153 research outputs found

    Transgenic miR156 Switchgrass in the Field: Growth, Recalcitrance and Rust Susceptibility

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    Sustainable utilization of lignocellulosic perennial grass feedstocks will be enabled by high biomass production and optimized cell wall chemistry for efficient conversion into biofuels. MicroRNAs are regulatory elements that modulate the expression of genes involved in various biological functions in plants, including growth and development. In greenhouse studies, overexpressing a microRNA (miR156) gene in switchgrass had dramatic effects on plant architecture and flowering, which appeared to be driven by transgene expression levels. Highexpressing lines were extremely dwarfed, whereas low and moderate-expressing lines had higher biomass yields, improved sugar release and delayed flowering. Four lines with moderate or low miR156 overexpression from the prior greenhouse study were selected for a field experiment to assess the relationship between miR156 expression and biomass production over three years. We also analysed important bioenergy feedstock traits such as flowering, disease resistance, cell wall chemistry and biofuel production. Phenotypes of the transgenic lines were inconsistent between the greenhouse and the field as well as among different field growing seasons. One low expressing transgenic line consistently produced more biomass (25%–56%) than the control across all three seasons, which translated to the production of 30% more biofuel per plant during the final season. The other three transgenic lines produced less biomass than the control by the final season, and the two lines with moderate expression levels also exhibited altered disease susceptibilities. Results of this study emphasize the importance of performing multiyear field studies for plants with altered regulatory transgenes that target plant growth and development

    The TcEG1 beetle (Tribolium castaneum) cellulase produced in transgenic switchgrass is active at alkaline pH and auto-hydrolyzes biomass for increased cellobiose release

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    Background Genetically engineered biofuel crops, such as switchgrass (Panicum virgatum L.), that produce their own cell wall-digesting cellulase enzymes would reduce costs of cellulosic biofuel production. To date, non-bioenergy plant models have been used in nearly all studies assessing the synthesis and activity of plant-produced fungal and bacterial cellulases. One potential source for cellulolytic enzyme genes is herbivorous insects adapted to digest plant cell walls. Here we examine the potential of transgenic switchgrass-produced TcEG1 cellulase from Tribolium castaneum (red flour beetle). This enzyme, when overproduced in Escherichia coliand Saccharomyces cerevisiae, efficiently digests cellulose at optima of 50 °C and pH 12.0. Results TcEG1 that was produced in green transgenic switchgrass tissue had a range of endoglucanase activity of 0.16–0.05 units (µM glucose release/min/mg) at 50 °C and pH 12.0. TcEG1 activity from air-dried leaves was unchanged from that from green tissue, but when tissue was dried in a desiccant oven (46 °C), specific enzyme activity decreased by 60%. When transgenic biomass was “dropped-in” into an alkaline buffer (pH 12.0) and allowed to incubate at 50 °C, cellobiose release was increased up to 77% over non-transgenic biomass. Saccharification was increased in one transgenic event by 28%, which had a concurrent decrease in lignin content of 9%. Histological analysis revealed an increase in cell wall thickness with no change to cell area or perimeter. Transgenic plants produced more, albeit narrower, tillers with equivalent dry biomass as the control. Conclusions This work describes the first study in which an insect cellulase has been produced in transgenic plants; in this case, the dedicated bioenergy crop switchgrass. Switchgrass overexpressing the TcEG1 gene appeared to be morphologically similar to its non-transgenic control and produced equivalent dry biomass. Therefore, we propose TcEG1 transgenics could be bred with other transgenic germplasm (e.g., low-lignin lines) to yield new switchgrass with synergistically reduced recalcitrance to biofuel production. In addition, transgenes for other cell wall degrading enzymes may be stacked with TcEG1 in switchgrass to yield complementary cell wall digestion features and complete auto-hydrolysis

    Spitzer Mid-infrared Spectroscopy of Ices Toward Extincted Background Stars

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    A powerful way to observe directly the solid state inventory of dense molecular clouds is by infrared spectroscopy of background stars. We present Spitzer/IRS 5-20 micron spectra of ices toward stars behind the Serpens and Taurus molecular clouds, probing visual extinctions of 10-34 mag. These data provide the first complete inventory of solid-state material in dense clouds before star formation begins. The spectra show prominent 6.0 and 6.85 micron bands. In contrast to some young stellar objects (YSOs), most (~75%) of the 6.0 micron band is explained by the bending mode of pure water ice. In realistic mixtures this number increases to 85%, because the peak strength of the water bending mode is very sensitive to the molecular environment. The strength of the 6.85 micron band is comparable to what is observed toward YSOs. Thus, the production of the carrier of this band does not depend on the energetic input of a nearby source. The spectra show large abundances of carbon monoxide and carbon dioxide (20-40% with respect to water ice). Compared to YSOs, the band profile of the 15 micron carbon dioxide bending mode lacks the signatures of crystallization, confirming the cold, pristine nature of these lines of sight. After the dominant species are removed, there are residuals that suggest the presence of minor species such as formic acid and possibly ammonia. Clearly, models of star formation should begin with dust models already coated with a fairly complex mixture of ices.Comment: 4 pages, 5 figures; To appear in Astrophysical Journal Letter

    The c2d Spitzer spectroscopic survey of ices around low-mass young stellar objects II: CO2

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    This paper presents Spitzer-IRS spectroscopy of the CO2 15.2 micron bending mode toward a sample of 50 embedded low-mass stars in nearby star-forming clouds, taken mostly from the ``Cores to Disks (c2d)'' Legacy program. The average abundance of solid CO2 relative to water in low-mass protostellar envelopes is 0.32 +/- 0.02, significantly higher than that found in quiescent molecular clouds and in massive star forming regions. It is found that a decomposition of all the observed CO2 bending mode profiles requires a minimum of five unique components. Roughly 2/3 of the CO2 ice is found in a water-rich environment, while most of the remaining 1/3 is found in a CO environment. Ground-based observations of solid CO toward a large subset of the c2d sample are used to further constrain the CO2:CO component and suggest a model in which low-density clouds form the CO2:H2O component and higher density clouds form the CO2:CO ice during and after the freeze-out of gas-phase CO. It is suggested that the subsequent evolution of the CO2 and CO profiles toward low-mass protostars, in particular the appearance of the splitting of the CO2 bending mode due to pure, crystalline CO2, is first caused by distillation of the CO2:CO component through evaporation of CO due to thermal processing to ~20-30 K in the inner regions of infalling envelopes. The formation of pure CO2 via segregation from the H2O rich mantle may contribute to the band splitting at higher levels of thermal processing (>50 K), but is harder to reconcile with the physical structure of protostellar envelopes around low-luminosity objects.Comment: Accepted for Ap

    An old disk still capable of forming a planetary system

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    From the masses of the planets orbiting the Sun, and the abundance of elements relative to hydrogen, it is estimated that when the Solar System formed, the circumstellar disk must have had a minimum mass of around 0.01 solar masses within about 100 astronomical units of the star. (One astronomical unit is the Earth–Sun distance.) The main constituent of the disk, gaseous molecular hydrogen, does not efficiently emit radiation from the disk mass reservoir, and so the most common measure of the disk mass is dust thermal emission and lines of gaseous carbon monoxide. Carbon monoxide emission generally indicates properties of the disk surface, and the conversion from dust emission to gas mass requires knowledge of the grain properties and the gas-to-dust mass ratio, which probably differ from their interstellar values. As a result, mass estimates vary by orders of magnitude, as exemplified by the relatively old (3–10 million years) star TW Hydrae, for which the range is 0.0005–0.06 solar masses. Here we report the detection of the fundamental rotational transition of hydrogen deuteride from the direction of TW Hydrae. Hydrogen deuteride is a good tracer of disk gas because it follows the distribution of molecular hydrogen and its emission is sensitive to the total mass. The detection of hydrogen deuteride, combined with existing observations and detailed models, implies a disk mass of more than 0.05 solar masses, which is enough to form a planetary system like our own

    Challenges in Designing a National Surveillance Program for Inflammatory Bowel Disease in the United States:

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    This review describes the history of US government funding for surveillance programs in IBD, provides current estimates of the incidence and prevalence of inflammatory bowel diseases (IBD) in the United States (US), and enumerates a number of challenges faced by current and future IBD surveillance programs. A rationale for expanding the focus of IBD surveillance beyond counts of incidence and prevalence, in order to provide a greater understanding of the burden of IBD, disease etiology and pathogenesis, is provided. Lessons learned from other countries are summarized, as well as potential resources that may be used to optimize a new form of IBD surveillance in the US. A consensus recommendation on the goals and available resources for a new model for disease surveillance are provided. This new model should focus upon “surveillance of the burden of disease,” including 1) natural history of disease and 2) outcomes and complications of the disease and/or treatments

    Identification of pediatric septic shock subclasses based on genome-wide expression profiling

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    <p>Abstract</p> <p>Background</p> <p>Septic shock is a heterogeneous syndrome within which probably exist several biological subclasses. Discovery and identification of septic shock subclasses could provide the foundation for the design of more specifically targeted therapies. Herein we tested the hypothesis that pediatric septic shock subclasses can be discovered through genome-wide expression profiling.</p> <p>Methods</p> <p>Genome-wide expression profiling was conducted using whole blood-derived RNA from 98 children with septic shock, followed by a series of bioinformatic approaches targeted at subclass discovery and characterization.</p> <p>Results</p> <p>Three putative subclasses (subclasses A, B, and C) were initially identified based on an empiric, discovery-oriented expression filter and unsupervised hierarchical clustering. Statistical comparison of the three putative subclasses (analysis of variance, Bonferonni correction, <it>P </it>< 0.05) identified 6,934 differentially regulated genes. K-means clustering of these 6,934 genes generated 10 coordinately regulated gene clusters corresponding to multiple signaling and metabolic pathways, all of which were differentially regulated across the three subclasses. Leave one out cross-validation procedures indentified 100 genes having the strongest predictive values for subclass identification. Forty-four of these 100 genes corresponded to signaling pathways relevant to the adaptive immune system and glucocorticoid receptor signaling, the majority of which were repressed in subclass A patients. Subclass A patients were also characterized by repression of genes corresponding to zinc-related biology. Phenotypic analyses revealed that subclass A patients were younger, had a higher illness severity, and a higher mortality rate than patients in subclasses B and C.</p> <p>Conclusion</p> <p>Genome-wide expression profiling can identify pediatric septic shock subclasses having clinically relevant phenotypes.</p
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