The bumblebee, Bombus terrestris, has played an important role as one of the alternative pollinators. Recently, pathogens and parasites affect the life span and fecundity of their host and been isolated from B. terristris. In order to detect viral infection in the field populations of B. terristris, we collected adults and isolated total RNA for reverse transcriptase-polymerase chain reaction (PCR). The PCR primers specific for several viruses such as deformed wing virus, Israel acute paralysis virus, Kashmir bee virus and black queen cell virus (BQCV) were newly designed and applied to gene amplification for cloning. Only BQCV was successfully amplified and sequenced, which suggests that BQCV may mainly infects the examined field population of B. terristris. To detect of capsid protein gene of BQCV, 4 selected regions were analyzed by primary PCR and 1 region was successfully amplified, which was further analyzed in quantitative real-time PCR (qRT-PCR). The qRT-PCR analysis demonstrated that BQCV was detected at concentrations as low as 0.1ng/μl total RNA. This result suggests that the detection via qRT-PCR can be applied for the rapid and sensitive diagnosis of BQCV infection in the field population as well as risk assessment of B. terristris.

본 시험은 사료용 옥수수의 검은줄오갈병에 대한 저항성 품종을 선발하기 위하여 2006년부터 2008년까지 전라북도고창에서 수행하였다. 국내외 사료용 옥수수 10품종을 1모작과 2모작으로 재배하여 품종에 따른 검은줄오갈병의 이병률과 수량성을 조사하였다. 그 결과 사료용 옥수수 품종에 따른 검은줄오갈병의 발병률이 차이가 있는 것으로 나타났다 (p<0.05). 검은줄오갈병에 대한 저항성의 정도를 4개의 그룹으로 분류하면, 매우 강한 그룹은 ‘광안옥’과 ‘광평옥’, 강한 그룹은 ‘P3156’과 ‘P3394’, 중간 그룹은 ‘청안옥’과 ‘P32P75’, 약한 그룹은 ‘NC7117’, ‘수원 19호’ ‘DK697’및 ‘GW6959’ 품종이었다. 검은줄오갈병 발생에 따른 사료용 옥수수의 건물수량은 품종 간에 차이가 있었으며(p<0.05), 검은줄오갈병 저항성 품종인 ‘광안옥’, ‘광평옥’,‘P3156’ 및 ‘P3394’ 품종은 이병성 품종보다 건물수량이14～26% 많았다. 따라서 검은줄오갈병 발생이 심한 지역에서 옥수수 재배를 할 때는 반드시 저항성 품종을 선택하는것이 중요하다

본 연구는 나리 인편자구 포장정식 시 재배시설에 따른 바이 러스 감염률 등을 조사하여 포장 구근생산체계를 확립하고자 하 였다. 맹아률은 비가림하우스 및 망실하우스에서 높았으며, 품종 별로는 ‘Yelloween’이 ‘Star Gazer’보다 높았고 ‘Yelloween’ 의 망실하우스에서 맹아률은 90.3%였다. 총생체중 및 구근특성 도 ‘Yelloween’이 ‘Star Gazer’보다 높았으며 ‘Yelloween’ 노 지 정식구에서 총생체중은 15.8g이었으며 구중은 10.7g으로 가장 양호하였다. 진딧물 비래의 최성기는 5월이었고 노지 38마리 에 비해 비가림 및 망실에서 8마리, 2마리로 월등히 낮았으며, 바이러스 이병률은 CMV가 LSV보다 높았으며 노지에서 CMV 이병률은 17.8%로 비가림 및 망실보다 4배 이상 높았다. Pyrethroid처리는 250mg • L-1이상에서 바이러스 방제효과가 있 었다.

구제역 바이러스는 소나 돼지 등의 우제류 가축에 경구 또는 흡입 노출 등의 다양한 노출 경로를 통해 심각한 감염성을 유발하는 생물학적 유해인자로 국내 축산업 운영에 있어 막대한 경제적 손실을 초래하는 병원균들 중 하나다. 구제역 발병에 대한 사전적 예방 관리를 도모할 수 있는 조치 방안 중 하나는 공기를 통해 전파되는 구제역 바이러스 확산에 대해 수학적 모델 적용을 통해 예측된 분석 결과에 따라 설정될 수 있다. 대기 확산 모델들은 일반적으로 구제역 바이러스의 주요 전파 경로인 경구 및 접촉 감염의 노출 시나리오를 예측할 수는 없으나, 상대적으로 예측도가 높은 대기 확산 모델의 경우 공기 전파에 대한 구제역 바이러스의 위해성 관리 방안을 결정하는 데 주요 역할을 담당할 수 있는 유용가능한 수단으로 여러 나라에서 활용되어 왔다. 구제역 바이러스 전파를 억제하기 위한 엄격한 관리 방안 중 다량의 가축 살처분 방법은 축산 농가들에게 심각한 경제적 손실을 초래하여 심각한 경우 새로운 자립의 여건마저도 상실시킬 수도 있다. 반면 낮은 수준의 대응 방안은 향후 구제역 발생에 따른 추가적 피해를 근원적으로 억제할 수 없는 역학적 한계에 봉착할 수 있다. 본 연구는 구제역 바이러스의 기본 특성 및 발생 모델 적용에 따른 공기 중 전파의 감염 위험성 평가 관련 선행 문헌들을 고찰하였다. 또한 1970년대 이후 구제역 바이러스의 공기 전파 경로를 예측하기 위해 여러 나라에서 실제 활용된 다양한 대기 확산 모델들의 적용 사례 및 장/단점을 비교 분석하였다.

Japanese encephalitis virus (JEV), a mosquito-borne pathogen, is the most important cause of viral encephalitis worldwide. The major vector mosquito of this virus, known as Culex tritaeniorynchus, is mainly live in paddy rice field in Korea. So Japanese encephalitis (JE) is a disease of rural areas. During urbanization, rice cultivated land has decreased also resulting the number of JE patients has decreased. However, recent outbreak of JE in Korea 2010, the patients distribution was not related with that of the mosquito. Therefore, this study was conducted to evaluate involving other mosquito species in the transmission of JEV. Five localities, JE reported in 5 years, have been selected to collect mosquitoes according their habitats (mountain, swap, cow shed, and downtown area) in 2012. Total 22,774 collected mosquitoes were pooled by species, date and site of collection. Of the 1,282 pools, eight (five were from Culex orientalis, one from Culex pipiens complex, and two from Aedes vexans) were found positive for flavivirus RNA in SYBR Green I-based real-time RT-PCR assay. The results of sequences blasting on NCBI showed that the flaviviruses from Cx. orientalis and Cx. pipiens complex were identified as JEV genotype V.

Insecticide resistance and activation of the metabolic detoxification enzymes of female Culex pipiens pallens by the blood meal were assessed using a micro-application bioassay and micro-plate enzyme activity assays. Four group of Cx. pipiens pallens were used, a susceptible non-engorging group at seven days after emerging, SNE7 Cp; a resistant non-engorging group at seven days after emerging, RNE7 Cp; a resistant engorged group at one day after blood feeding and a resistant engorged group at seven days after blood feeding, REG7 Cp. Insecticide resistance of Cx. pipiens pallans was increased by the blood feeding. Based on LC50 values, SNE7 Cp demonstrated >50 fold of higher susceptibility to all tested insecticides when compared with RNE7 Cp. RNE7 Cp showed higher susceptibility to all tested insecticides than REG1 Cp and REG7 Cp with a relative susceptibility LC50 (SRLC50) of 25.8 to 50.0 and 25.0 to 48.8. In micro-plate enzyme assays, the metabolic detoxification enzyme activity of Cx. pipiens pallans adult females was increased by the blood feeding. Activation of non-specific esterases (EST), glutathione-S-transferase (GST) and mixed function oxidase (MFO) in RNE7 were higher than in SNE7 and the all tested enzymes in REG1 and REG7 demonstrated significantly higher enzyme activation than RNE7, except for activation of GST in REG1. Activation of MFO in REG1 and REG7 were 209.4- and 74.6- fold higher than in REG7, respectively. Non-specific esterases (EST) and glutathione-S-transferase exhibited < 10 fold of higher Rr values. These results may be significant in terms of the criteria that are used to evaluate resistance, because blood fed female mosquitoes may show enhanced expression of the resistance phenotype, possibly allowing for earlier detection of insecticide resistance.

토마토 TYLCV는 병이 발생되면 약제나 다른 방법으로 방제가 어려우므로 내병성 품종의 개발이 반드시 필요하다. 특히, TYLCV 저항성 유전자 Ty-1,2,3(부분우성)을 한 개체 내로 집적을 시켜서 저항성이 증대된 품종의 개발이 필요하다. 토마토 TYLCV 저항성 11-TY1-AV 등 7계통을 대상으로 보독인 담배가루이를 이용하여 표현형을 조사하였다. 이병성 대비(11-TY5-AV; 슈퍼썬로드)와 F3 세대 3계통 및 F2 세대 4계통을 TYLCTHV 균주를 이용하여 접종하였다. 접종 13일 후에 이병성 대비가 100% 이병증상을 나타내었으며, 이때 11-TY4-AV는 48개체 중에서 23개체가 이병되었으며, 11-TY6-AV는 42개체 중에서 14개체, 11-TY7-AV는 48개체 중에서 14개체, 11-TY8-AV는 48개체 중에서 15개체가 이병 되었다. F2 세대 4 집단 모두 p>0.05 수준에서 TYLCV 접종 후 표현형이 정상적인 유전분리비(3:1)를 보이는 것을 확인 하였다. 또, TYLCV에 저항성인 동일한 자원을 부계로 사용하더라도 모계로 어떤 것을 사용 하느냐에 따라서 저항성 정도가 다르게 나타나는 것을 확인할 수 있었다.

Sweetpotato whitefly, Bemisia tabaci, is a vector of more than 100 plant-diseased viruses, as well as a serious pest of various horticultural plants. This species harbors a primary endosymbiont Portiera along with several secondary endosymbionts such as Cardinium and Hamiltonella. We investigated whether or not TYLCV acquisition alters the densities of endosymbionts in the body of B. tabaci using quantitative real-time PCR. Our results showed that the densities of both Cardinium and Hamiltonella, but not Portiera, increased upon acquisition of TYLCV. In addition, expression of GroEL, a molecular chaperone produced by Hamiltonella, was significantly upregulated in TYLCV-infected whiteflies. Our results suggest that endosymbionts may play an important role in TYLCV transmission mechanism within the body of B. tabaci.

The sweet potato whitefly, Bemisia tabaci (Gennadius, 1889) (Hemiptera: Aleyrodidae), is one of the most important agricultural pests by direct feeding and begomovirus transmission in the world. B. tabaci species complex consisting of about 30 species worldwidely has been mainly distinguished by molecular methods because of difficulties detecting morphological differences. In Korea, distribution of two exotic groups, the Mediterranean (Q biotype) and the Middle East-Asia Minor 1 (B biotype), were confirmed by comprehensive molecular methods in 2012. Of them, the Q biotype as a vector of Tomato yellow leaf curl virus (TYLCV) has caused significant crop yield losses. Tobacco leaf curl virus (TbLCV), one of four begomoviruses in Korea, was first reported in 2011. It damaged, although not frequent, to greenhouse tomatoes at Iksan, Gimje, Sunchang, and Jeju. The vector transmitting TbLCV, however, is not known yet. In this study, we sampled an unknown B. tabaci populations on leaves of the Japanese honeysuckle (Lonicera japonica) at Iksan in outdoor conditions. PCR analysis revealed that some populations were infected with TbLCV. We identified an unknown indigenous genetic group named as JpL and clarified its phylogenetic relationships based on nucleotide sequences of mitochondrial cytochrome oxidase I gene.

Among hemipteran insects which is the most important insect vector of plant viruses, small brown planthopper, Laodelphax striatellus, transmits the rice stripe virus (RSV) causing rice stripe disease. For effective control of RSV, it is important to understand interaction between RSV and L. striatellus. Therefore, in this study, expressed sequence tag (EST) databases were generated based on 454 GS-FLX pyrosequencing for comparative transcriptome analysis between nonviruliferous and RSV-viruliferous L. striatellus. By comparing the two EST libraries, we showed that 108 host genes were significantly up-regulated and 28 host genes were significantly down-regulated in viruliferous insects. Interestingly, genes encoding ribosomal proteins were mainly up-regulated in viruliferous L. striatellus, whereas genes related to translation were concentrated in the downregulated cohort. These RSV-dependently regulated genes may have important function in the behavior of planthopper or the transmission of RSV.

Nepal is endowed with rich diversity in land races of rice and also some wild rice species. Some of them are known for their resistance to insect and diseases, including rice hoppers and viruses. Nepal has so far released 68 improved rice varieties for different domains. With the introduction of improved varieties incidences of insect and diseases increased. Leafhoppers and planthoppers are among the rice insects of economic importance in Nepal. Till a decade ago, rice gundhi bug used to be the most important insect of rice. Now, hoppers have become more important. Of leafhoppers, green leafhopper (Nephotettix nigropictus), white leafhoppers (Cofena spectra) and zig zag leafhopper (Resilia dorsalis) are the common ones. Of them, N. nigropictus was found to transmit rice dwarf phytoreovirus, and N. virescens to transmit rice tungro bacilliform badna virus and rice tungro spherical waikavirus in Nepal. Of planthoppers, brown planthopper (BPH) (Nilaparvata lugens) and white-backed planthopper (WbPH) (Sogatella furcifera) are the major ones. Isolated outbreaks of BPH have been recorded from 1977 to recent past years and of WbPH in 1982. However, no virus diseases associated with BPH and WBPH have been recorded in Nepal though some yellow syndrome in large areas has been reported from BPH occurring area (Chitwan) for the last couple of years. It is worth to note that IPM-FFS (farmers’ field school) approach was adopted in 1997 in rice after the outbreak of BPH in Chitwan district. Farmers are practicing wetting and drying of rice fields for the management of BPH. Other management practices recommended include use of biopesticides, predators, light traps, chemicals and resistant varieties. With the objective of monitoring rice planthoppers and associated rice viruses Nepal in 2011 joined “Collaboration network for the management of migratory rice planthoppers and associated virus diseases of rice in Asia” - a regional project of Asian Food and Agriculture Cooperation Initiatives (AFACI), Korea.

In the past four years, outbreaks of acute respiratory diseases associated with canine influenza H3N2 viruses in dogs and cats have been reported in South Korea and China. For prevention of disease from spread of the disease and for administration of timely medical treatments, including countermeasures for quarantine, use of a rapid and highly sensitive detection method are important to detection of the causative viruses. This study was conducted in order to develop a real time RT-PCR for the H3N2 subtype. It was based on primers targeting the highly homologous sequences of matrix, hemagglutinin, and neuraminidase genes. The detection limit of real time RT-PCR was 10 copies/ul with matrix and hemagglutinin genes, and 1 copy with neuraminidase genes, respectively. This real time RT-PCR was as sensitive as virus isolation in 52 clinical samples. The detection system developed in this study might provide more rapid and highly sensitive results than commercial rapid kits based on immunochromatographic assay.

우리나라에서 가장 우점하는 BaYMV (Barley yellow mosaic virus)에 의한 국내 육성 청보리 품종의 저항성 정도에 따른 생육 및 수량 피해를 조사하였다. 바이러스 감염 검정에서 선우는 BaYMV와 BaMMV (Barley mild mosaicvirus)에 감염되어 있었으나 영양보리와 유연보리는 BaYMV에만 감염된 것으로 나타났다. 병징 발생정도에 따라 선우와 유연보리가 각각7~9와 5의 이병 정도를 보였으며 저항성 품종인 영양은 3정도의 저항성이 조사 되었다. 월동 후 초장과 분얼 경수의 생육을 조사한 결과품종별 저항성 정도에 따라 14.6~32.9%의 신장억제율을 보였으며, 생육 후기의 분얼 경수 단축률은 8.7~19.7%로 낮아지는 결과를 보였다.수량구성 요소의 피해를 건전포장 결과와 비교했을 때 중도 저항성과 감수성 품종에서 저항성 품종에 비해 간장에서 특히 큰 차이가 났으며, 수량 구성요소인 간장, 수수 및 수장 등을발병포장과 건전포장에서 수확한 재료에 대해조사하였다. 간장 단축 피해율을 조사한 결과영양보리는 14.5%가, 유연 및 선우보리는 각각24.8%와 42.7%의 신장 피해를 보였다. 수장과수수에 대한 조사 결과에서도 간장의 피해정도와 비슷한 결과를 보였다. 수장과 수수는 품종의 저항성 정도에 따라 각각 8.9~21.3%와 24.3~31.0%의 감소율을 보였다. 총체 건물 수량성을 조사한 결과 저항성 정도에 따라 건전 포장에 비해 21.6~58.0% 감소를 보여 바이러스 감염에 의해 유의적인 차이가 있었다. 특히, 감수성이 큰 선우보리에서는 건전포장의 8.0톤/ha에비해 58.0%의 감소 피해를 나타내었다. 종실수량은 약 30.0~61.2%의 감소율로 총체 수량의피해와 비슷한 결과였다. 감수성인 선우보리에서 감염포장에서 60% 이상의 종실 수량이 감소하여 가장 큰 피해정도를 나타내었다.

The influenza virus is an important respiratory risk affecting humans, and effective treatments are needed. Some oriental medicines are currently applied for treatment of common colds as well as influenza infection. Previous studies have reported that the therapeutic properties of MA-128 are effective for treatment of psoriasis antiasthmatic and atopic dermatitis. In this study, we investigated the therapeutic properties of the novel herbal medicine, MA-128, for treatment of influenza virus infection by oral administration. MA-128 is an active natural biological compound from herbal-marine origin. The results showed that oral administration of MA-128 in mice could confer a survival benefit against Type A influenza virus infection. Daily oral administration of MA-128 resulted in delayed death in infected mice for three days against mouse adapted H3N2 (A/Philippines/2/82). However, it protected more than 60% of mice from lethal infection of 2009 pandemic H1N1 (A/Korea/CJ01/2009) influenza virus. In addition, lung viral titers were significantly reduced at seven days post infection (~100 times) compared with mock-treated mice and viruses were cleared at 9 dpi only in the MA-128 treated groups. This study demonstrated the potential of the novel herbal medicine, MA-128, as an herbal remedy against influenza A viruses.

Ski protein is a nuclear transcription factor that does not bind DNA directly. Due to its unique binding properties with multiple factors, Ski could perform various roles in the regulation of both cellular proliferation and differentiation. We had previously reported that Ski protein is present in granulosa cells of atretic follicles, but not in preovulatory follicles, suggesting that Ski has a role in apoptosis of granulosa cells. The alternative fate of granulosa cells other than apoptosis is to differentiate to luteal cells; however, it is unknown whether Ski is expressed and has a role in granulosa cells undergoing luteinization. Thus, the aim of the present study was to examine whether the initiation of luteinization with luteinizing hormone (LH) directly regulates expression of Ski in the luteinized granulosa and luteal cells after ovulation by in vitro models. RT-PCR and real time PCR analysis respectively revealed that LH had no effect on c-Ski mRNA expression in the cultured granulosa cells regardless of LH treatment. Though Ski protein isabsent in granulosa cells of preovulatory follicle, its mRNA(c-Ski) was expressed and the level was unchanged even after LH surge. Taken together, these results demonstrated that Ski protein expression is induced in granulosa cells upon luteinization, and suggested that its expression is regulated post-transcriptionally. Moreover, expression of mRNA of Arkadia, an E3 ubiquitin ligases, in luteinizing granulosa cells in vivo was assessed by real time-PCR. The levels of Arkadia mRNA expression were unchanged during follicular growth and post ovulatory luteinization. These findings suggest that Ski protein level may be regulated during luteinization at translational and/or post-translational level but not by Arkadia.

Sloan-Kettering virus gene product of a cellular pro-oncogene c-Ski is an unique nuclear pro-onco protein and belongs to the Ski/Sno proto-oncogene family. Ski plays multiple roles in a variety of cell types, it can induce both oncogenic transformation and terminal muscle differentiation when expressed at high levels. Ski protein is implicated in proliferation/differentiation in a variety of cells. The alternative fate of granulosa cells other than apoptosis is to differentiate to luteal cells, however, it is unknown whether Ski is expressed and has a role in granulosa cells undergoing luteinization. Thus, the aim of this study was, by means of immunohistochemical methods, to locate Ski protein in the rat ovaries during ovulation and corpora lutea(CL) formation to predict the possible involvement of Ski in luteinization. In addition, to examine whether the initiation of luteinization with luteinizing hormone(LH) directly regulates expression of Ski in the luteinized granulosa and luteal cells after ovulation by in vivo models. In order to examine the expression pattern of Ski protein along with the progress of luteinization, follicular growth was induced by administration of equine chorionic gonadtropin to immature female rat, and luteinization was induced by human chorionic gonadtropin treatment to mimic luteinizing hormone(LH) surge. While no Ski-positive granulosa cells were present in preovulatory follicle, Ski protein expression was induced in response to LH surge, and was maintained after the formation of corpus luteum(CL). These results indicate that Ski is profoundly expressed in the luteinized granulosa cells and luteal cells of CL during luteinization, and suggest that Ski may play a role in luteinization of granulosa cells.

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.