• Title/Summary/Keyword: virus free

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Establishment of Mass Propagation System of Virus-Free Sweetpotato Plants and Conservation

  • Lee, Joon-Seol;Chung, Mi-Nam;Jeong, Byeong-Choon;Ahn, Young-Sup;Kim, Hag-Sin;Park, Jong-Suk;Bang, Jin-Ki
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.52 no.2
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    • pp.220-227
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    • 2007
  • Sweetpotato fields in Korea are highly infected with virus and virus like diseases that greatly diminish both yield and quality as indicated by field observations and laboratory tests. In order to solve this problem, there is an urgent need to produce and mass propagate virus-free planting materials for distribution to the farmers. These experiments were conducted, firstly, to determine the most appropriate culture media, nutrient solution, and cutting intervals to maintain growth and vigor of tissue cultured plantleta as mother plants for propagation in insect-proof greenhouse. And as a labor saving method, the production efficiency of plug trays for rapid propagation of stem cuttings as a source of planting materials was likewise evaluated. Results showed that plants grown in medium B supplied with 0.5 and 1.0 strength of MS nutrients had high growth rate, and 20-day cutting interval was the best. 72-plug tray was better than 128-plug. Secondly, it was to develop a technique for the production of first-generation seed roots using hydroponics cultivation system. The yield of virus-free plants propagated in the non-insect proof and open-field cultivation was 2,402 kg/10a, 6% higher than those in the insect-proof cultivation, and the rate of virus re-infection was 18% higher compared to 3.3% with insect-proof cultivation. Lastly, it was to investigate the growth performance of virus free plants in farmers' field. Differences were existed in the yield depending on the variety used, but virus free plants showed an increase of $6{\sim}24%$ over virus infected plants.

Growth Characteristics and Yield of Sweet Potato Cultivars between Virus-free and Farmer's Slips in Late Season Cultivation (고구마 바이러스 무병묘와 농가묘의 만기재배에서 품종 간 생육 및 수량특성)

  • Yoo, Kyoung-Ran;Lee, Seung-Yeob
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.58 no.1
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    • pp.43-49
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    • 2013
  • This work was conducted to obtain some information about stable production of high quality seed-tubers in the late season cultivation of virus-free sweet potato [Ipomoea batatas (L.) Lam.]. Growth characteristics and storage root yield between virus-free and farmer's slips in 9 cultivars were investigated using black-film vinyl mulching cultivation with $75{\times}25cm$ planting density on July 10. At 30 days after planting, vine length, vine diameter, number of node, and number of branch in virus-free slips were significantly increased than those in farmer's slips. The vine growth was significantly different among cultivars, and vine elongation was excellent in 'Kogeonmi', 'Shincheonmi', 'Shinhwangmi', 'Shinyulmi', and 'Yeonhwangmi' compared to the other cultivars. At 110 days after planting, vine length, vine diameter, number of node, number of branch, and fresh weight were significantly different among cultivars, but no significant differences between virus-free and farmer's slips were seen except number of node. Total yield in virus-free slips was increased by 12-49% among cultivars than that in farmer's slips. The mean yields between virus-free and farmer's slips were 1,625 kg/10a and 1,230 kg/10a, respectively, and it was significantly different between virus-free and farmer's slips. Percentage of marketable storage root in virus-free slips was 65.6%, and it was significantly higher than 57.8% in farmer's slips. Marketable yields ($40g{\leq}$) between virus-free and farmer's slips were 1,067 kg/10a and 710 kg/10a, respectively. Marketable yield in 'Shincheonmi', 'Shinyulmi' and 'Shinzami' was more than 1,300 kg/10a, and these cultivars showed to be highly adaptable for the late-season cultivation among 9 tested cultivars.

Effect of Virus Free Stocks of Sweetpotato Cultivated at Different Regions (지역을 달리하여 재배한 고구마 바이러스 무병묘 효과)

  • Yang, Jung-Wook;Kim, Jea-Myung;Lee, Hyeong-Un;Han, Seon-Kyeong;Lee, Joon-Seol;Nam, Sang-Sik;Chung, Mi-Nam;Song, Yeon Sang;Ahn, Seung-Hyun;Choi, In-Hu
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.60 no.1
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    • pp.54-62
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    • 2015
  • To analysis of virus free sweetpotato effect, 5 virus free sweetpotato and virus normal sweetpotato varieties were planted in 5 different regions at 2010 year. The average yields of virus free sweetpotato are showed different results according to regions. Sinjami which cultivated at Iksan were increased maximum 68% compare to normal. However, Sinjami which cultivated in Hamyang were decreased yield 11% compare to normal. Analysis of tuber formation ratio of Sinjami, Yenhwangmi, Singeonmi which cultivated in Nonsan were decreased tuber number compare to normal. However, 3 varieties were all increased on Average storage root weight and yield of marketable storage root. In the results of analysis of marketable storage root according to cultivated regions and varieties, all varieties except Sinjami which cultivated in Hamyang were increased yield. Also, quality of virus free sweetpotato were enhanced 7 to 9 compare to virus infected sweet potato which showing average 3. Contents of starch between virus free and virus infected sweetpotato were not affected by virus. Virus free sweetpotato were more increased starch products according to increased total production yield. Also, $Brix^{\circ}$(%) was not showing difference between virus free and virus infected sweetpotatoes. In this experiment, Virus free sweetpotato are enhanced production yields and quality. Therefore, we suggested that virus free sweetpotato is one of the methods to reduce damage by sweetpotato virus.

Production of Virus-Free Stocks from Citrus Plant by the Shoot-Tip Grafting and Heat Treatment (열처리와 Shoot-Tip Grafting에 의한 감귤 바이러스 무독묘 생산)

  • Kim Daehyun;Shim Hyekyung;Kwon Hyeogmo;Hyun Jaewook;Kim Kwangsik;Lee Jinkyung;Lee Sukchan
    • Journal of Plant Biotechnology
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    • v.32 no.1
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    • pp.45-50
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    • 2005
  • Virus-free stocks was produced by the combination of the heat treatment of virus infected plant and shoot-tip grafting (STS). To produce virus-free stocks, the plants infected with citrus viruses were used for virus-free stock production using the modified method of STG in thermotherapy at $40^{\circ}C$ for 16 hours in the light, and at $30^{\circ}C$ for 8 hours of darkness for 4 weeks. Trifoliate orange (P. trifoliata) were used as rootstock seedling for STG. Percentages of virus-free stocks against citrus tristeza virus (CTV), satsuma dwarf virus (SDV) and citrus tatter leaf virus (CTLV) were $75.7\%,\;100.0,\%\;82.6\%$ respectively. Shoot tip size for successful STG were as small as possible. Less than $0.3\;\cal{mm}$ of shoot tips gave the hight efficiency of virus free plants but survival rates were low. And, survival rate after shoot-tip culture was analyzed and the rates were dependant on the cultivars; Yuzu cultivar showed the hight survival rate ($74.6\%$) and early satsuma mandarin (Iwasagi) was $13.3\%$ as the lowest cultivar. But citrus trees were not succeed to grown, turned brown, and died.

Study on Persistent Infection of Japanese Encephalitis Virus Beijing-l Strain in Serum-free Sf9 Cell Cultures

  • Kim, Hun;Lee, Su-Jeen;Park, Jin-Yong;Park, Yong-Wook;Kim, Hyun-Sung;Kang, Heui-Yun;Hur, Byung-Ki;Ryu, Yeon-Woo;Han, Sang-In
    • Journal of Microbiology
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    • v.42 no.1
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    • pp.25-31
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    • 2004
  • Sf9 cells have obvious advantages for the conventional production technology of vaccine. They are useful tools for high concentration and large-scale cultures. Sf9 cells were grown to maximal concentration, 8${\times}$l0$\^$6/ cells/$m\ell$ in a 500$m\ell$ spinner flask, with a doubling time at the exponentially growing phase of 24.5 hours, using serum-free media. To explore the ability of Sf9 cells to be infected by the Japanese encephalitis (JE) virus Beijing-l strain, Sf9 cells were infected with the virus. By 4-5 days post-infection, 10-15 % of the Sf9 cells showed cytopathic effect (CPE), from granularity to the formation of syncytia and multinucleated giant cells continuously observed over a period of 35 days. Positive fluorescent reactions were detected in 30-40% of cells infected with the JE virus Beijing-l strain, and the uninfected Sf9 cells were completely negative. Virus particles, propagated in Sf9 and Vero cells, were concentrated by sedimentation on 40% trehalose cushions by ultracentrifugation, and showed identical patterns of viral morphogenesis. Complete virus particles, 40 to 50 nm in diameter, were observed, and JE virus envelope (E) proteins, at 53 kDa, were found in the western blot analysis to the anti-JE virus E protein monoclonal antibody and reacted as a magenta band in the same position to the glycoprotein staining. To evaluate whether the infectious virus was produced in Sf9 cells inoculated with the JE virus Beijing-l stain, Sf9 cells were inoculated with the virus, and sample harvested every 5 days. The titers of the JE virus Beijing-l strain rose from 1.0${\times}$l0$\^$5/ to 1.5${\times}$l0$\^$6/ pfu/$m\ell$. The infected Sf9 cells could be subcultured in serum-free medium, with no change in the plaque sizes formed by the JE virus Beijing-l strain in the plaque assay. It is suggested that the ability of the JE virus Beijing-l strain to infect Sf9 cells in serum-free media will provide a useful insect cell system, where the JE virus replication, cytopathogenicity and vaccine immunogen can be studied.

Effect of Virus-free Plant and Subsoiling Reversion Soil for Reduction of Injury by Continuous Cropping of Sweet Potato (고구마 연작장해 경감을 위한 바이러스 무병묘 재배와 심토반전 효과)

  • Song, Hae-Ahn;Kim, Kab-Cheol;Lee, Seung-Yeob
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.57 no.3
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    • pp.254-261
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    • 2012
  • To reduce the injury by continuous cropping of sweet potato (Ipomoea batatas (L.) Lam.), the farmer's plant and virus-free plant were cultivated with the density of $70{\times}25cm$ (June 10, 2011) in continuous cropping soil (CCS) and subsoiling reversion soil (SRS). Fertilizer was applied at the rates of 55-63-156 $kg\;ha^{-1}$ ($N-P_2O_5-K_2O$) and 10 $ton\;ha^{-1}$ of cattle manure in CCS, and it was applied the 50% increased cattle manure compost and nitrogen in DRS. Symptoms of viral infection were revealed in the farmer's plant at 30 days after planting, but there were no symptoms in virus-free plant. The yield of virus-free plant was more increased 15% and 10.5% than that of farmer's plant in DRS and CCS, respectively. The yield of sweetpotato in SRS was more increased 8.8% and 3.2% in farmer's plant and virus-free plant compared to CCS, respectively. In DRS, the rate of marketable tuber of virus-free plant was increased by 80% compared to the farmer's plant (60.1%). The virus-free plant was produced the tuber with more brilliant peel color and well-formed shape compared to the farmer's plant. The increased yield of virus-free plant and in SRS soil condition showed a positive relationship (p=0.05) with the number of leaf per plant at 30 days and the number of branch per plant at 120 days after planting. The results showed that the early growth after planting was very important for the development of storage root. Therefore, the deep-subsoil reversion and cultivation of virus-free plant could be reduced the injury by continuous cropping of sweet potato, and increased farm income.

Effects of Thermotherapy and Shoot Apical Meristem Culture, Antiviral Compounds for GLRaV-3 Elimination in Grapevines (열처리와 생장점 배양 및 항바이러스제 처리에 의한 포도 GLRaV-3의 무독화효과)

  • Kim, Hyun-Ran;Chung, Jae-Dong;Park, Jin-Woo;Choi, Yong-Mun;Yiem, Myoung-Soon
    • Journal of Plant Biotechnology
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    • v.30 no.2
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    • pp.155-160
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    • 2003
  • Grapevine leafroll-associated virus 3(GLRaV-3) is one of the most severe pathogens for viral diseases found in Korea. This study was conducted to establish the virus-free stock production system for the virus disease control. The effects of thermotherapy, merestem culture and chemotheratpy to eliminate the GLRaV-3 in gratevines were tested. Thermotherapy at 37$\pm$2$^{\circ}C$ for 6∼8 weeks combined with 0.5∼1.0mm size of meristem culture method was the most effective for virus elimination. Thermotherapy alone was not effective. In chemotheratpy, DHT and Amantadine (20, 40mg/L) treatment in medium was more effective than Ribavirin to eliminate the GLRaV-3 in grapevine. However, Ribavirin spraying to potted was not available for virus elimination. Therefore, virus-free stock production system using the thermotherapy combined with shoot apical meristem culture was the most effective in grapevine.

Detecting outliers in segmented genomes of flu virus using an alignment-free approach

  • Daoud, Mosaab
    • Genomics & Informatics
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    • v.18 no.1
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    • pp.2.1-2.11
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    • 2020
  • In this paper, we propose a new approach to detecting outliers in a set of segmented genomes of the flu virus, a data set with a heterogeneous set of sequences. The approach has the following computational phases: feature extraction, which is a mapping into feature space, alignment-free distance measure to measure the distance between any two segmented genomes, and a mapping into distance space to analyze a quantum of distance values. The approach is implemented using supervised and unsupervised learning modes. The experiments show robustness in detecting outliers of the segmented genome of the flu virus.

Study of Viral Effects of the Mycovirus (LeV) and Virus-Free Commercial Line in the Edible Mushroom Lentinula edodes

  • Kim, Jung-Mi;Song, Ha-Yeon;Yun, Suk-Hyun;Lee, Hyun-Suk;Ko, Han-Kyu;Kim, Dae-Hyuk
    • 한국균학회소식:학술대회논문집
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    • 2015.11a
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    • pp.37-37
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    • 2015
  • dsRNA was found in malformed cultures of Lentinula edodes strain FMRI0339, one of the three most popular sawdust cultivated commercial strains of shiitake, and was also found in healthy-looking fruiting bodies and actively growing mycelia. Cloning of the partial genome of the dsRNA revealed the presence of the RdRp sequence of a novel L. edodes mycovirus (LeV), and sequence comparison of the cloned amplicon showed an identical sequence to known RdRp genes of LeV found in strain HKA. The meiotic stability of dsRNA was examined by measuring the ratio of the presence of dsRNA among sexual monokaryotic progeny. More than 40% of the monokaryotic progeny still contained the dsRNA, indicating the persistence of dsRNA during sexual reproduction. Comparing the mycelia growth of monokaryotic progeny suggested that, although variations in the growth rate existed among progeny and virus infection was observed in highly actively growing progeny, there appeared to be a tendency toward a lower frequency of virus incidence in actively growing progeny. This study attempted to cure the edible mushroom L. edodes strain FMRI0339 of the L. edodes mycovirus (LeV) in order to obtain an isogenic virus-free fungal strain as well as a virus-infected strain for comparison. Mycelial fragmentation, followed by being spread on a plate with serial dilutions resulted in a virus-free colony. Viral absence was confirmed with gel electrophoresis after dsRNA-specific virus purification, Northern blot analysis, and PCR using reverse transcriptase (RT-PCR). Once cured, all of fungal cultures remained virus-free over the next two years. Interestingly, the viral titer of LeV varied depending on the culture condition. The titer from the plate culture showed at least a 20-fold higher concentration than that grown in the liquid culture. However, the reduced virus titer in the liquid culture was recovered by transferring the mycelia to a plate containing the same medium. In addition, oxygen-depleted culture conditions resulted in a significant decrease of viral concentration, but not to the extent seen in the submerged liquid culture. Although no $discernable phenotypic changes in colony morphology were observed, virus-cured strains showed significantly higher growth rates and mycelial mass than virus-infected strains. We were also explored effects of LeV on fruiting body formation and mushroom yield. The fruiting body formation yield of virus-free L. edodes was larger than virus-infected L. edodes. These results indicate that LeV infection has a deleterious effect on mycelial growth and fruiting body formation. In addition, we have been investigated host-parasite interaction between L. edodes and its mycovirus interaction to study viral mechanism by establishment of proteomics.

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Comparison of Virus Elimination Methods for Disease-free Seedlings of the Apple Dwarfing Rootstock (사과 왜성대목 무독묘 생산을 위한 바이러스 제거 방법 비교)

  • Kwon, Young Hee;Lee, Joung Kwan;Kim, Hee Kyu;Kim, Kyung Ok;Park, Jae Seong;Huh, Yoon Sun;Yoon, Yeo Joong
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.54-54
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    • 2019
  • Apple (Malus domestica) is one of the most economically important fruits in Korea. But virus infection has decreased sustainable production of apple and caused the serious problems such as yield loss and poor fruit quality. Virus or viroid infection including Apple chlorotic leaf spot virus (ACLSV), Apple stem pitting virus (ASPV), Apple stem grooving virus (ASGV), Apple mosaic virus (ApMV) and Apple scar skin viroid (ASSVd) has been also reported in Korea. In many cases, apple is infected with virus and viroid with no specific symptoms, the damage caused by the virus are unaware significantly. In our research, we tried to eliminate viruses in the rootstock for the disease-free seedlings of the apple dwarfing rootstock M.9 and M.26. The method of virus elimination was meristem culture, heat($37^{\circ}C$, 6weeks) treatment and chemistry($Ribavirin^{(R)}$) treatment. The analytical methods commonly used for the detection of virus is Enzyme-linked Immuno-Sorbent Assay(ELlSA) and Reverse Transcription-polymerase Chain Reaction(RT-PCR). RT-PCR method was more 30% sensitive than ELISA method. Efficiency of method eliminate virus appeared meristem method > heat treatment > chemistry treatment. The higher acquisition rate of disease-free seedlings is 30~40% on meristem treatment. In meristem treatment, the apple dwarfing rootstock M.9 gained infection ratio of ACLSV, ASPV and ASGV were 45%, 60% and 50% respectively. In the apple dwarfing rootstock M.26, infection ratio of ACLSV, ASPV and ASGV were 40%, 55%, 55%, respectively. Based on our results, it was found that most effective method of disease-free seedlings apple dwarfing rootstocks was by meristem treatment than heat method and chemistry treatment.

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