• Title/Summary/Keyword: Virus cultivation

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Survey of the Routes and Incidence of Viral Infection of Tomato and Paprika Growing in Greenhouses in Cherwon Province, Korea During 2015-2017 (2015-2017 철원군 시설재배 과채류(토마토와 파프리카)의 바이러스병 발생 현황 및 감염경로 조사)

  • Kil, HyungBae;Kang, Minji;Choi, Won-Seok;Kim, Joong-Il;Phan, Mi Sa Vo;Im, JiHui;Kim, MeeKyoung;Park, Mi-Ri
    • Research in Plant Disease
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    • v.24 no.2
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    • pp.145-152
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    • 2018
  • During 2015-2017, we surveyed the incidence of viral infections of tomato and paprika growing in greenhouses in Cherwon province, Korea. In 2015 and 2016, we collected leaves and fruits from tomato and paprika plants growing in greenhouses. We detected viruses in the samples collected using specific primer sets for Broad bean wilt virus 2 (BBWV2), Cucumber mosaic virus (CMV), Pepper mild mottle virus (PMMoV), Pepper mottle mosaic virus (PepMoV), and Tomato spotted wilt virus (TSWV). We detected PMMoV, CMV, and TSWV in the samples, and CMV and TSWV were the most prevalent. For the prevention of future viral diseases, we then surveyed the routes of infection by these viruses in tomato and paprika plants growing in greenhouses in Cherwon province in 2017. Leaf and fruit samples were collected from seedlings and crops two and four months after transplanting into greenhouses. As a result, we found that TSWV was transferred from seedlings to plants, and outbreaks of the virus occurred at the early stage of cultivation. On the other hand, we found that CMV was a virus indigenous to the soil of some towns in Cherwon province, and thus outbreaks of this virus occurred at the middle stage of cultivation.

The Incidence of Virus Diseases in Rehmannia glutinosa in Korea (국내 지황에 발생하는 바이러스병 발생 현황)

  • Kwon, Sun-Jung;Yoon, Ju-Yeon;Cho, In-Sook;Choi, Gug-Seoun
    • Research in Plant Disease
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    • v.25 no.1
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    • pp.38-42
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    • 2019
  • While rehmannia (Rehmannia glutinosa Libosch) was identified as a host of at least five viruses, including Rehmannia mosaic virus (ReMV), Youcai mosaic virus (YoMV), Broad bean wilt virus 2 (BBWV2), Plantago asiatica mosaic virus (PlAMV), and Rehmannia virus 1 (ReV1), viral incidence surveys have not been performed yet in rehmannia fields in Korea. In this study, we performed field surveys during 2017-2018 to investigate the incidence of 5 major viruses in rehmannia. A total of 145 symptomatic samples were collected from the rehmannia fields in major cultivation areas of Korea. Molecular diagnosis assays showed that all the collected leaf samples were infected with more than two viruses. Particularly, two species of Tobamovirus, ReMV and YoMV, were detected in all the samples. In addition, our analysis showed that the root stocks of 4 rehmannia cultivars were infected with at least two viruses. Since rehmannia is propagated by vegetative propagation, it is highly important to produce virus-free root stocks of rehmannia to control virus diseases in rehmannia.

Monitoring of Nervous Necrosis Virus (NNV) in the Broodstock of Seven Band Grouper Hyporthodus septemfasciatus (신경괴사증바이러스(Nervous Necrosis Virus, NNV) 모니터링을 통한 무감염 능성어(Hyporthodus septemfasciatus)친어의 선발)

  • Kim, Si-Woo;Kim, Wi-Sik;Seo, Han-Gill;Kim, Kyong Min;Oh, Myung-Joo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.5
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    • pp.527-533
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    • 2017
  • We investigated the infection of nervous necrosis virus (NNV) in seven band grouper Hyporthodus septemfasciatus broodstocks, which have been reared in aquaculture farms in South Korea during 2012-2014. To investigate the prevalence of NNV within the broodstock, egg, sperm, and blood were sampled in the spawning season. The egg and sperm samples were subjected to a nested reverse transcription (RT) polymerase chain reaction (PCR) assay to detect NNV and were inoculated on SSN-1 cells to culture the virus. Blood samples were used to detect antibodies against NNV using enzyme linked immunosorbent assay (ELSIA). Positive values from ELISA were found in 39 of 162 samples (24%) in 2012, and 13 of 28 samples (46%) in 2014. Additionally, 4 of 34 broodstocks (11%) investigated in 2013-2014 were determined to be carriers from the nested RT-PCR and in vitro cultivation. The broodstocks in which antibodies against NNV were detected by ELISA, or in which NNV was detected by the nested RT-PCR assay, posed a risk of vertical transmission of NNV. Therefore, it is necessary to select virus-free broodstocks in seed production to reduce the possibility of the vertical transmission of NNV.

Detection of Barley yellow mosaic virus from Soil Using Nested PCR (Nested PCR 기법을 이용한 토양으로부터 Barley yellow mosaic virus 검출)

  • Lee, Joong-Hwan;Son, Chang-Gi;Kwon, Joong-Bae;Nam, Hyo-Hun;Kim, Yeong-Tae;Lee, Bong-Choon;Shin, Dong-Bum
    • Research in Plant Disease
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    • v.23 no.1
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    • pp.65-68
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    • 2017
  • Barley yellow mosaic virus (BaYMV), which is transmitted by the root-inhabiting protist Polymyxa graminis, causes a soil-borne disease. In this study, we detected BaYMV from soil using two-step nested polymerase chain reaction (PCR). Specific primers based on a coat protein region of BaYMV segment RNA1 were used in the first round of amplification. Based on the sequenced amplicon, an inner primer was designed for the second round of amplification. A PCR product of 372 bp exhibited 98%-100% nucleotide sequence identity with the coat protein region of BaYMV segment RNA1. In this study, we propose an easy method for the detection of BaYMV from soil, may considerably assist in accurate fungus-transmitted virus diagnosis and subsequent disease forecasting. This is the first report on the detection of BaYMV from soil.

De novo genome assembly and single nucleotide variations for Soybean yellow common mosaic virus using soybean flower bud transcriptome data

  • Jo, Yeonhwa;Choi, Hoseong;Kim, Sang-Min;Lee, Bong Choon;Cho, Won Kyong
    • Journal of Applied Biological Chemistry
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    • v.63 no.3
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    • pp.189-195
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    • 2020
  • The soybean (Glycine max L.), also known as the soya bean, is an economically important legume species. Pathogens are always major threats for soybean cultivation. Several pathogens negatively affect soybean production. The soybean is also known as a susceptible host to many viruses. Recently, we carried out systematic analyses to identify viruses infecting soybeans using soybean transcriptome data. Our screening results showed that only few soybean transcriptomes contained virus-associated sequences. In this study, we further carried out bioinformatics analyses using a soybean flower bud transcriptome for virus identification, genome assembly, and single nucleotide variations (SNVs). We assembled the genome of Soybean yellow common mosaic virus (SYCMV) isolate China and revealed two SNVs. Phylogenetic analyses using three viral proteins suggested that SYCMV isolate China is closely related to SYCMV isolates from South Korea. Furthermore, we found that replication and mutation of SYCMV is relatively low, which might be associated with flower bud tissue. The most interesting finding was that SYCMV was not detected in the cytoplasmic male sterility (CMS) line derived from the non-CMS line that was severely infected by SYCMV. In summary, in silico analyses identified SYCMV from the soybean flower bud transcriptome, and a nearly complete genome of SYCMV was successfully assembled. Our results suggest that the low level of virus replication and mutation for SYCMV might be associated with plant tissues. Moreover, we provide the first evidence that male sterility might be used to eliminate viruses in crop plants.

Cultivation and Electron Microscopy of Bovine Leukemia Virus from Peripheral Blood Lymphocytes of Holstein-Friesian Dairy Cattle (젖소 말초혈액 림프구로부터 소백혈병 바이러스 배양 및 전자현미경적 관찰)

  • Yoon, Soon-Seek;Park, Jung-Won;Byun, Jae-Won;Kang, Mun-Il;Yoo, Han-Sang;Han, Hong-Ryul
    • Applied Microscopy
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    • v.35 no.1
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    • pp.23-30
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    • 2005
  • Many studies have been performed on the bovine leukemia virus (BLV) since bovine leukosis had been reported in 1968 in Korea. However, there was no report on the ultrastructural examination of BLV. An attempt to detect C-type viral particles in the cultured peripheral blood lymphocytes of Holstein-Friesian dairy cattle, was made to determine whether in vitro viral expression might be used as a reliable method to identify the cow which is likely to transmit BLV. In transmission electron microscopic (TEM) examination, the virus particles were found predominantly outside of the lymphocytes even though a few particles were also observed within the membrane bound cytoplasmic vacuoles. All of them were C-type particles consisting of a central, electron-dense core separated by a clear area from a limiting envelope with a unit membrane structure. Virus particles were easily detected in the lymphocyte which was cultured with medium supplemented with either T-lymphocyte mitogen (conconavalin A) or B-lymphocyte mitogen (lipopolysaccharide). Identical viral particles, although fewer, were also consistently present in the lymphocytes cultured with medium which was containing foetal bovine serum (FBS) only and which was containing neither FBS or mitogen. By contrast, no virus particle was detected in extensive examination of lymphocytes before culture. In conclusion, the BLV cultivation and detection methods established in this study could be used as a tool to identify and eliminate the cattle which can transmit the BLV.

Hepatitis C Virus Core Protein Sensitizes Cells to Apoptosis Induced by Anti-Cancer Drug

  • Kang, Mun-Il;Mong Cho;Kim, Sun-Hee;Kang, Chi-Dug;Kim, Dog-Wan
    • Journal of Microbiology
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    • v.37 no.2
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    • pp.90-96
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    • 1999
  • The core protein of the hepatitis C virus (HCV) is a multifunctional protein. The HCV core protein was reported to regulate cellular gene expression and transform primary rat embryo fibroblast cells. However, the role of the core protein in the pathogenesis of HCV-associated liver diseases is not well understood. To investigate the functional role of the core protein in cytophathogenicity, we have constructed stable expression systems of full length or truncated HCV core protein lacking the C-terminal hyderophobic domains and established HepG2 cell clones constitutively expressing the core protein. The full length core protein was localized in the cytoplasm and the C-terminal truncated core protein was localized in the nucleus. HepG2 cells expressing nuclear, truncated core protein showed elevated cell death during cultivation compared to untransfected cells and full length core-expressing cells. In the treatment with bleomycin, both cell clones expressing full length or truncated core protein appeared to be more sensitive to blemoycin than the parental HepG2 cells. These results suggest that the core protein may play a role in HCV pathogenesis promoting apoptotic cell death of infected cells.

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Viral Metatranscriptomic Analysis to Reveal the Diversity of Viruses Infecting Satsuma Mandarin (Citrus unshiu) in Korea

  • Hae-Jun Kim;Se-Ryung Choi;In-Sook Cho;Rae-Dong Jeong
    • The Plant Pathology Journal
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    • v.40 no.2
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    • pp.115-124
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    • 2024
  • Citrus cultivation plays a pivotal role, making a significant contribution to global fruit production and dietary consumption. Accurate identification of viral pathogens is imperative for the effective management of plant viral disease in citrus crops. High-throughput sequencing serves as an alternative approach, enabling comprehensive pathogen identification on a large scale without requiring pre-existing information. In this study, we employed HTS to investigate viral pathogens infecting citrus in three different regions of South Korea: Jejudo (Jeju), Wando-gun (Wando), and Dangjin-si (Dangjin). The results unveiled diverse viruses and viroids that exhibited regional variations. Notably, alongside the identification of well-known citrus viruses such as satsuma dwarf virus, citrus tatter leaf virus, and citrus leaf blotch virus (CLBV), this study also uncovered several viruses and viroids previously unreported in Korean citrus. Phylogenetic analysis revealed that majority of identified viruses exhibited the closest affilations with isolates from China or Japan. However, CLBV and citrus viroid-I-LSS displayed diverse phylogenetic positions, reflecting their regional origins. This study advances our understanding of citrus virome diversity and regional dynamics through HTS, emphasizing its potential in unraveling intricate viral pathogens in agriculture. Consequently, it significantly contributes to disease management strategies, ensuring the resilience of the citrus industry.

Growth Characteristics of an Attenuated Japanese Encephalitis Virus in a Monkey Kidney Cell (Vero) (베로 세포에 적응된 약독화 일본뇌염바이러스의 성장 특성)

  • 홍선표;정용주;문상범;신영철;이성희;김수옥
    • KSBB Journal
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    • v.13 no.3
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    • pp.231-237
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    • 1998
  • An attenuated Japanese encephalitis virus (JEV) clone SA-14-14-2(Vero) was obtained through successive adaptation of a primary cell adapted strain, SA-14-14-2(PDK) to Vero cell, a continuous line of monkey kidney cells. The virus titer reached above the 107 plaque forming unit (pfu) per mL of culture supernatant with 3 passages in Vero cells and was maintained close to this level in the further passages. The optimum temperature for the virus growth was $35^{\circ}C$. Growth pattern of the virus indicated that optimum time for the virus harvest is 4 days post infection and the virus accomplished rapid initial propagation even in medium containing no serum supplement. The roller bottle (RB) system and the spinner flask (SF) system using micro-carrier (Cytodex-1) for the JEV cultivation were explored. When RB, SF, and T-flask culture system were compared, there was no significant difference in virus yield. Furthermore, the results indicated that virus could be harvested multiple times from 3 days to 9 days post infection; neither severe cytopathic effect (CPE) in the infected cells nor the decrease in the titer was observed on duration of 9 days.

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Survey of Viral Diseases Occurrence on Major Crops in 2007 (2007년 우리나라 주요 작물 바이러스병 발생 상황)

  • Kim, Jeong-Soo;Lee, Su-Heon;Choi, Hong-Soo;Choi, Guk-Sun;Cho, Jeom-Deog;Chung, Bong-Nam
    • Research in Plant Disease
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    • v.14 no.1
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    • pp.1-9
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    • 2008
  • The severe damage induced by the important viruses of Rice stripe virus (RSV), Cucumber green mottle mosaic virus (CGMMV), Melon necrotic spot virus (MNSV), Tomato spotted wilt virus (TSWV) and Tomato bushy stunt virus (TBSV) was described on major crops in Korea. In 2007, the plot incidence rate of RSV was 100% on the precocious rice cultivars at the Western coastal provinces of Gyeonggido, Chungcheongnamdo, Jellabugdo and Jellanamdo, and Jejudo. RSV occurred in 2,441 ha with incidence rate of 70% over at 5 areas of Seocheon, Seosan, Boryung, Hongsung and Buyou in Chungcheongnamdo. At 4 areas of Buan, Gimje, Gunsan and Gochang in Jellabukdo, RSV occurred in 2.016 ha. CGMMV occurred on watermelon in 4.6 ha at Cheongyang area, and its outbreak was also 890 ha on oriental melon for 120 farmers with the incidence area of 23% against total cultivation areas of Seongju. MNSV was recorded firstly on watermelon in 2006 at Andong and it spread to 3 areas of Hapcheon, Gochang and Yanggu. TSWV occurred firstly at Danggin in Chungcheongnamdo in 2005. TSWV in 2006 spread to 6 areas; Taian, Hongsung and Seosan in Chungcheongnando, Namwon in Jellabukdo, and Sunchon and Kwangju in Jellanamdo. In 2007, TSWV covered 17 areas of western and southern parts; the 5 area including Taian in Chungcheongnamdo, Kwangju in Jellanamdo, Bucheon in Gyunggido, and so forth. TBSV was described firstly on table tomato at Sacheon in Kyungsangnamdo in 2004. TBSV occurred on cherry tomato at Chungju in 2006 and on table tomato at Busan area.