Transcriptome analysis of grain development in hexaploid wheat

Background: Hexaploid wheat is one of the most important cereal crops for human nutrition. Molecular understanding of the biology of the developing grain will assist the improvement of yield and quality traits for different environments. High quality transcriptomics is a powerful method to increase this understanding. Results: The transcriptome of developing caryopses from hexaploid wheat ( Triticum aestivum, cv. Hereward) was determined using Affymetrix wheat GeneChip (R) oligonucleotide arrays which have probes for 55,052 transcripts. Of these, 14,550 showed significant differential regulation in the period between 6 and 42 days after anthesis ( daa). Large changes in transcript abundance were observed which were categorised into distinct phases of differentiation ( 6 - 10 daa), grain fill ( 12 - 21 daa) and desiccation/maturation ( 28 - 42 daa) and were associated with specific tissues and processes. A similar experiment on developing caryopses grown with dry and/or hot environmental treatments was also analysed, using the profiles established in the first experiment to show that most environmental treatment effects on transcription were due to acceleration of development, but that a few transcripts were specifically affected. Transcript abundance profiles in both experiments for nine selected known and putative wheat transcription factors were independently confirmed by real time RT-PCR. These expression profiles confirm or extend our knowledge of the roles of the known transcription factors and suggest roles for the unknown ones. Conclusion: This transcriptome data will provide a valuable resource for molecular studies on wheat grain. It has been demonstrated how it can be used to distinguish general developmental shifts from specific effects of treatments on gene expression and to diagnose the probable tissue specificity and role of transcription factors.

In planta proteomics and proteogenomics of the biotrophic barley fungal pathogen blumeria graminis f. sp hordei

To further our understanding of powdery mildew biology during infection, we undertook a systematic shotgun proteomics analysis of the obligate biotroph Blumeria graminis f. sp. hordei at different stages of development in the host. Moreover we used a proteogenomics approach to feed information into the annotation of the newly sequenced genome. We analyzed and compared the proteomes from three stages of development representing different functions during the plant-dependent vegetative life cycle of this fungus. We identified 441 proteins in ungerminated spores, 775 proteins in epiphytic sporulating hyphae, and 47 proteins from haustoria inside barley leaf epidermal cells and used the data to aid annotation of the B. graminis f. sp. hordei genome. We also compared the differences in the protein complement of these key stages. Although confirming some of the previously reported findings and models derived from the analysis of transcriptome dynamics, our results also suggest that the intracellular haustoria are subject to stress possibly as a result of the plant defense strategy, including the production of reactive oxygen species. In addition, a number of small haustorial proteins with a predicted N-terminal signal peptide for secretion were identified in infected tissues: these represent candidate effector proteins that may play a role in controlling host metabolism and immunity. Molecular & Cellular Proteomics 8: 2368-2381, 2009.

In planta proteomics and proteogenomics of the biotrophic barley fungal pathogen blumeria graminis f. sp hordei

To further our understanding of powdery mildew biology during infection, we undertook a systematic shotgun proteomics analysis of the obligate biotroph Blumeria graminis f. sp. hordei at different stages of development in the host. Moreover we used a proteogenomics approach to feed information into the annotation of the newly sequenced genome. We analyzed and compared the proteomes from three stages of development representing different functions during the plant-dependent vegetative life cycle of this fungus. We identified 441 proteins in ungerminated spores, 775 proteins in epiphytic sporulating hyphae, and 47 proteins from haustoria inside barley leaf epidermal cells and used the data to aid annotation of the B. graminis f. sp. hordei genome. We also compared the differences in the protein complement of these key stages. Although confirming some of the previously reported findings and models derived from the analysis of transcriptome dynamics, our results also suggest that the intracellular haustoria are subject to stress possibly as a result of the plant defense strategy, including the production of reactive oxygen species. In addition, a number of small haustorial proteins with a predicted N-terminal signal peptide for secretion were identified in infected tissues: these represent candidate effector proteins that may play a role in controlling host metabolism and immunity. Molecular & Cellular Proteomics 8: 2368-2381, 2009.

The influence of lodgepole pine (Pinus contorta) provenance on the development and survival of larvae of the pine beauty moth Panolis flammea (Lepidoptera: Noctuidae)

Larvae of the pine beauty moth Panolis flammae (Denis & Schiffermuller) were reared in sleeve cages on five different seed origins (provenances) of pole stage Pinus contorta in the field in each of four years from 1985 to 1988. Survival varied significantly between the years. In those years when survival was high, significant differences between tree provenance were not found. However, between provenance significant differences were found in larval weight and stage of development. In the years when survival was low, the results seen in good years were reversed. Significant differences attributable to provenance were found but these were not reflected in significant differences between larval weight or development. In addition, there was a significant correlation between the proportion surviving and larval weight, which was not the case in those years where larval survival was high. The results are discussed in light of the pest status of P, flammea in Britain and in view of current silvicultural policies. The use of trees resistant to insect attack as part of an integrated pest management programme is highlighted and the need to coordinate laboratory and field studies so as to control for environmental variation discussed.

Temperature and the development rates of thrips: evidence for a constraint on local adaptation?

Typically, the relationship between insect development and temperature is described by two characteristics: the minimum temperature needed for development to occur (T-min) and the number of day degrees required (DDR) for the completion of development. We investigated these characteristics in three English populations of Thrips major and T tabaci [Cawood, Yorkshire (N53degrees49', W1degrees7'); Boxworth, Cambridgeshire (N52degrees15', W0degrees1'); Silwood Park, Berkshire (N51degrees24', W0degrees38')], and two populations of Frankliniella occidentalis (Cawood; Silwood Park). While there were no significant differences among populations in either T-min (mean for T major = 7.0degreesC; T tabaci = 5.9degreesC; F. occidentalis = 6.7degreesC) or DDR (mean for T major = 229.9; T tabaci = 260.8; F occidentalis = 233.4), there were significant differences in the relationship between temperature and body size, suggesting the presence of geographic variation in this trait. Using published data, in addition to those newly collected, we found a negative relationship between T-min. and DDR for F occidentalis and T tabaci, supporting the hypothesis that a trade-off between T-min and DDR may constrain adaptation to local climatic conditions.

An examination of actors that impact technology introduction to FM

Developing and implementing a technology for Facilities Management (FM) can be a complex process. This is particularly the case when a technology impacts on an organisation as a whole. There are often a number of relevant actors, internal and external to FM, who should be engaged. This engagement is guided by the strategy of the organisation which is led by top management decisions. Indeed, it is top management who have the final decision to implement a technology. Actors of top management and other relevant actors will have their own discourses toward the implementation of the technology based on how they foresee the technology befittingly benefitting the organisation. This paper examines actors who play a relevant and necessary part in supporting and implementing a technology to FM. It examines how an actor’s discourse toward the project inhibits or speeds up the implementation of a technology. The methods used for this paper are based on a two year case study in a FM department where a technology development was observed and interviews with key participants were conducted. Critical discourse analysis is used to analyse the data. Prominent discourses that emerge from the data are emphasised during the process of introducing the technology. This research moves beyond focusing purely on project successes but examines the difficulties and the hurdles that must be overcome to reach a successful technology implementation.

Technologies for biological containment of GM and Non-GM crops

International Perspective The development of GM technology continues to expand into increasing numbers of crops and conferred traits. Inevitably, the focus remains on the major field crops of soybean, maize, cotton, oilseed rape and potato with introduced genes conferring herbicide tolerance and/or pest resistance. Although there are comparatively few GM crops that have been commercialised to date, GM versions of 172 plant species have been grown in field trials in 31 countries. European Crops with Containment Issues Of the 20 main crops in the EU there are four for which GM varieties are commercially available (cotton, maize for animal feed and forage, and oilseed rape). Fourteen have GM varieties in field trials (bread wheat, barley, durum wheat, sunflower, oats, potatoes, sugar beet, grapes, alfalfa, olives, field peas, clover, apples, rice) and two have GM varieties still in development (rye, triticale). Many of these crops have hybridisation potential with wild and weedy relatives in the European flora (bread wheat, barley, oilseed rape, durum wheat, oats, sugar beet and grapes), with escapes (sunflower); and all have potential to cross-pollinate fields non-GM crops. Several fodder crops, forestry trees, grasses and ornamentals have varieties in field trials and these too may hybridise with wild relatives in the European flora (alfalfa, clover, lupin, silver birch, sweet chestnut, Norway spruce, Scots pine, poplar, elm, Agrostis canina, A. stolonifera, Festuca arundinacea, Lolium perenne, L. multiflorum, statice and rose). All these crops will require containment strategies to be in place if it is deemed necessary to prevent transgene movement to wild relatives and non-GM crops. Current Containment Strategies A wide variety of GM containment strategies are currently under development, with a particular focus on crops expressing pharmaceutical products. Physical containment in greenhouses and growth rooms is suitable for some crops (tomatoes, lettuce) and for research purposes. Aquatic bioreactors of some non-crop species (algae, moss, and duckweed) expressing pharmaceutical products have been adopted by some biotechnology companies. There are obvious limitations of the scale of physical containment strategies, addressed in part by the development of large underground facilities in the US and Canada. The additional resources required to grow plants underground incurs high costs that in the long term may negate any advantage of GM for commercial productioNatural genetic containment has been adopted by some companies through the selection of either non-food/feed crops (algae, moss, duckweed) as bio-pharming platforms or organisms with no wild relatives present in the local flora (safflower in the Americas). The expression of pharmaceutical products in leafy crops (tobacco, alfalfa, lettuce, spinach) enables growth and harvesting prior to and in the absence of flowering. Transgenically controlled containment strategies range in their approach and degree of development. Plastid transformation is relatively well developed but is not suited to all traits or crops and does not offer complete containment. Male sterility is well developed across a range of plants but has limitations in its application for fruit/seed bearing crops. It has been adopted in some commercial lines of oilseed rape despite not preventing escape via seed. Conditional lethality can be used to prevent flowering or seed development following the application of a chemical inducer, but requires 100% induction of the trait and sufficient application of the inducer to all plants. Equally, inducible expression of the GM trait requires equally stringent application conditions. Such a method will contain the trait but will allow the escape of a non-functioning transgene. Seed lethality (‘terminator’ technology) is the only strategy at present that prevents transgene movement via seed, but due to public opinion against the concept it has never been trialled in the field and is no longer under commercial development. Methods to control flowering and fruit development such as apomixis and cleistogamy will prevent crop-to-wild and wild-to-crop pollination, but in nature both of these strategies are complex and leaky. None of the genes controlling these traits have as yet been identified or characterised and therefore have not been transgenically introduced into crop species. Neither of these strategies will prevent transgene escape via seed and any feral apomicts that form are arguably more likely to become invasives. Transgene mitigation reduces the fitness of initial hybrids and so prevents stable introgression of transgenes into wild populations. However, it does not prevent initial formation of hybrids or spread to non-GM crops. Such strategies could be detrimental to wild populations and have not yet been demonstrated in the field. Similarly, auxotrophy prevents persistence of escapes and hybrids containing the transgene in an uncontrolled environment, but does not prevent transgene movement from the crop. Recoverable block of function, intein trans-splicing and transgene excision all use recombinases to modify the transgene in planta either to induce expression or to prevent it. All require optimal conditions and 100% accuracy to function and none have been tested under field conditions as yet. All will contain the GM trait but all will allow some non-native DNA to escape to wild populations or to non-GM crops. There are particular issues with GM trees and grasses as both are largely undomesticated, wind pollinated and perennial, thus providing many opportunities for hybridisation. Some species of both trees and grass are also capable of vegetative propagation without sexual reproduction. There are additional concerns regarding the weedy nature of many grass species and the long-term stability of GM traits across the life span of trees. Transgene stability and conferred sterility are difficult to trial in trees as most field trials are only conducted during the juvenile phase of tree growth. Bio-pharming of pharmaceutical and industrial compounds in plants Bio-pharming of pharmaceutical and industrial compounds in plants offers an attractive alternative to mammalian-based pharmaceutical and vaccine production. Several plantbased products are already on the market (Prodigene’s avidin, β-glucuronidase, trypsin generated in GM maize; Ventria’s lactoferrin generated in GM rice). Numerous products are in clinical trials (collagen, antibodies against tooth decay and non-Hodgkin’s lymphoma from tobacco; human gastric lipase, therapeutic enzymes, dietary supplements from maize; Hepatitis B and Norwalk virus vaccines from potato; rabies vaccines from spinach; dietary supplements from Arabidopsis). The initial production platforms for plant-based pharmaceuticals were selected from conventional crops, largely because an established knowledge base already existed. Tobacco and other leafy crops such as alfalfa, lettuce and spinach are widely used as leaves can be harvested and no flowering is required. Many of these crops can be grown in contained greenhouses. Potato is also widely used and can also be grown in contained conditions. The introduction of morphological markers may aid in the recognition and traceability of crops expressing pharmaceutical products. Plant cells or plant parts may be transformed and maintained in culture to produce recombinant products in a contained environment. Plant cells in suspension or in vitro, roots, root cells and guttation fluid from leaves may be engineered to secrete proteins that may be harvested in a continuous, non-destructive manner. Most strategies in this category remain developmental and have not been commercially adopted at present. Transient expression produces GM compounds from non-GM plants via the utilisation of bacterial or viral vectors. These vectors introduce the trait into specific tissues of whole plants or plant parts, but do not insert them into the heritable genome. There are some limitations of scale and the field release of such crops will require the regulation of the vector. However, several companies have several transiently expressed products in clinical and pre-clinical trials from crops raised in physical containment.

Change, rigidity & delay in the UK system of land-use development control

The British system of development control is time-consuming and uncertain in outcome. Moreover, it is becoming increasingly overloaded as it has gradually switched away from being centred on a traditional ‘is it an appropriate land-use?’ type approach to one based on multi-faceted inspections of projects and negotiations over the distribution of the potential financial gains arising from them. Recent policy developments have centred on improving the operation of development control. This paper argues that more fundamental issues may be a stake as well. Important market changes have increased workloads. Furthermore, the UK planning system's institutional framework encourages change to move in specific directions, which is not always helpful. If expectations of increased long-term housing supply are to be met more substantial changes to development control may be essential but hard to achieve.

Star/poverty space: the making of the ‘development celebrity’

What is it that gives celebrities the voice and authority to do and say the things they do in the realm of development politics? Asked another way, how is celebrity practised and, simultaneously, how does this praxis make celebrity, personas, politics and, indeed, celebrities themselves? In this article, we explore this ‘celebrity praxis’ through the lens of the creation of the contemporary ‘development celebrity’ in those stars working for development writ large in the so-called Third World. Drawing on work in science studies, material cultures and the growing geo-socio-anthropologies of things, the key to understanding the material practices embedded in and creating development celebrity networks is the multiple and complex circulations of the everyday and bespectacled artefacts of celebrity. Conceptualised as the ‘celebrity–consumption–compassion complex’, the performances of development celebrities are as much about everyday events, materials, technologies, emotions and consumer acts as they are about the mediated and liquidised constructions of the stars who now ‘market’ development.Moreover, this complex is constructed by and constructs what we are calling ‘star/poverty space’ that works to facilitate the ‘expertise’ and ‘authenticity’ and, thus, elevated voice and authority, of development celebrities through poverty tours, photoshoots, textual and visual diaries, websites and tweets. In short, the creation of star/poverty space is performed through a kind of ‘materiality of authenticity’ that is at the centre of the networks of development celebrity. The article concludes with several brief observations about the politics, possibilities and problematics of development celebrities and the star/poverty spaces that they create.

The new localism, anti-political development machines, and the role of planning consultants: lessons from London's South Bank

This paper draws on a study of the politics of development planning in London’s South Bank to examine wider trends in the governance of contemporary cities. It assesses the impacts and outcomes of so-called new localist reforms and argues that we are witnessing two principal trends. First, governance processes are increasingly dominated by anti-democratic development machines, characterized by new assemblages of public- and private-sector experts. These machines reflect and reproduce a type of development politics in which there is a greater emphasis on a pragmatic realism and a politics of delivery. Second, the presence of these machines is having a significant impact on the politics of planning. Democratic engagement is not seen as the basis for new forms of localism and community control. Instead, it is presented as a potentially disruptive force that needs to be managed by a new breed of skilled private-sector consultant. The paper examines these wider shifts in urban politics before focusing on the connections between emerging development machines and local residential and business communities. It ends by highlighting some of the wider implications of change for democratic modes of engagement and nodes of resistance in urban politics.

Ontogenetic Changes in the Bell Morphology and Kinematics and Swimming Behavior of Rowing Medusae: the Special Case of the Limnomedusa Liriope tetraphylla

Swimming animals may experience significant changes in the Reynolds number (Re) of their surrounding fluid flows throughout ontogeny. Many medusae experience Re environments with significant viscous forces as small juveniles but inertially dominated Re environments as adults. These different environments may affect their propulsive strategies. In particular, rowing, a propulsive strategy with ecological advantages for large adults, may be constrained by viscosity for small juvenile medusae. We examined changes in the bell morphology and swimming kinematics of the limnomedusa Liriope tetraphylla at different stages of development. L. tetraphylla maintained an oblate bell (fineness ratio approximate to 0.5-0.6), large velar aperture ratio (R(v) approximate to 0.5-0.8), and rapid bell kinematics throughout development. These traits enabled it to use rowing propulsion at all stages except the very smallest sizes observed (diameter = 0.14 cm). During the juvenile stage, very rapid bell kinematics served to increase Re sufficiently for rowing propulsion. Other taxa that use rowing propulsion as adults, such as leptomedusae and scyphomedusae, typically utilize different propulsive strategies as small juveniles to function in low Re environments. We compared the performance values of the different propulsive modes observed among juvenile medusae.

Evidence of fluoride oxidation for development of the electrochemically-induced crystallization observed in lead oxyfluoroborate glasses

This work reports on a distinct experimental procedure conceived to closely approach the question of development of crystallization in lead oxyfluoroborate glasses in the presence of an electric field. After proposing earlier that this phenomenon should involve occurrence of redox-type electrochemical reactions occurring at the electrodes. it was in fact recently shown that a direct contact of the glasses with both the cathode and anode revealed essential, provided that crystallization did not develop when ions migration to these electrodes became frustrated. The present study demonstrates that. even in Pt,Ag/Glass/YSZ:PbF(2)/Ag,Pt-type electrochemical cells subjected to electric field action, where YSZ:PbF(2) represents composite-like mixtures (formed by Y(2)O(3)-doped ZrO(2) and PbF(2)) placed between the glass and anode. crystallization was observable in given cases. In summary, supported by (micro)structural and electrical characterizations, clear evidence is provided here that, besides Pb(2+) reduction at the cathode, crystallization really involves simultaneous F(-) oxidation at the anode, completing thus the whole redox electrochemical reaction so far postulated. In these cases, F(-) migration to the anode was achievable following PbF(2) percolative-like paths through the YSZ:PbF(2) mixtures. (C) 2010 Elsevier B.V. All rights reserved.

Evaluation of a PVT Air Collector

Hybrid Photovoltaic Thermal (PVT) collectors are an emerging technology that combines PV and solar thermal systems in a single solar collector producing heat and electricity simultaneously. The focus of this thesis work is to evaluate the performance of unglazed open loop PVT air system integrated on a garage roof in Borlänge. As it is thought to have a significant potential for preheating ventilation of the building and improving the PV modules electrical efficiency. The performance evaluation is important to optimize the cooling strategy of the collector in order to enhance its electrical efficiency and maximize the production of thermal energy. The evaluation process involves monitoring the electrical and thermal energies for a certain period of time and investigating the cooling effect on the performance through controlling the air mass flow provided by a variable speed fan connected to the collector by an air distribution duct. The distribution duct transfers the heated outlet air from the collector to inside the building. The PVT air collector consists of 34 Solibro CIGS type PV modules (115 Wp for each module) which are roof integrated and have replaced the traditional roof material. The collector is oriented toward the south-west with a tilt of 29 ᵒ. The collector consists of 17 parallel air ducts formed between the PV modules and the insulated roof surface. Each air duct has a depth of 0.05 m, length of 2.38 m and width of 2.38 m. The air ducts are connected to each other through holes. The monitoring system is based on using T-type thermocouples to measure the relevant temperatures, air sensor to measure the air mass flow. These parameters are needed to calculate the thermal energy. The monitoring system contains also voltage dividers to measure the PV modules voltage and shunt resistance to measure the PV current, and AC energy meters which are needed to calculate the produced electrical energy. All signals recorded from the thermocouples, voltage dividers and shunt resistances are connected to data loggers. The strategy of cooling in this work was based on switching the fan on, only when the difference between the air duct temperature (under the middle of top of PV column) and the room temperature becomes higher than 5 °C. This strategy was effective in term of avoiding high electrical consumption by the fan, and it is recommended for further development. The temperature difference of 5 °C is the minimum value to compensate the heat losses in the collecting duct and distribution duct. The PVT air collector has an area of (Ac=32 m2), and air mass flow of 0.002 kg/s m2. The nominal output power of the collector is 4 kWppv (34 CIGS modules with 115 Wppvfor each module). The collector produces thermal output energy of 6.88 kWth/day (0.21 kWth/m2 day) and an electrical output energy of 13.46 kWhel/day (0.42 kWhel/m2 day) with cooling case. The PVT air collector has a daily thermal energy yield of 1.72 kWhth/kWppv, and a daily PV electrical energy yield of 3.36 kWhel /kWppv. The fan energy requirement in this case was 0.18 kWh/day which is very small compared to the electrical energy generated by the PV collector. The obtained thermal efficiency was 8 % which is small compared to the results reported in literature for PVT air collectors. The small thermal efficiency was due to small operating air mass flow. Therefore, the study suggests increasing the air mass flow by a factor of 25. The electrical efficiency was fluctuating around 14 %, which is higher than the theoretical efficiency of the PV modules, and this discrepancy was due to the poor method of recording the solar irradiance in the location. Due to shading effect, it was better to use more than one pyranometer.

Health system context and implementation of evidence-based practices-development and validation of the Context Assessment for Community Health (COACH) tool for low- and middle-income settings

Background: The gap between what is known and what is practiced results in health service users not benefitting from advances in healthcare, and in unnecessary costs. A supportive context is considered a key element for successful implementation of evidence-based practices (EBP). There were no tools available for the systematic mapping of aspects of organizational context influencing the implementation of EBPs in low- and middle-income countries (LMICs). Thus, this project aimed to develop and psychometrically validate a tool for this purpose. Methods: The development of the Context Assessment for Community Health (COACH) tool was premised on the context dimension in the Promoting Action on Research Implementation in Health Services framework, and is a derivative product of the Alberta Context Tool. Its development was undertaken in Bangladesh, Vietnam, Uganda, South Africa and Nicaragua in six phases: (1) defining dimensions and draft tool development, (2) content validity amongst in-country expert panels, (3) content validity amongst international experts, (4) response process validity, (5) translation and (6) evaluation of psychometric properties amongst 690 health workers in the five countries. Results: The tool was validated for use amongst physicians, nurse/midwives and community health workers. The six phases of development resulted in a good fit between the theoretical dimensions of the COACH tool and its psychometric properties. The tool has 49 items measuring eight aspects of context: Resources, Community engagement, Commitment to work, Informal payment, Leadership, Work culture, Monitoring services for action and Sources of knowledge. Conclusions: Aspects of organizational context that were identified as influencing the implementation of EBPs in high-income settings were also found to be relevant in LMICs. However, there were additional aspects of context of relevance in LMICs specifically Resources, Community engagement, Commitment to work and Informal payment. Use of the COACH tool will allow for systematic description of the local healthcare context prior implementing healthcare interventions to allow for tailoring implementation strategies or as part of the evaluation of implementing healthcare interventions and thus allow for deeper insights into the process of implementing EBPs in LMICs.