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We have developed a cleaved amplified polymorphic sequence (CAPS) marker with the aim of identifying the best species among the different wild Lilium species native to Korea for breeding purpose. The CAPS marker is based on single nucleotide polymorphisms (SNPs) in the nucleotide sequences of the internal transcribed spacer genes, which are widely used in the breeding of angiosperms. We cloned and sequenced the ITS regions of nuclear ribosomal genes from 11 Lilium species and deduced the phylogenetic relationships among them. Phylogenetic analysis based on the maximum likelihood method showed that Lilium species formed a monophyletic clade and were divided into two groups that linked to the two sections, Martagon and Sinomartagon, out of seven sections of Lilium. The lengths of the ITS1 and ITS2 regions in the wild native Lilium species were distinct, and nucleotide polymorphisms were distinguished in these regions. A conserved region in the sequence was found from position 330–340 in all the species. We analyzed the SNP sites and found a suitable restriction endonuclease, Eco52I, having six base pair recognition site, to be suitable for the development of the CAPS marker. Among the 11 Lilium species, only the amplicon from the ITS region of L. hansonii contained the restriction site and produced two distinct bands of around 623 and 268 bp upon digestion with Eco52I. The genotyping of the 11 wild Lilium species by the developed marker can be recommended for breeding programs as it provides an indirect selection of plants and can effectively differentiate wild native lilies. The method using CAPS marker is simple, quick, and highly reliable for identifying the best species for efficient breeding of Lilium.

A mapped Chebyshev pseudo-spectral method is developed as one of the Fourier-spectral approaches and solves nonlinear PDE systems for unsteady flows and dynamic aero-elastic problem in a given time interval, where the flows or elastic motions can be periodic, nonperiodic, or periodic with an unknown frequency. The method uses the Chebyshev polynomials of the first kind for the basis function and redistributes the standard Chebyshev–Gauss–Lobatto collocation points more evenly by a conformal mapping function for improved numerical stability. Contributions of the method are several. It can be an order of magnitude more efficient than the conventional finite difference-based, time-accurate computation, depending on the complexity of solutions and the number of collocation points. The method reformulates the dynamic aero-elastic problem in spectral form for coupled analysis of aerodynamics and structures, which can be effective for design optimization of unsteady and dynamic problems. A limit cycle oscillation (LCO) is chosen for the validation and a new method to determine the LCO frequency is introduced based on the minimization of a second derivative of the aero-elastic formulation. Two examples of the limit cycle oscillation are tested: nonlinear, one degree-of-freedom mass–spring–damper system and two degrees-of-freedom oscillating airfoil under pitch and plunge motions. Results show good agreements with those of the conventional time-accurate simulations and wind tunnel experiments.

An efficient production system for optically pure l- and d-lactic acid (LA) from Jerusalem artichoke tuber powder (JAP) was developed by metabolic engineering of Kluyveromyces marxianus. To construct LA-producing strains, the ethanol fermentation pathway of K. marxianus was redirected to LA production by disruption of KmPDC1 and expression of l- and d-lactate dehydrogenase (LDH) genes derived from Lactobacillus plantarum under the control of the K. marxianus translation elongation factor 1α promoter. To further increase the LA titer, the l-LA and d-LA consumption pathway of host strains was blocked by deletion of the oxidative LDH genes KmCYB2 and KmDLD1. The recombinant strains produced 130g/L l-LA and 122g/L d-LA by direct fermentation from 230g/L JAP containing 140g/L inulin, without pretreatment or nutrient supplementation. The conversion efficiency and optical purity were ≫>95% and ≫>99%, respectively. This system using JAP and the inulin-assimilating yeast K. marxianus could lead to a cost-effective process for the production of LA.

Fructans are polymers of fructose that are present as storage carbohydrates in various plants. Jerusalem artichoke (Helianthus tuberosus L.) contains a high amount of inulin. Two enzymes are involved in inulin biosynthesis. The sucrose:sucrose 1-fructosyltransferase (1-SST) enzyme mainly catalyzes the synthesis of 1-kestose from sucrose. In the next step, fructan:fructan 1-fructosyltransferase (1-FFT) catalyzes the synthesis of inulin from 1-kestose. In this study, the Ht1-SST and Ht1-FFT genes were isolated from Jerusalem artichoke and expressed in potato (Solanum tuberosum L.), either separately or together, via Agrobacterium-mediated transformation. Transgenic potato tubers overexpressing Ht1-SST accumulated 1-kestose to a high level (up to 3.36 mg/g), while tubers overexpressing both Ht1-SST and Ht1-FFT accumulated up to 3.14 mg/g short-chain inulin-type fructans, with the degree of polymerization (DP) ranging from 3 to 5, excluding high DP inulins. Transgenic potato plants accumulated fructo-oligosaccharides to a high level, following the fructan biosynthetic pathway of Jerusalem artichoke, and therefore present a high potential for the mass production of inulin through established potato breeding and cultivation methods.

Plant possesses particular Golgi-resident cyclophilin 21 proteins (CYP21s) and the catalytic isomerase activities have a negative effect on ABA signalling gene expression during early seedling development. Cyclophilins (CYPs) are essential for diverse cellular process, as these catalyse a rate-limiting step in protein folding. Although Golgi proteomics in Arabidopsis thaliana suggests the existence of several CYPs in the Golgi apparatus, only one putative Golgi-resident CYP protein has been reported in rice (Oryza sativa L.; OsCYP21-4). Here, we identified the Golgi-resident CYP21 family genes and analysed their molecular characteristics in Arabidopsis and rice. The CYP family genes (CYP21-1, CYP21-2, CYP21-3, and CYP21-4) are plant-specific, and their appearance and copy numbers differ among plant species. CYP21-1 and CYP21-4 are common to all angiosperms, whereas CYP21-2 and CYP21-3 evolved in the Malvidae subclass. Furthermore, all CYP21 proteins localize to cis-Golgi, trans-Golgi or both cis- and trans-Golgi membranes in plant cells. Additionally, based on the structure, enzymatic function, and topological orientation in Golgi membranes, CYP21 proteins are divided into two groups. Genetic analysis revealed that Group I proteins (CYP21-1 and CYP21-2) exhibit peptidyl prolyl cis–trans isomerase (PPIase) activity and regulate seed germination and seedling growth and development by affecting the expression levels of abscisic acid signalling genes. Thus, we identified the Golgi-resident CYPs and demonstrated that their PPIase activities are required for early seedling growth and development in higher plants.

Jerusalem artichoke (Helianthus tuberosus L.), a plant of the Asteraceae family, is widely cultivated for its multiple pharmacological properties and is being developed as a renewable feedstock, as well as a source of biofuels and biochemicals for industrial applications. Despite its nutritional benefits and economic potential, transcriptomic and genomic information is scarce. In the present study, we performed phenotypic characterization and RNA-Seq analysis of three Jerusalem artichoke cultivars, “Purple Jerusalem artichoke” (PJA), “Hindung Jerusalem artichoke” (HJA) and “Dafeng Jerusalem artichoke” (DJA). The cultivars exhibited obvious differences in their responses to drought, high salinity and oxidative stress, as well as morphological variations. The PJA cultivar had the highest concentration of anthocyanin, and the DJA cultivar had the strongest tolerance to environmental stresses among the three cultivars. Based on the three assembled transcriptomes, we identified 2435, 3283 and 3657 putative cultivar-specific transcripts from leaf and tuber tissues in cultivars PJA, HJA and DJA, respectively, which enlarges the pool of transcriptomes available for Jerusalem artichoke. We also detected 11,319, 13,190 and 12,717 potential cultivar-specific simple sequence repeats (SSRs) from the transcriptomic data for, PJA, HJA and DJA, respectively. In addition, five SSRs were identified as candidate molecular markers for cultivar identification, as determined by genomic PCR analysis. Our comparative RNA-Seq analysis and de novo transcriptome assemblies constitute a comprehensive transcriptome resource and provide essential sequence information for identifying Jerusalem artichoke cultivars. These results should therefore be useful for future gene discovery, molecular studies and agricultural improvement of this important non-model species.

Transgenic potato (Solanum tuberosum L. cv. Desiree) plants expressing components of a novel cyanobacterial photorespiratory glycolate catabolism pathway were developed. Transgenic plant expressing glcD1 (glycolate dehydrogenase I) gene was referred to as synGDH and transgenic plants expressing gcl (glyoxylate carboligase) and tsr (tartronic semialdehyde reductase) genes simultaneously were designated as synGT. Both synGDH and synGT plants showed stable gene transformation, integration and expression. Enhanced glyoxylate contents in synGDH plants were detected as compared to synGT and non-transgenic (NT) plants. Phenotypic evaluation revealed that synGDH plants accumulated 11 % higher dry weight, while, tuber weight was 38 and 16 % higher than NT and synGT, respectively. Upon challenging the plants in high temperature and high light conditions synGDH plants maintained higher Fv/Fm and showed less bleaching of chlorophyll as compared to synGT and NT plants. These results indicate that genetic transformation of complete pathway in one plant holds promising outcomes in terms of biomass accumulation to meet future needs for food and energy.

In our current study, to develop an efficient regeneration and transformation system of Helianthus tuberosus, we verified effects of plant growth hormones, growth conditions, explant type, and transformation conditions. Leaf segments from regenerated shoots showed higher regeneration efficiency on MS basal medium containing 1.0 mg L−1 zeatin under darkness than those from maintained in vitro plant. To carry out transformation, various parameters including plant materials, Agrobacterium cell density, immersion time, and Agrobacterium strains (leaf segment, OD600-0.6, 60 min, and Agrobacterium tumefaciens LBA4404 harboring binary vector pCAMBIA1301) have been determined. The putatively transformed H. tuberosus were screened by survival rate and beta-glucuronidase (GUS) histochemical assay following two cycles of 3.0 mg L−1 hygromycin selection at callus stage. The presence of the selectable marker gene hygromycin phosphotransferase and the GUS reporter gene with intron was then confirmed by genomic PCR, Southern blot, reverse transcriptase PCR (RT-PCR) and GUS histochemical assay. The method presented here could be helpful in genetic improvement of H. tuberosus through efficient shoot regeneration and stable Agrobacterium-mediated transformation.

High-throughput sequencing revealed a tentative new badnavirus infecting Codonopsis lanceolata, provisionally named Codonopsis vein clearing virus (CoVCV). The complete 8,112-nt CoVCV genomic DNA sequence (GenBank accession: MK044821) comprises three open reading frames (ORFs) encoding conserved domains, with typical features of badnaviruses. Additionally, BLASTn searches indicated the CoVCV genome sequence is most similar to the grapevine vein clearing virus (GVCV) genome (72% identity and 46% query coverage). Moreover, the polyprotein encoded in CoVCV ORF3 is most similar to the corresponding protein of GVCV, with 60% amino acid sequence identity (89% query coverage). These results suggest that CoVCV is a new member of the genus Badnavirus in the family Caulimoviridae.

Diverse HPV subtypes are responsible for considerable disease burden worldwide, necessitating safe, cheap, and effective vaccines. The HPV minor capsid protein L2 is a promising candidate to create broadly protective HPV vaccines, though it is poorly immunogenic by itself. To create highly immunogenic and safe vaccine candidates targeting L2, we employed a plant-based recombinant protein expression system to produce two different vaccine candidates: L2 displayed on the surface of hepatitis B core (HBc) virus-like particles (VLPs) or L2 genetically fused to an immunoglobulin capable of forming recombinant immune complexes (RIC). Both vaccine candidates were potently immunogenic in mice, but were especially so when delivered together, generating very consistent and high antibody titers directed against HPV L2 (>1,000,000) that correlated with virus neutralization. These data indicate a novel immune response synergy upon co-delivery of VLP and RIC platforms, a strategy that can be adapted generally for many different antigens.

Isoflavonoids are a class of biologically active natural products that accumulate in soybean ( Glycine max L.) seeds during development, play vital roles in plant defense, and act as phytoestrogens with important human health benefits. Plant cell suspension cultures represent an excellent source of biologically important secondary metabolites. We found that methyl jasmonate (MJ) treatment increased isoflavone production in soybean suspension cell cultures. To investigate the underlying mechanism, we examined the expression of structural genes ( CHS6, CHS7, CHI1, IFS1, IFS2, IFMaT, and HID) in the isoflavonoid biosynthesis pathways in soybean suspension cells under various abiotic stress conditions. MJ treatment had the most significant effect on gene expression and increased the production of three glycosidic isoflavones (daidzin, malonyldaidzin, and malonylgenistin), with the maximum total isoflavone production (∼10-fold increase) obtained on day 9 after MJ application. MJ treatment significantly increased total phenolic contents and upregulated isoflavonoid biosynthesis genes, shedding light on the underlying mechanism.

Sophorae Radix (Sophora flavescens Aiton) has long been used in traditional medicine in East Asia due to the various biological activities of its secondary metabolites. Endogenous contents of phenolic compounds (phenolic acid, flavonol, and isoflavone) and the main bioactive compounds of Sophorae Radix were analyzed based on the qualitative HPLC analysis and evaluated in different organs and at different developmental stages. In total, 11 compounds were detected, and the composition of the roots and aerial parts (leaves, stems, and flowers) was significantly different. trans-Cinnamic acid and p-coumaric acid were observed only in the aerial parts. Large amounts of rutin and maackiain were detected in the roots. Four phenolic acid compounds (benzoic acid, caffeic acid, ferulic acid, and chlorogenic acid) and four flavonol compounds (kaempferol, catechin hydrate, epicatechin, and rutin) were higher in aerial parts than in roots. To identify putative genes involved in phenolic compounds biosynthesis, a total of 41 transcripts were investigated. Expression patterns of these selected genes, as well as the multiple isoforms for the genes, varied by organ and developmental stage, implying that they are involved in the biosynthesis of various phenolic compounds both spatially and temporally.

Background Vancomycin-resistant enterococci (VRE) infections have become a major healthcare-associated pathogen problem worldwide. Nosocomial VRE infections could be effectively controlled by screening patients at high risk of harboring VRE and thereby lowering the influx of VRE into healthcare centers. In this study, we evaluated factors associated with VRE colonization in patients transferred to emergency departments, to detect patients at risk for VRE carriage. Methods This study was conducted in the emergency department of a medical college-affiliated hospital in Korea. Every patient transferred to the emergency department and admitted to the hospital from January to December 2016 was screened for VRE using rectal cultures. In this cross-sectional study, the dependent variable was VRE colonization and the independent variables were demographic and clinical factors of the patients and factors related to the transferring hospital. Patients were divided into two groups, VRE and non-VRE, and previously collected patient data were analyzed. Then we performed logistic regression analyses of characteristics that differed significantly between groups. Results Out of 650 patients, 106 (16.3%) had positive VRE culture results. Significant variables in the logistic analysis were transfer from geriatric long-term care hospital (adjusted odds ration [aOR]: 8.017; 95% confidence interval [CI]: 1.378–46.651), hospital days (4–7 days; aOR: 7.246; 95% CI: 3.229–16.261), duration of antimicrobial exposure (1–3 days; aOR: 1.976; 95% CI: 1.137–3.436), and age (aOR: 1.025; 95% CI: 1.007–1.043). Conclusion VRE colonization in patients transferred to the emergency department is associated primarily with factors related to the transferred hospitals rather than demographic and clinical characteristics., Graphical Abstract

Developing rice lines with various amylose contents is necessary to diverse usages of rice in terms of raw materials for processed food production, and thereby to promote rice consumption in Korea. A rice mutant line, 'Namil(SA)-dull1' was established through sodium azide mutagenesis on 'Namil', a non-glutinous Korean Japonica rice cultivar. Namil(SA)-dull1' had dull endosperm characteristics and the evaluated amylose content was 12.2%. A total of 94 F2 progenies from a cross between 'Namil(SA)-dull1' and 'Milyang23', a non-glutinous Tongil-type rice cultivar, was used for genetic studies on the endosperm amylose content. Association analyses, between marker genotypes of 53 SSR anchor markers and evaluated amylose contents of each 94 F2:3 seeds, initially localized rice chromosome 6 as the harboring place for the modified allele(s) directing low amylose content of 'Namil(SA)-dull1'. By increasing SSR marker density on the putative chromosomal region followed by association analyses, the target region was narrowed down 0.94 Mbp segment, expanding from 28.95 Mbp to 29.89 Mbp, on rice chromosome 6 pseu- domolecule. Among the SSR loci, RM7555 explained 84.2% of total variation of amylose contents in the F2 population. Further physical mapping on the target region directing low amylose content of 'Namil(SA)-dull1' would increase the breeding efficiency in developing promising rice cultivars with various endosperm characteristics.

The soilborne fungus Fusarium oxysporum f. sp. lilii (Fol) is a serious threat to all lily cultivars, especially infecting bulbs and flowers. It has become increasingly important to develop varieties resistant against the bulb rot disease. Genetic diversity of cultivars and reliable screening methods are required for this purpose. Here, an efficient in vitro screening system for evaluating resistance to Fol in 38 in vitro-grown lily plants was investigated. Various factors including culture conditions of Fol, inoculum density, appropriate plant materials, inoculation method and duration, and incubation period of plant materials after inoculation were combined to optimize the screening method. As a result, we suggest optimal conditions for an in vitro screening system for the selection of Fol-resistant lily cultivars as follows. Fol was grown on potato dextrose agar (PDA) medium for 6 days at 25°C in darkness and used as working inoculation. Spore suspensions were prepared (inoculum density: 1.0 × 10 4 spores‧mL -1 ), and then leaf segments 1.5 × 2.0 cm 2 were inoculated by dipping for 22 hours at 25°C in dark. Later, leaves were cultured on 0.6% agar plates at 25°C and 50% humidity with a photoperiod of 16 hours light/8 hours dark (fluence rate of 40 μmol・m -2 ・s -1 ) to examine the progress of bulb rot. After 7 days, disease levels were classified into indices 1 (no symptom) to 6 (serious bulb rot). Soil inoculation of Fol carried out with resistant or susceptible lily cultivars that had been selected through in vitro screening confirmed the reproducibility of results. Therefore, the in vitro screening method established in this study is efficient and reliable for selection of lily cultivars resistant against bulb rot disease. Additional key words: bulb rot, disease index, disease resistance, Lillium spp. *Corresponding author: hyuns@kribb.re.kr ※ Received 15 May 2015; Revised 6 July 2015; Accepted 16 July 2015. 본 연구는 농림축산식품부・해양수산부・농촌진흥청・산림청 Golden Seed 프로젝트 원예종자사업단의 지원에 의해 이루어진 것임. C 2015 Korean Society for Horticultural Science 서 언 우리나라의 백합은 2013년 통계 기준 전년 대비 7.3% 재 배면적이 감소하였으나 절화 수출액의 32.5%를 차지하는 등 화훼 수출산업에 있어서 중요한 위치를 차지하고 있다 (MAFRA, 2014). 주로 비닐하우스나 온실에서 재배되는 백 합은 그 재배환경으로 인해 뿌리나 구근의 부패병에 많이 노출되어 과다한 농약사용이 문제시되고 있는 것이 현실이 다(Lebiush-Mordechai et al., 2014). 구근으로 번식하는 백 합은 꽃 재배기간뿐만 아니라 구근의 증식, 보관 기간에도 Penicillium sp.와 Fusarium oxysporum f. sp. lilii(Fol)균에 인한 감염으로 절화나 종구의 생산량과 품질에 치명적인 저 하가 발생한다(Loffler and Rumine, 1991). 이 구근부패병은 주로 종구나 지하부 줄기에서 감염이 일어나는데, 연작지 사질토에서 발생이 심하고 주로 토양으로부터 전염이 된다. 구근표면에 갈색 부정형 병반이 생기며, 심하면 구근 전체 Korean J. Hortic. Sci. Technol. 33(6), December 2015 884 가 갈색으로 변하여 썩고, 계속 진전되면 인편이 탈락하게 된다(Komada, 1975). 외관상으로는 아랫잎부터 점차 떨어 져버리고 포기 전체가 죽어가기도 한다. 고사한 줄기의 지 제부위에는 병원체의 균사가 하얗거나 붉은색으로 자라나 있는 것을 볼 수 있다. 주로 정식 전에 토양해충방제를 철저 히 하고 작업 중에 상하지 않도록 하여 상처부위를 통한 침 입을 예방하는 등의 방법으로 피해 정도를 최소화하고 있다. 그러나 이러한 방법보다도 더욱 효율적으로 구근부패병을 방제하기 위해서는 구근부패병에 대한 저항성이 있는 품종 을 선별하여 재배하는 것이 중요하며, 이를 위하여 각 품종 의 구근부패병 저항성 정도를 진단하기 위한 간편하고 민감

Early-maturing Korean Japonica cultivars and Jungmo1024 were used as plant materials to evaluate hightemperature tolerance during grain filling stage. National Institute of Crop Science (NICS) in Korea developed Jungmo1024, a mutant line from Namil (wild type) treated by using sodium azide (SA) as mutagen. To evaluate high-temperature tolerance, all cultivars were exposed to high-temperature (day / night ) and ordinary temperature (day / night ) during grain filling stage. In these conditions, we performed compared evaluation of grain shape such as length and width and grain quality by using a 1625 Cervitec grain inspector. High-temperature during grain filling stage caused decrease of grain shape. In grain shape such as length and width, the decrease rate of width (average 6.3%) was higher than the decrease rate of length (average 1.3%). Hence, high-temperature affected width of grain than length of grain. In addition, high-temperature showed a decided difference in rate of head rice between ordinary temperature (average 76.3%) and high-temperature (average 13.3%).As a result, Taebong, Ungwang, Manan, and Jungmo1024 seemed relatively a decent high-temperature tolerance than other cultivars. Especially, Jungmo1024 seemed remarkable rate of head rice (average ) than other cultivars under high-temperature. It is considered that a genetic trait of Jungmo1024 can be useful to improve breeding for high-temperature tolerance.

A majority of menopausal women experience serious depressive symptom because they are disturbed by their menstruation stop and by multiple physiological symptoms. The purpose of this study is to analyze the related factors of depressive symptom of menopausal women in Korea. A multiple regression model was used to study in association with depressive symptom, controlling for socio-economic characteristics, health status and behavior characteristics, women`s health, and family support characteristics. For the data, Menopausal women were 4,680 peoples were selected for this study. PASW 18.0 was used to study in association with affecting factors of depressive symptom in the menopausal women The study shows that Education level, Marital status, Income, Employment status, Activity limitations, Perceived health status, Perceived body Image recognition, Current smoking, AUDIT, Frequency of induced abortion, Frequency of eating out, Breakfast with family, Lunch with family were associated with depressive symptom of menopausal women in Korea.

Introduction High-throughput RNA sequencing (RNA-Seq) studies demonstrate that Alter-native Splicing (AS) is a widespread mechanism that enhances transcriptome diversity, particularly in plants exposed to environmental stress. In an attempt to determine the transcriptome and AS patterns of cabbage inbred line "HO" under Heat Stress (HS), RNA-Seq was carried out using HS-treated and con-trol samples. Genome-wide analysis indicated that AS is differentially regulated in response to HS. The number of AS events markedly increased in HS-treated samples compared to the control. Conclusion We identified 1,864 genes, including Heat shock transcription factor (Hsf) and heat shock protein (Hsp) genes, that exhibited >4-fold changes in expression upon exposure to HS. The enriched Gene Ontology (GO) terms of the 1,864 genes included 'response to stress/abiotic stimulus/chemical stimulus', among, which the genes most highly induced by HS encode small Hsps and Hsf proteins. The heat-induced genes also showed an increased number of AS events under HS conditions. In addi-tion, the distribution of AS types was altered under HS conditions, as the level of Intron Retention (IR) decreased, whereas other types of AS increased, under these conditions. Severe HS-induced AS was al-so observed in Hsfs and Hsps, which play crucial roles in regulating heat tolerance. Our results support the notion that AS of HS-related genes, such as HsfA2 and HsfB2a, are important for heat stress adapta-tion in cabbage.

Carbohydrates are the primary energy source for plant development. Plants synthesize sucrose in source organs and transport them to sink organs during plant growth. This metabolism is sensitive to environmental changes in light quantity, quality, and photoperiod. In the daytime, the synthesis of sucrose and starch accumulates, and starch is degraded at nighttime. The circadian clock genes provide plants with information on the daily environmental changes and directly control many developmental processes, which are related to the path of primary metabolites throughout the life cycle. The circadian clock mechanism and processes of metabolism controlled by the circadian rhythm were studied in the model plant Arabidopsis and in the crops potato and rice. However, the translation of molecular mechanisms obtained from studies of model plants to crop plants is still difficult. Crop plants have specific organs such as edible seed and tuber that increase the size or accumulate valuable metabolites by harvestable metabolic components. Human consumers are interested in the regulation and promotion of these agriculturally significant crops. Circadian clock manipulation may suggest various strategies for the increased productivity of food crops through using environmental signal or overcoming environmental stress.

We previously isolated Nicotiana benthamiana matrix metalloprotease 1 (NMMP1) from tobacco leaves. The NMMP1 gene encodes a highly conserved, Zn-containing catalytic protease domain that functions as a factor in the plant’s defense against bacterial pathogens. Expression of NMMP1 was strongly induced during interactions between tobacco and one of its pathogens, Phytophthora infestans. To elucidate the role of the NMMP1 in defense of N. benthamiana against fungal pathogens, we performed gain-of-function and loss-of-function studies. NMMP1-overexpressing plants had stronger resistance responses against P. infestans infections than control plants, while silencing of NMMP1 resulted in greater susceptibility of the plants to the pathogen. This greater susceptibility correlated with fewer NMMP1 transcripts than the non-silenced control. We also examined cell death as a measure of disease. The amount of cell death induced by the necrosis-inducing P. infestans protein 1, PiNPP1, was dependent on NMMP1 in N. benthamiana. Potato plants overexpressing NMMP1 also had enhanced disease resistance against P. infestans. RT-PCR analysis of these transgenic potato plants revealed constitutive up-regulation of the potato defense gene NbPR5. NMMP1-overexpressing potato plants were taller and produced heavier tubers than control plants. We suggest a role for NMMP1in pathogen defense and development.

CYP21-4 is a novel Golgi-localized cyclophilin protein involved in oxidative stress tolerance. Here, we generated transgenic plants overexpressing AtCYP21-4 and OsCYP21-4 in potato and rice, respectively. The stems and roots of AtCYP21-4–overexpressing potato plants were longer than those of wild-type (WT) plants, which resulted in heavier tubers. In vitro tuberization in the transgenic potato also resulted in significantly greater tuber number and weight, as well as a shorter time to microtuber formation. Similarly, OsCYP21-4–overexpressing transgenic rice plants had higher biomass and productivity with longer early-stage internodes than the WT and higher seed weight. Immunoblot analysis with CYP21-4 antibody showed that these productivity-enhancing phenotypes were associated with high CYP21-4s protein expression. Anatomically, transgenic potato stems exhibited higher lignin content in xylem cells and thicker leaves. In addition, relative content of mannosidic glycoproteins per unit of total protein was above 20% in transgenic potato tubers and rice grains. Based on these findings, we propose that CYP21-4s are involved in the growth and development of plant vegetative and storage tissues via their effects on glycoprotein abundance or glycan processing in the Golgi apparatus. Thus, increasing CYP21-4s expression in crops could represent an alternative way to increase crop productivity and yield.

Responding to the need for recombinant acidic fibroblast growth factor in the pharmaceutical and cosmetic industries, we established a scalable expression system for recombinant human aFGF using transient and a DNA replicon vector expression in Nicotiana benthamiana. Recombinant human-acidic fibroblast growth factor was recovered following Agrobacterium infiltration of N. benthamiana. The optimal time point at which to harvest recombinant human acidic fibroblast growth factor expressing leaves was found to be 4 days post-infiltration, before necrosis was evident. Commassie-stained SDS-PAGE gels of His-tag column eluates, concentrated using a 10 000 molecular weight cut-off column, showed an intense band at the expected molecular weight for recombinant human acidic fibroblast growth factor. An immunoblot confirmed that this band was recombinant human acidic fibroblast growth factor. Up to 10 µg recombinant human-acidic fibroblast growth factor/g of fresh leaves were achieved by a simple affinity purification protocol using protein extract from the leaves of agroinfiltrated N. benthamiana. The purified recombinant human acidic fibroblast growth factor improved the survival rate of UVB-irradiated HaCaT and CCD-986sk cells approximately 89 and 81 %, respectively. N. benthamiana-derived recombinant human acidic fibroblast growth factor showed similar effects on skin cell proliferation and UVB protection compared to those of Escherichia coli-derived recombinant human acidic fibroblast growth factor. Additionally, N. benthamiana-derived recombinant human acidic fibroblast growth factor increased type 1 procollagen synthesis up to 30 % as well as reduced UVB-induced intracellular reactive oxygen species generation in fibroblast (CCD-986sk) cells.UVB is a well-known factor that causes various types of skin damage and premature aging. Therefore, the present study demonstrated that N. benthamiana-derived recombinant human acidic fibroblast growth factor effectively protects skin cell from UVB, suggesting its potential use as a cosmetic or therapeutic agent against skin photoaging.

Tissue-specific promoters for efficient expression of transgenes at specific times or in specific plant tissues can be applied to develop new transgenic plants. To exploit a promoter capable of driving strong expression in potato tubers, we isolated the promoter region of the laccase gene from potato (Solanum tuberosum L. cv. Desiree) and characterized its activity in transgenic Solanaceae plants, such as potato, tobacco and tomato. The ability of the laccase promoter to induce the β-glucuronidase (GUS) reporter was evaluated in independent transgenic potato lines and compared with that of the constitutive CaMV35S promoter. To determine the tissue specificity of expression in transgenic potato, GUS levels in shoot tips, leaves, stems, roots and tubers were measured by histochemical analysis. The laccase promoter conferred tuber-specific expression in transgenic potato regardless of the developmental stage, and there was no GUS reporter expression in leaves, stems or roots. Serial 5′ deletion analysis of the laccase promoter revealed that the tuber-specific regulatory elements might be scattered throughout the promoter. The laccase promoter responded weakly to salt stress, mannitol stress, and mechanical wounding but not to cold stress in the leaves and stems of transgenic potato. In transgenic tobacco, weak GUS expression driven by the laccase promoter was detected throughout the entire plant, whereas in transgenic tomato, GUS expression was detected only in the roots and seeds. Our data show that the laccase promoter represents a feasible candidate to drive high and preferential expression of genes in potato tubers.

Transgenic potato was generated to express recombinant 5 repeated β-amyloid (Aβ) peptides, potential antigens to be applied as a preventive accine for Alzheimer’s disease using Agrobacterium mediated transformation. The Aβ peptides were fused to the human IgG Fc fragment enhancing protein and KDEL, which is the endoplasmic reticulum (ER) retention signal (5Aβ-FcK). The 5Aβ-FcK, was expressed under the control of the duplicated 35S promoter. PCR analysis confirmed the presence of the transgene in several transgenic potato lines. Southern blot analysis showed only a single gene copy number in transgenic line 22, whereas multiple gene copy numbers were shown for transgenic lines 31 and 44. Northern blot analysis showed that line 22 had stronger mRNA levels when compared to lines 31 and 44. Immunoblot analysis confirmed that the 5Aβ-FcK protein was expressed in the transgenic potato plant. These results indicate that 5Aβ fused to Fc can be expressed in potato plants.

Transgenic plants have been tested as an alternative host for the production and delivery of experimentaloral vaccines. Here, we developed transgenic potatoes that express the major antigenic sites A and D of the glycoproteinS from transmissible gastroenteritis coronavirus (TGEV-S 0.7 ) under three expression vector systems. The DNAintegration and mRNA expression level of the TGEV-S 0.7 gene were confirmed in transgenic plants by PCR and northernblot analysis. Antigen protein expression in transgenic potato was determined by western blot analysis. Enzyme-linkedimmunosorbent assay results revealed that based on a dilution series of Escherichia coli-derived antigen, the transgenicline P-2 had TGEV-S 70. protein at levels that were 0.015% of total soluble proteins. We then examined theimmunogenicity of potato-derived TGEV-S 70. antigen in mice. Compared with the wild-type potato treated group andsynthetic antigen treated group, mice treated with the potato-derived antigen showed significantly higher levels ofimmunoglobulin (Ig) G and IgA responses.Keywords : oral administration, potato-derived antigen, transmissible gastroenteritis virus, transgenic plant

This study was carried out to obtain basic information about the improvement of corn breeding in Cambodia. Different varieties of Corn seeds; 4 varieties from Korea and 2 check varieties were sown in the CARDI field in wet season, which is located at 11.4 latitude and 104.8 longitudes. The study results are summarized as follows: The precipitation during the experimental period in 2012 was 790 mm which is higher compared in 2011, which is 538 mm. The rainfall in August to September 2012 is three times than in same periods in 2011. Minimum temperature, accumulative temperature and solar radiation in 2012 were the same in 2011. The maximum temperature in 2012 is 41.8 compared to 33.3 in 2011 (which is over threshold temperature 35oC for corn). Due to heavy rains, soil moisture was not maintained effectively. High rainfall at a maximum 43mm per hour affect the drainage and influenced, the growth and root development. Seed weight of Korean varieties (175 kg~242 kg/10 a) were less than that of check varieties, of Cambodia and Thailand. The result showed a positive correlation between yield and agronomic characteristics which include seed weight, silking day, stem height, ear height rate, number of ear per 100 plants, ear length, ear width and weight of 100grain. Therefore, when agronomic characteristics increase, the yield of corn also increases. The study also showed a positive correlation coefficient between root characters, such as root length, fresh and dry root weight, and agronomic characters. It is therefore concluded that when root becomes better, agronomic characters are also better and consequently seed weight also increases. Likewise, CP888 variety was the best in root characters, and produces high yield. In contrast with the Korean varieties, which are not good in root characters, hence the yield decreases. The result of Korean varieties experiments in wet season (Kwangpyeongok, Cheongdaok, Dapyeongok, Andaok) are poorer than that of CP888 in terms of root, growth and yield characteristics. Therefore, Korea varieties provide fewer yields than CP888. The causes are estimated to be poor root due to a lot of rainfall and high temperatures in wet season. Experimental result illustrates that Korean varieties cannot adapt in Cambodia. To respond to this issue, we should develop adaptive variety to the Cambodia weather condition in wet season.

The complete genome sequence of ligustrum virus A (LVA) from a Ligustrum obtusifolium Sieb. & Zucc. plant was determined. The genomic RNA has 8,525 nucleotides, excluding the poly(A) tail, and consists of six open reading frames typical of members of the genus Carlavirus, family Betaflexiviridae. Phylogenetic analysis of the viral replicase and coat protein (CP) indicated that LVA is closely related to daphne virus S and helenium virus. The replicase and CP of LVA shared 44.73–52.35 % and 25.39–62.46 % amino acid identity, respectively, with those of other carlaviruses. These results suggest that LVA is a member of a distinct carlavirus species.

Alternative splicing (AS) is an important molecular mechanism by which single genes can generate multiple mRNA isoforms. We reported previously that, in Oryza sativa, the cyclophilin 19-4 (OsCYP19-4.1) transcript was significantly upregulated in response to cold stress, and that transgenic plants were cold tolerant. Here we show that, under cold stress, OsCYP19-4 produces eight transcript variants by intron retention and exon skipping, resulting in production of four distinct protein isoforms. The OsCYP19-4 AS isoforms exhibited different cellular localizations in the epidermal cells: in contrast to OsCYP19-4.1, the OsCYP19-4.2 and OsCYP19-4.3 proteins were primarily targeted to guard and subsidiary cells, whereas OsCYP19-4.5, which consists largely of an endoplasmic reticulum (ER) targeting signal, was co-localized with the RFP-BiP marker in the ER. In OsCYP19-4.2, the key residues of the PPIase domain are altered; consistent with this, recombinant OsCYP19-4.2 had significantly lower PPIase activity than OsCYP19-4.1 in vitro. Specific protein-protein interactions between OsCYP19-4.2/3 and AtRCN1 were verified in yeast two-hybrid (Y2H) and bimolecular fluoresence complementation (BiFC assays), although the OsCYP19-4 isoforms could not bind each other. Based on these results, we propose that two OsCYP19-4 AS isoforms, OsCYP19-4.2 and OsCYP19-4.3, play roles linking auxin transport and cold stress via interactions with RCN1.

Introduction: High-throughput RNA sequencing (RNA-Seq) studies demonstrate that Alter-native Splicing (AS) is a widespread mechanism that enhances transcriptome diversity, particularly in plants exposed to environmental stress. In an attempt to determine the transcriptome and AS patterns of cabbage inbred line “HO” under Heat Stress (HS), RNA-Seq was carried out using HS-treated and con-trol samples. Genome-wide analysis indicated that AS is differentially regulated in response to HS. The number of AS events markedly increased in HS-treated samples compared to the control. Conclusion: We identified 1,864 genes, including Heat shock transcription factor (Hsf) and heat shock protein (Hsp) genes, that exhibited >4-fold changes in expression upon exposure to HS. The enriched Gene Ontology (GO) terms of the 1,864 genes included ‘response to stress/abiotic stimulus/chemical stimulus’, among, which the genes most highly induced by HS encode small Hsps and Hsf proteins. The heat-induced genes also showed an increased number of AS events under HS conditions. In addi-tion, the distribution of AS types was altered under HS conditions, as the level of Intron Retention (IR) decreased, whereas other types of AS increased, under these conditions. Severe HS-induced AS was al-so observed in Hsfs and Hsps, which play crucial roles in regulating heat tolerance. Our results support the notion that AS of HS-related genes, such as HsfA2 and HsfB2a, are important for heat stress adapta-tion in cabbage.

Erythropoietin (EPO) is a glycoprotein hormone that regulates red blood cell formation in mammals. It also plays an important role in wound healing and the brain’s response to neuronal injury. Recombinant EPO has been produced in various hosts including Escherichia coli, insect cells, and yeast. Plant systems are also useful for the production of recombinant proteins. In this study, we used Nicotiana benthamiana for the production of recombinant human EPO (rhEPO) using an Agrobacterium-mediated transient expression system. We evaluated the effect of repeated agroinfiltration and Agrobacterium density on the rhEPO production. In addition, the rhEPO expression vector was coinfiltrated with a vector expressing cucumber mosaic virus suppressor 2b (CMV2b), which suppresses posttranscriptional gene silencing. We found that rhEPO expression was increased approximately 5.5-fold in N. benthamiana leaves coinfiltrated with CMV2b. In contrast, neither Agrobacterium density nor the number of infiltrations influenced rhEPO expression.

This study was performed to develop a therapeutic diet against aging and obesity, using Yangha(Zingiber mioga R.). Before development of a therapeutic diet, we performed cell viability assay, analysis of general composition, macrominerals and antioxidantive activities of Yangha. Based on the findings from analyzing the results, mook using Yangha powder(0~ 20%) was processed, and tested for quality characteristics such as color values, sensory evaluation and mechanical properties. The result of cell viability assay of myoga, using liver cells, revealed that within the concentration range from 500 ㎍/㎖ to 10,000 ㎍/㎖, cell survivability increased in line with the concentration rate. Therefore, it will not be harmful to consume it as food. Regarding the normal substance of myoga, the water substance of myoga was 94%, which exceeds that of ginger and tumeric with 89% and 83%, respectively. As for crude protein, fat, carbon hydrates and ash, myoga contained less than the other two, which I think is due to the high water substance. Regarding the minerals, potassium had the highest contents among macrominerals of 234.74 ㎎%. As for the antioxidant test, hydroxy radical scavenging activity and superoxide radical scavenging activity were shown. As for the production of Yangha mook(Yangha powder levels were 0~ 20%) for quality characteristics, the more of the powder, the less the L, but the greater the a and b values. Also, for the material property, an increased amount of the powder, resulted in chewiness and springiness, but less gumminess in a correlated manner. However, there were no significant differences in the springiness and cohesiveness in relation to the powder. For the sensory test, jelly type ZM5 with 5% powder showed highest overall preference. According to the sensory test, based on the powder substance, the jelly with 5% powder showed the highest overall score, including preference.

This study is to investigate consumer`s purchasing behavior by types of retail store and to conduct Importance-Performance Analysis(IPA) to exhort adequate competitive strategy to the traditional market. The results of this study are as follows. First, the result shows that consumer spends more time and less money at the traditional market due to the difficulties of store searching by store arrangement and construction of the market. Second, this study suggest that the traditional market needs to change their competitive strategy in same level with the discount stores, the traditional market can concentrate their capability on specialization and uniqueness of their product to meet the consumer`s needs. Third, according to the result on quadrant(concentrate here) of IPA, traditional market is marked on ease of refund, convenience of after service and parking facility, and cleanliness of bathroom while the discount store and the department store are marked on high credibility level and quality of product.

An agarolytic bacterium, designated as strain M5c(T), was isolated from sea sand in Jeju Island, Korea. This isolate was Gram-negative, positive for catalase and oxidase, rod and motile by means of monotrichous flagella. Strain M5c(T) has translucent or dark ivory colonies, forms a dent on an agar plate under colonies, and grows in the presence of 1-12% (w/v) NaCl and at 10-37°C. This isolate hydrolyzes agar, alginic acid, carboxymethyl (CM)-cellulose and starch. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain M5c(T) can be considered as a species within the genus Gilvimarinus, being most closely related to Gilvimarinus chinensis QM42(T), with a 16S rRNA gene sequence similarity of 95.6%. The major cellular fatty acids were C16:1ω7c and/or iso-C15:0 2OH (33.5%), C16:0 (26.5%) and C18:1ω7c (14.1%). The DNA G+C content was 53.8 mol%. Based on these polyphasic data, strain M5c(T) should be classified as a novel species, for which the name Gilvimarinus agarilyticus sp. nov. is proposed. The type strain for the novel species is M5c(T) (= KCTC 23325(T) = NCAIM B 02425(T)).

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 dif-ferent 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 im-munization 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 anti-gens 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 devel-opment of oral vaccines.

Antibodies are powerful and versatile tools to play a critical role in the diagnosis and treatment of many diseases. Their application has been enhanced significantly with the advanced recombinant DNA and heterologonous expression technologies, allowing to produce immunotherapeutic proteins with improved biofunctional properties. However, with currently available technologies, mammalian cell-based therapeutic antibody production, as an alternative for production in humans and animals, is often not plentiful for passive immunotherapeutics in treatment of many diseases. Recently, plant expression systems for therapeutic antibodies have become well-established. Thus, plants have been considered to provide an attractive alternative production system for therapeutic antibodies, as plants have several advantages such as the lack of human pathogens, and low cost of upstream production and flexible scale-up of highly valuable recombi-nant glycoproteins. Recent adva nces in modification of post- translational processing for human-like glycosylation in transgenic plants will make it possible that plant can become a suitable protein expression system over the animal cell- based current production system. This review will discuss recent advances in plant expression technology and issues for their application to therapeutic antibody production.