The transformation of banana with potential virus resistance genes

One approach to reducing the yield losses caused by banana viral diseases is the use of genetic engineering and pathogen-derived resistance strategies to generate resistant cultivars. The development of transgenic virus resistance requires an efficient banana transformation method, particularly for commercially important 'Cavendish' type cultivars such as 'Grand Nain'. Prior to this study, only two examples of the stable transformation of banana had been reported, both of which demonstrated the principle of transformation but did not characterise transgenic plants in terms of the efficiency at which individual transgenic lines were generated, relative activities of promoters in stably transformed plants, and the stability of transgene expression. The aim of this study was to develop more efficient transformation methods for banana, assess the activity of some commonly used and also novel promoters in stably transformed plants, and transform banana with genes that could potentially confer resistance to banana bunchy top nanovirus (BBTV) and banana bract mosaic potyvirus (BBrMV). A regeneration system using immature male flowers as the explant was established. The frequency of somatic embryogenesis in male flower explants was influenced by the season in which the inflorescences were harvested. Further, the media requirements of various banana cultivars in respect to the 2,4-D concentration in the initiation media also differed. Following the optimisation of these and other parameters, embryogenic cell suspensions of several banana (Musa spp.) cultivars including 'Grand Nain' (AAA), 'Williams' (AAA), 'SH-3362' (AA), 'Goldfinger' (AAAB) and 'Bluggoe' (ABB) were successfully generated. Highly efficient transformation methods were developed for both 'Bluggoe' and 'Grand Nain'; this is the first report of microprojectile bombardment transformation of the commercially important 'Grand Nain' cultivar. Following bombardment of embryogenic suspension cells, regeneration was monitored from single transfom1ed cells to whole plants using a reporter gene encoding the green fluorescent protein (gfp). Selection with kanamycin enabled the regeneration of a greater number of plants than with geneticin, while still preventing the regeneration of non-transformed plants. Southern hybridisation confirmed the neomycin phosphotransferase gene (npt II) was stably integrated into the banana genome and that multiple transgenic lines were derived from single bombardments. The activity, stability and tissue specificity of the cauliflower mosaic virus 358 (CaMV 35S) and maize polyubiquitin-1 (Ubi-1) promoters were examined. In stably transformed banana, the Ubi-1 promoter provided approximately six-fold higher p-glucuronidase (GUS) activity than the CaMV 35S promoter, and both promoters remained active in glasshouse grown plants for the six months they were observed. The intergenic regions ofBBTV DNA-I to -6 were isolated and fused to either the uidA (GUS) or gfjJ reporter genes to assess their promoter activities. BBTV promoter activity was detected in banana embryogenic cells using the gfp reporter gene. Promoters derived from BBTV DNA-4 and -5 generated the highest levels of transient activity, which were greater than that generated by the maize Ubi-1 promoter. In transgenic banana plants, the activity of the BBTV DNA-6 promoter (BT6.1) was restricted to the phloem of leaves and roots, stomata and root meristems. The activity of the BT6.1 promoter was enhanced by the inclusion of intron-containing fragments derived from the maize Ubi-1, rice Act-1, and sugarcane rbcS 5' untranslated regions in GUS reporter gene constructs. In transient assays in banana, the rice Act-1 and maize Ubi-1 introns provided the most significant enhancement, increasing expression levels 300-fold and 100-fold, respectively. The sugarcane rbcS intron increased expression about 10-fold. In stably transformed banana plants, the maize Ubi-1 intron enhanced BT6.1 promoter activity to levels similar to that of the CaMV 35S promoter, but did not appear to alter the tissue specificity of the promoter. Both 'Grand Nain' and 'Bluggoe' were transformed with constructs that could potentially confer resistance to BBTV and BBrMV, including constructs containing BBTV DNA-1 major and internal genes, BBTV DNA-5 gene, and the BBrMV coat protein-coding region all under the control of the Ubi-1 promoter, while the BT6 promoter was used to drive the npt II selectable marker gene. At least 30 transgenic lines containing each construct were identified and replicates of each line are currently being generated by micropropagation in preparation for virus challenge.

Analytical modeling and sensitivity analysis for travel time estimation on signalized urban networks

This paper presents a model for estimation of average travel time and its variability on signalized urban networks using cumulative plots. The plots are generated based on the availability of data: a) case-D, for detector data only; b) case-DS, for detector data and signal timings; and c) case-DSS, for detector data, signal timings and saturation flow rate. The performance of the model for different degrees of saturation and different detector detection intervals is consistent for case-DSS and case-DS whereas, for case-D the performance is inconsistent. The sensitivity analysis of the model for case-D indicates that it is sensitive to detection interval and signal timings within the interval. When detection interval is integral multiple of signal cycle then it has low accuracy and low reliability. Whereas, for detection interval around 1.5 times signal cycle both accuracy and reliability are high.

Urban dynamic origin-destination matrices estimation

The aim of this paper is to explore a new approach to obtain better traffic demand (Origin-Destination, OD matrices) for dense urban networks. From reviewing existing methods, from static to dynamic OD matrix evaluation, possible deficiencies in the approach could be identified: traffic assignment details for complex urban network and lacks in dynamic approach. To improve the global process of traffic demand estimation, this paper is focussing on a new methodology to determine dynamic OD matrices for urban areas characterized by complex route choice situation and high level of traffic controls. An iterative bi-level approach will be used, the Lower level (traffic assignment) problem will determine, dynamically, the utilisation of the network by vehicles using heuristic data from mesoscopic traffic simulator and the Upper level (matrix adjustment) problem will proceed to an OD estimation using optimization Kalman filtering technique. In this way, a full dynamic and continuous estimation of the final OD matrix could be obtained. First results of the proposed approach and remarks are presented.

Estimation of travel time on urban networks with midlink sources and sinks

This paper presents a methodology for estimation of average travel time on signalized urban networks by integrating cumulative plots and probe data. This integration aims to reduce the relative deviations in the cumulative plots due to midlink sources and sinks. During undersaturated traffic conditions, the concept of a virtual probe is introduced, and therefore, accurate travel time can be obtained when a real probe is unavailable. For oversaturated traffic conditions, only one probe per travel time estimation interval—360 s or 3% of vehicles traversing the link as a probe—has the potential to provide accurate travel time.

Playpolis : transyouth and urban networking in Seoul

The overarching aim of this study is to create new knowledge about how playful interactions (re)create the city via ubiquitous technologies, with an outlook to apply the knowledge for pragmatic innovations in relevant fields such as urban planning and technology development in the future. The study looks at the case of transyouth, the in-between demographic bridging youth and adulthood in Seoul, one of the most connected, densely populated, and quickly transforming metropolises in the world. To unravel the elusiveness of ‘play’ as a subject and the complexity of urban networks, this study takes a three-tier transdisciplinary approach comprised of an extensive literature review, Shared Visual Ethnography (SVE), and interviews with leading industry representatives who design and develop the playscape for Seoul transyouth. Through these methodological tools, the study responds to the following four research aims: 1. Examine the sociocultural, technological, and architectural context of Seoul 2. Investigate Seoul transyouth’s perception of the self and their technosocial environment 3. Identify the pattern of their playful interaction through which meanings of the self and the city are recreated 4. Develop an analytical framework for enactment of play This thesis argues that the city is a contested space that continuously changes through multiple interactions among its constituents on the seam of control and freedom. At the core of this interactive (re)creation process is play. Play is a phenomenon that is enacted at the centre of three inter-related elements of pressure, possibility, and pleasure, the analytical framework this thesis puts forward as a conceptual apparatus for studying play across disciplines. The thesis concludes by illustrating possible trajectories for pragmatic application of the framework for envisioning and building the creative, sustainable, and seductive city.

Quantitative PCR assay of sewage-associated Bacteroides markers to assess sewage pollution in an urban lake in Dhaka, Bangladesh

This paper aimed to assess the magnitude of sewage pollution in an urban lake in Dhaka, Bangladesh by using Quantitative PCR (qPCR) of sewage-associated Bacteroides HF183 markers. PCR was also used for the quantitative detection of ruminant wastewater-associated CF128 markers along with the enumeration of traditional fecal indicator bacteria, namely, enterococci. The number of enterococci in lake water samples ranged from 1.1 x 104 to 1.9 x 105 CFU/100 ml of water. From the 20 water samples tested, 14 (70%) and 7 (35%) were PCR positive for the HF183 and CF128 markers, respectively. The numbers of the HF183 and CF128 markers in lake water samples were 3.9 x 104 to 6.3 × 107 and 9.3 x 103 to 6.3 x 105 genomic units (GU)/100 ml of water, respectively. The high numbers of enterococci and the HF183 markers indicate sewage pollution and potential health risks to those who use the lake water for non-potable purposes such as bathing and washing clothes. This is the first study that investigated the presence of microbial source tracking (MST) markers in Dhaka, Bangladesh where diarrhoeal diseases is one of the major causes of childhood mortality. The molecular assay as used in this study can provide valuable information on the extent of sewage pollution, thus facilitating the development of robust strategies to minimise potential health risks.

Investigation into travel of transit oriented development : school students’ travel survey of Kelvin Grove Urban Village

Travel surveys were conducted for collecting data related to school students’ travel at Kelvin Grove Urban Village (KGUV). Currently, KGUV has school students studying at grade 10 to 12. As a part of data collection process, travel surveys were undertaken for school students studying. This document contains the questionnaire form used to collect the demographic and travel data related to school students at KGUV. The surveys forms were hand delivered to the school and the responses were collected back via reply paid envelop provided with the questionnaire form.

Reconfiguring the higher education value chain

Forces of demand and supply are changing the dynamics of the higher education market. Transformation of institutions of higher learning into competitive enterprise is underway. Higher education institutions are seemingly under intense pressure to create value and focus their efforts and scarce funds on activities that drive up value for their respective customers and other stakeholders. Porter’s generic ‘value chain’ model for creating value requires that the activities of an organization be segregated in to discrete components for value chain analysis to be performed. Recent trends in higher education make such segregation possible. Therefore, it is proposed that the academic process can be unbundled into discrete components which have well developed measures. A reconfigured value chain for higher education, with its own value drivers and critical internal linkages is also proposed in this paper.

Urban growth monitoring techniques for sustainable development

This paper discusses the role of advance techniques for monitoring urban growth and change for sustainable development of urban environment. It also presents results of a case study involving satellite data for land use/land cover classification of Lucknow city using IRS-1C multi-spectral features. Two classification algorithms have been used in the study. Experiments were conducted to see the level of improvement in digital classification of urban environment using Artificial Neural Network (ANN) technique.

Lift@Weimar : sustainable interaction with food, technology, and the city (Workshop)

This workshop explores innovative approaches to understanding and cultivating sustainable food culture in urban environments via human-computer-interaction (HCI) design and ubiquitous technologies. We perceive the city as an intersecting network of people, place, and technology in constant transformation. Our 2009 OZCHI workshop, Hungry 24/7? HCI Design for Sustainable Food Culture, opened a new space for discussion on this intersection amongst researchers and practitioners from diverse backgrounds including academia, government, industry, and non-for-profit organisations. Building on the past success, this new instalment of the workshop series takes a more refined view on mobile human-food interaction and the role of interactive media in engaging citizens to cultivate more sustainable everyday human-food interactions on the go. Interactive media in this sense is distributed, pervasive, and embedded in the city as a network. The workshop addresses environmental, health, and social domains of sustainability by bringing together insights across disciplines to discuss conceptual and design approaches in orchestrating mobility and interaction of people and food in the city as a network of people, place, technology, and food.