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.

Effect of clear zone widths on run-off-road crash outcomes

Current guidelines on clear zone selection and roadside hazard management adopt the US approach based on the likelihood of roadside encroachment by drivers. This approach is based on the available research conducted in the 1960s and 70s. Over time, questions have been raised regarding the robustness and applicability of this research in Australasia in 2010 and in the Safe System context. This paper presents a review of the fundamental research relating to selection of clear zones. Results of extensive rural highway statistical data modelling suggest that a significant proportion of run-off-road to the left casualty crashes occurs in clear zones exceeding 13 m. They also show that the risk of run-off-road to the left casualty crashes was 21% lower where clear zones exceeded 8 m when compared with clear zones in the 4 – 8 m range. The paper discusses a possible approach to selection of clear zones based on managing crash outcomes, rather than on the likelihood of roadside encroachment which is the basis for the current practice. It is expected that this approach would encourage selection of clear zones wider than 8 m when the combination of other road features suggests higher than average casualty crash risk.

The role of organisational capacity in the knowledge-based transformation of Brisbane, Australia

Purpose - The cumulative impacts of the knowledge economy together with the emerging dominance of knowledge-intensive sectors, have led to an unprecedented period of socio-economic and spatial restructuring. As a result, the paradigm of knowledge-based urban development (KBUD) has emerged as a development strategy to guide knowledge-based economic transformation (Knight, 1995; Yigitcanlar, 2007). Notwithstanding widespread government commitment and financial investment, in many cases providing the enabling circumstances for KUBUD has proven a complicated task as institutional barriers remain. Researchers and practitioners advocate that the way organisations work and their institutional relationships, policies and programs, will have a significant impact on a regions capacity to achieve KBUD (Savitch, 1998; Savitch and Kantor, 2002; Keast and Mandell, 2009). In this context, building organisational capacity is critical to achieving institutional change and bring together all of the key actors and sources, for the successful development, adoption, and implementation of knowledge-based development of a city (Yigitcanlar, 2009). Design/methodology/approach - There is a growing need to determine the complex inter-institutional arrangements and intra-organisational interactions required to advance urban development within the knowledge economy. In order to design organisational capacity-building strategies, the associated attributes of good capacity must first be identified. The paper, with its appraisal of knowledge-based urban development, scrutinises organisational capacity and institutional change in Brisbane. As part of the discussion of the case study findings, the paper describes the institutional relationships, policies, programs and funding streams, which are supporting KBUD in the region. Originality/value - In consideration that there has been limited investigation into the institutional lineaments required to provide the enabling circumstances for KBUD, the broad aim of this paper is to discover some good organisational capacity attributes, achieved through a case study of Brisbane. Practical implications - It is anticipated that the findings of the case study will contribute to moving the discussion on the complex inter-institutional arrangements and intra-organisaational interactions required for KBUD, beyond a position of rhetoric.

The effect of flocculants, shear, vacuum and depithing on the formation of thin bagasse pulp pads

The effect of bentonite micro-particles and cationic polyacrylamide (CPAM) on the filtration properties of bagasse pulp was investigated under shearing conditions. CPAM improves retention but the bentonite addition level must be optimised for further improvements in retention. A Dynamic Drainage Jar (‘Britt Jar’) was modified to allow bagasse pulp slurry to be subjected to vacuum allowing a thin pulp pad to be formed. Bagasse pulp which had had the majority of the fine fibre removed prior to pulping drained more quickly than a conventional bagasse pulp when vacuum was not applied, however this situation was reversed when vacuum was used. The flocculants continue to improve fibre retention under vacuum and shear conditions but with reduced effectiveness.

Enhancing agrobacterium-mediated transformation efficiency of sugarcane : progress towards an efficient, genotype-independent method

Several key issues need to be resolved before an efficient and reproducible Agrobacterium-mediated sugarcane transformation method can be developed for a wider range of sugarcane cultivars. These include loss of morphogenetic potential in sugarcane cells after Agrobacterium-mediated transformation, effect of exposure to abiotic stresses during in vitro selection, and most importantly the hypersensitive cell death response of sugarcane (and other nonhost plants) to Agrobacterium tumefaciens. Eight sugarcane cultivars (Q117, Q151, Q177, Q200, Q208, KQ228, QS94-2329, and QS94-2174) were evaluated for loss of morphogenetic potential in response to the age of the culture, exposure to Agrobacterium strains, and exposure to abiotic stresses during selection. Corresponding changes in the polyamine profiles of these cultures were also assessed. Strategies were then designed to minimize the negative effects of these factors on the cell survival and callus proliferation following Agrobacterium-mediated transformation. Some of these strategies, including the use of cell death protector genes and regulation of intracellular polyamine levels, will be discussed.

Review : Surfers Paradise foreshore redevelopment masterplan - urban plaza zone

The Centre for Subtropical Design has prepared this submission to assist the Gold Coast City Council to finalise a plan and detailed design guidelines for the Urban Plaza Zone of Surfers Paradise Foreshore Redevelopment Masterplan which will create a public open space ‘alive’ with the quality appropriate to a place which is both a local centre and an international destination. This review has been informed by the two over-arching values identified as characteristics of a subtropical place and people’s connection to it:  A sense of openness and permeability, and  Engagement with the natural environment. The existing qualities of the foreshore area proposed as the Urban Plaza Zone, reflect these subtropical place values, and are integral to the Surfers Paradise identity:  Seamless visual and spatial access to the beach and sea,  Permeable interface between beach and built zones provided by beach planting and shade to sand by Pandanus,  A shade zone mediating beach and linear promenade, road and commercial zones, enabling a variety of social and visual experiences, on soft and hard finishes, and  A lively, constantly moving shared road and pedestrian way catering for events and day to day activities with visual access to beach and shaded areas. The Centre for Subtropical Design commends the Gold Coast City Council on preparing a plan for a public open space that is a contemporary departure from the adhoc basis of development that has occurred, in that it will make this area more accessible. However, the proposed plan seems to be working too hard in terms of ‘program’. While providing an identifiable interruption in the linear extent of the Foreshore, the lack of continuity of design in terms of both hardscaping (such as perpendicular paving elements) and softscaping (such as tree selections) may contribute to a lack of definition for the entire Foreshore as a place that mediates, along its length, between sea and land. Providing a hard edge to a beach character of soft and planted transitional elements needs to balance the proposed visual and physical barrier with the need for perceived and actual easy access. The Surfers Paradise identity needs strengthening through attention to planting for shade, materials, particularly selection of paving colours, and stronger delineation of the linear nature of the Foreshore. The Urban Plaza zone is an appropriate interruption to the continuous planting, however the link from the commercial zone overtakes the public and beach zone. A more seamless transition from shop to sea, better integration of the roadway and pedestrian zone and improved physical transition from concrete to sand is recommended. Built form solutions must be robust and designed with the subtropical design principles and the Surfers Paradise identity as underpinning parameters for a lasting and memorable public open space.

Factors affecting somatic embryogenesis and transformation in modern plant breeding

Somatic embryogenesis and transformation systems are indispensable modern plant breeding components since they provide an alternative platform to develop control strategies against the plethora of pests and diseases affecting many agronomic crops. This review discusses some of the factors affecting somatic embryogenesis and transformation, highlights the advantages and limitations of these systems and explores these systems as breeding tools for the development of crops with improved agronomic traits. The regeneration of non-chimeric transgenic crops through somatic embryogenesis with introduced disease and pest-resistant genes for instance, would be of significant benefit to growers worldwide.

Effect of spacing of reinforcement on the behaviour of partially grouted masonry shear walls

Partially Grouted Reinforced Masonry (PGRM) shear walls perform well in places where the cyclonic wind pressure dominates the design. Their out-of-plane flexural performance is better understood than their inplane shear behaviour; in particular, it is not clear whether the PGRM shear walls act as unreinforced masonry (URM) walls embedded with discrete reinforced grouted cores or as integral systems of reinforced masonry (RM) with wider spacing of reinforcement. With a view to understanding the inplane response of PGRM shear walls, ten full scale single leaf, clay block walls were constructed and tested under monotonic and cyclic inplane loading cases. It has been shown that where the spacing of the vertical reinforcement is less than 2000mm, the walls behave as an integral system of RM; for spacing greater than 2000mm, the walls behave similar to URM with no significant benefit from the reinforced cores based on the displacement ductility and stiffness degradation factors derived from the complete lateral load – lateral displacement curves.

Shear tests of Litesteel beams with web openings

This paper presents the details of experimental studies on the shear strength of a recently developed, cold-formed steel beam known as LiteSteel Beam (LSB) with web openings. The innovative LSB sections have the beneficial characteristics of torsionally rigid closed rectangular flanges combined with economical fabrication processes from a single strip of high strength steel. They combine the stability of hot-rolled steel sections with the high strength to weight ratio of conventional cold-formed steel sections. The LSB sections are commonly used as flexural members in the building industry. Current practice in flooring systems is to include openings in the web element of floor joists or bearers so that building services can be located within them. Shear behaviour of LSBs with web openings is more complicated while their shear strengths are considerably reduced by the presence of web openings. However, limited research has been undertaken on the shear behaviour and strength of LSBs with web openings. Therefore a detailed experimental study involving 26 shear tests was undertaken to investigate the shear behaviour and strength of different LSB sections. Simply supported test specimens of LSBs with an aspect ratio of 1.5 were loaded at midspan until failure. This paper presents the details of this experimental study and the results. Experimental results showed that the current design rules in cold-formed steel structures design codes (AS/NZS 4600) [1] are very conservative for the shear design of LSBs with web openings. Improved design equations have been proposed for the shear strength of LSBs with web openings based on experimental results from this study.

Shear strength of LiteSteel beams with web openings

This paper presents the details of numerical studies on the shear strength of a recently devel-oped, cold-formed steel channel beam known as LiteSteel Beam (LSB) with web openings. The LSB sections are commonly used as floor joists and bearers in residential, industrial and commercial buildings. In these ap-plications they often include web openings for the purpose of locating services. This has raised concerns over the shear capacity of LSB floor joists and bearers. Therefore experimental and numerical studies were under-taken to investigate the shear behavior and strength of LSBs with web openings. In this research, finite ele-ment models of LSBs with web openings in shear were developed to simulate the shear behavior of LSBs. It was found that currently available design equations are conservative or unconservative for the shear design of LSBs with web openings. Improved design equations have been proposed for the shear capacity of LSBs with web openings based on both experimental and numerical study results.

Shear strength and design of LiteSteel beams

This paper presents the details of a parametric study based on finite element analyses (FEA) and development of design rules for the shear strength of a recently developed, cold-formed steel channel beam known as LiteSteel Beam (LSB). The LSB sections are commonly used as flexural members in residential, in-dustrial and commercial buildings. In order to ensure safe and efficient designs of LSBs, many research stu-dies have been undertaken on the flexural behaviour of LSBs. However, no research has been undertaken on the shear behaviour of LSBs. Therefore a detailed investigation including both numerical and experimental studies was undertaken to investigate the shear behaviour of LSBs. Both the experimental and FEA parametric study results showed that the current design rules in cold-formed steel design codes are very conservative for the shear design of LSBs. New shear strength equations for LSBs were proposed based on the experimental and FEA parametric study results.

The course of bone healing is influenced by the initial shear fixation stability

Fracture healing is influenced by fixation stability and experimental evidence suggests that the initial mechanical conditions may determine the healing outcome. We hypothesised that mechanical conditions influence not only the healing outcome, but also the early phase of fracture healing. Additionally, it was hypothesised that decreased fixation stability characterised by an increased shear interfragmentary movement results in a delay in healing. Sixty-four sheep underwent a mid-shaft tibial osteotomy which was treated with either a rigid or a semi-rigid external fixator. Animals were sacrificed at 2, 3, 6 and 9 weeks postoperatively and the fracture callus was analysed using radiological, biomechanical and histological techniques. The tibia treated with semi-rigid fixation showed inferior callus stiffness and quality after 6 weeks. At 9 weeks, the calluses were no longer distinguishable in their mechanical competence. The calluses at 9 weeks produced under rigid fixation were smaller and consisted of a reduced fibrous tissue component. These results demonstrate that the callus formation over the course of healing differed both morphologically and in the rate of development. In this study, we provide evidence that the course of healing is influenced by the initial fixation stability. The semi-rigid fixator did not result in delayed healing, but a less optimal healing path was taken. An upper limit of stability required for successful healing remains unknown, however a limit by which healing is less optimal has been determined.

Zonal chondrocyte subpopulations reacquire zone-specific characteristics during in Vitro redifferentiation

Background: If chondrocytes from the superficial, middle, and deep zones of articular cartilage could maintain or regain their characteristic properties during in vitro culture, it would be feasible to create constructs comprising these distinctive zones. ----- ----- Hypothesis: Zone-specific characteristics of zonal cell populations will disappear during 2-dimensional expansion but will reappear after 3-dimensional redifferentiation, independent of the culture technique used (alginate beads versus pellet culture).----- ----- Study Design: Controlled laboratory study.----- ----- Methods: Equine articular chondrocytes from the 3 zones were expanded in monolayer culture (8 donors) and subsequently redifferentiated in pellet and alginate bead cultures for up to 4 weeks. Glycosaminoglycans and DNA were quantified, along with immunohistochemical assessment of the expression of various zonal markers, including cartilage oligomeric protein (marking cells from the deeper zones) and clusterin (specifically expressed by superficial chondrocytes).----- ----- Results: Cell yield varied between zones, but proliferation rates did not show significant differences. Expression of all evaluated zonal markers was lost during expansion. Compared to the alginate bead cultures, pellet cultures showed a higher amount of glycosaminoglycans produced per DNA after redifferentiation. In contrast to cells in pellet cultures, cells in alginate beads regained zonal differences, as evidenced by zone-specific reappearance of cartilage oligomeric protein and clusterin, as well as significantly higher glycosaminoglycans production by cells from the deep zone compared to the superficial zone.----- ----- Conclusion: Chondrocytes isolated from the 3 zones of equine cartilage can restore their zone-specific matrix expression when cultured in alginate after in vitro expansion.

The emerging management services of business transformation management

Business transformations are large-scale organizational change programs that, evidence suggests, are often unsuccessful. Our interest is in identifying the management capabilities required for the successful execution of these projects. We advance a service-oriented view of the enterprise, which suggests that different management services need to be identified and integrated in order to execute business transformation. In order to identify those management services that require integration, we conducted an exploratory empirical study of the demand for management services in US and Asia, and we show that two archetypes of management services exist in business transformation initiatives: transactional and transformational management services. We identify the relevant set of transactional and transformational services and discuss what the demand for these services implies for the execution of business transformations.