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The First Identified Citrus tristeza virus Isolate of Turkey Contains a Mixture of Mild and Severe Strains

  • Cevik, Bayram;Yardimci, Nejla;Korkmaz, Sava
    • The Plant Pathology Journal
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    • v.29 no.1
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    • pp.31-41
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    • 2013
  • The presence of Citrus tristeza virus (CTV) has previously been reported in citrus growing regions of Turkey. All serologically and biologically characterized isolates including I$\breve{g}$d${\i}$r, which was the first identified CTV isolates from Turkey, were considered mild isolates. In this study, molecular characteristics of the I d r isolate were determined by different methods. Analysis of the I$\breve{g}$d${\i}$r isolate by western blot and BD-RT-PCR assays showed the presence of MCA13 epitope, predominantly found in severe isolates, in the I$\breve{g}$d${\i}$r isolate revealing that it contains a severe component. For further characterization, the coat protein (CP) and the RNA-depen-dent RNA polymerase (RdRp) genes representing the 3' and 5' half of CTV genome, respectively, were amplified from dsRNA by RT-PCR. Both genes were cloned separately and two clones for each gene were sequenced. Comparisons of nucleotide and deduced amino acid sequences showed that while two CP gene sequences were identical, two RdRp clones showed only 90% and 91% sequence identity in their nucleotide and amino acid sequences, respectively, suggesting a mixed infection with different strains. Phylogenetic analyses of the CP and RdRp genes of I$\breve{g}$d${\i}$r isolate with previously characterized CTV isolates from different citrus growing regions showed that the CP gene was clustered with NZRB-TH30, a resistance breaking isolate from New Zealand, clearly showing the presence of severe component. Furthermore, two different clones of the RdRp gene were clustered separately with different CTV isolates with a diverse biological activity. While the RdRp-1 was clustered with T30 and T385, two well-characterized mild isolates from Florida and Spain, respectively, the RdRp-2 was most closely related to NZRB-G90 and NZRB-TH30, two well-characterized resistance breaking and stem pitting (SP) isolates from New Zealand confirming the mixed infection. These results clearly demonstrated that the I$\breve{g}$d${\i}$r isolate, which was previously described as biologically a mild isolate, actually contains a mixture of mild and severe strains.

Effects of Melatonin on the Meiotic Maturation of Mouse Oocytes in vitro (생쥐 난자의 체외 성숙에 미치는 Melatonin의 영향)

  • Ahn, Hee-Jin;Bae, In-Ha
    • Clinical and Experimental Reproductive Medicine
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    • v.31 no.3
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    • pp.155-168
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    • 2004
  • Objective: Melatonin, which is secreted by pineal gland play an important role in the regulation of ovarian function via seasonal rhythm and sleep in most mammals. It also has a role in the protection of cells by removing toxic oxygen free radicals brought about by metabolism. In the present study, effects of melatonin on the mouse oocyte maturation were examined using two different culture conditions provided with 5% or 21% oxygen concentration. Material and Method: Immature mouse oocytes were obtained from the ovarian follicles of $3{\sim}4$ weeks old ICR strain mice intraperitoneally injected with 5 I.U. PMSG 44 hour before. Under stereomicroscope, morphologically healthy oocytes with distinct germinal vesicle (GV) were liberated from the graafian follicles and collected using mouth-controlled micropipette. They were then cultured for 17 hour at $37^{circ}C$, 5% $CO_2$ and 21% $O_2$ (95% air) or 5% $CO_2$, 5% $O_2$ and 90% $N_2$. New modified Hank's balanced salt solution (New MHBS) was used as a culture medium throughout the experiments. Effects of melatonin were examined at a concentration of $0.0001{\mu}M$, $0.01{\mu}M$ or $1.0{\mu}M$. For the prevention of spontaneous maturation of immature oocytes during culture, dibutyryl cyclic AMP (dbcAMP) and/or hypoxanthine were included in the medium. Results: Under 21% oxygen condition, oocytes cultured in the presence of $0.01{\mu}M$ melatonin showed a significantly higher maturation rates, in terms of germinal vesicle breakdown (95.0% vs 89.0%) and polar body formation (88.1% vs 75.4%), compared to those cultured with $0.0001{\mu}M$ or $1.0{\mu}M$ melatonin. However, no difference was observed in oocytes cultured under 5% oxygen whether they were treated with melatonin or not. In the presence of $0.01{\mu}M$ melatonin, oocytes either cultured under 21% or 5% oxygen exhibited no difference in the polar body formation (85.6% vs 86.7%). However, in the absence of melatonin, oocytes cultured under 21% oxygen exhibited lower polar body formation (74.7%). When oocytes were cultured in the presence of dbcAMP alone or with varying concentrations of melatonin, those treated with both compounds always showed better maturation, i.e., germinal vesicle breakdown and polar body formation, compared to those cultured with dbcAMP alone. At the same concentration of melatonin, however, oocytes exposed to 21% oxygen showed poor maturation than those to 5% oxygen. Similar results were obtained from the experiments using hypoxanthine instead of dbcAMP. Conclusion: Based upon these results, it is suggested that melatonin could enhance the meiotic maturation of mouse oocytes under 21% oxygen concentration, and release oocytes from the meiotic arrest by dbcAMP or hypoxanthine regardless of the concentration of oxygen, probably via the removal of oxygen free radicals.

Carbon-induced reconstructions on W(110)

  • Kim, Ji-Hyeon;Rojas, Geoff;Anders, Axel;Kim, Jae-Seong
    • Proceedings of the Korean Vacuum Society Conference
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    • 2010.02a
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    • pp.362-362
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    • 2010
  • Today, vast attention has been paid to periodic arrays of nanostructures due to their potential for applications such as memory with huge storage density. Such application requires large-scale fabrication of well ordered nano-sized structures. One of the most widely used methods for the ordered nanostructures is lithography. This top-down process, however, has the limit to reduce size. Here the promising alternative is the self-organization of ordered nano-sized structures such as large scale 2d carbon-induced reconstructions on W(110). In the present study, we report on the first well-resolved atomic resolution STM studies of the well-known R($15{\times}3$) and R($15{\times}12$) carbon induced reconstruction of the W(110). From the atomic image of R($15{\times}3$) for different values of tunneling gap resistance, we can tell there are no missing atoms in unit cells of R($15{\times}3$) and some atomic displacements are substantial from the clean W(110), even though not all the imaged position of atoms correspond to tungsten, but may include those of carbon. We are considering two cases; First case is related to lattice deformation, or top layer of W(110) is deformed in the process of relief of strain caused by random inserting of carbon atoms possibly in the interstitial position. In the second case, R($15{\times}3$) unit cell results from a coincidence lattice between clean W(110) substrate and tungsten carbide overlayer which has rectangular atomic arrangement and giving R($15{\times}3$) coincidence lattice. beta-W2C showing rectangular unit cell should be a candidate. Further, we report on new reconstructions. Unlike the well-known R($15{\times}12$) consisting of two parts, two inner structures between two "Backbone" structures. The new reconstruction, which we found for the first time, contains more parts between the "Backbone"s. Sometimes we can observe the reconstruction consists of only inner parts without "Backbone" parts. Thus, the observed reconstruction can be built by constructing of two types of "Lego"-like block. Moreover, the rectangle shape of "Backbone" transform to parallelogram-like shape over time, the so-called wavy-R($15{\times}12$). Adsorption of hydrogen can be the reason for this transformation.

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Isolation of Hypervirulent Agrobacterium spp from Korea and Application for Transformation of Tobacco (한국산 고감염 Agrobacterium spp의 분리 및 연초의 형질전환에 이용)

  • 양덕춘;정재훈;이정명
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.3
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    • pp.207-217
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    • 1998
  • Total of 78 strains were characterized based on the morphological characteristics of colonies isolated on Schroth, and New & Kerr's media for selection of hypervirulent wild-type Agrobacterium spp from galls, hairy root-like process and soil of Populus, Malus, Salix and Diopyros in Korea. Among them, 48 strains were able to induce tumors in carrot disc. Hypervirulent A. tumefaciens SP101 and SM042 were identified as biotype 1 and biotype 2, respectively, These strains formed fast growing, larger tumors as compared to those induced by other strains. The binary vector pGA643 with kanamycin resistant gene was mobilized from E. coli MC100 into A. tumefaciens strain SM042 isolated from soil, and/or disarmed vector PC2760 using a triparental mating method with E. coli HB101/pRK2013, and transconjugants, A. tumefaciens SM643 and PC643 were obtained in minimal media containing kanamycin and tetracycline. Tobacco tissues were cocultivated with conjugant Agrobacterium and then transferred to selective medium with 2,4-D and kanamycin to induce the transformants. Calli were formed more efficiently in cocultivation with A. tumefaciens SM643 than that with A. tumefaciens PC643. Most of calli transformed with A. tumefaciens PC643 were friable and regenerated into normal plantlets, while the calli transformed with A. tumefaciens SM643 were compact, hard, and mixed with friable calli. The friable calli formed normal shoots, while compact calli did not form shoots but only grew to typical compact tumor calli. When the shoots formed directly from tobacco stems without callus induction after transformation by A. tumefaciens SM643 with wild-type Ti-plasmid, normal transformed plants can be induced without using disarmed Ti-plasmid.

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Development of New Strains of Wolfiporia cocos for Sclerotium Formation by 2-Way Cross-Breeding (이원교배에 의해 균핵 형성하는 복령 균주 개발)

  • Ka, Kang-Hyeon;Kim, Suyeon;Park, Mi-Jeong;Jeong, Yeun Sug;Ryoo, Rhim;Jang, Yeongseon;Choi, Jong-Woon;Kim, Seong Hwan
    • The Korean Journal of Mycology
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    • v.49 no.3
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    • pp.405-412
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    • 2021
  • Wolfiporia cocos is an important medicinal fungus that has been used in regions of Northeast Asia including Korea, Japan, and China. W. cocos is classified in Korea into two types (red bokryeong and white bokryeong) based on the internal colors (yellow orange-pale pink and white) of the sclerotium. Generally, the W. cocos type cultivated on farms produces white sclerotium. In this study, we endeavored to select strains that form sclerotium in sawdust medium using 2-way cross-breeding among two cultivated strains and three wild strains. Monospores were isolated from the fruiting bodies of cultivated and wild strains on potato dextrose agar. Thirty-nine strains of 338 hybrid strains isolated formed sclerotia with white or yellow colors upon culture for 3 months in Pinus densiflora sawdust medium. Selection for sclerotium forming strains using sawdust culture follows a very simple and easy procedure that is presented for the first time in this paper. We plan to test selected strains in the field to aid in developing new varieties for the future.

Development and Evaluation of a Next-Generation Sequencing Panel for the Multiple Detection and Identification of Pathogens in Fermented Foods

  • Dong-Geun Park;Eun-Su Ha;Byungcheol Kang;Iseul Choi;Jeong-Eun Kwak;Jinho Choi;Jeongwoong Park;Woojung Lee;Seung Hwan Kim;Soon Han Kim;Ju-Hoon Lee
    • Journal of Microbiology and Biotechnology
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    • v.33 no.1
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    • pp.83-95
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    • 2023
  • These days, bacterial detection methods have some limitations in sensitivity, specificity, and multiple detection. To overcome these, novel detection and identification method is necessary to be developed. Recently, NGS panel method has been suggested to screen, detect, and even identify specific foodborne pathogens in one reaction. In this study, new NGS panel primer sets were developed to target 13 specific virulence factor genes from five types of pathogenic Escherichia coli, Listeria monocytogenes, and Salmonella enterica serovar Typhimurium, respectively. Evaluation of the primer sets using singleplex PCR, crosscheck PCR and multiplex PCR revealed high specificity and selectivity without interference of primers or genomic DNAs. Subsequent NGS panel analysis with six artificially contaminated food samples using those primer sets showed that all target genes were multi-detected in one reaction at 108-105 CFU of target strains. However, a few false-positive results were shown at 106-105 CFU. To validate this NGS panel analysis, three sets of qPCR analyses were independently performed with the same contaminated food samples, showing the similar specificity and selectivity for detection and identification. While this NGS panel still has some issues for detection and identification of specific foodborne pathogens, it has much more advantages, especially multiple detection and identification in one reaction, and it could be improved by further optimized NGS panel primer sets and even by application of a new real-time NGS sequencing technology. Therefore, this study suggests the efficiency and usability of NGS panel for rapid determination of origin strain in various foodborne outbreaks in one reaction.

Breeding a new variety of the white winter mushroom, Flammulina velutipes 'Baekwoon' (흰색 팽이버섯 신품종 '백운' 육성)

  • Ji-Hoon Im;Minji Oh;Minseek Kim;Youn-Lee Oh
    • Journal of Mushroom
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    • v.21 no.4
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    • pp.241-246
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    • 2023
  • Flammulina velutipesis a popular edible mushroom in South Korea, accounting for approximately 30% of the total mushroom export in the country. Despite its significant position in the domestic mushroom industry, approximately 65% of the cultivated F. velutipes are foreign varieties. To secure variety rights and enhance competitiveness in the export market, there is a need to develop domestic varieties with stable production and excellent shelf life. We focused on breeding a new variety called 'Baekwoon' through mon-mon crossing, using 'Baekseung' and an international collection strain (KMCC02260) as parents. Baekwoon exhibited faster mycelial growth rates at 15 ℃ and 25 ℃ and higher mycelial growth across four tested media compared to that of the control variety. Under bottle cultivation, Baekwoon had a mycelial cultivation period that was shorter by approximately 2 days compared to that of the control. The yield of Baekwoon was 228.0±10.9 g, 8.3% more than that of the control. Furthermore, Baekwoon displayed distinct morphological features characterized by a larger pileus and thicker stipe compared to that of the control variety. Genetic tests confirmed that Baekwoon is genetically different from both parental strains and the control variety. It is expected that the newly developed F. velutipes variety, Baekwoon, will be evaluated for its adaptation in different regions and actively promoted in the field, ultimately increasing the market share of domestic varieties.

Breeding of a new cultivar of Lentinula edodes 'Charmjon' (표고버섯 신품종 '참존' 육성 및 특성)

  • Ji-Hoon Im;Youn-Lee Oh;Minji Oh;Minseek Kim;Kab-Yeul Jang
    • Journal of Mushroom
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    • v.21 no.4
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    • pp.261-265
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    • 2023
  • We aimed to develop outstanding domestic varieties suitable for both columnar and cylindrical-shape substrates, aiming to replace foreign varieties. and bred a high-quality new strain named 'Charmjon', using genetic resources collected from Japan and China. The optimal cultivation temperature for Charmjon's mycelial growth was found to be 25℃, and its mycelial growth at 15℃ and 25℃ was superior to the control variety. In terms of mycelial growth characteristics based on the substrate, Charmjon exhibited excellent mycelial strength on PDA medium compared to the control variety. Through columnar and cylindrical-shape substrates cultivation, we assessed the quantity and morphological characteristics of the fruiting bodies. The results confirmed that Charmjon can be produced stably using both cultivation methods, and it showed higher yields and individual weights than the control variety. In addition, the color of the pileus was notably darker, and the shape of the pileus varied depending on the cultivation method. The test of genetic diversity revealed that Charmjon has distinct genetic characteristics compared to the control varieties.

Comparison of Endo-, Exo-Cellular Enzyme Activity for New Strains of Hypsizygus marmoreus (느티만가닥버섯의 신품종에 대한 endo-, exo-cellular 효소 활성도의 비교)

  • Lee, Chang-Yun;Song, Ho-Sung;Ro, Hyeon-Su;Woo, Ju-Ri;You, Young-Hyun;Kim, Jong-Guk
    • Journal of Life Science
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    • v.22 no.6
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    • pp.837-843
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    • 2012
  • This study was carried out to investigate the morphological and physiological characteristics of six new cultivars of Hypsizygus marmoreus (Hm) and measure endo-, exo-cellular enzyme-specific activity. The domestic wild stain (Hm3-10) and commercial strain in Japan (Hm1-1) were mated by crossing monokaryon mycelia. We gained 58 strains from one of 400 crosses through the $1^{st}$ cultivation experiment, and selected six strains from one of 58 strains through the $2^{nd}$ cultivation experiment. When six of the selected new strains were grown during several spawn culture periods (60, 70, 80, 90, and 100 days), a spawn culture period of more 80 days was considered to be excellent as being shorter than 19~20 days. Therefore, we determined the period of spawn culture as 80 days. Three strains such as Hm15-3, Hm15-4, and Hm17-5 showed an excellent result. When endo-cellular enzyme activity measured eight strains, we obtained a result of that specific activity of ${\alpha}$-amylase at the highest as 73.9~102.2 unit/mg protein, and chitinase is lower than ${\alpha}$-amylase at 8.1~13.1 unit/mg protein. When exo-cellular enzyme activity measured eight strains, we determined the result of that specific activity of ${\alpha}$-amylase is the highest at 5,292~1,184 unit/mg protein, and CMCase and xylanase were 1,140~245 unit/mg protein, 94~575 unit/mg protein, compared to each other. However, the enzyme activity of ${\beta}$-glucosidase and chitinase is low.

A Research on Private apparel Brand's Product Strategy in Discounted Stores. (할인점의 의류PB 상품전략에 관한 연구)

  • Choi, Sung-Sik;Kim, Pan-Jin;Lee, Sang-Youn
    • The Journal of Industrial Distribution & Business
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    • v.2 no.2
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    • pp.25-38
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    • 2011
  • After the financial crisis, what has been the rapidly growth of large supermarkets, stores, and restaurants linked to concerns that have already reached saturation point, but the new opening large supermarkets is expected to continue into the future. The major supermarkets are continue to grow outward but growth slowed. And that is expected to continue differentiation of the product, acceptance the customer needs, acquiring high margin of sales products. Then the ongoing development of PB brand is to be positioned effective marketing strategy for overcoming the period of slow growth. In addition, big three local supermarkets continue to launch a clothing PB brand, expansion and operation strategy for the situation and based on this study and the success of the domestic large-Mart's PB and PB identifying problem and the need for differentiation and profit for the successful strategy is to discuss in this study. This research looks at the concept of major market's private brand, the strategy, the success example and the prospects, and views the globally rapid-growing private brands, not only having the limited role of distributing the products as retailers, but also having a control of the distribution channel as a manufacturing company. World's major advanced distribution companies, to differentiate their companies' products and increase the profitability, are putting a lot of efforts into private brand products, and there are many good examples that are globalizing, externally expanding, and creating high financial results. In this research, we presented three major domestic discount stores as examples to show that there is a need for a differentiated private brand management strategy in the saturated discount store industry in Korea. Also, we aim to provide a new product strategy for the future that has been saturated with discount stores to the limit, by providing suggestions that private brand products can be used as weapons with the strongest competiveness in the retail industry through pursuing store differentiations from thorough market analysis and product researches, meeting the customers' needs, and obtaining high margins. PB products, particularly clothing design, a thorough market analysis and product development trends and customer needs to reflect the acquisition of High margin differentiated powerful products and sustainable growth through the stores, large supermarkets, congested, a new breakthrough that can give a good opportunity to provide implications discount stores, new product strategy based on ways to limit proposed. This study discount the major three companies studied, the less strain is a generalization. In the future, domestic and local discount store brand PB, SPA brand that the multinational comparative analysis of the value of the PB expansion strategy centered on clothing, additional studies will be needed.

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