eggs were collected from the uterus of A. suum. The eggs of A. suum developed to embryonated eggs, which can infect humans, in 20℃-30℃, but not 15℃ and 35℃, when cultured at different temperatures. The eggs developed to embryonated eggs after drying for 0-24h when cultured after different drying times. Effect of soakage different salt solution and times showed that the eggs developed to embryonated eggs after soakage at 0-25% salt solution for 0-24h. For eliminating the parasite eggs attached leafy vegetables, it was efficient to soak at salt solution for 5 minutes and washing 5 times with water.
187.
2009.05
구독 인증기관·개인회원 무료
Nam Sook Park, Han Seok Kang, Seon Ku Kim, Yong Gyun Kim, Byung Uuk Cho, Teak Soon Shin, Keun Ki Kim, Hyun Chul Park, Hong Joo Son, Hong Gu Lee, Sook Jae Sea, Hong Ja Kim, Eunju Park, Gyeong Im Jeon, Hyun Jung Lee, Uram Park, Sang Mong Lee
A full genomic DNA microarray technique was employed to investigate the effects of Dongchunghacho on aortal and hepatic gene expression in apolipoprotein E knockout mice fed a high-fat/high-cholesterol diet. Male 8- week - old ApoE-/- mice were randomly divided into two groups, control(high cholesterol group; HC) and supplementation of Dongchunghacho (SD). All of the mice were fed a high-fet/high cholesterol diet with or without Dongchunghacho supplemented by 1% for 6 weeks. At first, lipid profile of the Dongchunghacho was measured by biochemical analysis. No differences were observed in serum triglyceride and total cholesterol levels between the two groups. Antigenotoxic effect of the Dongchunghacho was measured by the single cell gel electrophoresis assay (Comet assay) and quantified as % fluorescence in tail. Dongchunghacho supplementation decreased significantly leukocytic DNA damage and also there was a tendency of reduction in hepatic DNA damage in Dongchunghacho group compared with the control group. In up regulated genes in liver and aorta of the mice, genes with 0 to 2- fold difference in expression level between the two group (HD and SD) was very much more in liver than in aorta, on the contrary, those with 2-fold to 16-flod difference increased greatly rather in aorta than in liver. Also, almost the same results were observed in down regulated genes in liver and aorta between the two groups. These results suggested that supplementation of Dongchunghacho might be helpful in preventing leukocytic DNA damage induced by high fat diet, and has a more crucial roles in aortal gene expression.
188.
2009.05
구독 인증기관·개인회원 무료
Microsporidia are obligate fungal intracellular parasites of all animal taxa. Among them the genus Nosema (Nosematidae) is known as the most common entomopathogen. Of these parasites, the ribosomal organization is one of the most pronounced molecular characteristics. One type is the normalarrangement of small subunit (SSU)-internal transcribed spacer (ITS)-large subunit (LSU) in the DNA sequence order. The other is the reverse arrangement of LSU-ITS-SSU. The latter is assigned to be the ‘true’ Nosema in the Nosema/Vairimorpha clade. However, we found that the SSU sequence of a strain of Nosema species having the normal arrangement of its rRNA sequence seemed to be more closely related to the ‘true’ Nosemagroup. Consequently we have further analyzed the complete sequence of rRNA. The results imply that there might be arecombination event in its rRNA evolution and/or the strain may form a novel group near the ‘true’ Nosema group. Interestingly both SSU and LSU of the ‘true’ Nosema and others may be under different selection pressure. We have also found that the size of ITS is distinct between the ‘true’ Nosema and other microsporidian species within the Nosema/Vairimorpha clade. This feature should be a useful diagnostic tool to distinguish the ‘true’ Nosema from others in the clade.
189.
2009.05
구독 인증기관·개인회원 무료
트랜스페린은 척추동물 및 무척추 동물에 존재하는 다양한 기능을 가진 철 결합 단백질 중 하나이다. 현재까지 선천성 면역에서 트랜스페린의 역할이 제 안되었음에도 불구하고 감염기간동안 트랜스페린의 면역학적 역할은 보고된 바 없다. 본 연구에서는 누에로부터 트랜스페린(이하 BmTf)을 분리한 후 그 특성을 분석하였다. BmTf 프로모터 영역의 염기서열 분석을 통해 면역기능 조절을 담당하는 전사인자인 Nuclear factor κB(NF-κB)의 결합부위가 다수 존 재하고, 바이러스, 세균, 곰팡이 등 다양한 병원체와 반응한 누에에서 그 발현 량이 크게 증가하므로 선천성면역 관련 기능이 있음을 추정할 수 있었다. 또 한, 배큘로바이러스 발현계를 이용하여 생산된 재조합 BmTf 단백질은 이온 부착능력과 다양한 그람 양성 및 음성 세균에 대하여 항균활성을 나타냄을 확인하였다. 이상의 결과를 통해 BmTf는 곤충 체내에 병원체 감염시 병원체 제거에 중요한 역할을 하는 곤충의 선천성 면역관련 기능을 수행함을 확인할 수 있었다.
190.
2009.05
구독 인증기관·개인회원 무료
A multiplex polymerase chain reaction (PCR) was developed for the simultaneous detection and differentiation among Nosema apis and Nosema ceranae in honeybee. Three sets of primers were selected from different genomic sequences to specifically amplify a 831 bp amplicon within the SSU rRNA gene, specific for both N. apis and N. ceranae (MSSR primer); a 375 bp amplicon within the SSU rRNA gene, specific for N. apis (NA primer); and a 1,131 bp amplicon within SSU rRNA gene, specific for N. ceranae (NC primer). Using the primers in conjunction (multiplex PCR) we were able to N. apis and N. ceranae and to differentiate between them. The sensitivity of this PCR assay was approximately 102 spores per milliliter. We proposed that the multiplex PCR was sensitive, specific and rapid tool that can serve as a useful differential diagnostic tool for detecting N. apis and N. ceranae in honeybee.
191.
2009.05
구독 인증기관·개인회원 무료
Sweetpotato whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae), is a serious pest of many economically important crops. The insect has developed resistance to chemical insecticides. Therefore, the development of microbial agent is necessary. Among the several entomopathogenic fungi, Lecanicillium lecanii Btab01 which has high insecticidal activity was carried out this experiments. To develop mass culture, we subcultured L. lecanii Btab01 on PDA, TSA, SDA+Y, RA and GSA media at 25℃ incubator to select the optimal solid culture medium. Hyphal growth was measured every 3 or 4 days. L. lecanii Btab01 grew fastest in RA, followed GSA, SDA+Y, PDA and TSA. L. lecanii Btab01 was cultured on PDB, TSB, SDB+Y, RB, GSB media at 25℃, 180rpm shaking incubator to select the optimal liquid medium. Spore germination was measured by spread plate method every 12 or 24 hours. Spore germination appeared 7.8×108 CFU/ml after 4 days in RB, followed GSB (5.5×108 CFU/ml), SDB+Y (2.7×108 CFU/ml), TSB (1.7×108 CFU/ml) and PDB (0.6×108 CFU/ml).
192.
2009.05
구독 인증기관·개인회원 무료
벼멸구(Nilaparvata lugens)는 한국에서 월동하지 못하고 이주해 오는 종으로 서, 매년 한국의 벼 재배에 큰 손실을 주고 있다. 한국의 지역별로 채집된 벼 멸구의 유전적 변이 및 다양성을 알아보고자 마이크로새털라이트를 이용하여 실험을 수행하였다. 한국의 다섯 지역, 곤양, 광양, 진주, 남해, 남원에서 벼멸구를 각각 채집하 였고, 마이크로새털라이트 5개의 위치(18, 27, 65, 67, 76)를 기준으로 분석하 였다. 그 결과, 3개의 위치(27, 65, 67)에서 높은 유의성을 확인할 수 있었고, 한국의 벼멸구 개체 간에도 유전적으로 차이가 있음을 알 수 있었다. 마이크로새털라이트 마커를 이용한 방법은 벼멸구의 근원지 및 이동을 결 정하기 위한 실질적인 방법이 될 것으로 생각되며, 이러한 방법을 통해 비래 해충인 벼멸구의 개체군 다양성을 분석함으로서 발생 근원지와 이동경로를 파악할 수 있을 것으로 생각된다.
193.
2009.05
구독 인증기관·개인회원 무료
Jong Yul Roh, Yong Wang, Qin Liu, Xueying Tao, Jae Young Choi, Hee Jin Shim, Hong Guang Xu, Soo Dong Woo, Byung Rae Jin, Yeon Ho Je
Plasmid capture system (PCS) was developed for easy cloning and manipulation of circular double-stranded DNA from various sources. Recently, we improved PCS system (named PCS-LZ) to clone relatively large-sized DNA molecules (30-150 kb). PCS-LZ donor consists of a Mini-F replicon and a kanamycin resistance marker between Tn7L and Tn7R regions. Both replicon and marker gene of PCS-LZ donor are transferred into target plasmid DNAs by in vitro transposition and the transposed DNAs can replicate in E. coli cells by transformation. White/blue screening by LacZ expression is also available to avoid backgrounds. Up to now, we acquired various circular DNA clones from four sources such as plasmids of B. thuringiensis, bacteriophage genome isolated from B. thuringiensis, genome segments of Cotesia glomerata bracovirus, and polymorphic genomes of Autographa californica nucleopolyhedrovirus. Among them, interestingly, the genome clones of bacteriphage (Ph1-3) were screened from the PCS transposition with plasmids of B. thuringiensis 1-3 strain. The genome of Ph1-3 was fully sequenced (46517 bp) and open reading frames were analyzed. In accordance with this genome finding, the phage particles and its DNA were confirmed from the supernatant of B. thuringiensis 1-3. Ph1-3 showed infectivity to B. thuringiensis type strains such as subsp. galleriae, entomocidus, and morrisoni. Based on these results, we screened the existence of phage in B. thuringiensis type strains by PCR with terminase small subunit-specific primers. Ten of 67 type strains showed PCR products and their sequence similarity was more than 70%. Conclusively, we expect this PCS-LZ system would be a powerful tool for genomic analysis and mutagenesis study at the area of invertebrate pathology and further its application will be enlarged to the vertebrate pathology area.
194.
2009.05
구독 인증기관·개인회원 무료
현재 주요 해충의 방제제로서 주목받고 있는 곤충병원성곰팡이의 이용을 위해 곰팡이 감염 이병충과 전국의 토양시료로 부터 곤충병원성 곰팡이를 분 리하고 그 분포를 조사하였다. 곤충병원성곰팡이의 효율적인 분리를 위해 기 보고된 선택배지의 조성을 바탕으로 가장 효율적인 선택배지 조성을 결정하 고, 선택배지를 이용하여 11종의 사충으로부터 Beauveria 속 9종과 Metarhizium 속 2종을 그리고 토양으로부터는 213개의 후보군을 1차 선발하고 배양한 후 그 형태학적 증식상으로 2차 분리하여 93개의 곤충병원성곰팡이 시료를 선별하 였다. 서식처별 분포는 산림 41개소 중 14개소(34.1%), 논 49개소 중 12개소 (24.5%), 과수 24개소 중 5개소(20.8%), 하천 87개소 중 18개소(20.7%), 밭 127 개소 중 23개소(18.1%), 수목 117개소 중 21개소(17.9%)에서 곤충병원성곰팡이 가 검출되었다. 이렇게 분리되어진 곤충병원성곰팡이는 여러 가지 다양한 해 충 방제를 위해 효율적으로 사용될 수 있을 것으로 기대된다
195.
2009.05
구독 인증기관·개인회원 무료
최근 친환경농업의 보급에 따른 유기농업의 확산으로 유기농 배추재배농가 포장에서 도둑나방의 발생이 급증하고 있으나 국내의 경우 등록된 유효약제 로 방제가 어렵다. 이러한 점에서 도둑나방의 효과적인 친환경적 방제를 위해 서는 곤충바이러스를 이용한 살충제의 개발이 시급하며 본 연구에서는 국내 에서 분리된 MabrNPV-K1의 전체게놈 특성과 더불어 도둑나방유충 영기별 병 원성 그리고 바이러스의 대량생산을 위한 적정 온도 및 증식 개체군의 크기 를 결정하고자 하였다. MabrNPV-K1 DNA를 제한효소로 처리하여 DNA단편을 관찰하고 패턴을 분석한 결과 평균 약 150 kb크기를 보여 기존에 보고된 MabrNPV DNA의 크기와 유사한 것으로 확인되었다. 또한 MabrNPV-K1의 영 기별, 온도별 병원성을 검정한 결과 영기가 낮고 온도가 높을수록 높은 살충 률을 보였고, 제제화를 위한 대량생산의 기초자료로 활용될 수 있는 다각체 생산량은 도둑나방 유충 3령 30℃ 조건에서 104 PIBs/larva 로 접종하였을 때 가장 효율적인 생산량을 보였다.
196.
2009.05
구독 인증기관·개인회원 무료
A novel recombinant baculovirus, NeuroBactrus, was constructed to develop an improved baculovirus insecticide with additional beneficial properties such as higher insecticidal activity and recovery to wild-type baculovirus. For this, Bacillus thuringiensis crystal protein gene (cry1-5) was introduced into Autographa californica nucleopolyhedrovirus (AcMNPV) genome by fusion of polyhedrincry1- 5-polyhedrin under the control of poyhedrin gene promoter. In the opposite direction of this fusion gene, an insect-specific neurotoxin gene (AaIT) under the control of early promoter from Cotesia plutellae bracovirus was introduced by fusion of orf603 partial fragment. Western hybridization and confocal microscopy revealed that AaIT neurotoxin and Polyhedrin-Cry1-5-Polyhedrin fusion protein expressed by the NeuroBactrus and that the fusion protein occluded into the polyhedra. In addition, the fusion protein was activated as about 65 kDa of crystal protein when treated with trypsin. The NeuroBactrus showed high level of insecticidal activity against Plutella xylostella larvae and significant reduction in median lethal time (LT50) against Spodoptera exigua larvae compared to those of wild-type AcMNPV. Re-recombinants derived from the NeuroBactrus, NBt-Del5 (deleted cry1-5), NBt-DelA (deleted AaIT) and NBt-Del5A (deleted cry1-5 and AaIT; wild-type baculovirus) were generated in serial passages in vitro and in vivo. These results suggested that the NeuroBactrus could be transferred to wild-type baculovirus along with serial passages by the homologous recombination between two polyhedrin genes and two partial orf603 fragments.
197.
2009.05
구독 인증기관·개인회원 무료
The genome of a granulovirus isolated from the tobacco cutworm, Spodoptera litura, was completely sequenced. The nucleotide sequence of the Spodoptera litura granulovirus (SlGV) genome was 124,121 bp long, with a 61.2% A+T content and contained 133 putative open reading frames (ORFs) of 150 nucleotides or larger. The 133 putative ORFs covered 86.3% of the genome. Among these, 29 ORFs were conserved in most completely sequenced baculovirus genomes, 35 were granuloviruses (GVs)-specific, and 60 were present in some NPVs and/or GVs. Especially, we proved that there were 9 SlGV-specific ORFs by RT-PCR. When the phylogenic relationship was analyzed using the nucleotide sequence of granulin gene, SlGV was most closely related to Trichoplusia ni granulovirus (TnGV) and Xestiac-nigrum granulovirus (XcGV) which were belonged to Type-I granulovirus. Comparative analysis of gene organization of the SlGV genome with those of other baculoviruses were carried out using blast matrix and gene order diagram.
198.
2009.05
구독 인증기관·개인회원 무료
Qin Liu, Jong Yul Roh, Yong Wang, Jae Young Choi, Hee Jin Shim, Hong Guang Xu, Xueying Tao, Byung Rae Jin, Yeon Ho Je
Bacillus thuringiensis 1-3 (Bt 1-3), belonging to subsp. aizawai (H7), showed different characteristics in plasmid profiles and had cry2A gene in addition to cry1Aa, cry1Ab, cry1C and cry1D. This strain exhibited dual insecticidal activity against Aedes aegypti as well as Plutella xylostella. Recently, we improved the donor-s of plasmid capture system (PCS) by inserting attB sites including lacZ between transposable elements (designated as pPCS-Troy), to construct E.coli-Bt shuttle vector. Through in vitro transposition with total plasmids DNA of Bt 1-3, 53 clones were acquired and their range of sizes were approximately 10 kb. Based on the sequence analysis, they were classified in 4 groups showing similarity with 4 known plasmids, pGI1, pGI2, pGI3 and pBMB175, respectively. One of pGI3-like clones was fully sequenced and its open reading frames were analyzed. As a donor for construction of shuttle vector, pDonr-attPEm vector harboring erythromycin resistant gene between attP sites was constructed. Through BP recombination with pPCS-Troy-cloned Bt plasmids and pDonr-attPEm, erythromycin resistant gene was transposed to Bt plasmids. This scheme proposes that in vitro transposition using pPCS-Troy and BP recombination using pDonr-attPEm can easily construct novel shuttle vectors with any Bt plasmids and this combined procedure can introduce foreign gengs into various circular DNA molecular.
199.
2009.05
구독 인증기관·개인회원 무료
작은뿌리파리(Bradysia difformis)의 유충, 번데기 및 성충에 의한 Pythium spp.과 Fusarium oxysporum 균의 전반을 조사하기 위하여 두 균을 증식한 PDA 배지상에 작은뿌리파리 유충, 번데기 및 성충을 접종하여 Pythium spp.과 Fusarium oxysporum을 먹이거나, 접촉시킨 2일 후 유충과 번데기는 0.6% sodium hypochlorite와 0.1% Triton-X 100의 혼합액에서 1분 동안 표면 살균을 하고 새로 만든 PDA 배지에 올려 놓은 다음 colony수를 조사한 결과 각각 26.7개, 18.0개였다. 그리고 번데기는 전반을 하지 못하였으며, 성충은 Fusarium oxysporum만 5.7개의 colony를 형성하였다. 작은뿌리파리 유충을 육묘용 트레 이에 0, 1, 2마리씩 접종하고 Pythium spp.을 106 농도로 접종한 결과 수박, 오 이, 참외, 메론, 애호박에서 각각 유충 2마리를 접종했을때 60.0%, 43.3%, 40.0%, 13.3%, 26.7%로 수박에서 가장 발병률이 높았다. 육묘용 트레이에 작 은뿌리파리 유충을 0, 1, 2마리씩 접종하고 106 농도로 Fusarium oxysporum을 접종하여 발병률을 조사한 결과 수박, 오이, 참외, 메론, 애호박에서 각각 46.7%, 33.3%, 30.0%, 10.0%, 20.0%의 병이 발생하였다. Pythium spp.과 Fusarium oxysporum 106 농도를 각 트레이에 방사한 후 작은뿌리파리 성충을 방사하여 병 전반을 알아본 결과 Pythium spp.은 수박에서 23.3%, Fusarium oxysporum도 수박에서 20.0%의 병 발병률을 보여 가장 높았다. 한편 형광성단백질(GFP, Green Fluorescencee Protein)를 삽입한 Fusarium oxysporum 균을 PDA배지에서 배양한 다음 3일 동 안 작은뿌리파리의 유충을 넣어서 균을 먹인 후 형광현미경에서 유충을 촬영 한 결과 균을 섭식하는 것을 입증할 수 있었다.
200.
2009.05
구독 인증기관·개인회원 무료
Among bee venom proteins, phospholipase A2 (PLA2) is critical one of bee venom components to defend against predators intruders. In this study, PLA2 gene from cDNA libarary using the venom glands of Bombus ignitus worker bees(BiVn-PLA2) was cloned and characterized. BiVn-PLA2 spans 2211 bp and consists of three introns and four exons encoding 180 amino acid residues. BiVn-PLA2 shares high levels of identity with a bumblebee, B. terristris (89% protein sequence identity), B. pennsylvanicus (88%), and a honey bee, Apis mellifera (53%). Northern blot analysis revealed that BiVn-PLA2 is expressed in venom gland, indicating that BiVn-PLA2 is one of the venom components of B. ignitus. To determine BiVn-PLA2 of venom components from venom sac, N-terminal amino acid sequencing of a putative BiVn-PLA2 (the purified 18 kDa) was performed by Edman degradation. The N-terminal amino acid sequencing of the 18 kDa protein was coincident with the N-terminal amino acid residues of the mature BiVn-PLA2 and the 18 kDa protein catalysed the hydrolysis of DBPC subs trate[1-O-(6-Dabcyl-aminohexanoyl)-2-O-(12-(5-B ODIPY-entanoyl) aminododecanoyl)-sn-glyceryl phosphatidylcholine] that is a sensitive fluorogenic probe for PLA2 activation. Western blot analysis revealed that BiVn-PLA2 is expressed in the venom gland, stored in the venom sac, and then emitted throughout sting apparatus. Finally, to test BiVn-PLA2 toxicity, BiVn-PLA2 was adjusted to a insect cell (Sf9) at different concentrations (1-30 μg/2×105 cells). The apoptotic cell death assay results showed that the cell survival decreased with increasing concentrations (1-30 μg/2×105 cells).
