• Journal of Microbiology
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• v.34 no.3
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• pp.263-263
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• 1996

Infection of fish cells with IHNV resulted in gradual increase in cytosolic free $Ca^{2+}$ concentration $([Ca^{2+}]_i)$ in CHSE, gradual decrease in $[Ca^{2+}]_i$ in FHM, and no significant change in RTG cells. The degree of $[Ca^{2+}]_i$ increase or decrease was dependent on the amount of infectious virus, and these $[Ca^{2+}]_i$ variations were maximal at 16 hours after virus infection (p. i.) in both cell lines. When the fish cells were infected with inactivated IHNV, evident variation in $[Ca^{2+}]_i$ was not observed. Thus, infectivity of IHNV appears to correlate with changes in $[Ca^{2+}]_i$ in virus-infected cells. These IHNV-induced $[Ca^{2+}]_i$ changes were partially blocked by cycloheximide, but not affected by cordycepin. It seems to be that virus-induced $Ca^{2+}$ variations were more related with protein synthesis than RNA synthesis. Various $Ca^{2+}$ related drugs were used in search for the mechanisms of the $[Ca^{2+}]_i$, changes following IHNV infection of CHSE cells. Decreasing extracellular $Ca^{2+}$ concentration or blocking $Ca^{2+}$ influx from extracellular media inhibited the IHNV-induced increase in $[Ca^{2+}]_i$, in CHSE cells. Similar results were obtained with intracellular $Ca^{2+}$ blockers. Thus it is suggested that both the extracellular and the intracellular $Ca^{2+}$ sources are important in IHNV-induced $[Ca^{2+}]_i$ increase in CHSE cells.

• The Plant Pathology Journal
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• v.36 no.1
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• pp.76-86
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• 2020

Cucumber mosaic virus (CMV) is damaging to the growth and quality of lettuce crops in Lanzhou, China. Recently, however, for the first time an isolate of lettuce necrotic yellows virus (LNYV) has been detected in lettuce crops in China, and there is concern that this virus may also pose a threat to lettuce production in China. Consequently, there is a need to develop a rapid and efficient detection method to accurately identify LNYV and CMV infections and help limit their spread. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays were developed to detect the nucleoprotein (N) and coat protein (CP) genes of LNYV and CMV, respectively. RT-LAMP amplification products were visually assessed in reaction tubes separately using green fluorescence and gel electrophoresis. The assays successfully detected both viruses in infected plants without cross reactivity recorded from either CMV or LNYV or four other related plant viruses. Optimum LAMP reactions were conducted in betaine-free media with 6 mM Mg²⁺ at 65℃ for LNYV and 60℃ for 60 min for CMV, respectively. The detection limit was 3.5 pg/ml and 20 fg/ml using RT-LAMP for LNYV and CMV plasmids, respectively. Detection sensitivity for both RT-LAMP assays was greater by a factor of 100 compared to the conventional reverse transcription polymerase chain reaction assays. This rapid, specific, and sensitive technique should be more widely applied due to its low cost and minimal equipment requirements.

• Korean Journal of Veterinary Research
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• v.39 no.6
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• pp.1141-1150
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• 1999

Renal failure is one of the main causes of economic impacts in the poultry industry and complex syndrome with different severity of clinical signs caused by multiple nephropathogenic factors such as infectious bronchitis viral infection and excess salt and calcium in diet. To evaluate the correlation between severity of renal failure and the causative nephropathogenic factors, one-day-old specific pathogen free chickens were treated with either single causative factor or multiple causative factors described as above. Each group was designed as control for non-treated control, IB for infectious bronchitis virus (IB virus) infection, IBHNa for IB virus infection with high diet salt, IBHCa for IB virus infection with high diet calcium, IBHNC for IB virus infection with high diet salt and calcium, HNa for high diet salt, HCa for high diet calcium and HNC for high diet salt and calcium. Chickens were inoculated with IB virus at 1-day-old and remained on their respective diets until 21 day of age. Plasma $Na^+$, $Cl^-$, BUN, creatinine, calcium and uric acid values were examined. The results obtained were as follows ; IB virus and high dietary calcium combined treatment showed elevated plasma uric acid. BUN and creatinine values were not characteristic on chicken renal failure. But plasma uric acid values were increased according to renal lesion. Hypercalcemia and hyperuricemia did not induce urate deposition and mineralization in the kidney.

• 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.

• The Plant Pathology Journal
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• v.26 no.2
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• pp.194-197
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• 2010

Most orchids are propagated by tissue culture. To survey the viral infection of tissue cultured Orchids, total RNA was extracted from in vitro Cymbridium and Phalaenopsis spp. collected from companies producing tissue-cultured orchids, and RT-PCR analysis was conducted with primer pairs specific to Cymbidium mosaic virus (CymMV) and Odontoglossum ring spot virus(ORSV), which are infecting wide range of orchid genera. The bulb size of Cymbidium infected with CymMV and ORSV was compared with healthy one at 10 months after planting in vitro orchids in the glasshouse. The CymMV or ORSV infection in 97 Cymbidium and 55 Phalaenopsis plants was 84.5 and 89.1 %, respectively. Mixed infection was found in 52.6 and 47.3% of Cymbidium and Phalaenopsis tested, whereas virus-free orchids were 15.5 and 10.9%, respectively. The CymMV and ORSV reduced the bulb size by 2.7-50% depending on the cultivars of Cymbidium. The both viruses caused yellowing, mottle and mosaic with or without necrosis in 4 Cymbidium cultivars.

• The Plant Pathology Journal
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• v.36 no.5
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• pp.509-514
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• 2020

Satsuma dwarf virus (SDV) seriously damages citrus production by reducing the quality and yield of fruit. To avoid contamination with SDV, mother trees are checked to be SDV-free in advance of nursery tree distribution. In this study, we compared an immunochromatographic assay (ICA) kit with double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) for large-scale diagnosis of SDV in orchardgrown trees in Shizuoka Prefecture, Japan. The two methods gave conflicting results for 11 of 1,705 samples, all of which were negative by DAS-ELISA but positive by ICA. The samples scored as positive by either DAS-ELISA or ICA were analyzed by reverse transcription polymerase chain reaction and all were confirmed to be positive. These results validate the use of ICA as a screening method for large-scale diagnosis. Strain discrimination revealed that 16 of 22 isolates belonged to SDV, while citrus mosaic virus (CiMV) infection only and co-infection (SDV and CiMV) were in a minority.

• Journal of agriculture & life science
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• v.43 no.6
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• pp.105-109
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• 2009

Kyungsan nursery complex which has a vast area for the production of various species of fruit tree stocks is in a high demand of virus-free saplings. Apple tree stocks, the most important products, urgently need more rapid and reliable viral diagnosis. In this study, a bead beater was tested because of convenience in dealing with large number of samples. Also, industrial glass bead abrasive (0.4 mm in diameter) at very low cost was used in a disposable way. For bead beater-aided RNA extraction from apple stem tissues, the guanidine thiocyanate method was confirmed to be very reliable. Silca membrane filter tube in connection to vacuum filtering device was strongly suggested for simplifying RNA capture and washing steps. Apple virus detection was confirmed by RT-PCR.

• Research in Plant Disease
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• v.28 no.2
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• pp.92-97
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• 2022

Apple stem grooving virus (ASGV) is a high-risk viral pathogen that infects many types of fruit trees, especially pear and apple, and causes serious economic losses across the globe. Thus, rapid and reliable detection assay is needed to identify ASGV infection and prevent its spread. A reliable reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed, optimize, and evaluated for the coding region of coat protein of ASGV in pear leaf. The developed RT-LAMP facilitated the simple screening of ASGV using visible fluorescence and electrophoresis. The optimized reaction conditions for the RT-LAMP were 63℃ for 50 min, and the results showed high specificity and 100-fold greater sensitivity than the reverse transcription polymerase chain reaction. In addition, the reliability of the RT-LAMP was validated using field-collected pear leaves. Furthermore, the potential application of paper-based RNA isolation, combined with RT-LAMP, was also evaluated for detecting ASGV from field-collected samples. These assays could be widely applied to ASGV detection in field conditions and to virus-free certification programs.

• Journal of Plant Biotechnology
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• v.44 no.4
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• pp.379-387
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• 2017

Various sizes (0.2 ~ 1.2 mm) and developmental stages (referred to as Stage 1 ~ 3) of apical and lateral meristems were excised, together or separately, directly from dormant buds of apple 'Hongro'. They were mixed infected by Apple scar skin viroid (ASSVd), Apple chlorotic leaf spot virus (ACLSV), Apple stem pitting virus (ASPV) and Apple stem grooving virus (ASGV), which are major viruses attacking apples. A total of 31 callus lines (> 10 mm in diameter) were obtained by culturing the explants on Murashige and Skoog (MS) medium supplemented with 3% sucrose, 3.0 mg/L benzyladenine (BA) and 0.1 mg/L indole-3-butyric acid (IBA), and they were subjected to RT-PCR analysis for virus detection. A high rate of virus elimination (expressed as the percentage of calli that did not amplify during RT-PCR, i.e., RT-PCR negative calli per total number of calli obtained) was achieved for ACLSV (100%), ASSVd (93.7%), and ASPV (93.7%), whereas it was only 25.8% for ASGV. ASPV was detected in the presence of 2 ~ 3 bracts. Simultaneous virus elimination of ASSVd, ASPV, ACLSV, and ASGV occurred during the meristem culture, in which the early stages of the dormant buds (Stage 1) were used, because ASGV was mostly eliminated during that stage. The results of the present study will be valuable for the production of virus-free apple trees.

• Journal of Bio-Environment Control
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• v.23 no.4
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• pp.376-382
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• 2014

This work was conducted to investigate the variation of growth and yield among three generations ($TC_0$, $TC_1$, and $TC_2$) in the field cultivation of virus-free sweetpotato (Ipomoea batatas) plants. Virus-free generations of three cultivars ('Matnami', 'Shinhwangmi', and 'Yeonhwangmi') were cultivated with $75{\times}25cm$ planting density on May 20th, covered with black vinyl film. At 30 days after planting, vine growth in $TC_0$, $TC_1$, and $TC_2$ was significantly increased as compared to the farmer's plant, and vine length in $TC_0$ showed the highest growth among treatments. At harvesting time after 120 days, vine diameter, number of node, and number of branch in $TC_0$, $TC_1$, and $TC_2$ were more increased than farmer's plant, but were not statistically significant. Fresh weight of shoot in $TC_0$, $TC_1$, and $TC_2$ was significantly increased as compared to the farmer's plant, but was not statistically significant among generations or cultivars. Number of tuber per plant and mean weight of tuber in $TC_0$ and $TC_1$ showed significant increasement, but that in $TC_2$ did not show significant difference as compared to the farmer's plant. Weight of tuber per plant in $TC_0$, $TC_1$, and $TC_2$ was significantly increased as compared to the farmer's plant. Marketable yield, percentage of marketable tuber, and percentage of small tuber (40 to 200g) in $TC_0$, $TC_1$, and $TC_2$ was significantly increased as compared to the farmer's plant. The large tuber over 300g showed the lowest percentage in $TC_0$. Marketable yield in $TC_2$ was significantly decreased as compared to $TC_0$, and was not significantly different as compared to the farmer's plant. Marketable yield in 'Matnami' was highest among cultivars. From this results, Farmers are required to renew every three years to maintain the yield and quality of virus-free plants. However, the exchange period of virus-free plants is desirable to renew every 2 or 3 years according to the degree of virus reinfection.