남부지역에서 고구마 바이러스 무병묘의 정식시기 및 재배 기간에 따른 괴근 수량 및 씨고구마 생산량을 조사한 결과는 다음과 같다. 1. 바이러스 무병묘 정식시기에 따라 씨고구마로 사용하기에 알맞은 100~300 g 크기의 괴근 생산량은 6월 중순 정식 재배에서 ‘신율미’ 2,364 kg/10a, ‘풍원미’ 2,625 kg/10a 수준이었다. 2. 바이러스 무병묘를 7월 상순에 정식하여 재배하여도 씨고구마로 사용하기에 알맞은 100~300 g 크기의 괴근 수량은 ‘신율미’ 2,043, ‘진홍미’ 1,799, ‘다호미’ 1,390, ‘풍원미’ 1,985 kg/10a 정도가 생산되었다. 3. 고구마 바이러스 무병묘 정식 후 110일, 120일, 그리고 130 일 재배 시 씨고구마 수량은 ‘풍원미’ 1,605, 1,907, 1,834 kg/ 10a, ‘호감미’ 1,816, 1,771, 2,137 kg/10a 수준으로 생산되었다. 4. 수확 전 채취한 고구마 잎에서 SPLCV, SPFMV, SPVG, SPLV 등 4종 바이러스 이병 정도를 검정한 결과 정식시기, 재배기간 등 품종에 따라 약간 차이는 있으나 낮게 검출되어 다음해 씨고구마 종자로 문제가 없다고 판단하였다. 5. 바이러스 무병묘 대량 증식에는 시설과 노력이 많이 소요되고 증식량이 적어 면적 확대에 어려움이 있기 때문에 무병씨고구마를 생산하면 면적 확대에 유리하다. 무병묘가 소량 일 경우는 계속 증식하면서 7월 상순까지 본밭에 정식하여도 씨고구마 생산이 가능하였다. 금후에는 무병묘로 생산한 연차 간 씨고구마에 대해 품종별 수량과 품질 평가 후 씨고구마의 적정 갱신 주기를 설정하고자 한다.

In the case of foot-and-mouth disease (FMD), there is a great deal of impact on the national economy due to the disposal of diseases, the cost of disease control such as vaccination, reduction of productivity, and restriction of international trade of livestock products. Therefore, appropriate diagnostic methods for sensitive, accurate and rapid identification of virus serotypes are continuously required in terms of early prevention of FMD. This study was conducted to confirm the feasibility of immuno-PCR diagnostic method for the more sensitive detection of Korean FMD virus (FMDV). We synthesized a partial FMD type A viral gene. Protein antigen, monoclonal and polyclonal antibodies of FMDV were cloned, expressed and purified and then magnetic particles were attached to polyclonal antibodies and and oligomers to monoclonal antibodies for the immnuno-PCR. We confirmed the antigen-antibody and oligomer reaction using ELISA, Western blot, and real-time PCR. These results show that Korean FMDV can be detected by using difference of Ct values between positive group and negative group using immuno-PCR.. The results of this study also suggest that this technique will be the basis of the diagnosis method to detect Korean FMDV more sensitively in the future.

낭충봉아부패병에 저항성인 신품종 토종벌을 농업과학원에서 시험재료로 제공받아 일반 농가에서 사육중인 일반벌과 비교하였다. 꿀벌 수밀력을 조사한 결과 일벌 개체당 수밀력은 비저항성 계통보다 저항성계통이 15% 높았으며, 화분채집량은 저항성계통에서는 358mg, 비저항성 계통은 330.3mg으로 저항성계통이 비저항성계통보다 8.4%의 높은 화분채집력을 보였다. 질병저항성을 알아보기 위해 봉개유충 청소행동 평가에서 8시간 후 저항성계통에 서는 51.3%, 비저항성계통에서는 50.7%이었으며, 24시간 후에는 두계통 모두 98.7~100%의 사충 제거율을 보였다. 봉군발육조사에서는 저항성 계통의 경우, 일벌, 유충, 번데기의 수가 비저항성계통에 비하여 41%, 7%, 73% 로 발육이 우수하였다.

Honeybees are inevitably threatened by various pathogens including Sacbrood virus (SBV) and Galleria mellonella. Recently, RNA interference (RNAi) has been suggested as a promising strategy for suppression of honey bee viruses. Also, Bacillus thuringiensis (Bt) has been widely applied for the control of lepidopteran pests such as G. mellonella. In this study, it was intended to develop dsRNA production platform using Bt. For this, the pHT1K-SBV vp1 vector which transcribes sense and anti-sense SBV vp1 gene under control of Cyt1Aa sporulation-dependent promoter was introduced into Bt strain NT0423 expressing Cry1-types toxins. SBV replication was suppressed in the worker A. cerana ingested dsRNA produced from the Bt transformant. Crystal proteins from the Bt transformant showed high level of insecticidal activity against 4th instar larvae of G. mellonella.

The baculovirus-insect cell expression system has been widely used method for the recombinant protein expression. The present study has several limitation. In this study, we constructed vectors consisting of transcriptional enhanced factor and promoter that improve the expression level. To confirm the usefulness of these vector system, Human papillomavirus (HPV) VLPs have been expressed by baculovirus hyper expression system. HPV VLPs were purified using a CaptoTM Core 700 (GE Healthcare Life Sciences) chromatography approach. Baculovirus hyper expression system production efficiency was influenced by the HPV VLPs production. HPV VLPs vaccination to BALB/c mice induced the generation of antibody confirmed by ELISA. This study could provide improvements on the vaccine production for the development of VLP vaccines high expression of useful heterologous proteins.

South Korea has over 0.38 million of managed honey bee (Apis cerana) colonies before 2009 years ago, which produce the highest quantity of honey in the Korea; however, almost colony (90%) were collapsed by Korean Sacbrood Virus (KSBV) in South Korea. Korean Sacbrood Virus (KSBV) is the pathogen of A. cerana Sacbrood disease, which poses a serious threat to honeybee A. cerana, and tends to cause bee colony and even the whole apiary collapse. Colony collapse of A. cerana was first reported on the Pyeong-Chang of the South Korea in 2009. Several scientists and governments has been tried research for cure the sacbrood disease in A. cerana colony by medicines and management techniques. Unfortunately, The sacbrood disease dosen’t improve. So, we were developed a better breed of A. cerana for resistance of sacbrood virus by selection and then artificial insemination. A. cerana breeding technique was first successful applied with A. cerana in Korean. Queens was grafted from sacbrood resistance line and then it was growing in sacbrood disease colony that was survived 100%. Altogether selected 18 queens were artificially inseminated and 2,000 drones of A. cerana in Korea was used to evaluate amount of semen collection. We are select two scabrood resistance A. cerana line (R and H). R line be used for rearing the Queen. Drone was reared in H line colony. The RH hybrid were not infected sacbrood virus even spread sacbrood virus (2×106). RH colonies have very excellent hygienic behavior, brood, and sacbrood disease resistance activity.

We monitored the population of Haemaphysalis longicornis, in Andong, Gyungbuk from April to November, 2018. Among total 2,994 ticks collected for 8 months, H. longicornis 1,677(56%), H. spp Larva 1,074(35.9%), H. flava 213(7.1%), Ixodes nipponensis 30(1.0%) were identified. In addition, considering the environment, 1,727(57.7%), 907(30.3%), 192(6.4%) and 168(5.6%) ticks were collected in the grassland, graves, copse, mountain path, respectively. In the pathogen diagnosis with PCR using SFTS virus specific primers, positive viruses were detected in H. longicornis, H. flava and H. spp from June to October. The minimum field infection rate of June, July, August, September and October were 0.4%, 0.8%, 1.2%, 0.8% and 2.3%. respectively.

Sacbrood virus (SBV) caused significant colony collapse in Korean Apis cerana. Considering that hygienic behavior in honey bees confers colony-level resistance against brood diseases, we utilized this trait for selecting A. cerana colonies. In addition, the brood survival rate was evaluated after colonies were SBV-inoculated. Over four selective generations, dead brood removal and brood survivorship in selected colonies were higher than those in the unselected colonies (P < 0.01, 99.3 vs. 89.9% for removal of pin-killed pupae; P < 0.01, 99.0 vs. 63.9% for removal of SBV-killed larvae; and P < 0.01, 70.0 vs. 9.2% for brood survivorship). Following SBV-inoculation, selected colonies showed an increase in the number of surviving pupae and adults, whereas unselected colonies collapsed mostly. Our results confirm the feasibility of selecting SBV-resistant A. cerana.

Bemisia tabaci is a species complex consist of about 40 cryptic species. This species is also a vector of at least 100 begomovirus including Tomato yellow leaf curl virus (TYLCV). TYLCV transmitted by B. tabaci in a circulative propagative manner but its molecular mechanism is remain unclear. We investigate the transcriptional response of B. tabaci MED to TYLCV infection using next generation sequencing (NGS) analysis. In total 16757 differentially expressed genes between viruliferous and non-viruliferous whiteflies were analyzed. Among them 289 transcripts found significant, where 116 up-regulated and 173 down-regulated. Our results reveal the whitefly-TYLCV relationship and provide important molecular information about virus transmission of vector insects.

Among the various expression systems used for foreign protein expression, baculovirus expression system (BES) has the high level of post-translational modification ability such as glycosylation, folding and disulfide bonding. BES is widely used now in the production of VLPs because it is possible for the efficient multi-gene expression. However, there are not many cases of VLPs being manufactured through BES. Therefore, in this study, three improvements were made to increase the productivity of VLP through BES. A new heper enhanced expression vector was constructed to increase the expression of structural proteins of virus-like particles, and baculovirus bacmid was modified to increase production time. In addition, an easy purification system was constructed to efficiently purify VLP, and finally the construction of BES optimized for VLP production was completed.

미세방울 디지털 PCR(Droplet digital PCR, ddPCR) 법은 정량 PCR 법과 같이 형광물질을 사용하여 정량분석이 가능하다는 점은 유사하지만, PCR 반응액을 만든 후 이 반응액을 바로 PCR에 사용하지 않고 수 만 개의 동일한 크기의 미세방울(droplet)로 구획(partitioning)한 후에 PCR 반응을 수행하는 점이 기존의 PCR 법과 가장 큰 차이라고 볼 수 있다. 이렇게 구획된 나노리터 크기의 미세방울 중에서 형광신호가 검출되는 미세방울과 검출되지 않은 미세방울의 수를 계수하여 프아송 분포(Poisson distribution) 계산에 적용하면 표준검량선 없이 목적 유전자의 절대정량 분석이 가능하다. 곤충이 매개하는 질병 바이러스의 경우 소량의 바이러스 감염 여부를 확인하기 위해서 다수의 곤충 유전자를 확인해야 하는데, 이와 같이 미세방울 형태의 ddPCR을 이용하면 기존 Real-time PCR 법에 비해 극소량의 목적 유전자를 높은 민감도로 검출할 수 있으며, PCR 저해요소(inhibitor)에도 큰 영향을 받지 않는다는 장점이 있다. 또한 미세방울 방식의 디지털 PCR을 이용하면 다중 PCR 분석이 가능하여 1개의 시료에서 다양한 질병매개 바이러스를 검출할 수 있다.

진딧물은 감자에 다양한 바이러스병을 옮기는 대표적인 매개충이다. 따라서 감자밭으로 날아오는 진딧물을 방제하는 것은 매우 중요하다. 비래진딧물의 종류, 비래시기와 비래량, 바이러스 보독률 및 진딧물의 겨울철 월동기주를 구명하기 위하여 강원도 고랭지의 씨감자 생산지역 3곳 을 중심으로 진딧물 비래양상을 조사하였고, 비래초기부터 6월 하순까지 날아오는 진딧물의 잎말림바이러스(PLRV) 보독 여부를 조사하였다. 또한 진딧물의 이동방향을 따라 57종의 수목류 가지와 수피를 채집하여 월동중인 진딧물 알의 존재 여부를 조사하였다. 강릉 왕산 지역과 홍천 내 면 지역의 여름기주 비래최성기는 모두 6월 중순이었고 평창 횡계 지역은 5월 하순으로 나타났다. 겨울기주로 날아가는 비래최성기는 3개 지역 모두 10월 상순이었다. 비래진딧물의 2.8%가 PLRV를 갖고 있었으며, 진딧물 종류별 보독률은 복숭아혹진딧물 15.4%, 감자수염진딧물 9.1% 였다. 시기별로는 5월 하순부터 PLRV 보독 진딧물이 비래하였고 6월 중순에 비래한 진딧물의 바이러스 보독률이 10.4%로 가장 높게 나타났다. 고로쇠나무(Acer pictum subsp. mono (Maxim.)), 당단풍나무(Acer pseudosieboldianum (Pax)) 등 17종의 수목류에서 진딧물알이 월동하였다. 월동 진딧물은 복숭아혹진딧물(Myzus persicae), 홉사마귀진딧물(Phorodon humuli) 등 14종으로 동정되었다. 특히 자작나무진딧물(Betula platyphylla var. japonica), 단풍알락진딧물(Yamatocallis hirayamae) 등은 아직까지 감자에서 바이러스를 옮기는지 여부는 알려진 바 없지만, 넓 은 지역에 걸쳐 분포하는 기주식물에서 월동하기 때문에 앞으로 바이러스 매개 능력에 대한 연구가 필요하다.