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We report a hexagonal GaN nanorod-based two-dimensional photonic crystal (PhC) slab for phosphor-conversion white light emitting devices analyzed by three-dimensional finite-difference time-domain simulation; this slab has a broad reflection band at yellow wavelength with low Fabry-Pérot background at normal incidence. For practical use as a wavelength-selective reflector, a buffer layer under the PhC slab is employed to sustain the nanorods in the PhC slab. However, we observed that the buffer layer placed below the slab destroys the broad reflection band due to evanescent coupling of electromagnetic field in the slab and the buffer layer. By introducing small-sized base pillars between the slab and the buffer layer, we could decouple the interaction between the slab and the buffer layer and maintain the broad reflection band without any unexpected dips. Since this GaN nanorod-based PhC slab is designed for practical light emitting devices by considering dielectric and transparent conducting layers, this structure is directly applicable for developing hybrid white light emitting devices having both an (active) blue-color-emitting nanorod emitters and a (passive) normal reflector of phosphor emission.

Zoysiagrass (Zoysia japonica Steud.) is an important turfgrass species used in golf courses and athletic fields. However, zoysiagrass is susceptible to large patch disease caused by Rhizoctonia solani AG2-2 (IV). Chitinases are pathogen-related (PR) proteins induced by viruses, bacteria, and fungi that hydrolyze chitin. Recently, we isolated a class II chitinase gene (Zjchi2) from zoysiagrass. The purified recombinant Zjchi2 showed broad-spectrum activity against various fungi, including R. solani AG2-2 (IV). In the current study, we generated transgenic zoysiagrass overexpressing Zjchi2 and then verified the resistance of transgenic plants to R. solani AG2-2 (IV). Polymerase chain reaction and Southern blot hybridization showed the integration of transgenes in zoysiagrass genomes and constitutive expression of Zjchi2, respectively. Antifungal activity was enhanced significantly in the transgenic zoysiagrass compared with wild-type plants. To our knowledge, this report is the first on the antifungal activity of a class II chitinase in transgenic zoysiagrass.

한국형 잔디에서는 다른 병에 비해 진전 속도가 빠르고 주로 뿌리에서부터 발병하여 잔디를 고사시키고 발병 후 구제하기 매우 어려운 라이족토니아잎마름병(라지패취)이 큰 문제로 대두되고 있다. 라이족토니아잎마름병(라지패취)은 Rhizoctonia solani AG2-2 (IV)병원균에 의해 발생하는데, 이 병원균에 강한 내병성 들잔...

This study aims to establish a system for the rapid discrimination of Zoysia species using metabolite fingerprinting of FT-IR spectroscopy combined with multivariate analysis. Whole cell extracts from leaves of 19 identified Zoysia japonica, 6 identified Zoysia sinica, and 38 different unidentified Zoysia species were subjected to Fourier transform infrared spectroscopy (FT-IR). PCA (principle component analysis) and PLS-DA (partial least square discriminant analysis) from FT-IR spectral data successfully divided the 25 identified turf grasses into two groups, representing good agreement with species identification using molecular markers. PC (principal component) loading values show that the region of the FT-IR spectra are important for the discrimination of Zoysia species. A dendrogram based on hierarchical clustering analysis (HCA) from the PCA and PLS-DA data of turf grasses showed that turf grass samples were divided into Zoysia japonica and Zoysia sinica in a species-dependent manner. PCA and PLS-DA from FT-IR spectral data of Zoysia species identified and unidentified by molecular markers successfully divided the 49 turf grasses into Z. japonica and Z. sinica. In particular, PLS-DA and the HCA dendrogram could mostly discriminate the 47 Z. japonica grasses into two groups depending on their origins (mountainous areas and island area). Considering these results, we suggest that FT-IR fingerprinting combined with multivariate analysis could be applied to discriminate between Zoysia species as well as their geographical origins of various Zoysia species.

Zoysiagrass (Zoysia Japonica Steud.) is used frequently in golf courses and athletic fields. However, Zoysiagrass suffers from large-patch disease caused by Rhizoctonia solani AG2-2 (IV), which results in physical and economic loss. In this study, two full-length chitinase genes encoding pathogen-related proteins were isolated from zoysiagrass. Structural and expression analyses of these genes were carried out. The two isolated chitinases were classified into class Ib (Zjchi1) and class II (Zjchi2). Zjchi1 and, Zjchi2 expression was high in root and stolen and was induced in seedlings by Rhizoctonia solnai AG2-2 (IV) infection. To assess their antifungal activity, the two chitinases were overexpressed in Escherichia coli and purified using Ni2+ and glutathione affinity column chromatography. The purified recombinant chitinases showed broad-spectrum antifungal activity against Rhizoctonia solnai AG2-2 (IV), Rhizoctonia solnai AG-1 (IA), Rhizoctonia cerealis, Botrytis cinerea, Fusarium culmorum, Fusarium graminearum and Trichoderma reesei.

Leaf discs from ‘Yeobong’ and ‘Maehyang’ strawberry plants were used as explants for transformation. The Agrobacterium tumefaciens strain EHA105 harboring the monellin gene under the control of the CaMV 35S promoter was used in co-cultivation experiments. The frequencies of callus formation and plant regeneration from leaf explants after co-cultivation in ‘Yeobong’ were higher than those of ‘Maehyang’. These transgenic plants showed normal growth patterns and flowering. PCR and Southern hybridization confirmed that 1 to 2 copies of the monellin gene were integrated into genome of the transgenic strawberry plants. Northern blot analysis confirm that the transcripts were expressed in transgenic strawberry plants. Although long-term subcultured transgenic strawberry plants showed a phenomenon to escape the transgene, the transformation system established in this study provides new opportunities for genetic improvement of strawberry plants.

This study assessed the effect of supplementation of novel transgenic phytase on growth performance and bone mineralization in Korean native broiler chickens. The experiment was designed using four dietary groups: those with a diet supplemented with (A) recombinant phytase, (B) transgenic phytase from the plant Lemna minor, (C) or wild-type L. minor as well as (D) a control group that was supplemented with commercially available feed. Three hundred 1-day-old Korean native broiler chicks were used and divided into these four dietary treatment groups having three replicates of 25 birds each (n = 75). The results showed increases in growth performance and bone mineralization in Groups B and C; compared with Groups A and D. Hematological analyses revealed notable contrasts in erythrocyte sedimentation rate, red blood cell count, and hemoglobin levels among the experimental groups, whereas no impacts of dietary treatment were observed on total eosinophil, lymphocyte, heterophil, monocyte, and basophil levels. The relative expression profiling of candidate genes showed that the genes involved in growth response, meat quality, and P–Ca metabolism were significantly highly expressed in the phytase-supplemented groups. Hence, it is suggested that dietary supplementation with transgenic phytase plant L. minor for enhancing growth performance is a promising new approach in the broiler feed industry. To the best of our knowledge, we report here the most comprehensive analysis using a broiler model that provides a workable platform for further research on the cost-effective production of feed with different compositions that might be beneficial in the livestock feed industry.

A full-length phytase gene (phy) of Aspergillus nidulans was amplified from the cDNA library by polymerase chain reaction (PCR), and it was introduced into a bacterial expression vector, pET-28a. The recombinant protein (rPhy-E, 56 kDa) was overexpressed in the insoluble fraction of Escherichia coli culture, purified by Ni-NTA resin under denaturing conditions and injected into rats as an immunogen. To express A. nidulans phytase in a plant, the full-length of phy was cloned into a plant expression binary vector, pPZP212. The resultant construct was tested for its transient expression by Agrobacterium-infiltration into Nicotiana benthamiana leaves. Compared with a control, the agro-infiltrated leaf tissues showed the presence of phy mRNA and its high expression level in N. benthamiana. The recombinant phytase (rPhy-P, 62 kDa) was strongly reacted with the polyclonal antibody against the nonglycosylated rPhy-E. The rPhy-P showed glycosylation, two pH optima (pH 4.5 and pH 5.5), an optimum temperature at 45 similar to 55 degrees C, thermostability and broad substrate specificities. After deglycosylation by peptide-N-glycosidase F (PNGase-F), the rPhy-P significantly lost the phytase activity and retained 1/9 of the original activity after 10 min of incubation at 45 degrees C. Therefore, the deglycosylation caused a significant reduction in enzyme thermostability. In animal experiments, oral administration of the rPhy-P at 1500 U/kg body weight/day for seven days caused a significant reduction of phosphorus excretion by 16% in rat feces. Besides, the rPhy-P did not result in any toxicological changes and clinical signs.

The haustorium is a specific organ for penetration of host tissue in parasitic plants. To better understand the molecular mechanisms of the establishment of host-parasite relationships using gene expression profiles, early stage haustorium germination from mature mistletoe seeds was examined. We have generated a cDNA library that is an excellent source of expressed sequence tags (ESTs) specifically related to haustorium development and host penetration. We analyzed 4,771 ESTs derived from a mistletoe (Viscum album) haustorium cDNA library. Cluster analysis identified 727 contigs (23.88%) and 2,317 singletons (76.12%), yielding a total of 3,044 unique genes. Among these genes, 1,760 clones (57.8%) showed significant similarity to known genes; the remaining 1,284 clones were novel. Annotation of genes with Gene Ontology indicated that the most abundant ESTs reveals that defense against biotic / abiotic stresses, primary metabolic processes, cell wall loosening and modification is critical for haustorium development and establishment of the host-parasite connection. Especially, it is likely that xyloglucan endotransglycosylases (XETs), glucanase, expansins and other cell wall hydrolases cooperate in cell wall modification during the stages of host-parasite connection. Functional expression of full-length forms corresponding to target ESTs was applied to characterize haustorium genes. This EST information will help gene discovery and characterization and provide promising targets for genetic engineering. The EST analysis presented here represents the first reported transcriptome data for V. album var. coloratum. Although the number of ESTs analyzed may not be sufficient to completely elucidate the mechanism of the host-parasite connection, the integrated approaches reported here represent an essential step toward understanding haustorium development in parasitic plants and provide a valuable resource for defining molecular mechanisms in the host-parasite connection.

An efficient in vitro protocol has been established for somatic embryogenesis and plantlet conversion of Korean wild ginseng (Panax ginseng Meyer). Wild-type and mutant adventitious roots derived from the ginseng produced calluses on Murashige and Skoog (MS) medium supplemented with 0.5 mg/L 2,4-dichlorophenoxyacetic acid and 0.3 mg/L kinetin; 53.3% of the explants formed callus. Embryogenic callus proliferation and somatic embryo induction occurred on MS medium containing 0.5 mg/L 2,4-dichlorophenoxyacetic acid. The induced somatic embryos further developed to maturity on MS medium with 5 mg/L gibberellic acid, and 85% of them germinated. The germinated embryos were developed to shoots and elongated on MS medium with 5 mg/L gibberellic acid. The shoots developed into plants with well-developed taproots on one-third strength Schenk and Hildebrandt basal medium supplemented with 0.25 mg/L 1-naphthaleneacetic acid. When the plants were transferred to soil, about 30% of the regenerated plants developed into normal plants.

Future technologies require faster data transfer and processing with lower loss. A photonic diode could be an attractive alternative to the present Si-based electronic diode for rapid optical signal processing and communication. Here, we report highly asymmetric photonic diode behavior with low scattering loss, from tapered core-shell quantum well semiconductor rods that were fabricated to have a large gradient in their bandgap energy along their growth direction. Local laser illumination of the core-shell quantum well rods yielded a huge contrast in light output intensities from opposite ends of the rod.

Luminescence and carrier dynamics of GaN/InGaN multi-quantum-well coaxial nanorods (MCNRs) were studied by means of photoluminescence (PL), cathodoluminescence (CL) and time-resolved PL (TRPL). The PL of as-grown MCNRs showed an intense blue emission together with broad emission at longer wavelengths. CL measurements of several single MCNRs attributed the broad emission to the wetting layer frequently observed in vapor–liquid–solid-grown nanorods. Non-single exponential intensity decays were observed by TRPL, which were ascribed to the In fluctuation in the InGaN alloy. Radiative and non-radiative lifetimes were then calculated via a stretched exponential model. An ultrafast carrier lifetime in the range of a few tens of picoseconds along with a high internal quantum efficiency (IQE) of about 59% resulted. The ultrafast carrier lifetimes were attributed to the improvement in the carrier collection efficiency due to the radial heterostructuring of GaN with InGaN shells, while the high IQE implied that carriers were mostly recombined radiatively. This study reveals that the coaxial growth of InGaN with GaN nanorods resulted in an ultrafast carrier lifetime and a luminescence efficiency that could be controlled by adjusting the growth temperature gap between the GaN and the InGaN.

White light emitting InGaN nanostructures hold a key position in future solid-state lighting applications. Although many suggested approaches to form group III-nitride vertical structures have been reported, more practical and cost effective methods are still needed. Here, we present a new approach to GaN/InGaN core-shell nanostructures at a wafer level formed by chemical vapor-phase etching and metal-organic chemical vapor deposition. Without a patterning process, we successfully obtained high quality and polarization field minimized In-rich GaN/InGaN core-shell nanostructures. The various quantum well thicknesses and the multi-facets of the obelisk-shaped core-shell nanostructures provide a broad spectrum of the entire visible range without changing the InGaN growth temperature. Due to their high crystal quality and polarization field reduction, the core-shell InGaN quantum wells show an ultrafast radiative recombination time of less than 200 ps and uniformly high internal quantum efficiency in the broad spectral range. We also investigated the important role of polarization fields in the complex recombination dynamics in InGaN quantum wells.

In Korea, there are debris-flow disasters induced by typhoon and localized torrential rainfall annually. There are particularly extensive debris-flow disasters in Gangwon-do because of its geomorphological characteristics; the extensive coverage of mountainous region, steep slope, and shallow soil. In this paper, we constructed a GIS database about topological characteristics of debris-flow basin in Gangwon-do by years of field survey. Also, we conducted frequency analysis based on this database with the digital forest type map and the digital soil map. We analyzed frequencies of debris-flow by simple count for topological characteristics, whereas we analyzed by considering an area ratio based on GIS for physiognomic and geologic characteristics. We used slope, aspect, width, depth and destruction shapes for analysis about topological characteristics of debris-flow basin. Also we used attributes of forest physiognomy, diameter, age, and density about physiognomic characteristics, and i n terms of geologic characteristics, we used attributes of drainage class, effective soil depth, subsoil properties, subsoil grave content, erosion class, parent material of soil, and topsoil properties. In consequence, we figured out topographic, forest physiognomic, and geologic characteristics of debris-flow basin. This result is applicable to establish a rational disaster prevention policy as a fundamental information.

A dislocation-eliminating chemical control method for high-quality GaN nanostructures together with various types of InGaN quantum well structures are demonstrated using a chemical vapor-phase etching technique. Unlike chemical wet etching, chemical vapor-phase etching could efficiently control the GaN and form various shapes of dislocation-free and strain-relaxed GaN nanostructures. The chemically controlled GaN nanostructures showed improved crystal quality due to the selective etching of defects and revealed various facets with reduced residual strain via the facet-selective etching mechanism. These structural properties derived excellent optical performance of the GaN nanostructures. The chemical vapor-phase etching method also showed possibilities of the fascinating applications for high-efficiency InGaN quantum well structures, such as InGaN quantum well layer on void embedded GaN layer, InGaN quantum well embedded GaN nanostructure, and InGaN/GaN core/shell nanostructure.

Photoinhibition is the inhibition of photosynthesis by excessive light resulting in the reduction of plant growth. Exposure to additional stress factors during exposure to light increases the potential for photoinhibitory effects. Reversible photoinhibition is indicative of a protective mechanism aimed at dissipating excess light energy, while irreversible photoinhibition indicates damage to the photosynthetic systems. The present review summarizes the physiological mechanisms of photoinhibition and discusses the interaction between light and other stress factors. In addition, some of the features and strategies that help plants avoid or restrict the occurrence of photoinhibition are analyzed. Most of these defense mechanisms are associated with the dissipation of excessive energy such as heat. Therefore, these mechanisms would regulate the carbon available to the plant by the output ratio of ATP/NADPH to the stressful environmental conditions. Understanding these mechanisms can help avoid plant cell death and increase plant productivity.

This study describes conditions for the mass production of mutant hairy root lines by co-cultivation with A. rhizogenes harboring the activation tagging vector pHC7. Various sources of explants were subjected to genetic transformation with A. rhizogenes to determine optimum conditions and cultivar for the highest frequency of hairy root formation on explants. Hairy root formation also were investigated in transformed hairy roots grown in various culture media. Finally, a total of approximately 2,500 lines of hairy root mutants were produced in this study. A managing system for metabolomics in hairy root lines also were established. These hairy root lines will be useful to determine functions of genes relating biosynthesis pathway of secondary metabolites.

Electrically driven quantum dot, wire, and well hybrid light-emitting diodes are demonstrated by using nanometer-sized pyramid structures of GaN. InGaN quantum dots, wires, and wells are formed at the tops, edges, and sidewalls of pyramids, respectively. The hybrid light-emitting diodes containing low-dimensional quantum structures are good candidates for broad-band highly efficient visible lighting sources.

With the purpose of improving ginsenoside content in adventitious root cultures of Korean wild ginseng (Panax ginseng Meyer), the roots were treated with different dosages of γ-ray (5, 10, 25, 50, 75, 100, and 200 Gy). The growth of adventitious roots was inhibited at over 100 Gy. The irradiated adventitious roots showed significant variation in the morphological parameters and crude saponin content at 50 to100 Gy. Therefore, four mutant cell lines out of the propagation of 35 cell lines treated with 50 Gy and 100 Gy were selected on the basis of phenotypic morphology and crude saponin contents relative to the wild type control. The contents of 7 major ginsenosides (Rg 1 , Re, Rb 1 , Rb 2 , Rc, Rf, and Rd) were determined for cell lines 1 and 3 from 100 Gy and lines 2 and 4 from 50 Gy treatments. Cell line 2 showed more secondary roots, longer length and superior growth rate than the root controls in flasks and bioreactors. Cell line 1 showed larger average diameter and the growth rate in the bioreactor was comparable with that of the control but greater in the flask cultured roots. Cell lines 1 and 2, especially the former, showed much more ginsenoside contents than the control in flasks and bioreactors. Therefore, we chose cell line 1 for further study of ginsenoside contents. The crude saponin content of line 1 in flask and bioreactor cultures increased by 1.4 and 1.8-fold, respectively, compared to the control. Total contents of 7 ginsenoside types (Rg 1 , Re, Rb 1 , Rb 2 , Rc, Rf, and Rd) increased by 1.8 and 2.3-fold, respectively compared to the control. Crude saponin and ginsenoside contents in the bioreactor culture increased by about 1.4-fold compared to that the flask culture.

Transgenic zoysiagrass ( Zoysia japonica Steud.) expressing the bar gene inserted in the plant genome has been generated previously through Agrobacterium tumefaciens - mediated transformation. The GM zoysiagrass (event: JG21) permits efficient management of weed control of widely cultivated zoysiagrass fields, reducing the frequency and cost of using various herbicides for weed control. Now we have carried out the environmental risk assessment of JG21 prior to applying to the governmental regulatory agency for the commercial release of the GM turf grass outside of test plots. The morphological phenotypes, molecular analysis, weediness and gene flow from each test plot of JG21 and wild-type zoysiagrasses have been evaluated by selectively analyzing environmental effects. There were no marked differences in morphological phenotypes between JG21 and wild-type grasses. The JG21 retained its stable integration in the host plant in T 1 generation, exhibiting a 3:1 segregation ratio according to the Mendelian genetics. We confirmed the copy number (1) of JG21 by using Southern blot analysis, as the transgenic plants were tolerant to ammonium glufosinate throughout the culture period. From cross-fertilization and gene flow studies, we found a 9% cross-pollination rate at the center of JG21 field and 0% at distances over 3 m from the field. The JG21 and wild-type zoysiagrass plants are not considered “weed” because zoysiagrasses generally are not dominant and do not spread into weedy areas easily. We assessed the horizontal gene transfer (HGT) of the transgene DNA to soil microorganisms from JG21 and wild-type plants. The bar gene was not detected from the total genomic DNA extracted from each rhizosphere soil of GM and non-GM Zoysia grass fields. Through the monitoring of JG21 transgene’s unintentional release into the environment, we found no evidence for either pollen mediated gene flow of zoysiagrass or seed dispersal from the test field within a 3 km radius of the natural habitat.

Members of the APETALA1 (AP1)/FRUITFULL (FUL)-like gene family of MADS-box genes play important roles in controlling the development of floral organs. To understand the molecular mechanisms of floral development in orchid, we isolated and characterized a Phalaenopsis AP1/FUL-like gene, PhalFUL. The results of phylogenetic analysis indicated that PhalFUL is in the monocots group of AP1/FUL-like gene. PhalFUL transcripts were detected in the flower buds, but not in vegetative organs. Moreover, in situ hybridization experiments revealed PhalFUL hybridization signals in all floral organ primordia at a very early stage of floral development, and continued expression in the column of whorls 3 and 4 until late developmental stages. These expression patterns were similar to those of the FUL-like genes in Arabidopsis (FUL) and Antirrhinum (DEFH28), suggesting that the PhalFUL is similar in function to FUL and DEFH28.

Arabidopsis thaliana dynamin-like protein 1E (ADL1E) is known to regulate mitochondrial elongation. The adl1e mutant has no morphological phenotype, and the growth and photosynthetic activity of the mutant are similar to those of the wild type. Leaf O2 uptake, which is supported by mitochondrial activity in the dark, is increased 1.7-fold by mutation in adl1e gene. The ATP content in the dark of guard and mesophyll cell protoplasts (GCPs and MCPs, respectively) was 2.5- to 4-fold higher in GCPs of the mutant and the wild type, and increased upon the addition of glucose in both genotypes. Oligomycin, an inhibitor of mitochondrial ATPase, suppressed ATP synthesis in both GCPs and MCPS isolated from adl1e plants, indicating that mutant had higher mitochondrial activity. The stomatal apertures of mutant and wild-type plants were then analyzed in vitro. In the light, the stomata of both genotypes showed similar patterns of opening. However, in the dark response, the stomata of the adl1e mutant closed faster than did those of the wild type. Oligomycin severely inhibited dark-induced stomatal closure in both cell types. The results suggest that stomatal closure in the dark is governed by cytosolic ATP concentration, which is stimulated by mitochondrial activity.

We assessed whether the cow1 mutant defects are associated with growth of Tos17 and T-DNA insertional rice in blue light (BL). Growth of oscow1 mutants which encoded a member of the YUCCA protein family was retarded in BL. Root to shoot ratios of the mutants were reduced about 2 times lower in the absence of NAA and about 2.5 times lower in the presence of NAA; the shoot growth was not significantly changed by NAA addition. Photosynthetic activity of the mutants was however inhibited in high light. Pigment analysis showed significant difference between wild-type (Chl a:b = 3.02) and mutants (3.84). Carotenoid contents of the mutants were also decreased considerably, implying the involvement of cow1 in pigment formation. These findings lead us to suggest that the growth retardation of oscow1 mutant plants by BL results from the difference of photosynthetic activity in part.

The rice CHLH gene encodes the Mg2+-chelatase H subunit, which is involved in chlorophyll biosynthesis. Growth of the chlorophyll-deficient oschlh mutant is supported by mitochondrial activity. In this study, we investigated the activity of mitochondrial respiration in the illuminated leaves during oschlh seedling development. Growth of mutant plants was enhanced in the presence of 3% sucrose, which may be used by mitochondria to meet cellular energy requirements. ATP content in these mutants was, however, significantly lowered in light conditions. Low cytosolic levels of NADH in illuminated oschlh mutant leaves further indicated the inhibition of mitochondrial metabolism. This down-regulation was particularly evident for oxidative stress-responsive genes in the mutant under light conditions. Hydrogen peroxide levels were higher in oschlh mutant leaves than in wild-type leaves; this increase was largely caused by the impairment of the expression of the antioxidant genes, such as OsAPX1, OsRAC1, and OsAOXc in knockout plants. Moreover, treatment of mesophyll protoplasts with ascorbic acid or catalase recovered ATP content in the mutants. Taken together, these results suggest that the light-mediated inhibition of mitochondrial activity leads to stunted growth of CHLH rice seedlings.

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

Activation tagging that uses T-DNA vectors con-taining multimerized transcriptional enhancers from the cau-liflower mosaic virus (CaMV) 35S gene is a powerful tool to determine gene function in plants. This approach has been successfully applied in screening various types of mutations and cloning the corresponding genes. We generated an acti-vation tagged hairy root pool of ginseng ( Panax ginseng C.A. Meyer) in an attempt to isolate genes involved in the bio-synthetic pathway of ginsenoside (triterpene saponin), which is known as the major active ingredient of the root. Quanti-tative and qualitative variation of ginsenoside in activation tagged hairy root lines were profiled using LC/MS. Metabolic profiling data enabled selection of a specific hairy root line which accumulated ginsenoside at a higher level than other lines. The relative expression level of several genes of tri-terpene biosynthetic pathway in the selected hairy root line was determined by real time RT-PCR. Overall results suggest that the activation tagged ginseng hairy root system described in this study would be useful in isolating genes involved in a complex metabolic pathway from genetically intractable plant species by metabolic profiling.

High frequency plant proliferation system via direct frond regeneration of endemic duckweed plants Lemna paucicostata and Spirodela polyrhiza was established. Fronds of L. paucicostata and S. polyrhiza were able to multiply half-strength MS basal medium without plant growth regulators. However, addition of BA at a range of 1 to 3 mg/L was more effective than high concentration of BA treatments for fronds proliferation. Also half-strength MS salts was suitable for the fronds proliferation. Increase of salts concentration had inhibitory effect on fronds proliferation. Also the frequency of callus formation from fronds of L. paucicostata was 3.3%, when they cultured onto 1/2 MS medium supplemented with 1 mg/L of BA. Similarly the frequency of callus formation from S. polyrhiza was very low. After subculture of white globular structures derived from fronds of L. paucicostata, numerous globular somatic embryos and calluses were developed onto the surface of fronds. However these somatic embryos did not fully develop into normal plants when transferred to 1/2 MS basal medium. Therefore direct frond regeneration system was more efficient for mass proliferation of L. paucicostata and S. polyrhiza. The plant regeneration system of L. paucicostata and S. polyrhiza established in this study, might be applied to mass proliferation and genetic transformation for molecular breeding.

The earthworm fibrinolytic enzyme, which belongs to a group of serine proteases with strong fibrinolytic activity, has been used as an oral drug for prevention and treatment of thrombosis in East Asia. Fibrizyme is a fibrinolytic enzyme isolated from the earthworm Eisenia andrei. Here we report genetic engineering of tobacco plastids with stable integration of the fibrizyme gene into the tobacco chloroplast genome. A plastid transformation vector was constructed by introducing various regulatory elements into fibrizyme cDNA. This vector was delivered by particle bombardment into tobacco leaf explants and plastid-transformed plants were subsequently regenerated into whole plants through several rounds of selection. We confirmed stable integration of the fibrizyme gene into the tobacco plastid genome by PCR and Southern blot analyses. Northern and Western blot analyses revealed that mRNA and protein of recombinant fibrizyme were highly expressed in transformed tobacco plants.

Carrot (Daucus carota) somatic embryogenesis has been extensively used as an experimental system for studying embryogenesis. In maturing zygotic embryos, abscisic acid (ABA) is involved in acquisition of desiccation tolerance and dormancy. On the other hand, somatic embryos contain low levels of endogenous ABA and show desiccation intolerance and lack dormancy, but tolerance and dormancy can be induced by exogenous application of ABA. In ABA-treated carrot embryos, some ABA-inducible genes are expressed. We isolated the Daucus carota bZIP1 (DcBZ1) gene encoding a G-box binding factor-type basic region/leucine zipper (GBF-type bZIP) factor from carrot somatic embryos. The expression of DcBZ1 was detected in embryogenic cells, non-embryogenic cells, somatic embryos, developing seeds, seedlings, and true leaves. Notably, higher expression was detected in embryogenic cells, true leaves, and seedlings. The expression of DcBZ1 increased in seedlings and true leaves after ABA treatment, whereas expression was not affected by differences in light conditions. During the development of zygotic and somatic embryos, increased expression of DcBZ1 was commonly detected in the later phase of development. The recombinant DcBZ1 protein showed specific binding activity to the two ABA-responsive element-like motifs (motif X and motif Y) in the promoter region of the carrot ABA-inducible gene according to results from an electrophoretic mobility shift assay. Our findings suggest that the carrot GBF-type bZIP factor, DcBZ1, is involved in ABA signal transduction in embryogenesis and other vegetative tissues.

Laccase (EC 1.10.3.2) is a small group of enzymes that catalyze the oxidation of a broad range of phenolic compounds including hazardous and recalcitrant pollutants in the environment. This study attempted to develop an efficient system for production of a recombinant laccase by chloroplast genetic transformation of tobacco. Chloroplast transformation vector was constructed and introduced into the tobacco chloroplast genome using particle bombardment. Chloroplast-transformed plants were subsequently regenerated. PCR and southern blot analyses confirmed stable integration of the laccase gene into the chloroplast genome. Northern blot analysis revealed that mRNA of the laccase gene was highly expressed in chloroplast-transformed plants.

겨우살이의 캘러스 형성 및 종자 발아 와 haustorium 발달에 요구되는 환경요인 및 생장조절제의 영향을 조사하였다. 아울러 종자의 인위부착을 통한 식물체 재생 체계를 확립하였다. 겨우살이 조직관찰 결과 종자에는 1개 또는 2개의 막대모양의 접합자배가 존재하며 종자의 외부에 위치한 유근 부위에 주로 세포분열이 왕성한 세포층이 존재하며 반대로 자엽은 종자 내부에 위치하며 상대적으로 분열능을 가진 세포수가 감소하였다. 겨우살이 종자 발아 와 haustorium 발달에 요구되는 여러 요인 중에서 광의 요구도가 절대적이었다. 본 실험에 조사된 다른 배양 환경요인 및 생장조절제들은 광의 효과를 대체하지 못하였으며 ethylene의 경우 발아촉진 효과가 3배 이상 증가함을 알 수 있었다. 여러 조직 중에서 오로지 겨우살이 화아로부터 캘러스 형성이 가능하였으며 캘러스 형성빈도는 $0.1\;mgl^{-1}$ IAA가 첨가된 B5 배지에 배양 시 27.3%로 가장 높았다. 본 연구에서 확립된 겨우살이 캘러스 배양 및 종자 인위 부착을 통한 식물체 재생체계는 겨우살이의 유용물질 생산 연구 및 대량증식 연구분야에 활용이 가능할 것으로 기대된다. 【Effects of growth regulators and culture conditions on seed germination, haustorium development, and callus formation of Korean mistletoe (Viscum album var. coloratum (Kom.) Ohwi) were described. Histological examination showed that seed of V. album contained one or two zygotic embryos with rod shape, and actively dividing cells were mainly distributed in radicle region rather than cotyledon of zygotic embryo. The most significant factor for seed germination and haustorium development of V. album was the requirement of the light. Various growth regulators examined in this study failed to substitute the effect of the light on seed germination. The frequency of callus formation was highest at 27.3% when flower buds were cultured onto B5 medium containing $0.1\;mgl^{-1}$ IAA. Explants from other organs were recalcitrant in forming calluses. Culture conditions described in this study could be applied for production of useful metabolites and multiplication of V. album in future.】

In order to generate transgenic radish (Raphanus sativus L., cv. Jin Ju Dae Pyong), hypocotyl explants were cultured on Murashige and Skoog medium containing 4 mg l−1 AgNO3, 5 mg l−1 acetosyringone, 4 mg l−1 6-benzyladenine, and 3 mg l−1 α-naphthaleneacetic acid in addition to either 10 mg l−1 hygromycin or 100 mg l−1 paromomycin after co-cultivation with disarmed Agrobacterium tumefaciens harboring a plant expression binary vector. Explants co-cultivated with A. tumefaciens GV3101 harboring pCAMBIA1301 and A. tumefaciens EHA101 harboring pPTN290 produced putative transgenic adventitious shoots at frequencies of 0.26% and 0.18%, respectively. Northern blot analysis revealed the gus gene transcript was detected in 8 regenerated plants which confirmed their genetic transformation. The transgenic plants were grown to maturity after vernalization in a greenhouse and appeared morphologically normal. Progeny analysis of independent transgenic plants demonstrated that the gus gene was transmitted in a Mendelian pattern in 3 lines, indicating a single copied gene was incorporated into the genome.

본 연구는 아그로박테리움 공동배양법을 이용한 기능획득 인삼 모상근의 대량생산을 위한 조건 확립에 대한 것이다. 일반적으로, 인삼과 같이 형질전환을 통한종자의 확보가 어려운 식물에서는 loss-of-function을 이용한 기능유전체 연구에 한계가 있다. 한편, 유전자의 기능을 활성화시키는 방법 (gain-of-function)인 activation tagging 기술은 이러한 문제점을 극복할 수 있는 대안이 될 수 있으며, Agrobacterium rhizogenes를 이용한 모상근 생산 시스템은 대량의 돌연변이체를 안정적으로 용이하게 얻을 수 있다는 점에서 최적의 시스템이라고 할 수 있다. 본 연구에서는 activation-tagging된 효율적인 형질전환 모상근 생산에 있어서의 최적의 아그로박테리움 균주 및 인삼조직, 배지조성 등에 대한 조건을 확립하였으며, 다양한 배지에서의 형질전환 모상근의 생장률 및 분지율, 표현형 등을 조사하였다. 엽병 절편을 activation-tagging vector pKH01을 가지고 있는 A. rhizogenes R1000와 공동배양하였을 때 배양 4주후 85.9%의 빈도로 모상근이 생산되었다. 모상근의 최대 생장과 분지도를 나타내는 배양조건을 조사한 바 엽병절편을 1/2 SH 배지에서 4주 배양하였을 때 왕성하게 생장하였으며 2.6의 분지도를 보여주었다. 최종적으로 1,989개체의 독립적인 형질전환 모상근 line을 생산하였으며, 이들 모상근 line은 인삼 진세노사이드 생합성 관련 유전자의 발굴 및 기능해석에 유용하게 쓰일 것이다. 【This study describes conditions for the mass production of activation-tagged mutant hairy root lines of ginseng by cocultivation with Agrobacterium rhizogenes. Because it is not currently possible to produce progeny from transgenic ginseng, a loss-of-function approach for functional genomics cannot be appliable to this species. A gain-of-function approach is alternatively the choice and hairy root production by cocultivation of A. rhizogenes would be most practical to obtain a large number of mutants. Various sources of explants were subjected to genetic transformation with various strains of A. rhizogenes harboring the activation-tagging vector pKH01 to determine optimum conditions for the highest frequency of hairy root formation on explants. Petiole explants cocultivated with A. rhizogenes R1000 produced hairy roots at a frequency of 85.9% after 4 weeks of culture. Conditions for maximum growth or branching rate of hairy roots were also investigated by using various culture media. Petiole explants cultured on half strength Schenk and Hildebrandt medium produced vigorously growing branched roots at a rate of 2.6 after 4 weeks of culture. A total of 1,989 lines of hairy root mutants were established in this study. These hairy root lines will be useful to determine functions of genes for biosynthesis of ginsenosides.】

When whole cell extracts are subjected to proton nuclear magnetic resonance spectroscopy (1H NMR), metabolite profiles are generated that contain overlapping signals of the majority of compounds within the extract. In order to determine whether pattern recognition based on the metabolite profiles of higher plants is able to genetically discriminate between plants, we analyzed leaf samples of eight cultivars ofCatharanthus roseus by1H NMR. Hierarchical dendrograms, based on the principal component analysis of the1H NMR total, aliphatic carbohydrate and aromatic region data, revealed possible relationships between the cultivars. The dendrogram based on the aromatic region data was in general agreement with the genetic relationships determined by conventional DNA fingerprinting methods. Secologanin and polyphenols were assigned to the signals of the1H NMR spectra, and contributed most profoundly to the discrimination between cultivars. The overall results indicate that the genetic relationships betweenC. roseus cultivars are reflected in the differences of the aromatic compounds in the leaves.

Ginseng ESTs allowed us to identify an unknown transcript which is highly abundant in rhizomes and seeds. We called the cDNA ginseng-specific abundant protein (GSAP), and identified three homologues, GSAP1, GSAP2, and GSAP3. GSAP cDNAs encode a small polypeptide consisting of 121 or 117 amino acids, and GSAP3 shows 87.6% amino acid sequence homology with GSAP1. GSAP transcripts were detected in most plant tissues, but GSAP3 is highly expressed in seeds, and is up-regulated under stressed conditions, water deficit. GSAP3-GFP fusion protein is located in the cell wall when expressed in onion epidermis cells. The transgenic Arabidopsis seedlings which over-expressed GSAP3 grew faster than those of the wild-type plant on the medium containing 300 mM mannitol and 100 mM NaCl. GSAP3 may play a role in altering the characteristics of the cell wall to allow for more tolerance of water deficit stress under abiotic stress conditions.

The hybrid nature of exciton polaritons opens up possibilities for developing a new concept nonlinear photonic device (e.g., polariton condensation, switching, and transistor) with great potential for controllability. Here, we proposed a novel type of polariton system resulting from strong coupling between a two-dimensional exciton and whispering gallery mode photon using a core–shell GaN/InGaN hexagonal wire. High quality, nonpolar InGaN multiple-quantum wells (MQWs) were conformally formed on a GaN core nanowire, which was spatially well matched with whispering gallery modes inside the wire. Both high longitudinal-transverse splitting of nonpolar MQWs and high spatial overlap with whispering gallery modes lead to unprecedented large Rabi splitting energy of ∼180 meV. This structure provides a robust polariton effect with a small footprint; thus, it could be utilized for a wide range of interesting applications.

Human serum albumin (HSA) is the most abundant protein in plasma and is the most often used intravenous protein in many human therapies. However, HSA is currently extracted only from plasma because commercially feasible recombinant expression systems are not available. This study attempted to develop an efficient system for recombinant HSA production by chloroplast transformation of tobacco. A HSA cDNA was isolated from a cDNA library constructed with human liver tissue. Chloroplast transformation vectors were constructed by introducing various regulatory elements to HSA regulatory sequences. Vectors were delivered by particle bombardment into leaf explants and chloroplast-transformed plants were subsequently regenerated into whole plants. Southern blot analysis confirmed that the HSA cDNA was incorporated between rps12 and orf70B of the chloroplast genome as designed. Western blot analysis revealed that hyper-expression and increasing the stability of HSA were achieved by modification of the regulatory sequences using the psbA5`UTRs in combination with elements of the 14 N-terminal amino acids of the GFP and the FLAG tag. However, only plants transformed with the vector containing all of these elements were able to accumulate HSA.

Fourier transform - infrared spectroscopy (FT-IR) provides biochemical profiles containing overlapping signals from a majority of the compounds that are present when whole cell extracts are analyzed. We attempted to determine the ripeness of strawberry fruit flesh by FT-IR. Fruit ripeness was divided into four developmental stages based on fruit skin color: `yellow-green`, `pink-green`, `pink`, and `red` stages. Principal component analysis of FT-IR data of inside fruit flesh extracts clustered samples of four different developmental stages into three discrete groups: (1) `yellow-green` group, (2) `pink-green` group, and (3) `pink` and `red` group. The most remarkable difference between four different developmental stages was found in the carbohydrate fingerprint region of the FT-IR spectrum, indicating that differences in carbohydrate compounds represented the ripeness of strawberry fruit. Overall results indicate that FT-IR in combination with PCA enables discrimination of the ripeness of strawberry fruit flesh.

Phytosulfokine-α (PSK-α), a peptidyl plant growth factor that was originally isolated from conditioned medium derived from asparagus mesophyll cell culture, stimulates cell proliferation in some plants in suspension culture. However, the physiological relationship between phytohormones and this growth factor in plants is not well understood. Using carrot ( Daucus carota L.) non-embryogenic cell culture, we investigated the stimulatory effects of PSK-α on the cell proliferation induced by different combinations and concentrations of auxin [2,4-dichlorophenoxyacetic acid (2,4-D), indole-3-acetic acid (IAA), or naphthaleneacetic acid (NAA)] and PSK-α. Cell proliferation was not stimulated by PSK-α at high auxin concentrations, but was promoted at low auxin concentrations. There was no proliferation in the absence of auxin. These results indicate that PSK-α stimulates cell proliferation when cell division activity is low, and that auxin is required for this process. We also evaluated PSK-α production using medium derived from non-embryogenic cell culture with high or low concentrations of auxin (2,4-D, IAA or NAA) in a competition ELISA system. More PSK-α was produced at high auxin concentrations than at low auxin concentrations. These results suggest that auxin participates in PSK-α production.

We present group III-nitride semiconductor nanostructures grown on various types of GaN-based pyramid, annular, columnar, and tapered structures, together with their applications in broadband light emitting devices, unidirectional photonic diodes, and single photon sources.

We report the measurement of the (r13, r33) Pockels electro-optic coefficients in a GaN thin film grown on a Si(111) substrate. The converse piezoelectric (d33) and electro-absorptive coefficients are simultaneously determined. Single crystalline GaN epitaxial layers were grown with a AlGaN buffer layer by metal organic chemical vapor deposition, and their structural and optical properties were systematically investigated. The electro-optic, converse piezoelectric, and electro-absorptive coefficients of the GaN layer are determined using an original method. A semi-transparent gold electrode is deposited on the top of the GaN layer, and an alternating voltage is applied between top and bottom electrodes. The coefficients are simultaneously and analytically determined from the measurement of the electric-field-induced variation ΔR(θ) in the reflectivity of the Au/GaN/buffer/Si stack, versus incident angle and light polarization. The method also enables to determine the GaN layer polarity. The results obtained for a Ga-face [0001] GaN layer when using a modulation frequency of 230 Hz are for the electro-optic coefficients r13 = +1.00 ± 0.02 pm/V, r33 = +1.60 ± 0.05 pm/V at 633 nm, and for the converse piezoelectric coefficient d33 = +4.59 ± 0.03 pm/V. The value measured for the electro-absorptive variation at 633 nm is Δko/ΔE = +0.77 ± 0.05 pm/V.

A key issue in a single photon source is fast and efficient generation of a single photon flux with high light extraction efficiency. Significant progress toward high-efficiency single photon sources has been demonstrated by semiconductor quantum dots, especially using narrow bandgap materials. Meanwhile, there are many obstacles, which restrict the use of wide bandgap semiconductor quantum dots as practical single photon sources in ultraviolet-visible region, despite offering free space communication and miniaturized quantum information circuits. Here we demonstrate a single InGaN quantum dot embedded in an obelisk-shaped GaN nanostructure. The nano-obelisk plays an important role in eliminating dislocations, increasing light extraction, and minimizing a built-in electric field. Based on the nano-obelisks, we observed nonconventional narrow quantum dot emission and positive biexciton binding energy, which are signatures of negligible built-in field in single InGaN quantum dots. This results in efficient and ultrafast single photon generation in the violet color region.

International audience; We report here the experimental results on a field effect refractive index change into gallium nitride (GaN) structures. This effect is characterized through the common prism-coupling technique with the application of a vertical direct-current electric field. Surface plasmon propagation was used to increase the sensitivity of the electro-optic measurements. We have obtained a large refractive index variation for GaN epilayer, around 1.4×10−2 at 1.55μm wavelength. In order to understand the origin of the index modulation, we have conducted a scanning transmission electron microscopy analysis and discussed the influence of threading dislocations density acting as traps and thermo-optic effect. According to recent works, we observed experimentally the optical response of a non-linear electro-optic effect on GaN on Si(111) substrate and estimated a Kerr coefficient of about 2.14×10−16m2V−2.

A good justification for gallium nitride on silicone is the potential for integrated optoelectronic circuits and for the low cost bring by growth of GaN on a large size wafers. Actually, the application interest for GaN/Si is power electronics. This work focused on the optimization of the growth process for GaN/Si and the relation between the structure and the optical properties. Using the guided wave prism coupling technique, we have fully established the index dispersion of GaN at room temperature and its temperature dependence in the wavelength range 0.4 to 1.5µm. We report a slightly low temperature dependence. Results demonstrated excellent waveguide properties of GaN on silicon with optical propagation loss below 1dB/cm. We compared trhe results on Si with those on sapphire. This opens a real opportunity of future device using this technology.