Polycystic ovarian syndrome (PCOS) is a heterogeneous syndrome associated with follicle growth arrest, dysregulated sex hormone profile, hyperthecosis and insulin resistance. Chemerin, a novel adipokine, is associated with obesity and metabolic syndrome. Although obese women and in PCOS subjects have elevated plasma chemerin levels, whether and how chemerin is involved in the regulation of follicular growth/steroidogenesis and pathogenesis of PCOS is unknown. Our objective is to better understand the complex regulatory mechanisms involved in the control of these processes and gain insights in their dysregulation in the pathogenesis of PCOS. We hypothesize that: (a) hyperandrogenism induces small and medium antral follicle growth arrest and ovarian structural changes, resulting from granulosa cell and oocyte apoptosis and theca cell survival, and (b) chemerin regulates follicular growth and steroidogenesis and contributes to the pathogenesis of PCOS. Using immature rats (day 13～15 for follicle culture and day 21～24 for granulosa cells culture) and a chronically androgenized rat model [dihydrotestosterone (DHT); 83 μg daily, day 21～105] which recapitulates the reproductive and metabolic phenotypes of human PCOS, we have examined the granulosa cell expression patterns of chemerin and its receptor CMKLR1 and their steroidogenic and follicle growth capability. DHT treatment resulted in decreased follicle numbers in preantral to preovulatory stages and absence of corpus luteum, but increased numbers of condensed atypical follicles. Atypical follicles, constituted predominantly of theca cells, exhibited high expression of calpain and down‐regulation of the cytoskeletal protein substrates vimentin, fodrin and β‐tubulin. Granulosa cell aromatase expression was significantly down‐regulated, a response accompanied by increased activated caspase‐3 content and DNA fragmentation. While PTEN levels were considerably higher in granulosa cells in the PCOS rats than controls, phospho‐Akt (Ser473) content was lower. In addition, DHT also activated granulosa cell caspase‐3, decreased XIAP, PARP and phospho‐Akt contents and induced apoptosis in vitro, responses that could be attenuated by forced expression of XIAP. These findings are consistent with our hypothesis that dysregulated follicular growth in PCOS is associated with changes in follicular growth dynamics and follicle cell fate, a consequence of dysregulated interactions of pro‐survival (p‐Akt, XIAP, PARP) and proapoptotic (calpain, PTEN, caspase‐3) modulators in a cell‐specific manner. Chemerin and CMKLR1 were expressed in granulosa cells and negatively regulated by gonadotropin in vivo and in vitro. Serum and ovarian chemerin levels in DHT‐treated rats were elevated, and associated with arrested early antral follicular growth, remodeling of the follicle wall and decreased expression of p450 side‐chain cleavage enzyme (p450- scc), aromatase and hydroxysteroid dehydrogenases. Recombinant chemerin inhibited FSH ‐ induced estradiol secretion in granulosa cells from DHT‐treated rats in vitro. Chemerin also suppressed basal and FSH‐ and GDF9‐induced follicle growth and estradiol/ progesterone production in preantral follicle cultures. Moreover, chemerin suppressed FSH‐induced p450scc/aromatase expression and progesterone/estradiol secretion in immature rat granulosa cells in vitro. These studies demonstrate that chemerin is a novel negative regulator in FSH‐induced follicular growth and steroidogenesis and support the notion that the dysregulation of chemerin expression and function contributes to pathogenesis of PCOS. Our observations also suggest that this chronically androgenized rat model may be useful not only for studies on the long term effects of androgen on folliculogenesis, but also on the pathophysiology of PCOS. * This work was supported by grants from the Canadian Institutes of Health Research (CIHR; MOP‐119381) and the World Class University (WCU) program through the Ministry of Education, Science and Technology funded by the National Research Foundation of Korea (R31‐10056).

In the transcriptome surveys of Laodelphax striatellus, several cDNA sequences showed a high level of similarities to the insect picorna-like virus genomes. Interestingly, there was no sequence similarity between picorna-like virus sequences from the RSV-viruliferous and those from the non-viruliferous L. striatellus. Picorna-like virus from the non-viruliferous L. striatellus was a geographical isolate of Himetobi P virus (HiPV). The genome of the HiPV was 9,272 nt in length excluding the poly(A) tail and contained two open reading frames (ORFs), which were separated by a 176 nt intergenic region that functions as an internal ribosome entry site (IRES). The 5' ORF encodes the non-structural proteins and the 3' ORF encodes the capsid proteins. The partial genomic RNA of the picorna-like virus from the RSV-viruliferous L. striatellus, LsPV-2, was 8,769 nt in length excluding the poly(A) tail and contained a single, large open reading frame (nt 1–8,535) encoding a 2,845 aa polyprotein. In terms of sequence similarity, identity, and genome organization, LsPV-2 resembled insect picornalike viruses belonging to the family Iflaviridae. A phylogenetic analysis based on RNA-dependent RNA polymerase (RdRp) sequence showed that LsPV-2 was most closely related to the deformed wing virus (DWV). The HiPV and LsPV-2 were incompatible each other in L. striatellus, suggesting that these two picorna-like viruses may have important functions in transmission of the RSV.

Plasmids from Bacillus thuringiensis have been implicated in pathogenicity as they carry the genes responsible for different types of diseases in mammals and insects. B. thuringiensis serovar mogi of a novel serogroup (H3a3b3d), which showed mosquitocidal activity against Anopheles sinensis and Culex pipiens pallens, was isolated from fallen leaves in Mungyeong city, Republic of Korea. In contrast to the complicated plasmid profiles of B. thuringiensis H3 serotype strains, the B. thuringiensis serovar mogi contained only megaplasmid (> 30 MDa) on which the toxin genes were occasionally located. Sequence analysis using 454-pyrosequencing revealed that the megaplasmid harbored at least seven putative cry genes, showing about 84%, 75%, 73%, 58%, 84%, 39% and 75% similarities in amino acid sequences with Cry27Aa, Cry19Ba, Cry20-like, Cry56Aa, Cry39ORF2, Cry8Ba and Cry40ORF2, respectively. These cry genes were cloned to the Escherichia coli-B. thuringiensis shuttle vector, pHT1K, and then introduced into an acrystalliferous B. thuringiensis Cry-B strain for further molecular characterization. This novel 3a3b3d type strain, B. thuringiensis serovar mogi, could be used as a good resource for studying unknown mosquitocidal cry genes.

ORF11 (ac11) of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is a highly conserved baculovirus gene whose homologs are found in all lepidoteran Group I NPV, but its function is unknown so far. To determine the role of ac11 in baculovirus life cycle, ac11 knock-out mutant, Ac11KO, was constructed using the plasmid capture system (PCS). Real-Time PCR analysis showed that ac11 transcript was first detected at 6 h post-infection (p.i.) and accumulated to maximum at 48 h p.i., indicating that ac11 is belong to late gene. When the genomic DNA of Ac11KO was transfected into Sf9 cells, viral replication was restricted to a cell transfected originally. While viral transmission of the Ac11KO was not observed in Sf9 cells, production of budded virus (BV) in Sf9 cells transfected with Ac11KO was observed by transmission electron microscopy (TEM). These results suggest that the ac11 is essential for AcMNPV to produce infective BV.

인공야간조명에 유인된 곤충은 대부분 그대로 죽게 되고, 생태계 내에서 1차 또는 2차 소비자의 역할을 하는 곤충의 개체군 감소로 인한 생태계 피해는 매우 심각하다고 할 수 있다. 본 연구를 통해 각 조명의 곤충 유인특성을 알아내고 주변생태에 영향을 덜 미치는 조명을 제안하고자 한다. 조사는 2011년 6월에서 8월에 걸쳐 인공야간조명과 이격된 총 5곳의 산지에서 30W의 백열등, 형광등, 할로겐등, 삼파장등, LED등을 이용한 첫 번째 유인실험을 5회 반복 실시하였고, 50W의 수은등, 나트륨등, 메탈할라이드등을 이용한 두 번째 유인실험을 총 6회 반복 실시하였다. 전등 주변 가로 세로 1m X 1m에서 유인된 곤충을 전량 채집하여 동정 및 종수와 개체수를 계수하였다. 군집분석에는 우점도, 균등도, 풍부도, 다양도를 이용하였으며, 사용된 모든 조명에 대해서는 조도와 UV-A량을 측정하였다. 실험결과 인공조명의 특성에 따른 곤충유인특성을 분석해 보면 첫 번째 실험은 조도는 가장 낮지만 UV-A의 측정값이 가장 큰 형광등에서 가장 많은 수의 곤충이 유인되었으며, 조도는 가장 높지만 UV-A의 측정값이 0인 LED등에서 현저하게 적은 수의 곤충이 유인 되었다. 다양도와 풍부도는 UV-A 그래프와 유사한 형태이고 형광등에서 가장 높고 LED에서 가장 낮았으며, 균등도에는 큰 차이가 없었다. 두 번째 실험에서 수은등이나 메탈할라이드등의 곤충유인율과 다양도, 풍부도, 균등도에는 큰 차이가 없으며, 조도가 가장 높지만 UV-A의 측정값이 현저히 작은 나트륨에서 적은 유인률과 낮은 다양도, 풍부도, 균등도를 나타냈다. 결론적으로 인공야간조명은 생태계에 악영향을 끼치며, 설치해야하는 경우 LED등이나 나트륨등과 같이 동일한 와트(W)내에서 조도는 밝지만 UV-A 방출량이 적은 조명을 제안한다.

As one of the subfamilies of Crambidae, Spilomelinae comprises about 3767 species in the world (Solis and Maes, 2002), and this subfamily is characterized by the following characteristics: Chaetosema absent, antenna filiform, labial palpus and proboscis well developed, forewing with R3 and R4 stalk at base, or R2, R3 and R4 stalked, but R5 single, 2A and 1A form a loop; hindwing Sc+R1 stalked with Rs; male genitalia with gnathos absent or rudimentary, but uncus well developed, shape various. Cambodia is a country that bordered by Thailand, Laos, Vietnam and Gulf of Thailand in Southeast Asia. The total area is 181,035 km2. It is also a country with high biodiversity, of them, 212 mammal species, 536 bird species, 240 reptile species, 850 freshwater fish species, and 435 marine fish species, but only a few studies about the fauna of Lepidoptera there. Accordingly, to survey the diversity of Lepidoptera is of great significance for systematic study in Cambodia. In this research, we started our survey from 2009, up to now, we got numerous Pyraloidea from Cambodia, in this study, we identified 48 species belonging to 36 genera of Spilomelinae, most of them are reported for the first time in Cambodia, and some specimen could not be identified which we will report in future. All the materials examined come from the collection of University of Incheon.

Plasmids from Bacillus thuringiensis have been implicated in pathogenicity as they carry the genes responsible for different types of diseases that in mammals and insects. A novel serogroup (H3a3b3d), B. thuringiensis strain K4 which showed mosquitocidal activity against Anopheles sinensis and Culex pipiens pallens, was isolated from fallen leaves in Mungyeong city, Republic of Korea. In contrast to the complicated plasmid profiles of B. thuringiensis H3 serotype strains, the strain K4 (designated as serovar mogi) had only one large plasmid (>200kb) on which the toxin genes were occasionally located. A 454 pyrosequencing was used for the complete sequencing of the large plasmid. The sequence analysis showed that k4 plasmid had at least seven putative cry genes, ending up to showing 84%, 75%, 73%, 58%, 84%, 39% and 75% homology with Cry27Aa, Cry19Ba, Cry20-like, Cry56Aa, Cry39ORF2, Cry8Ba and Cry40ORF2 toxins in amino acids, respectively. This novel 3a3b3d type strain, B. thuringiensis serovar mogi, can be used as a good resource for studying unknown mosquitocidal cry genes. The E. coli-B. thuringiensis shuttle vector, pHT1K was used to clone these cry genes for characterization. In each clone, the level of transcription and production of crystal proteins will be investigated in near the future.

Among 154 putative ORFs of Autographa californica multiple nucleopolyhedrovirus (AcMNPV), ac78 and ac79 are highly conserved genes in baculovirus, but their functions in the virus life cycle have been unknown so far. To determine their roles in AcMNPV replication, knockout mutants, ac78KO and ac79KO, were constructed using the plasmid capture system (PCS). Real-Time PCR analysis showed that both of ac78 and ac79 transcripts were first detected at 6 hours post-infection, and accumulated to maximum at 24 hours post-infection, suggesting that both of ac78 and ac79 are belong to late gene. When the genomic DNA of ac78KO was transfected into Sf9 cells, viral replication was restricted to a single cell infection. These results demonstrated that the ac78 play an important role in BV production, and therefore is essential for AcMNPV to mount a successful infection. Whereas Sf9 cells infected with the ac79KO showed normal viral symptoms such as rounding and swelling, OBs were not observed from majority of infected cells. These results suggested that the ac79 might play an important role in OB production.

덕적군도는 인천시 옹진군 서남쪽 약 82km 지점에 있는 덕적도를 비롯한 소야도, 문갑도, 선갑도, 선미도, 백아도, 굴업도, 울도 등으로 구성되어 있으며, 대체로 해안선의 굴곡이 심하고 곳곳에 소만입이 발달해 있다. 도서생물 지리학적으로 섬은 육지와 전혀 다른 혹은 독립된 생태적 특징을 가진다. 도서지역에서는 한정된 공간과 먹이자원의 한계가 있는 특성상, 종의 침입에 대한 저항성이 내륙보다 상대적으로 낮기 때문에 안정화 단계의 종 군집이 유지되지 않고, 여러 종들이 불안정한 군집을 이루며, 정착과 소멸을 반복하는 등의 변화가 심한 구조를 갖는 과정에서 종 다양성이 높게 나타나는 경향(Webb and Vermaat, 1990)이 있다. 본 연구는 덕적군도 도서지역의 곤충상을 파악하여 덕적군도 주변 섬들의 곤충상 비교를 통해, 도서지역의 곤충 다양성에 대한 기초자료를 제공하고, 덕적군도의 주요도서인 백아도, 굴업도, 지도 및 선갑도의 곤충상을 판단하여 보다 객관적인 자료를 제시하는데 목적이 있다. 조사는 총 4회에 걸쳐 덕적군도 도서를 대상으로 정량조사 및 정성조사를 병행 실시하였다. 조사결과 덕적군도 내의 4개 도서에서 총 11목 71과 272종이 확인되었다. 조사 기간이 여름에만 한해 있던 것을 감안하면 실제 서식종은 더 많을 것으로 판단된다. 법적 보호종은 굴업도에서 멸종위기야생동물II급에 해당하는 애기뿔소똥구리와 왕은점표범나비 2종이 확인되었으며, 백아도에서 멸종위기 야생동물II급에 해당하는 물장군이 확인되어 총 3종이 확인되었다.

Bacillus thuringiensis (Bt) strain K4 was isolated from fallen leaves which had been collected at a forest stand in Mungyeong city, Republic of Korea. The flagellated vegetative cells of Bt K4 were agglutinated with the H3 reference antiserum among 55 reference H-antisera. In a further test to identify subfactors, 3b and 3d monospecific antisera were reactive to the cells, followed up with introducing a novel serogroup of 3a3b3d, designated as serovar mogi. The strain K4 had mosquitocidal activity against Dipteran larvae, Anopheles sinensis and Culex pipiens pallens, with no Lepidopteran toxicity observed. The SDS-PAGE profile of K4 crystal protein, ovoidal-shaped, included several bands ranging from 30-75 kDa. Four putative peptides, Cry19Ba, Cry40ORF2, Cry27Aa and Cry20Aa were detected from the bands by a nano-LC-ESI-IT MS analysis. Through a thermal asymmetric interlaced PCR, cry19Ba, cry40ORF2 and cry27Aa genes were partially cloned from K4 strain. Three cry genes were further found in the strain by a 454 pyrosequencing, ending up to showing 58%, 39% and 84% homology in amino acids with Cry56Aa, Cry8Ba and Cry39ORF2 toxins, respectively. This novel 3a3b3d type strain, B. thuringiensis subsp. mogi, can be used as a good resource for studying unknown mosquitocidal cry genes.

The genetic diversity of 16 Ganoderma strains was investigated by rDNA-ITS sequencing. Alignment analysis showed that whole length of internal transcribed spacer(ITS1+ITS2) was 500bp and with 139 variation sites (accounting for 23.3％, ITS1 was 66 and ITS2 was 73), 337 conserved sites (accounting for 72.2％), 59 informative sites (accounting for 9.88％), 86 conversion sites (G-A, C-T), 13 transversion site(C-G, T-A). The ratio of transition and transversion in ITS1 was higher than that in ITS2, and the variable sites of ITS2 were more than those of ITS1. The genetic distance among 16 Ganoderma strains is from 0 to 0.121. The genetic distance between G. lipsiense and F-1 was 0, and the genetic distance between Heizhi 02 and Huizhou, Jingda, G. capense was 0.121, 0.117 and 0.120, respectively. The 16 Ganoderma strains were classed into 4 groups. The biggest group is comprised of 12 strains, including Xinzhou, Huizhou, Jingda, 902, F-1, Xianzhi, Meiluo, Taishan, G. applanatum, 05, G. luteomarginatum. The G. atrum and G. sinense were clustered into one group. The G. capense and Zhongzhi was independent group, respectively. These results showed that there were some genetic difference among groups, and there was lower genetic diversity among strains in same groups.

Four Ganoderma lucidum strains, Chizhi 05, Jingda, Huizhou and Xinzhou, were screened out as hybrid parent in order to establish G. lucidum cross breeding system that based on protoplast monokaryogenesis method. Monokaryotic strains of each parental strains were obtained and mating type of each monokaryotic strains were determined. One to three monokaryotic strains that have different mating types were mated, and hybrids were identified by clamp connection observation and antagonist response. The results showed that the number of monokaryon came from Chizhi 05, Jingda, Huizhou and Xinzhou was 9, 14, 40 and 38, respectively. Only one mating type was obtained from Jingda, and two mating types were obtained from the other three strains, Chizhi 05, Huizhou and Xinzhou, respectively. Chi-square test showed that the ratio of two mating types of the three strains was 1:1. Fourteen monokaryotic strains of different mating types from 4 parental strains were select as a cross- breeding materia, and 17 hybrids were obtained, which were identified by clamp connection observation and antagonist response. This study proclaimed that the practicality of the hybridization breeding of G. lucidum by protoplast monokaryogenesis method.

Rice black-streaked dwarf virus (RBSDV), a member of the genus Fijivirus within the family Reoviridae, is the causative agent of maize rough dwarf and rice black-streaked dwarf diseases, both of which can lead to severe yield losses in east Asia. Although molecular approaches such as RT-PCR have potential for detection and diagnosis of this virus infections, their impact on high throughput certification is still limited. Therefore, the development of an antibody-based assay for rapid and effective diagnosis of RBSDV is preferable. In this study, we collected RBSDV from rice with rough dwarf disease and its complete nucleotide sequences of 10 genomic segments encoding 12 non-overlapping ORFs were determined. Among 12 ORFs, ORF1, 2 and 12 showed high level of similarities with the RdRp, major core protein and major outer shell protein, respectively. These ORFs were expressed as polyhedrin fusion protein or full-length soluble protein using baculovirus expression system for the preparation of specific antibody against RBSDV, which could be useful for the detection and diagnosis of this virus.

We isolated two baculoviruses, Spodoptera litura granulovirus (SlGV) and S. litura nucleopolyhedrovirus (SlNPV) in the dead larvae of S. litura. The granule of SlGV were ovoidal shape with an approximate measure of 240-340 nm×140-180 nm, and each granule contained one single rod-shape virion with a mean size of 180-200 nm×20-40 nm. Whereas, the polyhedra of SlNPV were irregular in shape with a approximate diameter of 1.0-1.5 ㎛, and numerous virions comprised of the multinucleocapsid were contained in each polyhedra. The major component of occlusion bodies produced by SlGV and SlNPV were about 29 and 30 kDa, respectively. When the phylogenic relationship between these viruses were analyzed using the nucleotide sequences of granulin gene from SlGV and polyhedrin gene from SlNPV, they were not closely related to each other. We also found that the two viruses showed similar insecticidal activity against 2nd instar larvae of Spodotera litura in terms of dose-response, but SlGV showed much longer LT50 than that of SlNPV. The two baculoviruses might be cooperatively be applied as biological control agent for the control of S. litura

Autographa californica nucleopolyhedrovirus (AcMNPV) has a large doublestrand DNA genome of approximately 134 kbp and harbors 156 open reading frames (ORFs). To elucidate DNA replication cascade of AcMNPV, we developed a novel baculovirus genome that can be maintained in Escherichia coli as a plasmid and can infect susceptible lepidopteran insect cells. This genome, named bAc-MK, contains a mini-F replicon and a kanamycin resistance marker. Using a convenient Tn7 transposon-based system, pPCS-S, 55 single ORF-truncated mutants were generated by random insertion into bAc-MK genome. These single ORF-truncated mutants were independently transfected into Sf9 cells, 16 of them were found affecting viral replication since they defected in producing polyhedra. Furthermore, to verify the pathogenicity of the single ORF-truncated mutants, the remaining 39 mutants were subjected to bioassay to Spodoptera exigua 3rd instar larvae. Among them, ac9-, ac49-, ac103- and ac105-knockout mutants showed higher mortality compared to that of bAc-MK. These results suggested that these ORFs could be related to pathogenicity of AcMNPV.

The Bacillus thuringiensis strain K4 was isolated from fallen leaves, sampled in a forest region of the city of Mungyeong, Korea. The flagellated vegetative cells of B. thuringiensis strain K4 were agglutinated with the H3 reference antiserum and further, agglutinated with 3b and 3d monospecific antisera but non-reactive for 3c and 3e factor sera. These results create a new serogroup with flagellar antigenic structure of 3a3b3d, designated serovar mogi. The strain K4 showed high activity against dipteran larvae, Anopheles sinensis and Culex pipiens pallens while no lepidopteran toxicity. It produced a single ovoidal-shaped parasporal crystal whose SDS-PAGE protein profile consisted of several bands ranging from 75 to 30 kDa. Through the protein identification by nano-LC-ESI-IT MS analysis, the putative peptides of Cry19Ba, Cry40ORF2, Cry27Aa and Cry20Aa were detected. In contrast to the plasmid profile of B. thuringiensis H3 serotype strains, the strain K4 contained only a large plasmid (~100 kb) and we cloned partial cry27Aa, cry19Ba and cry40ORF2 genes from it by thermal asymmetric interlaced PCR. Sequencing analysis showed 87%, 88% and 88% homologous with known cry27Aa, cry19Ba and cry40ORF2 genes, respectively. The new type strain, B. thuringiensis subsp. mogi (H3a3b3d) will be a good resource for new mosquitocidal cry genes.

Entomopathogenic fungi are widely available as biological control agents for controlling insect pests in agriculture and forestry. The fungal culture broth contains various pathogenesis-related components such as blastospores, mycelium and insecticidal enzymes such as chitinase, Pr1- and Pr2-proteases, which have been reported to play an important role in penetrating insect cuticles. In this study, we tried to evaluate the utility of culture broth from Beauveria bassiana SFB-205 to control lepidopteran pests. High level of insecticidal activity correspond to over 90% of mortality were observed when the culture broth of B. bassiana SFB-205 was inoculated to the Spodoptera litura larvae together with the B. thuringiensis K1. The freeze-dried culture broth showed synergistic effects in insecticidal activity against larvae of S. exigua and S. litura when treated with corresponding baculoviruses, SeNPV and SlNPV. Active ingredient of the B. bassiana SFB-205 culture broth was identified to chitinase, which have truncated form by insertional mutation compared to previously reported chitinases.

Bacillus thuringiensis 1-3 (Bt 1-3) which was isolated from a Korean soil sample showed high insecticidal activity against Aedes aegypti as well as Plutella xylostella. The isolate was determined to belong to ssp. aizawai (H7) type by an H antiserum agglutination test and produced bipyramidal-shaped crystal proteins with a molecular weight of 130 kDa. PCR analysis with cry gene specific primers showed that Bt 1-3 contained cry1Aa, cry1Ab, cry1C, cry1D and cry2A gene, differing from spp. aizawai (reference strain) which contains cry1Aa, cry1Ab, cry1C and cry1D. We modified the plasmid capture system (PCS) to clone plasmid from Bt 1-3 through in vitro transposition. Fifty-three clones were acquired and their sizes were approximately 10 kb. Based on the sequence analysis, they were classified according to similarities with four known Bt plasmids, pGI3, pBMB175, pGI1 and pGI2, respectively. One of pGI3-like clones, named as pBt1-3, was fully sequenced and its 20 putative open reading frames (ORFs), Rep-protein, double-strand origin of replication (dso), single-strand origin of replication (sso), have been identified. The structure of pBt1-3 showed high similarity with pGI3 which is one of rolling-circle replication (RCR) group VI family.