Seed Transmission of Tomato yellow leaf curl virus in White Soybean (Glycine max)

길의준,박정호,최홍수,김창석,이석찬 한국식물병리학회 2017 Plant Pathology Journal Vol.33 No.4

Tomato yellow leaf curl virus (TYLCV) infection of the common bean (Phaseolus vulgaris) has been reported, but soybean (Glycine max) has not previously been identified as a TYLCV host. Five cultivars of white soybean were agro-inoculated using an infectious TYLCV clone. At 30 days post-inoculation, they showed infection rates of 25% to 100%. Typical TYLCV symptoms were not observed in any inoculated plants. To examine whether TYLCV was transmitted in soybean seeds, DNA was isolated from bundles of five randomly selected seeds from TYLCV-inoculated soybean plants and amplified with a TYLCV-specific primer set. With the exception of one bundle, all bundles of seeds were verified to be TYLCV-infected. Virus dissemination was also confirmed in three of the 14 bunches. Viral replication was also identified in seeds and seedlings. This is the first report demonstrating that soybean is a TYLCV host, and that TYLCV is a seed-transmissible virus in white soybean.

Phylogenetic Characterization of Tomato chlorosis virus Population in Korea: Evidence of Reassortment between Isolates from Different Origins

이예지,길의준,곽해련,김미경,서장균,이석찬,최홍수 한국식물병리학회 2018 Plant Pathology Journal Vol.34 No.3

Tomato chlorosis virus (ToCV) is a whitefly-transmitted and phloem-limited crinivirus. In 2013, severe interveinal chlorosis and bronzing on tomato leaves, known symptoms of ToCV infection, were observed in greenhouses in Korea. To identify ToCV infection in symptomatic tomato plants, RT-PCR with ToCV-specific primers was performed on leaf samples collected from 11 tomato cultivating areas where ToCV-like symptoms were observed in 2013 and 2014. About half of samples (45.18%) were confirmed as ToCV-infected, and the complete genome of 10 different isolates were characterized. This is the first report of ToCV occurring in Korea. The phylogenetic relationship and genetic variation among ToCV isolates from Korea and other countries were also analysed. When RNA1 and RNA2 are analysed separately, ToCV isolates were clustered into three groups in phylogenetic trees, and ToCV Korean isolates were confirmed to belong to two groups, which were geographically separated. These results suggested that Korean ToCV isolates originated from two independent origins. However, the RNA1 and RNA2 sequences of the Yeonggwang isolate were confirmed to belong to different groups, which indicated that ToCV RNA1 and RNA2 originated from two different origins and were reassorted in Yeonggwang, which is the intermediate point of two geographically separated groups.

Honey Bee Viruses: An Ongoing History of Discovery

권민혁,정철의,길의준 한국양봉학회 2023 韓國養蜂學會誌 Vol.38 No.2

Numerous threats to honey bees‘ health exist, including climate change, pesticides, diseases, and pests, and their combined actions pose greater risks than a single factor. Viruses have been investigated for roughly 100 years since the first bee virus detection, Sacbrood virus from the United States in 1913. So far, seven viruses have been routinely included in honey bee health monitoring and surveillance systems in Korea, but new viruses have lately been found. However, understanding of viruses as a threat to honey bees remains insufficient. Although some of the threats to honey bee health posed by viruses and other factors have been identified, research is still in its initial step. In this paper, we have reviewed the diversity of honey bee viruses, virulence, mode of transmission, and potential molecular biological analysis approaches.

국내에서 발생하는 Apple scar skin viroid 분리주에 대한 계통분석

조강희,김인수,길의준,박서준,김세희,최인명,김대현 한국식물병리학회 2015 식물병연구 Vol.21 No.4

To identify genome sequences of Apple scar skin viroid (ASSVd) isolates in Korea, the field survey was performed from ‘Hongro’ apple orchards located in eight sites in South Korea (Bongwha, Cheongsong, Dangjin, Gimchoen, Muju, Mungyeong, Suwon, and Yeongwol). ASSVd was detected by RT-PCR and PCR fragments were cloned into cloning vector. Full-length viral genomes of eight ASSVd isolates were sequenced and compared with 21 isolates reported previously from Korea, India, China, Japan and Greece. Eight isolates in this study showed 92.2-99.7% nucleotide sequence identities with those reported previously. Phylogenetic analysis showed that seven isolates reported in this study belong to the same group distinct from other groups. 국내에서 재배되는 사과 ‘홍로’에 발생하는 Apple scar skin viroid병의 유전자 계통분석을 하기 위하여 바이로이드 증상이나타나는 8개 지역(봉화, 청송, 당진, 김천, 무주, 문경, 수원, 영월)의 농가에서 수집한 시료를 RT-PCR 방법을 이용하여 바이로이드 검정을 실시하였다. 검출된 바이로이드의 클로닝과 유전자 염기서열 분석을 통해 8종의 분리주들을 확인하였고 한국, 인도, 중국, 일본 및 그리스에서 보고된 21종의 분리주와 염기서열을 비교하였다. 8개의 분리주들의 핵산 서열은 기존에보고된 분리주들과 92.2-99.7%의 상동성을 나타냈다. 계통분석을 통해 본 연구에서 보고한 7종의 분리주들은 기존의 것들과 독립된 그룹에 속하는 것으로 나타났다.

시설재배지 고추를 가해하는 총채벌레류와 TSWV 유전자 서열 변이

김철영,최두열,강정훈,아흐메드샤비르,길의준,권기면,이관석,김용균 한국응용곤충학회 2021 한국응용곤충학회지 Vol.60 No.4

시설재배지를 대상으로 고추 정식 이후 황색 끈끈이트랩으로 총채벌레 발생을 모니터링하였다. 아울러 토마토반점위조바이러스(Tomato spotted wilt virus: TSWV)가 유발하는 고추 칼라병을 유관으로 조사하였다. 고추 정식 직후(3월 말) 낮은 밀도로 대만총채벌레(Frankliniella intonsa)가 트랩에 포획되었으며 4월 중순부터는 꽃노랑총채벌레(Frankliniella occidentalis)도 발견되었다. 이후 5월부터는 두 종이 전체 총채 벌레의 98% 이상을 차지하였고, 이 가운데 대만총채벌레가 꽃노랑총채벌레보다 다소 많은 발생 밀도를 보였다. 전체 총채벌레의 발생 피크를 보면 5월 중순에 낮은 피크를 기점으로 6-7월에 발생 최성기를 보였다. 이후 총채벌레의 발생은 급격하게 감소하였다. 포획된 꽃노랑총채벌레의 암수 비율이 일정하지 않았는데 이는 이 곤충의 특이적 단성생식 가능성으로 이에 대한 실험적 증거를 제공하였다. 지역간 꽃노랑총채벌레의 유전적 거리를 COI 서열로 비교한 결과 원거리에서 채집한 꽃노랑총채벌레 집단과는 차이를 보였지만 안동지역 내에서 발생한 꽃노랑총채벌레는 COI 서열에서 높은 유사성을 보였다. 이들 주요 두 종의 총채벌레가 전파할 것으로 추정되는 고추 칼라병이 일부 시설재배지를 중심으로 발견되었으며 항혈청 및 분자진단을 통해 확인되었다. 더불어 감염 고추에서 채집된 꽃노랑총채벌레에서도 분자진단을 통해 TSWV를 검출하였다. 감 염 TSWV의 게놈 구조를 비교하기 위해 기능성 단백질을 갖는 NSS, N, NSM의 유전자 서열을 분석하였다. 서로 다른 지역별 이들 유전자는 다수의 점돌연변이가 존재하였고 이들 가운데는 아미노산 서열 차이를 초래하는 오류 돌연변이를 포함하였다. 추출된 TSWV를 비보독충 꽃노랑 총채벌레에 섭식 처리한 유충과 성충 모두에서 감염으로 일어났으나, 유충에게서만 바이러스 증식이 일어나는 것을 확인하였다. Thrips infesting hot peppers were monitored in greenhouses using yellow sticky traps. In addition, the hot peppers infected with tomato spotted wilt virus (TSWV) were observed during the monitoring period. The flower thrips (Frankliniella intonsa) were initially trapped at a low density just after transplanting seedlings of hot peppers at late March. The western flower thrips (Frankliniella occidentalis) were trapped after mid April. These two thrips represented more than 98% of the total thrips attracted to the traps after May, in which F. intonsa showed higher occurrence frequency than F. occidentalis. The total number of thrips had two peaks at mid May with a small and short-term peak and at June-July with a large and long-term peak. The trapped thrips exhibited inconsistent sex ratios, suggesting a seasonal parthenogenesis. Different geographical populations were varied in cytochrome oxidase I sequences, in which local populations in Andong shared a high sequence similarity. TSWV-infected hot peppers, which might be mediated by these two thrips species, were observed and confirmed by an immunoassay kit and a molecular diagnosis using RT-PCR. In addition, the TSWV was detected in F. occidentalis collected from the infected hot peppers. Three open reading frames (NSS, N, and NSM) of the isolated TSWV genomes were sequenced and showed multiple point mutations containing missense mutations among geographical variants. When the isolated TSWV was fed to nonvirulent thrips of F. occidentalis, the virus was detected in both larvae and adults. However, the viral replication occurred in larvae, but not in adults.

Therapeutic Strategy for the Prevention of Pseudorabies Virus Infection in C57BL/6 Mice by 3D8 scFv with Intrinsic Nuclease Activity

이건섭,조승찬,Phuong Mai Hoang,김동준,이용준,길의준,변성준,김택균,김대현,김성한,이석찬 한국분자세포생물학회 2015 Molecules and cells Vol.38 No.9

3D8 single chain variable fragment (scFv) is a recombinant monoclonal antibody with nuclease activity that was originally isolated from autoimmune-prone MRL mice. In a previous study, we analyzed the nuclease activity of 3D8 scFv and determined that a HeLa cell line expressing 3D8 scFv conferred resistance to herpes simplex virus type 1 (HSV- 1) and pseudorabies virus (PRV). In this study, we demonstrate that 3D8 scFv could be delivered to target tissues and cells where it exerted a therapeutic effect against PRV. PRV was inoculated via intramuscular injection, and 3D8 scFv was injected intraperitoneally. The observed therapeutic effect of 3D8 scFv against PRV was also supported by results from quantitative reverse transcription polymerase chain reaction, southern hybridization, and immunohistochemical assays. Intraperitoneal injection of 5 and 10 μg 3D8 scFv resulted in no detectable toxicity. The survival rate in C57BL/6 mice was 9% after intramuscular injection of 10 LD50 PRV. In contrast, the 3D8 scFv-injected C57BL/6 mice showed survival rates of 57% (5 μg) and 47% (10 μg). The results indicate that 3D8 scFv could be utilized as an effective antiviral agent in several animal models.

시설 고추재배지 총채벌레 연중 발생 및 주요 총채벌레의 차등 해충성

김철영,최두열,이동현,Falguni Khan,권기면,함은혜,박정준,길의준,김용균 한국응용곤충학회 2022 한국응용곤충학회지 Vol.61 No.2

Andong is a place to culture the great amount of hot pepper (Capsicum annuum L.) in Korea. This study reports a yearly occurrence (March 31~October 25, 2021) of thrips infesting the hot pepper in Andong. Thrips caught to yellow sticky traps were diagnosed by morphological characters and showed two dominant species: Frankliniella occidentalis and F. intonsa. During this period, a total of 107,874 thrips were caught and included F. occidentalis at about 82%, F. intonsa at about 17%, and the other thrips at about 0.3%. There were two main peaks at May~June and at September~October, respectively, in which the total number of thrips was higher in the second peak and most were F. occidentalis. Interestinly, a low level of thrips occurred during July~August was observed and explained by their susceptibility to high temperatures. A laboratory experiment by exposing thrips to high temperatures showed that thrips were susceptible to temperatures higher than 35°C and not tolerant to 45°C for 1 h. Indeed, high temperatures higher than 45°C were recorded in the greenhouses in Andong during July~August. F. occidentalis was more tolerant to the high temperatures than F. intonsa. On the other hand, the thrips showed the highest occurrence peak at July~August in hot pepper-culturing greenhouse in Kangwon, where the average temperatures were mostly lower than those of Andong and no high temperatures higher than 45°C were recorded during July~August. A viral disease caused by tomato spotted wilt virus (TSWV) was observed in the hot peppers cultured in Andong greenhouses. Multiplex PCR was used to detect the virus along with identification of thrips. With a high record of about 30%, the virus-infected thrips were detected during all the monitoring period. The virulent thrips were identified to be only F. intonsa. These results suggest that F. occidentalis gives a direct damage especially during harvesting period with their high populations while F. intonsa gives indirect damage by transmitting TSWV. 전국 최대 규모의 고추(Capsicum annuum L.) 재배지는 안동이다. 이 지역을 중심으로 본 연구는 시설재배지 고추를 가해하는 총채벌레의 연중(2021년 3월31일~10월25일) 발생을 보고한다. 황색트랩을 이용하여 포획한 총채벌레를 유관으로 동정한 결과 꽃노랑총채벌레(Frankliniella occidentalis)와 대만총채벌레(F. intonsa)가 우점종으로 나타났다. 전체 포획충은 107,873마리였으며 이 가운데 꽃노랑총채벌레가 약 82%, 대만총채벌레가 약 17% 그리고 기타 총채벌레가 약 0.3% 를 차지하였다. 연중 전체적으로 2회 총채벌레 발생 피크를 보였다. 첫 번째 발생피크는 5-6월에 나타났고, 두 번째 발생 피크는 9월 이후에 일어났다. 발생규모는 첫번째보다는 두 번째 발생 피크에서 높았으며 대부분은 꽃노랑총채벌레가 차지하였다. 흥미롭게 7-8월에 이들 총채벌레의 발생이 매우 낮았는데 이들 주요 총채벌레가 고온에 대한 높은 감수성으로 기인되었다. 실내 고온 노출실험은 총채벌레들이 35°C 이상에서 고온 감수성을 보여 45°C에서는 1시간 노출을 견디지 못하였다. 실제로 안동지역 7-8월 시설재배지는 최고온도가 45°C 이상을 기록하였다. 이 가운데 꽃노랑총채벌레가 대만총채벌레에 비해 낮은 고온 감수성을 보였다. 반면에 안동에 비해 기온이 낮은 강원지역의 시설 고추 재배지에서는 7-8월에 오히려 최대 총채벌레 발생피크를 나타냈으며, 이 시기 최고온도는 45°C 이하를 기록하였다. 조사한 안동지역 고추에서는 TSWV (tomato spotted wilt virus)에 의해 발병되는 바이러스병이 발생하였다. 다중 PCR 검정법으로 이 바이러스 보독 유무 및 대상 총채벌레를 동시에 분석하였다. 이 결과 바이러스 보독충 비율은 최대 30%를 기록하며 연중 지속적으로 검출되었는데 이들 모두는 대만총채벌레로 판명되었다. 이상의 결과는 시설 고추재배지에서 꽃노랑총채벌레는 고추 수확기에 최성기를 이루며 고추에 직접피해를 줄 가능성이 높으며, 대만총채벌레는 높은 바이러스 보독율로 고추의 간접피해를 유발할 위험성이 있다는 것을 각각 제시하고 있다.

제주도 감귤원 및 키위 재배온실 발생 잡초 분포조사

정영재(Chung Young Jae),조명숙(Jo Myeong Sook),길의준(Kil Eui-Jun),변희성(Byeon Hee Seong),이인용(Lee In-yong),김창석(Kim Chang Seok) 한국잡초학회 2015 한국잡초학회 별책(학술대회 초록집) Vol.35 No.2

제주도지역 발생 밭잡초 분포조사

정영재(Chung Youngjae),김용성(Kim Yong-sung),조명숙(Jo Myeong Sook),김선무(Kim Sun Moo),오정현(Oh Jeong Hyeon),김동준(Kim Dong-Jun),길의준(Kil Eui-Joon),이인용(Lee In-yong),김창석(Kim Chang Seok) 한국잡초학회 2014 한국잡초학회 별책(학술대회 초록집) Vol.34 No.2