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Screening tests in which 30 species of succulent plants were inoculated with A136 or C58 strains of Agrobacterium tumefaciens led to selecting genus Kalanchoe for further study of Agrobacterium-mediated transformation. Regeneration studies showed that 4.55 µM Thidiazuron (TDZ) combined with 2.15 µM α-naphtaleneacetic acid (NAA) significantly proliferated callus development from Kalanchoe pinnata leaf explants in Murashige and Skoog (MS) medium. Subsequently, 4.55 µM TDZ significantly enhanced shoot formation from the green and compact callus. NAA (5.37 µM) accelerated root growth leading to establishment of transgenic plants in soil. A gene encoding the fluorescent protein, ZsGreen1, was cloned into a plasmid vector harboring a constitutive figwort mosaic virus (FMV) 35S promoter, a kanamycin resistance gene, and a NOS-terminator. The plasmids were delivered into the genome of K. pinnata by A. tumefaciens strain C58 (ABI). The ZsGreen1-transgenic plants exhibited very strong, green fluorescence from the callus, leaves, stems, roots, and flowers visible through a green emission filter (GFP, 480/510 nm, excitation/emission) under excitation by ultra-violet light. Transcription analysis by sqRT-PCR and qPCR confirmed the stable integration of the ZsGreen1 in the genome of K. pinnata. The FMV 35S promoter conferred uniform expression throughout whole plant organ. ZsGreen1 was proven to be a stable marker for gene expression in genetic transformation of K. pinnata. The ZsGreen1-transgenic Kalanchoe pinnata mediated by Agrobacterium tumefaciens exhibited strong, green fluorescence from the callus, leaves, stems, roots and flowers under the control of the figwort mosaic virus (FMV) 35S promoter.

Fluorescent proteins (FT) have become essential, biological research tools. Many novel genes have been cloned from a variety of species and modified for effective, stable, and strong expression in transgenic organisms. Although there are many applications, FT expression has been employed most commonly at the cellular level in plants. To investigate FT expression at the whole-plant level, particularly in flowers, petunia ‘Mitchell Diploid’ [MD (Petunia ×hybrida)] was genetically transformed with seven genes encoding FTs: DsRed2, E2Crimson, TurboRFP, ZsGreen1, ZsYellow1, rpulFKz1, or aeCP597. Each gene was cloned into a pHK-DEST-OE vector harboring constitutive figwort mosaic virus 35S promoter and NOS-terminator. These plasmids were individually introduced into the genome of MD by Agrobacterium tumefaciens–mediated transformation. Shoot regeneration efficiency from the cocultured explants ranged from 8.3% to 20.3%. Various intensities of red, green, and yellow fluorescence were detected from TurboRFP, ZsGreen1, and ZsYellow1-transgenic flowers, respectively, under ultraviolet light for specific excitation and emission filters. More than 70% of plants established from the regenerated shoots were confirmed as transgenic plants. Transgenic ZsGreen1 petunia generated strong, green fluorescence in all flower organs of T0 plants including petals, stigmas, styles, anthers, and filaments. Most of the chromophores were localized to the cytoplasm but also went into the nuclei of petal cells. There was a positive linear relationship (R2 = 0.88) between the transgene expression levels and the relative fluorescent intensities of the ZsGreen1-transgenic flowers. No fluorescence was detected from the flowers of DsRed2-, E2Crimson-, rpulFKz1-, or aeCP597-transgenic petunias even though their gene transcripts were confirmed through semiquantitative reverse transcriptase-polymerase chain reaction. T1 generation ZsGreen1 plants showed green fluorescence emission from the cotyledons, hypocotyls, and radicles, which indicated stable FT expression was heritable. Four homozygous T2 inbred lines were finally selected. Throughout this study, we demonstrated that ZsGreen1 was most suitable for generating visible fluorescence in MD flowers among the seven genes tested. Thus, ZsGreen1 may have excellent potential for better utility as a sensitive selectable marker.

Petunia × hybrida cv ‘Mitchell Diploid’ floral volatile benzenoid/phenylpropanoid (FVBP) biosynthesis ultimately produces floral volatiles derived sequentially from phenylalanine, cinnamic acid, and p-coumaric acid. In an attempt to better understand biochemical steps after p-coumaric acid production, we cloned and characterized three petunia transcripts with high similarity to p-coumarate 3-hydroxylase (C3H), hydroxycinnamoyl-CoA:shikimate/quinate hydroxycinnamoyl transferase (HCT), and caffeoyl shikimate esterase (CSE). Transcript accumulation of PhC3H and PhHCT was highest in flower limb tissue during open flower stages. PhCSE transcript accumulation was also highest in flower limb tissue, but it was detected earlier at initial flower opening with a bell-shaped distribution pattern. Down regulation of endogenous PhC3H transcript resulted in altered transcript accumulation of many other FVBP network transcripts, a reduction in floral volatiles, and the emission of a novel floral volatile. Down regulation of PhHCT transcript did not have as large of an effect on floral volatiles as was observed for PhC3H down regulation, but eugenol and isoeugenol emissions were significantly reduced on the downstream floral volatiles. Together these results indicate that PhC3H is involved in FVBP biosynthesis and the reduction of PhC3H transcript influences FVBP metabolism at the network level. Additional research is required to illustrate PhHCT and PhCSE functions of petunia.

Coleus (Plectranthus scutellarioides) is an attractive and popular ornamental plant with propagation mainly achieved through vegetative cuttings. For commercial purposes, it is of interest to enhance the speed of establishment while maintaining high quality. Light quality has been shown to influence adventitious root development, so these experiments examined the effect of narrow-bandwidth light treatments on root growth and overall plant quality for seven coleus cultivars with vegetative cuttings in potting soil and one cultivar with shoot tip in vitro cultures onto Murashige and Skoog (MS) agar medium. During the 28 days of the propagation period, the cuttings grown under narrow-bandwidth red light (R; 663.4 nm at peak) more than doubled in the adventitious root number compared with those under blue light (B; 445.7 nm at peak) and green light (G; 530.0 nm at peak) in five cultivars. R light also increased fresh weight of the cuttings by 55.6% more than G light. In comparison, the cuttings grown under G light yielded significantly lower root and shoot dry mass than other light treatments. R light cuttings showed more dry mass content (9.63%) than those under white light (W; 437.4 nm and 559.5 nm at peak) and G light (7.85% and 5.86%, respectively). A positive correlation (R2 = 0.598, P < 0.001) was found between the formation of adventitious roots and gained fresh weight of cuttings. R light made the reddish color of leaves significantly stronger in most cultivars, whereas the cuttings exposed to G light became less vivid compared with other light conditions. When the shoot tips were propagated in vitro onto MS medium, R light treatment initiated the root development more rapidly than other lights, with significantly greater rooting rate (20.0% and 63.6%, respectively) at day 5 and 10. The shoot tips under R light also formed significantly more roots (12.3 per cutting) than those grown under narrow-bandwidth B light (5.8 per cutting). The shoot tips showed browning at an early stage and newly emerged leaves grew very compactly under B light. The combination of red and green light (R+G) increased more than twice as much roots and dry mass compared with W light. In addition, the R+G light led to morphological changes, including larger leaves and longer petioles and internodes than those in other light treatments. The exposure to R+G+B and B light made the shoots very compact for the 28 days of in vitro culture period and significantly increased the chlorophyll contents resulting in dark green leaves.

Polyphenol oxidase (PPO) is a well-characterized enzyme that is conserved across plant species and is responsible for catalyzing the rapid polymerization of o-quinones that results in tissue browning. As these oxidation reactions result in significant reductions in food crop commercial viability, nutritional content, and general desirability, an opportunity exists to improve a variety of crops via genetic regulation of PPO expression. The main objective of this project was to clone the DNA sequence of genes encoding PPO. Using mRNA as a template to synthesize DNA, two putative PPO sequences were cloned from Ocimum basilicum and Ocimum americanum, two basil species of commercial relevance. A secondary objective of this project was to insert a cloned PPO sequence into an RNAi vector for the purpose of making transgenic basil plants with reduced PPO expression. A hairpin RNAi construct was successfully developed to reduce the expression of both PPO genes as their DNA sequences were highly homologous. The next step for this project will depend on the development of transformation and regeneration protocols for these species, which have proven to be difficult. Once the RNAi PPO constructs are introduced into basil cells and successfully regenerated into plants, reduced browning of basil leaf tissue when it is wounded or cut is expected.

This study examined monthly feeding rates and grazing impact on phytoplankton biomass, as well as diel feeding rhythms of four key copepod species in a tidally well mixed estuary (Asan Bay, Korean Peninsula). Monthly ingestion rates estimated based on gut pigment analysis were closely associated with their peak densities, but not with phytoplankton biomass, implying high ingestion may be related to reproductive output for population growth. The three smaller copepods, Acartia hongi, Acartia pacifica and Paracalanus parvus, showed feeding preference for smaller phytoplankton (

In this study, hMSCs encapsulated in a fibrin hydrogel containing heparinized NPs loaded with TGF-β3 (100 ng/ml), or TGF-β3 (100 ng/ml) alone, were subjected to growth factor release and denaturation tests at one, two and four weeks in in vitro culture systems. Additionally, stem cell differentiation was assessed via RT-PCR, real-time quantitative PCR (qPCR), histology, and immunohistochemical assays. In the in vivo studies with nude mouse, when transplanted into nude mice, hMSCs embedded in fibrin hydrogels survived and proliferated more readily in those samples containing TGF-β3-loaded NPs, or TGF-β3 alone, compared to those containing only NPs or the fibrin hydrogel alone. Additionally, RT-PCR, real-time qPCR, histology, Western blotting, and immunohistochemistry analyses revealed that chondrocyte-specific extracellular matrix (ECM) genes and their proteins were expressed at high levels by hMSCs embedded in hydrogels containing TGF-β3-loaded NPs. Finally, the results observed in the rabbit animal model treated with hMSCs embedded in a fibrin hydrogel containing TGF-β3-loaded NPs were also evaluated by the RT-PCR, real-time qPCR, histology, Western blotting, and immunohistochemistry analyses. The in vitro and in vivo results indicated that transplanted hMSCs together with TGF-β3 may constitute a clinically efficient method for the regeneration of hyaline articular cartilage.

Hybrid Lilium (common name lily) cultivars are among the top produced domestic fresh cut flowers and potted plants in the US today. Many hybrid Lilium cultivars produce large and showy flowers that emit copious amounts of volatile molecules, which can negatively affect a consumer's appreciation or limit use of the plant product. There are few publications focused on the biochemistry, genetics, and/or molecular regulation of floral volatile biosynthesis for Lilium cultivars. In an initial pursuit to provide breeders with molecular markers for floral volatile biosynthesis, a total of five commercially available oriental and oriental-trumpet hybrid Lilium cultivars were selected for analytical characterization of floral volatile emission. In total, 66 volatile molecules were qualified and quantitated among all cultivars. Chemical classes of identified volatiles include monoterpene hydrocarbons, monoterpene alcohols and aldehydes, phenylpropanoids, benzenoids, fatty-acid-derived, nitrogen-containing, and amino-acid-derived compounds. In general, the floral volatile profiles of the three oriental-trumpet hybrids were dominated by monoterpene hydrocarbons, monoterpene alcohols and aldehydes, while the two oriental hybrids were dominated by monoterpene alcohols and aldehydes and phenylpropanoids, respectively. Tepal tissues (two petal whirls) emitted the vast majority of total volatile molecules compared to the reproductive organs of the flowers. Tepal volatile profiles were cultivar specific with a high degree of distinction, which indicates the five cultivars chosen will provide an excellent differential genetic environment for gene discovery through comparative transcriptomics in the future. Cloning and assaying transcript accumulation from four floral volatile biosynthetic candidates provided few immediate or obvious trends with floral volatile emission.

Arbuscular mycorrhizal (AM) symbiosis can confer increased host resistance to drought stress, although the effect is unpredictable. Since AM symbiosis also frequently increases host resistance to salinity stress, and since drought and salinity stress are often linked in drying soils, we speculated that the AM influence on plant drought response may be partially the result of AM influence on salinity stress. We tested the hypothesis that AM-induced effects on drought responses would be more pronounced when plants of comparable size are exposed to drought in salinized soils. In two greenhouse experiments, several water relations characteristics were measured in sorghum plants colonized by Glomus intraradices (Gi), Gigaspora margarita (Gm) or a mixture of AM species, during a sustained drought following exposure to salinity treatments (NaCl stress, osmotic stress via concentrated macronutrients, or soil leaching). The presence of excess salt in soils widened the difference in drought responses between AM and nonAM plants in just two instances. Days required for plants to reach stomatal closure were similar for Gi and nonAM plants exposed to drought alone, but with exposure to combined NaCl and drought stress, stomates of Gi plants remained open 17-22% longer than in nonAM plants. Promotion of stomatal conductance by Gm occurred with exposure to NaCl/drought stress but not with drought alone or with soil leaching before drought. In other instances, however, the addition of salt tended to nullify an AM-induced change in drought response. Our findings confirm that AM fungi can alter host response to drought but do not lend much support to the idea that AM-induced salt resistance might help explain why AM plants can be more resilient to drought stress than their nonAM counterparts.

There is growing appreciation of arbuscular mycorrhizal effects on soil properties and their potential consequences on plant behavior. We examined the possibility that mycorrhizal soil may directly influence plant water relations. Using wild-type and noncolonizing bean mutants planted into soils previously produced using mycorrhizal or nonmycorrhizal sorghum plants, we partitioned mycorrhizal influence on stomatal conductance and drought resistance into soil and root components, testing whether effects of mycorrhizal fungi occurred mostly via mycorrhization of roots, mycorrhization of soil, or both. The mutation itself had no effect on any water relations parameter. Colonization by Gigaspora margarita Gerdemann & Trappe and Glomus intraradices Schenck & Smith had appreciable effects on leaf water potential at the lethal point and on osmotic adjustment, relative to nonmycorrhizal plants of comparable size. Mycorrhizal effects on drought resistance were attributable to an effect on the plant itself rather than to an effect of mycorrhizal soil. Mycorrhizal effects on stomatal conductance were attributable to mycorrhization of both roots and soil, as well as to mycorrhization of roots alone. Surprisingly, merely growing in a mycorrhizal soil resulted in promotion of stomatal conductance of nonmycorrhizal plants in both amply watered and droughted plants. Mycorrhizal effects on droughted plants did not appear to be related to altered soil water retention properties, as Gigaspora margarita and Glomus intraradices altered the soil's moisture characteristic curve only slightly.Key words: arbuscular mycorrhizal symbiosis, bean, drought, Gigaspora margarita, Glomus intraradices, stomatal conductance.

This report contains the first description of the genomic structure for a sulfotransferase (ST). The gene (ASTIV) encodes rat hepatic aryl ST IV, also known as tyrosine-ester ST (EC 2.8.2.9). A phage genomic clone containing 70% of the 3' AST gene coding sequence was isolated after screening a rat genomic library with an ASTIV cDNA. The remaining 5' sequence was determined from a PCR product obtained from rat genomic DNA and ASTIV cDNA-specific primers. ASTIV spans 3.5 kb and contains eight exons and seven introns. The fourth intron of this gene contains sequences homologous to rodent B1 repetitive elements and an Alu repeat found in rat. An alignment of the primary structures of ten different ST revealed several conserved regions, as well as a putative binding site for the cofactor for enzymatic sulfation reactions, 3'-phosphoadenosine-5'-phosphosulfate.

In prior studies we learned that colonization of soil can be as important as colonization of roots in determining mycorrhizal influence on the water relations of host plants. Here we use a path analysis modeling approach to test (a) whether quantity of hyphae in soil contributes to variations in stomatal behavior and soil drying, and (b) whether soil colonization or root colonization has a stronger influence on these stomatal and soil drying responses. Experiments were performed on Sorghum bicolor and Cucurbita pepo, with soils and roots colonized by a mixture of Glomus intraradices and Gigaspora margarita. Soil colonization generally made more significant contributions to stomatal conductance than did root colonization. Soil colonization did not make significant direct contributions to soil water potential measures (soil water potential at stomatal closure or soil drying rate), whereas root colonization did contribute a potentially important path to each. The findings further support a role for mycorrhization of the soil itself in contributing to the regulation of stomatal behavior of host plants.

Human alpha2-antiplasmin (alpha2AP), also known as alpha2-plasmin inhibitor, is the major inhibitor of the proteolytic enzyme plasmin that digests fibrin. There are 2 N-terminal forms of alpha2AP that circulate in human plasma: a 464-residue protein with Met as the N-terminus, Met-alpha2AP, and a 452-residue version with Asn as the N-terminus, Asn-alpha2AP. We have discovered and purified a proteinase from human plasma that cleaves the Pro12-Asn13 bond of Met-alpha2AP to yield Asn-alpha2AP and have named it antiplasmin-cleaving enzyme (APCE). APCE is similar in primary structure and catalytic properties to membrane-bound fibroblast activation protein/seprase for which a physiologic substrate has not been clearly defined. We found that Asn-alpha2AP becomes cross-linked to fibrin by activated factor XIII approximately 13 times faster than native Met-alpha2AP during clot formation and that clot lysis rates are slowed in direct proportion to the ratio of Asn-alpha2AP to Met-alpha2AP in human plasma. We conclude that APCE cleaves Met-alpha2AP to the derivative Asn-alpha2AP, which is more efficiently incorporated into fibrin and consequently makes it strikingly resistant to plasmin digestion. APCE may represent a new target for pharmacologic inhibition, since less generation and incorporation of Asn-alpha2AP could result in a more rapid removal of fibrin by plasmin during atherogenesis, thrombosis, and inflammatory states.

Mycorrhizal symbiosis can modify plant response to drying soil, but little is known about the relative contribution of soil vs. root hyphal colonization to drought resistance of mycorrhizal plants. Foliar dehydration tolerance, characterized as leaf and soil water potential at the end of a lethal drying episode, was measured in bean plants (Phaseolus vulgaris) colonized by Glomus intraradices or by a mix of arbuscular mycorrhizal fungi collected from a semi-arid grassland. Path analysis modeling was used to evaluate how colonization rates and other variables affected these lethal values. Of several plant and soil characteristics tested, variation in dehydration tolerance was best explained by soil hyphal density. Soil hyphal colonization had larger direct and total effects on both lethal leaf water potential and soil water potential than did root hyphal colonization, root density, soil aggregation, soil glomalin concentration, leaf phosphorus concentration or leaf osmotic potential. Plants colonized by the semi-arid mix of mycorrhizal fungi had lower lethal leaf water potential and soil water potential than plants colonized by G. intraradices. Our findings support the assertion that external, soil hyphae may play an important role in mycorrhizal influence on the water relations of host plants.

Colonization of roots and soil by arbuscular mycorrhizal (AM) fungi sometimes promotes stomatal conductance (g s) of the host plant, but scientists have had difficulty predicting or manipulating the response. Our objective was to test whether the magnitude of AM influence on g s is related to environmental conditions: irradiance, air temperature or leaf temperature. Stomatal conductances of two groups of uncolonized sorghum plants were compared to g s of plants colonized by Glomus intraradices (Gi) or Gigaspora margarita (Gm) in 31 morning and afternoon periods under naturally varying greenhouse conditions. Stomatal conductance of Gi and Gm plants was often markedly higher than g s of similarly sized nonAM plants. AM promotion of g s was minimal at the lowest irradiances and lowest air and leaf temperatures, but was substantial at intermediate irradiance and temperatures. AM promotion was again low or absent at the highest irradiances and temperatures. Magnitude of AM promotion of g s was not a function of absolute g s. Promotion of g s by Gi and Gm was remarkably similar. Differing phosphorus fertilization did not affect g s.

Krabbe's disease (globoid cell leukodystrophy) is a progressive cerebral degenerative disease of infancy characterized by severe myelin loss and the presence of globoid bodies in the white matter. Previous studies have suggested that psychosine is the causative agent for the pathogenesis of Krabbe's disease. In the present study, we investigated psychosine-induced injury and cell death of oligodendrocytes in enriched cultures of oligodendrocytes prepared from 3-week-old rat brain. The psychosine concentration sufficient to induce 50% cell death in oligodendrocytes was 30 micrograms/ml in the medium containing serum and 10 micrograms/ml in the serum-free medium. When oligodendrocytes were exposed to psychosine in the presence of phorbol esters, insulin, insulin-like growth factor I (IGF-I), demethylsulfoxide, or serum albumin, the survival of oligodendrocytes was greatly increased. These results indicate that psychosine cytotoxicity against oligodendrocytes is blocked by phorbol esters, insulin, and IGF-I through activation of protein kinase-C, by dimethylsulfoxide through activation of beta-galactosidase, and by albumin through its binding to psychosine.

Colonization of roots by arbuscular mycorrhizal (AM) fungi can increase host resistance to drought stress, although the effect is unpredictable. Since AM symbiosis also frequently increases host resistance to salt stress, and since drought and salt stress are often linked in drying soils, we speculated that the AM influence on plant drought response may be linked to AM influence on salt stress. We tested the hypothesis that AM-induced effects on drought responses would be more pronounced when plants of comparable size are exposed to drought in salinized soils. In two greenhouse experiments, several water relations characteristics were measured in sorghum plants colonized by Glomus intraradices, Gigaspora margarita, or a mixture of AM species during a sustained drought following exposure to salt treatments (NaCl stress, osmotic stress, or soil leaching). The presence of excess salt in soils widened the difference in drought responses between AM and non-AM plants in just two instances: days needed for plants to reach stomatal closure, and promotion of stomatal conductance. In other instances, the addition of salt tended to nullify an AM-induced change in drought response; e.g., an AM effect on the decline in leaf or soil water potential required to cause stomatal closure disappeared when soils were salinized. Our findings confirm that AM fungi can alter host response to drought but do not lend much support to the idea that AM-induced salt resistance might help explain why AM plants can be more resilient to drought stress than their non-AM counterparts.

The influence of calcium (Ca++) nutrition on the growth and root tissue electrolyte leakage (EL) of carrot (Daucus carota) was investigated using a hydroponic culture system. Seedlings of `Navajo' carrot were grown for 10 weeks with roots submersed in hydroponic nutrient solutions containing 0, 0.1, 1, 2, 4, or 8 meq/L Ca++. The nutrient solution was replenished weekly with its pH maintained at 5.8 for the entire experimental period. The tap root lengths increased as solution Ca++ concentration increased. The diameter and fresh and dry weights of the tap roots increased as Ca++ concentration increased up to 4 meq/L, and then decreased at 8 meq/L Ca++. The root and petiole concentrations of sugar, potassium, and nitrate were unaffected by changes in nutrient solution Ca++ levels. The tissue EL, when tested for the stored roots, decreased as solution Ca++ concentration increased (r = 0.602). Results of this experiment suggest that calcium nutrition is essential for maintaining cell wall integrity in hydroponically grown carrot roots.

The biomass yield, transpiration rate, and chlorophyll contents in Cymbidium goeringii plants grown under various light, temperature, and humidity conditions were investigated. Two-year-old plants potted in pine-bark medium were grown for 12 weeks during the summer months in polyethylene film-covered mini-greenhouses having four different environmental conditions: a) closed house (CH) with high humidity (95.1% RH), high light (800 μmol·m–2·s–1) and high temperature (37.5 °C), b) ventilated house (VH) with low humidity (41.4% RH), high light (800 μmol·m–2·s–1), and medium temperature (31.5 °C), c) shaded closed house (SCH) with high humidity (91.0% RH), low light (110 μmol·m–2·s–1) and medium temperature (33.3 °C), and d) shaded ventilated house (SVH) with medium humidity (61.5% RH), low light (110 μmol·m–2·s–1) and low temperature (30.5 °C). Plants grown in CH produced leaf chlorosis with 50% shorter leaves and 40% lower relative growth rate (7.9 mg/g fresh weight per day) compared to plants grown in SVH. Cymbidium plants grown in SCH or SVH showed higher leaf and root dry weights as compared to those grown in CH or VH. Leaf chlorophyll-a and -b contents as well as carbohydrate levels were the highest in plants grown in SVH, indicating the benefits of shading and ventilation. The rate of transpiration showed a quadratic response to increasing levels of leaf temperature (r2 = 0.81), wind velocity (r2 = 0.82), and vapor pressure deficit (VPD, r2 = 0.91). Regression analysis revealed that the maximum transpiration rate occurred at 25.4 °C leaf temperature, 2.1 m/s wind velocity, and 2.3 kPa VPD in this experiment.

The narrow-leaved purple coneflower (Echinacea angustifolia) produces echinacin and related compounds in the root, which are known to have immune and curative properties against viral, fungal, and bacterial infections. In recent years, cultivation of this species has increased in response to growing market demand for natural medicinal remedies. The objective of this study was to determine the influence of gibberellic acid and light on the germination of E. angustifolia seeds. Seeds soaked for 24 h in 0, 1, 5, 10, 50, 100, 250, 500 or 1000 mg/L GA3 solution were germinated on Whatman #1 filter paper inside petri dishes at 22 °C with or without light (80 μmol·m-2·s-1) for 21 days. The seeds germinated poorly in dark with the final percent germination range from 10% (GA3 1000 mg/L) to 36% (GA3 250 mg/L). Under light, seed germination showed a quadratic response (r = 0.84) to GA3 concentration. Percent germination exceeded 90% at 10, 50, and 100 mg/L GA3 with the mean time (T50) to germinate varying at 10.5, 11.7, and 13.3 days, respectively, under light. Seed germination under light was 3. In general, seed germination was best when treated with 10 or 50 mg/L GA3 under light. Results of this research may well be used in enhancing seed germination during field establishment of E. angustifolia.

Metabolomics, the comprehensive analysis of metabolites in a biological system, provides detailed information about the biochemical/physiological status of a biological system, and about the changes caused by chemicals. Metabolomics analysis is used in many fields, ranging from the analysis of the physiological status of genetically modified organisms in safety science to the evaluation of human health conditions. In toxicology, metabolomics is the -omics discipline that is most closely related to classical knowledge of disturbed biochemical pathways. It allows rapid identification of the potential targets of a hazardous compound. It can give information on target organs and often can help to improve our understanding regarding the mode-of-Action of a given compound. Such insights aid the discovery of biomarkers that either indicate pathophysiological conditions or help the monitoring of the efficacy of drug therapies. The first toxicological applications of metabolomics were for mechanistic research, but different ways to use the technology in a regulatory context are being explored. Ideally, further progress in that direction will position the metabolomics approach to address the challenges of toxicology of the 21st century. To address these issues, scientists from academia, industry, and regulatory bodies came together in a workshop to discuss the current status of applied metabolomics and its potential in the safety assessment of compounds. We report here on the conclusions of three working groups addressing questions regarding 1) metabolomics for in vitro studies 2) the appropriate use of metabolomics in systems toxicology, and 3) use of metabolomics in a regulatory context.