• Journal of Plant Biotechnology
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• v.6 no.1
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• pp.9-13
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• 2004

Transient uidA expression was used to optimize parameters required for biolistic transformation of suspension cells of Easter lily, Lilium longiflourm. Maximum uidA expression occurred following bombardment with gold particles as compared to tungsten. A 3hr pre-treatment of suspension cells with 0.125M osmoticum resulted in a 1.5X increase in uidA expression. A helium pressure of 1550 psi combined with a particle travelling distance of 6cm resulted in maximum uidA expression as compared to either 1100, 1200, or 1800 psi. Transient transformation resulted in up to 493 uidA expressing cells/Petri plate. For stable transformation suspension cells of Lilium longiflorum, were co-bombarded with plasmid DNA containing cucumber mosaic virus (CMV) replicase under the rice actin (Act1) promoter and either the bar or PAT genes under the cauliflower mosaic virus (CaMV 355) promoter. Ten regenerated plants contained the transgene as analyzed by PCR, and two of the ten plants were confirmed to contain the transgene by Southern hybridization. The two transgenic plants were independent transformants, one containing the bar gene and the other both the CMV replicase and bar genes. Plants were sprayed at the rosette stage and found to be resistant to 1000 mg/L of phosphinothricin (Trade name-Ignite) indicating expression of the bar gene throughout the leaves when bar was under control of the CaMV 35S promoter.

• The Plant Pathology Journal
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• v.16 no.6
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• pp.297-305
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• 2000

Weeds are widely grown in the field and are infected by many viruses. A survey was conducted to identify viruses infecting weeds in Korea. Virus-infected weed samples including Rorippa indica (L.) Hiern, R. islandica (Oed.) Bord, Crepidiastrum denticulatum (Houtt.) Pak & Kawanno, Achyranthes japonica (Miq.) Nakai, and Chrysanthemum boreale (Makino) Makino were collected in Kyonggi Province. These weeds were grown in the greenhouse and were isolated on 10 test plants. Several virus isolates were isolated fron infected tissues and were further studied by host range assay, serological test, electron microscopy (EM), reverse transcription-polymerase chain reaction (RT-PCR) and sequencing. Each isolated virus strain was mechanically transmitted to weeds and various hosts including Nicotiana spp., Brassica spp., Vigna unguiculata, Capsicum annuum, and Cucumis sativus and showed systemic mosaic, vein clearing, necrosis, mottle, malformation, chlorosis, and/or death of host plants in some cases. Each virus was then purified using infected leaves and observed by EM. From these results three viruses were isolated and identified as Turnip mosaic virus (TuMV), Broad bean wilt virus (BBWV), and Cucumber mosaic virus (CMV). RT-PCR using virus-specific oligonucleotide primers and the cloning were conducted to determine the nucleotide sequences of coat proteins of the three viruses their amino acid sequence were deduced. The amino acid sequence homologies were about 92.7 to 99.7%, 96.2 to 97.7%, and 93.9 to 98.6% to other reported TuMV, BBWV, and CMV strains, respectively. These results suggest that many weeds may serve as primary inoculum source of diseases caused by TuMV, BBWV, CMV and that the management of these viral diseases can be achieved through weed control.

• Pediatric Infection and Vaccine
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• v.30 no.3
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• pp.173-179
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• 2023

Complete DiGeorge syndrome (cDGS) refers to DGS with profound T cell deficiency. Herein, we present the case of an infant with cDGS suffering from refractory cytomegalovirus (CMV) infection and who was treated with CD45RA+ depleted lymphocyte infusion. The patient was diagnosed with cDGS by fluorescence in situ hybridization which verified 22q11.2 deletion and as well as by the observed profound T cell deficiency (CD3+ T cells 69/μL, CD4+ T cells 7/μL). On the 45th day of age, CMV viremia was first detected with a plasma viral load (VL) of 120,000 IU/mL. Ganciclovir treatment effectively reduced VL post 56 days of treatment; however, VL subsequently rebounded. A CMV UL97 phosphotransferase M460V mutation conferring ganciclovir resistance emerged and foscarnet was incorporated. Despite this, high titers of CMV viremia (VL 2,820,000 IU/mL) and CMV retinitis were complicated. To restore T cell immunity and treat refractory CMV infection, CD45RA+ depleted CMV-specific lymphocytes from the patient's father were infused twice on the 196th and 207th days after birth. After receiving the second infusion, a decline in CMV VL was observed, with a decrease to 87,100 IU/mL by the tenth day following infusion, despite the failure in maintaining T cell increase. The patient died of Pneumocystis jirovecii pneumonia and Elizabethkingia meningoseptica sepsis on the 222nd day after birth. CD45RA+ depleted lymphocyte infusion may be a therapeutic option for refractory CMV disease in cDGS patients.

• Journal of Life Science
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• v.18 no.10
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• pp.1395-1399
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• 2008

We have recently reported the PRC1 promoter as a promoter candidate to control expression of transcriptionally targeted genes for breast cancer gene therapy. We tested whether the PRC1 promoter could be also applied for the lung cancer gene therapy. In the transient transfection assay with naked plasmids containing the luciferase fused to the PRC1 promoter, the promoter showed little activity in the normal lung cell line, MRC5. However, in the lung cancer A549 cells, PRC1 showed approximately 30-fold activation which was similar to the survivin promoter, the gene whose promoter has been already reportedas a candidate for the gene therapy of lung cancer. In viral systems, the PRC1 promoter showed approximately 75% and 66% of transcriptional activity compared to the CMV promoter in the adeno-associated virus (AAV) and the adenovirus (AV) systems, respectively. However, the PRC1 promoter in either AAV or AV showed approximately 20% activity compared to the CMV promoter in the normal lung cells. In addition, human lung tumor xenograft mice showed that the PRC1 promoter activity was as strong as the CMV activity in vivo. Taken together, these results suggested that PRC1 might be a potential promoter candidate for transcriptionally targeted lung cancer gene therapy.

• Research in Plant Disease
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• v.14 no.1
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• pp.71-75
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• 2008

An isolate of Peanut stunt virus (PSV), named as Tr-PSV, was isolated from white clover (Trifolium repens L) showing mosaic symptom. Tr-PSV systemically infected all plants tested in the Nicotiana spp. and induced local lesions on inoculated leaves of Chenopodium amaranticolor. However, Tr-PSV induced typical mosaic symptoms as ER-PSV on Vigna unguiculata 5 to 6 days after inoculation, while Fny-CMV used as a control virus of Cucumovirus produced local lesions on inoculated leaves. In dsRNA analysis, Tr-PSV consisted of four dsRNAs, but satellite RNA was not detected. The cDNA of coat protein gene of Tr-PSV was amplified by RT-PCR using a Cucumovirus-specific single pair primers that designed to amplify a DNA fragment of approximately 950 bp. By restriction mapping analysis using RFLP of the RT-PCR products and by serological properties of gel diffusion test, Tr-PSV belongs to a typical member of PSV subgroup I. This is the first report on the occurrence of PSV in white clover in Korea.

• Korean Journal of Plant Resources
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• v.36 no.5
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• pp.527-539
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• 2023

Pepper (Capsicum annuum L.) is an important vegetable and spice crop that has been cultivated worldwide. Pepper fruits have unique taste and aroma, providing a variety of antioxidants and compounds important for human health, which makes a high economic value. In addition, there is a high demand for new pepper varieties, according to consumer's preference. However, pepper is a recalcitrant plant for in vitro tissue and organ differentiation and plant regeneration, which makes it difficult to develop demanded varieties using newly developed technologies such as genetic engineering and gene editing. In this study, tissue culture and regeneration conditions were investigated using seven pepper varieties that were obtained from the core-collection of Seoul National University. We observed callus and bud induction and shoot formation using several media composition composed of different cytokinins and auxin concentrations. As a result, it was found that there were differences in callus induction and shoot formation of each variety depending on the hormone composition, and the highest regeneration was shown when the medium containing Zeatin Riboside and the petioles of seedlings were used. In particular, out of seven pepper varieties, CMV980 exhibited a higher regeneration efficiency (approximately 48%) than other varieties, followed by Yuwolcho. Therefore, this study provides CMV980 and Yuwolcho as good candidates that can be used for pepper transformation, which might contribute to the development of various varieties through gene editing technology in the future.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.54 no.4
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• pp.384-389
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• 2009

Germination characteristics of Chinese milk vetch(CMV) seeds produced from Hanam Province, China and Milyang, Korea were investigated to give basic information on the stability of seedling establishment in the CMV cultivation. The germination percentage of the imported CMV seed from China varied according to importation year and seed collection site ranging from 79~95%. The germination of black colored seed coat was lower than the light green colored ones and germination by seed weight was not significantly different. Although the seed germination was lower under dark than in the presence of light, it was not significantly different. The germination of the imported CMV seed slightly declined to only less than 6% after one-year of storage under natural environment conditions but it significantly decreased after two years. However, when the seed was stored at the $5^{\circ}C$, the seed germination was the same as after two years of storage. On the other hand, fresh CMV seed produced in Milyang, Korea had only 8% germination due to seed coat dormancy but the germination increased to 73~85% after breaking seed dormancy after a year of storage. The high germination percentage of 72~82% was still maintained even after 27 months of seed storage unlike the CMV seed produced from China. These results indicate that CMV seeds do not require light for germination and the seed from China should be used within one-year after importation while the seeds produced from Korea can be used even after two years from harvest for stable CMV seedling establishment in the CMV-rice cropping system.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.54 no.4
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• pp.351-356
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• 2009

Growth, milled rice yield and edible quality of rice in naturally reseeded Chinese milk vetch(CMV)-rice cropping system was compared with those in rice mono cropping on silty loam soil in Milyang from 2006-2008. Practicing natural reseeding technology recorded high CMV reseeding stand ranging from 565-805 plants $m^{-2}$ and resulting in the production of 13.0-17.0 kg N/10a from the CMV plant biomass which is greater than the recommendation rate of 9 kgN/10a. The plant height of rice plant grown in natural reseeding field is shorter at tillering stage but it was similar to the rice mono cropping at later stage. Dry matter production had similar trend to plant height. On the other hand, the leaf color in naturally reseeded CMV-rice cropping system was similar to the rice mono cropping up to panicle heading stage but it was high at mature stage, indicating that the nitrogen was provided by the CMV decomposition until later stage of rice. The yield components such as culm number $m^{-2}$ was greater and 1,000-brown rice weight was heavier than those of rice mono cropping but the ripened grain ratio was lower in naturally reseeded CMV-rice cropping system. Milled rice yield of naturally reseeded CMV-rice cropping system was similar to that of rice mono cropping. However, head rice percentage of milled rice was lower due to low ripened grain ratio. This result indicates that natural CMV reseeding technology can completely replace chemical fertilizer in CMV-rice cropping system.

• Korean Journal of Soil Science and Fertilizer
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• v.44 no.2
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• pp.221-227
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• 2011

Applications of green manures generally improve the soil quality in rice paddy in part through restructuring of soil microbial communities. To determine how different green manures affect soil microbial communities during the early stages of rice growth, fatty acid methyl ester (FAME) profiles were used to the effects of different management practices: 1) conventional farming (CF), 2) no-treatment (NT), 3) Chinese milk vetch (CMV), 4) green barley (GB), and 5) triticale in paddy field. With applications of green manures, soil organic matter was significantly higher than CF, while soil Na concentration was significantly lower compared with CF (p<0.05). Total soil microbial biomass of CMV was higher (p<0.05) than NF by approximately 31%. The highest ratio of monounsaturated fatty acid to saturated fatty acid was found in the GB plot, followed by CMV and triticale compared with CF (p<0.05), possibly indicating that microbial stress was less in GB and CMV plots. Populations of Gram-negative bacteria and arbuscular mycorrhizal fungi also were significantly higher green manures than CF (p<0.05). Our findings suggest that GB should be considered as optimum green manure for enhancing soil microbial community at an early growing stage in paddy field.

• Molecules and Cells
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• v.38 no.12
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• pp.1079-1085
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• 2015

Originally, activins were identified as stimulators of FSH release in reproduction. Other activities, including secondary axis formation in development, have since been revealed. Here, we investigated the influence of activin ${\beta}_A$ on the body, including the gastro-intestinal (GI) tract. Initially, the activin ${\beta}_A$ protein was detected in the serum proportional to the amount of pCMV-rAct plasmid injected. The induced level of activin ${\beta}_A$ in muscle was higher in female than male mice. Subsequent results revealed that stomach and intestine were severely damaged in pCMV-rAct-injected mice. At the cellular level, loss of parietal cells was observed, resulting in increased pH within the stomach. This phenomenon was more severe in male than female mice. Consistent with damage of the stomach and intestine, activin ${\beta}_A$ often led to necrosis in the tip of the tail or foot, and loss of body weight was observed in pCMV-rAct-injected male but not female mice. Finally, in pCMV-rAct-injected mice, circulating activin ${\beta}_A$ led to death at supraphysiological doses, and this was dependent on the strain of mice used. Taken together, these results indicate that activin ${\beta}_A$ has an important role outside of reproduction and development, specifically in digestion. These data also indicate that activin ${\beta}_A$ must be controlled within a narrow range because of latent lethal activity. In addition, our approach can be used effectively for functional analysis of secreted proteins.