Genetic transformation of the bacterium Bacillus firmus for GFP expression and confocal laser microscopy analysis

Bacillus firmus I-1582 is a bacterium with nematicidal effects on root-knot nematodes, a group of plant parasites pertaining to the genus Meloidogyne. This bacterium is able to colonize the plant rhizosphere of some economically important crops and protect them from nematode parasites by reducing their juvenile hatching, their only infective stage. In this final degree project B.firmus was transformed with a green fluorescent protein gene. To transform the bacterium it was necessary the previous protoplast formation using lysozyme and the addition of polyethylene glycol. The plasmid containing this gene, pAD43-25, was cloned and extracted from an E.coli ECE166 culture. Once the plasmid was stable, the transformant was inoculated in tomato roots and nematode eggs. The samples were analyzed after 48 hours at 35°C with confocal laser microscopy to watch the distribution of the bacteria and the biofilm formation, which was fluorescent too. In the taken images is shown that the bacterium was not able to colonize the root neither penetrate the eggs. Although it seems that some of the bacterial cytoplasmic products can cross the egg wall.Bacillus firmus I-1582 es un bacteri amb efectes nematicides en nematodes formadors d'agalles, un grup de paràsits de plantes que pertanyen al gènere Meloidogyne. Aquest bacteri es capaç de colonitzar la rizosfera de plantes d'alguns cultius econòmicament importants i protegir-los dels nematodes paràsits reduint la emergència dels juvenils. En aquest treball de fi de grau B.firmus va ser transformat amb un gen de la proteïna verd fluorescent. Per transformar el bacteri va ser necessària la prèvia formació de protoplasts utilitzant lisozima i la adició de polietilenglicol. El plasmidi que conté el gen, pAD43-25 va ser clonat i extret d'un cultiu d'E.coli ECE166. Un cop el plasmidi era estable, els transformants van ser inoculats a arrel de tomàquet i ous de nematodes. Les mostres van ser analitzades després de 48 hores a 35°C amb microscòpia làser confocal per observar la distribució dels bacteris i la formació de biofilm, el qual també era fluorescent. A les imatges presses s'aprecia que el bacteri no ha estat capaç de colonitzar l'arrel y que tampoc ha penetrat als ous. No obstant sí que sembla que alguns productes citoplasmàtics del bacteri poden travessar la paret de l'ou.Bacillus firmus I-1582 es una bacteria con efectos nematicidas en nematodos formadores de agallas, un grupo de parásitos de plantas pertenecientes al género Meloidogyne. Esta bacteria es capaz de colonizar la rizosfera de plantas de algunos cultivos económicamente importantes y protegerlos de los nematodos parásitos reduciendo la emergencia de juveniles, su única etapa infectiva. En este trabajo de fin de grado B.firmus fue transformado con un gen de la proteína verde fluorescente. Para transformar la bacteria fue necesaria la previa formación de protoplastos usando lisozima y la adición de polietilenglicol. El plásmido que contiene el gen, pAD43-25, fue clonado y extraído de un cultivo de E.coli ECE166. Una vez el plásmido era estable, los transformantes fueron inoculados en raíz de tomate y huevos de nematodos. Las muestras fueron analizadas después de 48 horas a 35°C con microscopía láser confocal para observar la distribución de las bacterias y la formación de biofilm, el cual también era fluorescente. En las imágenes tomadas se aprecia que la bacteria no ha sido capaz de colonizar la raíz y que tampoco ha penetrado en los huevos. No obstante sí que parece que algunos productos citoplasmáticos de la bacteria puedan atravesar la pared del huevo

Assessing Future Water Availability in Arid Regions Using Composition and Salience of Decision Criteria

Water resources development options are usually selected on a least-cost basis. While economic considerations are dominant in choosing projects, there are also a mix of other factors including social demands, political expediency, social equity, and environmental considerations that impact final decisions and development of water supply systems. Understanding local priorities in water resource management decisions can allow for forming expectations of future regional water availability. In this research, we propose that future water availability in arid regions may be assessed by considering key projects that have been identified or planned by regional experts. Using Multi-Criteria Decision Analysis methods as a framework to organize set of decision criteria and their relative salience, the likelihood of selection (and development) of a project can be determined and used to form expectations of future regional water availability. We use this approach in a case study for Jordan, and find that large-scale desalination projects—that have been in the planning books for decades—are now most likely to be pursued and implemented in the country. Finally, we discuss strengths, limitations, and the general applicability of this method for assessing future water availability in other arid regions

The Future Costs of Nuclear Power Using Multiple Expert Elicitations: Effects of RD&D and Elicitation Design

Characterization of the anticipated performance of energy technologies to inform policy decisions increasingly relies on expert elicitation. Knowledge about how elicitation design factors impact the probabilistic estimates emerging from these studies is, however, scarce. We focus on nuclear power, a large-scale low-carbon power option, for which future cost estimates are important for the design of energy policies and climate change mitigation efforts. We use data from three elicitations in the USA and in Europe and assess the role of government research, development, and demonstration (RD&D) investments on expected nuclear costs in 2030. We show that controlling for expert, technology, and design characteristics increases experts' implied public RD&D elasticity of expected costs by 25%. Public sector and industry experts' cost expectations are 14% and 32% higher, respectively than academics. US experts are more optimistic than their EU counterparts, with median expected costs 22% lower. On average, a doubling of public RD&D is expected to result in an 8% cost reduction, but the uncertainty is large. The difference between the 90th and 10th percentile estimates is on average 58% of the experts' median estimates. Public RD&D investments do not affect uncertainty ranges, but US experts are less confident about costs than Europeans

Spin Seebeck effect in Y-type hexagonal ferrite thin films

Spin Seebeck effect (SSE) has been investigated in thin films of two Y-hexagonal ferrites Ba$_2$Zn$_{2}$Fe$_{12}$O$_{22}$ (Zn2Y) and Ba$_2$Co$_{2}$Fe$_{12}$O$_{22}$ (Co2Y) deposited by a spin-coating method on SrTiO$_3$(111) substrate. The selected hexagonal ferrites are both ferrimagnetic with similar magnetic moments at room temperature and both exhibit easy magnetization plane normal to $c$-axis. Despite that, SSE signal was only observed for Zn2Y, whereas no significant SSE signal was detected for Co2Y. We tentatively explain this different behavior by a presence of two different magnetic ions in Co2Y, whose random distribution over octahedral sites interferes the long range ordering and enhances the Gilbert damping constant. The temperature dependence of SSE for Zn2Y was measured and analyzed with regard to the heat flux and temperature gradient relevant to the SSE signal.Comment: 7 pages, 9 figure

International Workshop on Research, Development, and Demonstration to Enhance the Role of Nuclear Energy in Meeting Climate and Energy Challenges

Dramatic growth in nuclear energy would be required for nuclear power to provide a significant part of the carbon-free energy the world is likely to need in the 21st century, or a major part in meeting other energy challenges. This would require increased support from governments, utilities, and publics around the world. Achieving that support is likely to require improved economics and major progress toward resolving issues of nuclear safety, proliferation-resistance, and nuclear waste management. This is likely to require both research, development, and demonstration (RD&D) of improved technologies and new policy approaches. To gather information on the RD&D needs for the future of nuclear energy, the future cost and performance of nuclear technologies, and on the major barriers to large-scale deployment of nuclear energy, a team of researchers at Harvard University and the Fondazione Eni Enrico Mattei (FEEM) conducted two coordinated surveys of nuclear experts. The surveys asked experts how much they would recommend that their governments spend on nuclear energy RD&D; what progress in cost and performance might be expected by 2030 if those recommendations were followed; and what other factors might constrain or promote future nuclear energy growth. Leading experts from the United States (U.S.) and the European Union (E.U.) participated in this expert elicitation surveys during the summer and fall of 2010. In April 2011, the FEEM and Harvard teams held a workshop in Venice, Italy with a subset of the participating E.U. and U.S. experts to present and discuss the results of the elicitations, in an effort to understand where there is consensus and where the most important disputes and uncertainties lie. Given the Fukushima nuclear accident in Japan, the meeting opened with a discussion of the significance of that event for the future of nuclear power, and of the main lessons learned

High mountain dwarf shrubs under climate change: effects of in situ experimental warming, CO2 enrichment and drought on their growth and functioning = Subarbusts d'alta muntanya davant el canvi climàtic. Efectes de l'escalfament, l'enriquiment de CO2 i la sequera experimentals in situ en el seu creixement i funcionament

[eng] High-elevation and high-latitude ecosystems are expected to be especially vulnerable to global change. Dwarf shrub species cover extensive areas in high mountain ecosystems, and warmer temperatures and land-use change have favoured their expansion. Co-occurring dwarf shrub species may respond differently to climate change, which can have major effects on their interactions, growth and performance, and ultimately lead to drastic changes in plant communities. In this Thesis, warming, CO2 enrichment and drought experiments have been carried out at the ecotone between the subalpine and alpine belts in different study sites across the Pyrenees and the Alps. The objective was to assess how these factors may influence dwarf shrub species interactions, growth, xylem anatomy and C allocation dynamics. Results of this Thesis show species-specific responses to the treatments, and these responses differed in magnitude and direction depending on the treatment applied. Vaccinium myrtillus responded positively to increased temperatures, whereas the co-occurring Vaccinium uliginosum and Empetrum hermaphroditum showed no response. Vaccinium myrtillus growth and functioning was similar across stands differing in neighbouring shrub species and we did not find evidence of alterations in plant-plant interactions with warming. Vaccinium myrtillus responded positively to CO2 enrichment with an increased stem basal area and a lagged increased vessel lumen size and xylem-specific hydraulic conductivity, but hydraulic efficiency decreased with warming. Although there was a correlation between growth and accumulated hydraulic conductivity, the growth-anatomical relationship changed with soil warming and CO2 enrichment, and growth stimulation occurred without immediate accompanying changes in the xylem anatomy. We did not find major effects of the experimental summer drought on V. myrtillus water potential and its allocation of recently assimilated C. However, V. uliginosum showed evidence of mild stress, since its stomatal conductance, water potential and speed of C transfer to belowground organs were reduced under drought. All these results suggest that climate change acts differently on co-occurring species, which may lead to future changes in plant community composition, structure and function.[cat] Els ecosistemes d'elevades altituds i latituds són especialment vulnerables al canvi global. Les espècies subarbustives poden cobrir àrees extenses en zones d'alta muntanya, i l'augment de la temperatura i els canvis en l'ús del sòl n'afavoreixen l'expansió. Les espècies subarbustives que coexisteixen podrien respondre de manera diferent al canvi climàtic, cosa que podria tenir grans efectes en les seves interaccions, així com en el seu creixement i el seu funcionament i, en última instància, comportar canvis dràstics en les comunitats vegetals. En aquesta Tesi Doctoral s'han dut a terme experiments d'escalfament, d’augment de la concentració de CO2 de l’aire i de sequera a l'ecotò entre els estatges subalpí i alpí en diferents zones del Pirineu i dels Alps. L'objectiu era comprendre com aquests factors afecten les interaccions entre espècies subarbustives, el seu creixement, l’anatomia del xilema i la distribució de carboni (C). Els resultats d'aquesta Tesi mostren respostes específiques als tractaments. A més, aquestes respostes diferien en magnitud i direcció en funció del tractament aplicat. Vaccinium myrtillus va respondre positivament a l'escalfament, mentre que les espècies coexistents Vaccinium uliginosum i Empetrum hermaphroditum no van mostrar cap resposta. El creixement i funcionament de V. myrtillus era similar en claps amb diferent composició d'espècies arbustives i, contràriament al resultat de nombrosos estudis previs, no es van trobar evidències d'alteracions en les interaccions planta-planta amb l'escalfament. Vaccinium myrtillus va respondre positivament a l'augment de CO2 amb un augment de l'àrea basal i un augment retardat en la mida dels vasos conductors i la conductivitat hidràulica específica. Tanmateix, l'eficiència hidràulica es va reduir amb l'escalfament. Tot i que es va trobar una correlació entre el creixement i l’anatomia en termes de conductivitat hidràulica acumulada en V. myrtillus, aquesta relació va canviar amb els tractaments d'escalfament i augment de CO2, i el creixement va augmentar sota aquests tractaments sense que es produïssin canvis immediats en l'anatomia del xilema que l'acompanyessin. La sequera experimental no va tenir grans efectes en el potencial hídric ni en la distribució de C recentment assimilat de V. myrtillus. Tanmateix, V. uliginosum va presentar lleugers signes d'estrès amb la sequera, ja que la seva conductància estomàtica i el seu potencial hídric es van veure reduïts, així com també la velocitat de transferència de C cap a òrgans subterranis. Tots aquests resultats suggereixen que el canvi climàtic actua de diferent manera sobre espècies coexistents, cosa que podria comportar canvis en la composició de les comunitats, així com en la seva estructura i funcionament en el futur

Palaeoenvironmental and diagenetic reconstruction of a closed-lacustrine carbonate system - the challenging marginal setting of the Miocene Ries Crater Lake (Germany)

Chemostratigraphic studies on lacustrine sedimentary sequences provide essential insights on past cyclic climatic events, on their repetition and prediction through time. Diagenetic overprint of primary features often hinders the use of such studies for palaeoenvironmental reconstruction. Here the potential of integrated geochemical and petrographic methods is evaluated to record freshwater to saline oscillations within the ancient marginal lacustrine carbonates of the Miocene Ries Crater Lake (Germany). This area is critical because it represents the transition from shoreline to proximal domains of a hydrologically closed system, affected by recurrent emergent events, representing the boundaries of successive sedimentary cycles. Chemostratigraphy targets shifts related to subaerial exposure and/or climatic fluctuations. Methods combine facies changes with δ13C–δ18O chemostratigraphy from matrix carbonates across five closely spaced, temporally equivalent stratigraphic sections. Isotope composition of ostracod shells, gastropods and cements is provided for comparison. Cathodoluminescence and back‐scatter electron microscopy were performed to discriminate primary (syn‐)depositional, from secondary diagenetic features. Meteoric diagenesis is expressed by substantial early dissolution and dark blue luminescent sparry cements carrying negative δ13C and δ18O. Sedimentary cycles are not correlated by isotope chemostratigraphy. Both matrix δ13C and δ18O range from ca −7·5 to +4·0‰ and show clear positive covariance (R = 0·97) whose nature differs from that of previous basin‐oriented studies on the lake: negative values are here unconnected to original freshwater lacustrine conditions but reflect extensive meteoric diagenesis, while positive values probably represent primary saline lake water chemistry. Noisy geochemical curves relate to heterogeneities in (primary) porosity, resulting in selective carbonate diagenesis. This study exemplifies that ancient lacustrine carbonates, despite extensive meteoric weathering, are able to retain key information for both palaeoenvironmental reconstruction and the understanding of diagenetic processes in relation to those primary conditions. Also, it emphasizes the limitation of chemostratigraphy in fossil carbonates, and specifically in settings that are sensitive for the preservation of primary environmental signals, such as lake margins prone to meteoric diagenesis

Transforming U.S. Energy Innovation

The United States and the world need a revolution in energy technology—a revolution that would improve the performance of our energy systems to face the challenges ahead. A dramatic increase in the pace of energy innovation is crucial to meet the challenges of: • Energy and national security, to address the dangers of undue reliance on dwindling supplies of oil increasingly concentrated in some of the most volatile regions of the world, and to limit the connection between nuclear energy and the spread of nuclear weapons; • Environmental sustainability, to reduce the wide range of environmental damages due to energy production and use, from fine particulate emissions at coal plants, to oil spills, to global climate disruption; and • Economic competitiveness, to seize a significant share of the multi-trillion-dollar clean energy technology market and improve the balance of payments by increasing exports, while reducing the hundreds of billions of dollars spent every year on importing oil. In an intensely competitive and interdependent global landscape, and in the face of large climate risks from ongoing U.S. reliance on a fossil-fuel based energy system, it is important to maintain and expand long-term investments in the energy future of the U.S. even at a time of budget stringency. It is equally necessary to think about how to improve the efficiency of those investments, through strengthening U.S. energy innovation institutions, providing expanded incentives for private-sector innovation, and seizing opportunities where international cooperation can accelerate innovation. The private sector role is key: in the United States the vast majority of the energy system is owned by private enterprises, whose innovation and technology deployment decisions drive much of the country’s overall energy systems. Efficiently utilizing government investments in energy innovation requires understanding the market incentives that drive private firms to invest in advanced energy technologies, including policy stability and predictability. The U.S. government has already launched new efforts to accelerate energy innovation. In particular, the U.S. Department of Energy is undertaking a Quadrennial Technology Review to identify the most promising opportunities and provide increased coherence and stability. Our report offers analysis and recommendations designed to accelerate the pace at which better energy technologies are discovered, developed, and deployed, and is focused in four key areas: • Designing an expanded portfolio of federal investments in energy research, development, demonstration (ERD&D), and complementary policies to catalyze the deployment of novel energy technologies; • Increasing incentives for private-sector innovation and strengthening federal-private energy innovation partnerships; • Improving the management of energy innovation institutions to maximize the results of federal investments; and • Expanding and coordinating international energy innovation cooperation to bring ideas and resources together across the globe to address these global challenges

Aprendizaje colaborativo/competitivo mediante gamificación, a través del uso integrado de un mando wiimote como pizarra digital interactiva, un sistema virtual de respuesta en el aula, y dispositivos móviles con acceso a internet

Integración de pizarra digital, una aplicación informática propia como sistema de respuesta (clicker) y otras aplicaciones no propias para facilitar la evaluación continua, aumentar la motivación y rendimiento de los estudiantes a un coste muy bajo