Agrobacterium-mediated transformation of indica rice cultivars using binary and superbinary vectors

The Agrobacterium-mediated transformation system was extended to two indica cultivars: a widely cultivated breeding line IR-64 and an elite basmati cultivar Karnal Local. Root tips and shoot tips of seedlings, and scutellar-calli derived from mature seeds showed high-efficiency Agrobacterium tumefaciens infection and stable transformation. In addition to the superbinary vector pTOK233 in Agrobacterium strain LBA4404, almost equally high levels of transformation were achieved with a relatively much smaller (13.1 kb) binary vector (pCAMBIA1301) in a supervirulent host strain AGL1. In both cases, as well as in both cultivars, while 60–90% of the infected explants produced calli resistant to the selectable agent hygromycin, 59–75% of such calli tested positive for GUS. A high level (400 μM) of acetosyringone in the preinduction medium for Agrobacterium and a higher level (500 μM) in the cocultivation medium was necessary for an enhancement in transformation frequency of the binary vector to levels comparable to a superbinary. Hygromycin-resistant calli could be produced from all the explants used. Transformants could be regenerated for both cultivars using the superbinary and binary vector, but only for calli of scutellar origin. In addition to the molecular confirmation of hpt and gus gene transfer and transcription, absence of gene sequences outside the transferred DNA (T-DNA) region confirmed absence of any long T-DNA transfer.

Qualitative inquiry as transformation and agency : the black sheep and workplace mobbing

This chapter discusses an action research project into the lived experience of the workplace mobbing phenomenon. The action research methodology is based on the exemplarian model (Coenen & Khonraad, 2003) from the Netherlands Group. This model requires positive outcomes for those immersed in the problem to reduce the adversity of their circumstances. The findings challenge the psychological perspective of the existing bullying literature that tends to focus on individual behaviour. This research, undertaken over a three year period with 212 participants, identified the dysfunctional nature of public sector bureaucracies and the power gained through gossip and rumour as some of the key emergent themes to explain the workplace mobbing problem. In addition, resistance, conscientisation, and agency were identified as the key to transformation for those targeted. The discussion focuses on the crystallisation phase of the exemplarian model where the participants identified themselves as the Black Sheep and adopted the motto that “a black sheep is a biting beast” (Bastard, 1565 or 6-1618, p. 90), reflecting a sense of empowerment, individual agency, and a sense of humour in dealing with the serious yet seemingly absurd reality of their situations. The identity of the Black Sheep was consolidated when the group organised a 2 day conference with over 200 attendees to discuss how best to prevent workplace mobbing. This self-affirming action was a proactive step towards metaphorically “biting back” at the problem. A number of positive outcomes were achieved including the conference with over 200 attending leading to national media coverage across Australia and additional interviews with magazines, newspapers, and radio.

Development of a transformation system for sorghum (Sorghum bicolor L.)

Sorghum (Sorghum bicolor (L.) Moench) is the world’s fifth major cereal crop and holds importance as a construction material, food and fodder source. More recently, the potential of this plant as a biofuel source has been noted. Despite its agronomic importance, the use of sorghum production is being constrained by both biotic and abiotic factors. These challenges could be addressed by the use of genetic engineering strategies to complement conventional breeding techniques. However, sorghum is one of the most recalcitrant crops for genetic modification with the lack of an efficient tissue culture system being amongst the chief reasons. Therefore, the aim of this study was to develop an efficient tissue culture system for establishing regenerable embryogenic cell lines, micropropagation and acclimatisation for Sorghum bicolor and use this to optimise parameters for genetic transformation via Agrobacterium-mediated transformation and microprojectile bombardment. Using five different sorghum cultivars, SA281, 296B, SC49, Wray and Rio, numerous parameters were investigated in an attempt to establish an efficient and reproducible tissue culture and transformation system. Using immature embryos (IEs) as explants, regenerable embryogenic cell lines (ECLs) could only be established from cultivars SA281 and 296B. Large amounts of phenolics were produced from IEs of cultivars, SC49, Wary and Rio, and these compounds severely hindered callus formation and development. Cultivar SA281 also produced phenolics during regeneration. Attempts to suppress the production of these compounds in cultivars SA281 and SC49 using activated charcoal, PVP, ascorbic acid, citric acid and liquid filter paper bridge methods were either ineffective or had a detrimental effect on embryogenic callus formation, development and regeneration. Immature embryos sourced during summer were found to be far more responsive in vitro than those sourced during winter. In an attempt to overcome this problem, IEs were sourced from sorghum grown under summer conditions in either a temperature controlled glasshouse or a growth chamber. However, the performance of these explants was still inferior to that of natural summer-sourced explants. Leaf whorls, mature embryos, shoot tips and leaf primordia were found to be unsuitable as explants for establishing ECLs in sorghum cultivars SA281 and 296B. Using the florets of immature inflorescences (IFs) as explants, however, ECLs were established and regenerated for these cultivars, as well as for cultivar Tx430, using callus induction media, SCIM, and regeneration media, VWRM. The best in vitro responses, from the largest possible sized IFs, were obtained using plants at the FL-2 stage (where the last fully opened leaf was two leaves away from the flag leaf). Immature inflorescences could be stored at 25oC for up to three days without affecting their in vitro responses. Compared to IEs, the IFs were more robust in tissue culture and showed responses which were season and growth condition independent. A micropropagation protocol for sorghum was developed in this study. The optimum plant growth regulator (PGR) combination for the micropropagation of in vitro regenerated plantlets was found to be 1.0 mg/L BAP in combination with 0.5 mg/L NAA. With this protocol, cultivars 296B and SA281 produced an average of 57 and 13 off-shoots per plantlet, respectively. The plantlets were successfully acclimatised and developed into phenotypically normal plants that set seeds. A simplified acclimatisation protocol for in vitro regenerated plantlets was also developed. This protocol involved deflasking in vitro plantlets with at least 2 fully-opened healthy leaves and at least 3 roots longer than 1.5 cm, washing the media from the roots with running tap water, planting in 100 mm pots and placing in plastic trays covered with a clear plastic bag in a plant growth chamber. After seven days, the corners of the plastic cover were opened and the bags were completely removed after 10 days. All plantlets were successfully acclimatised regardless of whether 1:1 perlite:potting mix, potting mix, UC mix or vermiculite were used as potting substrates. Parameters were optimised for Agrobacterium-mediated transformation (AMT) of cultivars SA281, 296B and Tx430. The optimal conditions were the use of Agrobacterium strain LBA4404 at an inoculum density of 0.5 OD600nm, heat shock at 43oC for 3 min, use of the surfactant Pluronic F-68 (0.02% w/v) in the inoculation media with a pH of 5.2 and a 3 day co-cultivation period in dark at 22oC. Using these parameters, high frequencies of transient GFP expression was observed in IEs precultured on callus initiation media for 1-7 days as well as in four weeks old IE- and IF-derived callus. Cultivar SA281 appeared very sensitive to Agrobacterium since all tissue turned necrotic within two weeks post-exposure. For cultivar 296B, GFP expression was observed up to 20 days post co-cultivation but no stably transformed plants were regenerated. Using cultivar Tx430, GFP was expressed for up to 50 days post co-cultivation. Although no stably transformed plants of this cultivar were regenerated, this was most likely due to the use of unsuitable regeneration media. Parameters were optimised for transformation by particle bombardment (PB) of cultivars SA281, 296B and Tx430. The optimal conditions were use of 3-7 days old IEs and 4 weeks old IF callus, 4 hour pre- and post-bombardment osmoticum treatment, use of 0.6 µm gold microparticles, helium pressure of 1500 kPa and target distance of 15 cm. Using these parameters for PB, transient GFP expression was observed for up to 14, 30 and 50 days for cultivars SA281, 296B and Tx430, respectively. Further, the use of PB resulted in less tissue necrosis compared to AMT for the respective cultivars. Despite the presence of transient GFP expression, no stably transformed plants were regenerated. The establishment of regenerable ECLs and the optimization of AMT and PB parameters in this study provides a platform for future efforts to develop an efficient transformation protocol for sorghum. The development of GM sorghum will be an important step towards improving its agronomic properties as well as its exploitation for biofuel production.

ACE research vignette 020: Entrepreneurial becoming - a self-induced transformation

This series of research vignettes is aimed at sharing current and interesting research findings from our team of international Entrepreneurship researchers. In this vignette, Dr Marcello Tonelli and Associate Professor Carol Dalglish consider the delivery of entrepreneurial education through experiential learning in a developing context.

An improved method to detect damage using modal strain energy based damage index

This paper presents two novel concepts to enhance the accuracy of damage detection using the Modal Strain Energy based Damage Index (MSEDI) with the presence of noise in the mode shape data. Firstly, the paper presents a sequential curve fitting technique that reduces the effect of noise on the calculation process of the MSEDI, more effectively than the two commonly used curve fitting techniques; namely, polynomial and Fourier’s series. Secondly, a probability based Generalized Damage Localization Index (GDLI) is proposed as a viable improvement to the damage detection process. The study uses a validated ABAQUS finite-element model of a reinforced concrete beam to obtain mode shape data in the undamaged and damaged states. Noise is simulated by adding three levels of random noise (1%, 3%, and 5%) to the mode shape data. Results show that damage detection is enhanced with increased number of modes and samples used with the GDLI.

Innovative damage assessment of steel truss bridges using modal strain energy correlation

As a part of vital infrastructure and transportation network, bridge structures must function safely at all times. Bridges are designed to have a long life span. At any point in time, however, some bridges are aged. The ageing of bridge structures, given the rapidly growing demand of heavy and fast inter-city passages and continuous increase of freight transportation, would require diligence on bridge owners to ensure that the infrastructure is healthy at reasonable cost. In recent decades, a new technique, structural health monitoring (SHM), has emerged to meet this challenge. In this new engineering discipline, structural modal identification and damage detection have formed a vital component. Witnessed by an increasing number of publications is that the change in vibration characteristics is widely and deeply investigated to assess structural damage. Although a number of publications have addressed the feasibility of various methods through experimental verifications, few of them have focused on steel truss bridges. Finding a feasible vibration-based damage indicator for steel truss bridges and solving the difficulties in practical modal identification to support damage detection motivated this research project. This research was to derive an innovative method to assess structural damage in steel truss bridges. First, it proposed a new damage indicator that relies on optimising the correlation between theoretical and measured modal strain energy. The optimisation is powered by a newly proposed multilayer genetic algorithm. In addition, a selection criterion for damage-sensitive modes has been studied to achieve more efficient and accurate damage detection results. Second, in order to support the proposed damage indicator, the research studied the applications of two state-of-the-art modal identification techniques by considering some practical difficulties: the limited instrumentation, the influence of environmental noise, the difficulties in finite element model updating, and the data selection problem in the output-only modal identification methods. The numerical (by a planer truss model) and experimental (by a laboratory through truss bridge) verifications have proved the effectiveness and feasibility of the proposed damage detection scheme. The modal strain energy-based indicator was found to be sensitive to the damage in steel truss bridges with incomplete measurement. It has shown the damage indicator's potential in practical applications of steel truss bridges. Lastly, the achievement and limitation of this study, and lessons learnt from the modal analysis have been summarised.

A rhetorical study of in-flight real estate advertisements as a potential site of ethical transformation in Chinese cities

Real estate markets in Chinese cities are in transition. Advertising for new developments in these markets often reflects changing city aspirations and branding rather than environmental and social experience. This paper investigates real estate marketing as a site of potential ethical transformation of values related to new urban development. It uses Kenneth Burke’s rhetorical analysis as an approach to coding real estate representations from in-flight magazine advertisements as a means of capturing environmental and social viewpoints in China during 2008 - 2009. Both Chinese and foreign participants coded representations into four code modalities. These were based on anthropocentric - non-anthropocentric environmental orientations and nationalistic - universal social orientations. The results suggested that new developments in China are more likely to be understood as based on environmental resource use for continued national economic expansion rather than for a more sustainable world. Emerging patterns in coded representations have opened up the possibility of greater social choices that were however difficult to unambiguously decode from Chinese real estate advertising. From this it is concluded that it may take some time before real estate demand shifts in response to representations of Chinese eco-cities being promoted by Chinese policy makers in the 2000s.

The digital transformation of 21st century news journalism

This presentation will deal with the transformations that have occurred in news journalism worldwide in the early 21st century. I will argue that they have been the most significant changes to the profession for 100 years, and the challenges facing the news media industry in responding to them are substantial, as are those facing journalism education. It will develop this argument in relation to the crisis of the newspaper business model, and why social media, blogging and citizen journalism have not filled the gap left by the withdrawal of resources from traditional journalism. It will also draw upon Wikileaks as a case study in debates about computational and data-driven journalism, and whether large-scale "leaks" of electronic documents may be the future of investigative journalism.

Ability of modal analysis to detect osseointegration of implants in transfemoral amputees : a physical model study

Owing to the successful use of non-invasive vibration analysis to monitor the progression of dental implant healing and stabilization, it is now being considered as a method to monitor femoral implants in transfemoral amputees. This study uses composite femur-implant physical models to investigate the ability of modal analysis to detect changes at the interface between the implant and bone simulating those that occur during osseointegration. Using electromagnetic shaker excitation, differences were detected in the resonant frequencies and mode shapes of the model when the implant fit in the bone was altered to simulate the two interface cases considered: firm and loose fixation. The study showed that it is beneficial to examine higher resonant frequencies and their mode shapes (rather than the fundamental frequency only) when assessing fixation. The influence of the model boundary conditions on the modal parameters was also demonstrated. Further work is required to more accurately model the mechanical changes occurring at the bone-implant interface in vivo, as well as further refinement of the model boundary conditions to appropriately represent the in vivo conditions. Nevertheless, the ability to detect changes in the model dynamic properties demonstrates the potential of modal analysis in this application and warrants further investigation.

Re-made in Asia : transformation across Asian markets and popular culture

The thesis is an examination of how Japanese popular culture products are remade (rimeiku). Adaptation of manga, anime and television drama, from one format to another, frequently occurs within Japan. The rights to these stories and texts are traded in South Korea and Taiwan. The ‘spin-off’ products form part of the Japanese content industry. When products are distributed and remade across geographical boundaries, they have a multi-dimensional aspect and potentially contribute to an evolving cultural re-engagement between Japan and East Asia. The case studies are the television dramas Akai Giwaku and Winter Sonata and two manga, Hana yori Dango and Janguru Taitei. Except for the television drama Winter Sonata these texts originated in Japan. Each study shows how remaking occurs across geographical borders. The study argues that Japan has been slow to recognise the value of its popular culture through regional and international media trade. Japan is now taking steps to remedy this strategic shortfall to enable the long-term viability of the Japanese content industry. The study includes an examination of how remaking raises legal issues in the appropriation of media content. Unauthorised copying and piracy contributes to loss of financial value. To place the three Japanese cultural products into a historical context, the thesis includes an overview of Japanese copying culture from its early origins through to the present day. The thesis also discusses the Meiji restoration and the post-World War II restructuring that resulted in Japan becoming a regional media powerhouse. The localisation of Japanese media content in South Korea and Taiwan also brings with it significant cultural influences, which may be regarded as contributing to a better understanding of East Asian society in line with the idea of regional ‘harmony’. The study argues that the commercial success of Japanese products beyond Japan is governed by perceptions of the quality of the story and by the cultural frames of the target audience. The thesis draws on audience research to illustrate the loss or reinforcement of national identity as a consequence of cross-cultural trade. The thesis also examines the contribution to Japanese ‘soft power’ (Nye, 2004, p. x). The study concludes with recommendations for the sustainability of the Japanese media industry.