• The Plant Pathology Journal
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• v.30 no.3
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• pp.254-260
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• 2014

Potato Rpi-blb2 encodes a protein with a coiled-coil-nucleotide binding site and leucine-rich repeat (CC-NBSLRR) motif that recognizes the Phytophthora infestans AVRblb2 effector and triggers hypersensitive cell death (HCD). To better understand the components required for Rpi-blb2-mediated HCD in plants, we used virus-induced gene silencing to repress candidate genes in Rpi-blb2-transgenic Nicotiana benthamiana plants and assayed the plants for AVRblb2 effector. Rpi-blb2 triggers HCD through NbSGT1-mediated pathways, but not NbEDS1- or NbNDR1-mediated pathways. In addition, the role of salicylic acid (SA), jasmonic acid (JA), and ethylene (ET) in Rpi-blb2-mediated HCD were analyzed by monitoring of the responses of NbICS1-, NbCOI1-, or NbEIN2-silenced or Rpi-blb2::NahG-transgenic plants. Rpi-blb2-mediated HCD in response to AVRblb2 was not associated with SA accumulation. Thus, SA affects Rpi-blb2-mediated resistance against P. infestans, but not Rpi-blb2-mediated HCD in response to AVRblb2. Additionally, JA and ET signaling were not required for Rpi-blb2-mediated HCD in N. benthamiana. Taken together, these findings suggest that NbSGT1 is a unique positive regulator of Rpi-blb2-mediated HCD in response to AVRblb2, but EDS1, NDR1, SA, JA, and ET are not required.

• Plant Biotechnology Reports
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• v.4 no.1
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• pp.49-52
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• 2010

A DNA extraction method using Chelex 100 is widely used for bacteria, Chlamydomonas, and animal cell lines, but only rarely for plant materials due to the need for additional time-consuming and tedious steps. We have modified the Chelex 100 protocol and successfully developed a rapid and simple method of DNA extraction for efficient PCR-based detection of transgenes from a variety of transgenic plant and algal species. Our protocol consists of homogenizing plant tissue with a pestle, boiling the homogenized tissue in a microfuge tube with 5% Chelex 100 for 5 min, and centrifuging the boiled mixture. The supernatant, which is used for PCR analysis, was able to successfully amplify transgenes in transgenic tobacco, tomato, potato, Arabidopsis, rice, strawberry, Spirodela polyrhiza, Chlamydomonas, and Porphyra tenera. The entire DNA extraction procedure requires <15 min and is therefore comparable to that used for bacteria, Chlamydomonas, and animal cell lines.

• The Plant Pathology Journal
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• v.37 no.2
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• pp.182-193
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• 2021

The transcription factor SHE1 was identified as an interacting partner with the cucumber mosaic virus (CMV) 1a protein in the yeast two-hybrid system, by a pull-down assay, and via bimolecular fluorescent complementation. Using fluorescent-tagged proteins and confocal microscopy, the CMV 1a protein itself was found distributed predominantly between the nucleus and the tonoplast membrane, although it was also found in speckles in the cytoplasm. The SHE1 protein was localized in the nucleus, but in the presence of the CMV 1a protein was partitioned between the nucleus and the tonoplast membrane. SHE1 expression was induced by infection of tobacco with four tested viruses: CMV, tobacco mosaic virus, potato virus X and potato virus Y. Transgenic tobacco expressing the CMV 1a protein showed constitutive expression of SHE1, indicating that the CMV 1a protein may be responsible for its induction. However, previously, such plants also were shown to have less resistance to local and systemic movement of tobacco mosaic virus (TMV) expressing the green fluorescent protein, suggesting that the CMV 1a protein may act to prevent the function of the SHE1 protein. SHE1 is a member of the AP2/ERF class of transcription factors and is conserved in sequence in several Nicotiana species, although two clades of SHE1 could be discerned, including both different Nicotiana species and cultivars of tobacco, varying by the presence of particular insertions or deletions.

• Journal of Plant Biotechnology
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• v.37 no.3
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• pp.250-261
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• 2010

The expression of vaccine antigens in transgenic plants has the potential to provide a convenient, stable, safe approach for oral vaccination alternative to traditional parenteral vaccines. Over the past two decades, many different vaccine antigens expressed via the plant nuclear genome have elicited appropriate immunoglobulin responses and have conferred protection upon oral delivery. Up to date, efforts to produce antigen proteins in plants have focused on potato, tobacco, tomato, banana, and seed (maize, rice, soybean, etc). The choice of promoters affects transgene transcription, resulting in changes not only in concentration, but also in the stage tissue and cell specificity of its expression. Inclusion of mucosal adjuvants during immunization with the vaccine antigen has been an important step towards the success of plant-derived vaccines. In animal and Phase I clinical trials several plant-derived vaccine antigens have been found to be safe and induce sufficiently high immune response. Future areas of research should further characterize the induction of the mucosal immune response and appropriate dosage for delivery system of animal and human vaccines. This article reviews the current status of development in the area of the use of plant for the development of oral vaccines.

• Journal of Microbiology and Biotechnology
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• v.17 no.1
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• pp.130-137
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• 2007

A disease resistance related gene, MbR7, was identified in the wild apple species, Malus baccata. The MbR7 gene has a single open reading frame (ORF) of 3,288 nucleotides potentially encoding a 1,095-amino acid protein. Its deduced amino acid sequence resembles the N protein of tobacco and the NL27 gene of potato and has several motifs characteristic of a TIR-NBS-LRR R gene subclass. Ectopic expression of MbR7 in Arabidopsis enhanced the resistance against a virulent pathogen, Pseudomonas syringae pv. tomato DC3000. Microarray analysis confirmed the induction of defense-related gene expression in 35S::MbR7 heterologous Arabidopsis plants, indicating that the MbR7 gene likely activates a downstream resistance pathway without interaction with pathogens. Our results suggest that MbR7 can be a potential target gene in developing a new disease-resistant apple variety.

• Research in Plant Disease
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• v.8 no.2
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• pp.78-83
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• 2002

Tobacco diseases have not been recorded until 1900s in Korea, where tobacco plants were introduced at early 1700s. Practical researches on the disease have been conducted since mid 1960s. Major ten tobacco diseases were mosaic caused by tobacco mosaic virus·potato virus Y·cucumber mosaic virus, bacterial wilt, hollow stalk, wild fire caused by angular leaf spot strain, black shank, brown spot, powdery mildew and fusarium wilt. But their annual occurrences were varied according to changes of tobacco varieties and their cultivating practices. As no useful chemicals, several biological tactics have been developed to control the viral or bacterial diseases that give significant economic damages on sustainable crop yield, but not practicable to field farming condition yet. Transgenic tobacco plants containing foreign disease resistant genes have been developed by current bio-technology, but not released to farmers yet. Though some disease-resistant tobacco varieties have been developed by the conventional breeding technology and currently used by farmers, their disease controlling efficacy have been diminished by occurrence of the new strain or race. Future research on tobacco diseases has been focused on technical development to produce high quality tobacco with less production cost, which leads Korean tobacco industry to keep its competence against foreign industry and decreasing overall market.

• Journal of Plant Biotechnology
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• v.30 no.4
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• pp.405-410
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• 2003

Environmental stress is the major limiting factor in plant productivity. In order to evaluate the stress tolerance of potato plants, leaf discs of two potato cultivars, Atlantic and Superior, were subjected to various stress conditions of high temperature, methyl viologen, H2O2, or $H_2O$$_2$. When potato leaf discs were exposed to high temperature at 37$^{\circ}C$ for 84 hr, Atlantic plants, a cultivar with high sensitivity to heat stress, showed about 20％ higher membrane damage than Superior plants. When exposed to 2$\mu$M methyl violgen (MV), a superoxide generating non-selective herbicide, for 36 hr, Atlantic plants also showed about 38％ higher membrane damage than Superior plants, and were more susceptible up to 10$\mu$M MV concentration tested. On treatment with 0.75M NaCl, Atlantic plants also had about 45％ less chlorophyll contents in leaf discs than Superior plants. There was, however, no difference in chlorophyll content of two cultivars at higher NaCl concentrations. The effect of $H_2O$$_2$ on the two cultivars was mixed. At low $H_2O$$_2$ concentration (25 mM) , Superior plants were more susceptible to $H_2O$$_2$stress after 36 hr. However, at high $H_2O$$_2$ concentration (100 mM), Atlantic plants exhibited higher susceptibility after 36 hr. The results indicate that in vitro leaf discs reflecting the whole plants in this study will be useful for selection and characterization of elite transgenic potato plants with enhanced tolerance to environmental stress.

• The Plant Pathology Journal
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• v.27 no.2
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• pp.138-147
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• 2011

Xylogone ganodermophthora, an ascomycetous fungus, is known to cause yellow rot in the cultivated mushroom Ganoderma lucidum. In this study, we investigated the dissemination of this fungal pathogen in G. lucidum grown in cultivation houses. To determine the role of ascospores produced by X. ganodermophthora in disease development, we constructed a green fluorescent protein-labeled transgenic strain. This X. ganodermophthora strain produced a number of ascomata in the tissues of oak logs on which G. lucidum had been grown and on the mushroom fruit bodies. However, the ascospores released from the ascomata were not able to germinate on water agar or potato dextrose agar. Moreover, less than 0.1% of the ascospores showed green fluorescence, indicating that most ascospores of X. ganodermophthora were not viable. To determine the manner in which X. ganodermophthora disseminates, diseased oak logs were either buried in isolated soil beds as soil-borne inocula or placed around soil beds as air-borne inocula. In addition, culture bottles in which G. lucidum mycelia had been grown were placed on each floor of a five-floor shelf near X. ganodermophthora inocula. One year after cultivation, yellow rot occurred in almost all of the oak logs in the soil beds, including those in beds without soil-borne inocula. In contrast, none of the G. lucidum in the culture bottles was infected, suggesting that dissemination of X. ganodermophthora can occur via the cultivation soil.

• Journal of Plant Biotechnology
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• v.1 no.1
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• pp.20-30
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• 1999

In order to modify lipid production of Perilla qualitatively as well as quantitatively by genetic engineering, genes involved in carbon metabolism were isolated and characterized. These include acyl-ACP thioesterases from Perilla frutescens and Iris sp., four different $\beta$-ketoacyl- ACP synthases from Perilla frutescens, and two $\Delta$15 a-cyl-ACP desaturases(Pffad7, pffad3). Δ15 acyl-ACP desa turase (Δ15-DES) is responsible for the conversion of linoleic acid (18:2) to $\alpha$-linolenic acid (ALA, 18:3). pffad 3 encodes Δ15 acyl-desaturase which is localized in ER membrane. On the other hand, Pffad7 encodes a 50 kD plastid protein (438 residues), which showed highest sequence similarity to Sesamum indicum fad7 protein. Northern blot analysis revealed that the Pffad7 is highly expressed in leaves but not in roots and seeds. And Pffad3 is expressed throughout the seed developmental stage except very early and fully mature stage. We constructed Pffad7 gene under 355 promoter and Pffad3 gene under seed specific vicillin promoter. Using Pffad7 construct, Perilla, an oil seed crop in Korea, was transformed by Agrobacterium leaf disc method. $\alpha$-linolenic acid contents increased in leaves but decreased in seeds of transgenic Perilla. Currently, we are transforming Perilla with Pffad3 construct to change Perilla seed oil composition. We isolated three ADP-glucose pyrophosphorylase (AGP) genes from Perilla immature seed specific cDNA library. Nucleotide sequence analysis showed that two of three AGP (Psagpl, Psagp2) genes encode AGP small subunit polypeptides and the remaining (Plagp) encodes an AGP large subunit. PSAGPs, AGP small subunit peptide, form active heterotetramers with potato AGP large subunit in E. coli expressing plant AGP genes.

• Proceedings of the Korean Society of Tobacco Science Conference
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• 1997.10a
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• pp.134-152
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• 1997

Plant viruses of tobacco including tobacco mosaic virus (TMV) and potato virus Y (PVY) cause severe economic losses in leaf-tobacco production. Cultural practices do not provide sufficient control against the viruses. Use of valuable resistant cultivars is most recommendable for the control of the viruses. However, conventional breeding programs are not always proper for the development of virus-resistant plants mostly owing to the frequent lack of genetic sources and introduction of their unwanted properties. Therefore, we tried to develop virus-resistant tobacco plants by transforming commercial tobacco cultivars, NC 82 and Burley 21, with coat protein (CP) or replicase (Nlb) genes of TMV and PVY necrosis strain (PVY-VN) with or without untranslated region (UTR) and with or without mutation. Each cDNA was cloned and inserted in plant expression vectors with 1 or 2 CaMV 35S promotors, and introduced into tobacco leaf tissues by Agrobacterium tumefaciens LBA 4404. Plants were regenerated in kanamycin-containing MS media. Regenerated plants were tested for resistance to TMV and PVY In these studies, we could obtain a TMV-resistant transgenic line transformed with TMV CP and 6 genetic lines with PVY-VN cDNAs out of 8 CP and replicase genes. In this presentation, resistance rates, verification of gene introduction in resistant plants, stability of resistance through generations, characteristics of viral multiplication and translocation in resistant plants, and resistance responses relative to inoculum potential and to various PVY strains will be shown. Yield and quality of leaf tobacco of a promising resistant tobacco line will be presented.