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.

Science education policy-making : eleven emerging issues

The Perth Declaration on Science and Technology Education of 2007 expresses strong concern about the state of science and technology education worldwide and calls on governments to respond to a number of suggestions for establishing the structural conditions for their improved practice. The quality of school education in science and technology has never before been of such critical importance to governments. There are three imperatives for its critical importance. The first relates to the traditional role of science in schooling, namely the identification, motivation and initial preparation of those students who will go on to further studies for careers in all those professional fi elds that directly involve science and technology. A suffi cient supply of these professionals is vital to the economy of all countries and to the health of their citizens. In the 21st century they are recognised everywhere as key players in ensuring that industrial and economic development occurs in a socially and environmentally sustainable way. In many countries this supply is now falling seriously short and urgently needs to be addressed. The second imperative is that sustainable technological development and many other possible societal applications of science require the support of scientifically and technologically informed citizens. Without the support and understanding of citizens, technological development can all too easily serve short term and sectional interests. The longer term progress of the whole society is overlooked, citizens will be confused about what should, and what should not be supported, and reactive and the environment will continue to be destroyed rather than sustained. Sustainable development, and the potential that science and technology increasingly offers, involves societies in ways that can often interact strongly, with traditional values, and hence, making decisions about them involve major moral decisions. All students need to be prepared through their science and technology education to be able to participate actively as persons and as responsible citizens in these essential and exciting possibilities. This goal is far from being generally achieved at present, but pathways to it are now more clearly understood. The third imperative derives from the changes that are resulting from the application of digital technologies that are the most rapid, the most widespread, and probably the most pervasive influence that science has ever had on human society. We all, wherever we live, are part of a global communication society. Information exchange and access to it that have been hitherto the realm of the few, are now literally in the hands of individuals. This is leading to profound changes in the World of Work and in what is known as the Knowledge Society. Schooling is now being challenged to contribute to the development in students of an active repertoire of generic and subject-based competencies. This contrasts very strongly with existing priorities, in subjects like the sciences that have seen the size of a student’s a store of established knowledge as the key measure of success. Science and technology education needs to be a key component in developing these competencies. When you add to these imperatives, the possibility that a more effective education in science and technology will enable more and more citizens to delight in, and feel a share in the great human enterprise we call Science, the case for new policy decisions is compellingly urgent. What follows are the recommendations (and some supplementary notes) for policy makers to consider about more operational aspects for improving science and technology education. They are listed under headings that point to the issues within each of these aspects. In the full document, a background is provided to each set of issues, including the commonly current state of science and technology education. Associated with each recommendation for consideration are the positive Prospects that could follow from such decision making, and the necessary Prerequisites, if such bold policy decisions are to fl ow, as intended, into practice in science and technology classrooms.

Velocity and Concentration field of discharge from a boat propeller

Two-stroke outboard boat engines using total loss lubrication deposit a significant proportion of their lubricant and fuel directly into the water. The purpose of this work is to document the velocity and concentration field characteristics of a submerged swirling water jet emanating from a propeller in order to provide information on its fundamental characteristics. Measurements of the velocity and concentration field were performed in a turbulent jet generated by a model boat propeller (0.02 m diameter) operating at 1500 rpm and 3000 rpm. The measurements were carried out in the Zone of Established Flow up to 50 propeller diameters downstream of the propeller. Both the mean axial velocity profile and the mean concentration profile showed self-similarity. Further, the stand deviation growth curve was linear. The effects of propeller speed and dye release location were also investigated.

Gender

The OED informs us that “gender” has at its root the Latin genus, meaning “race, kind,” and emerges as early as the fifth century as a term for differentiating between types of (especially) people and words. In the following 1500 years, gender appears in linguistic and biological contexts to distinguish types of words and bodies from one another, as when words in Indo-European languages were identified as masculine, feminine, or neuter, and humans were identified as male or female. It is telling that gender has historically (whether overtly or covertly) been a tool of negotiation between our understandings of bodies, and meanings derived from and attributed to them. Within the field of children’s literature studies, as in other disciplines, gender in and of itself is rarely the object of critique. Rather, specific constructions of gender structure understandings of subjectivity; allow or disallow certain behaviors or experiences on the basis of biological sex; and dictate a specific vision of social relations and organization. Critical approaches to gender in children’s literature have included linguistic analysis (Turner-Bowker; Sunderland); analysis of visual representations (Bradford; Moebius); cultural images of females (Grauerholz and Pescosolido); consideration of gender and genre (Christian-Smith; Stephens); ideological (Nodelman and Reimer); psychoanalytic (Coats); discourse analysis (Stephens); and masculinity studies (Nodelman) among others. In the adjacent fields of education and literacy studies, gender has been a sustained point of investigation, often deriving from perceived gendering of pedagogical practices (Lehr) or of reading preferences and competencies, and in recent years, perceptions of boys as “reluctant readers” (Moss). The ideology of patriarchy has primarily come under critical scrutiny 2 because it has been used to locate characters and readers within the specific binary logic of gender relations that historically subordinated the feminine to the masculine. Just as feminism might be broadly defined as resistance to existing power structures, a gendered reading might be broadly defined as a “resistant reading” in that it most often reveals or contests that which a text assumes to be the norm.

Poster : Understanding interactions between autonomous animal control and temperament when cattle are subjected to virtual fencing applications

Virtual fencing has the potential to control grazing livestock. Understanding and refi ning the cues that can alter behaviour is an integral part of autonomous animal control. A series of tests have been completed to explore the relationship between temperament and control. Prior to exposure to virtual fencing control the animals were scored for temperament using fl ight speed and a sociability index using contact logging devices. The behavioural response of 30, Belmont Red steers were observed for behavioural changes when presented with cues prior to receiving an electrical stimulation. A control and four treatments designed to interrupt the animal’s movement down an alley were tested. The treatments consisted of sound plus electrical stimulation, vibration plus electrical stimulation, a visual cue plus electrical stimulation and electrical stimulation by itself. The treatments were randomly applied to each animal over fi ve consecutive trials. A control treatment in which no cues were applied was used to establish a basal behavioural pattern. A trial was considered completed after each animal had been retained behind the cue barrier for at least 60 sec. All cues and electrical stimulation were manually applied from a laptop located on a portable 3.5 m tower located immediately outside the alley. The electric stimulation consisted of 1.0 Kv of electricity. Electric stimulation, sound and vibration along with the Global Position System (GPS) hardware to autonomously record the animal’s path within the alley were recorded every second.

Exercise-induced energy expenditure : implications for exercise prescription and obesity

Objective: Walking is commonly recommended to help with weight management. We measured total energy expenditure (TEE) and its components to quantify the impact of increasing exercise-induced energy expenditure (ExEE) on other components of TEE. Methods: Thirteen obese women underwent an 8-week walking group intervention. TEE was quantified using doubly labeled water, ExEE was quantified using heart rate monitors, daily movement was assessed by accelerometry and resting metabolic rate was measured using indirect calorimetry. Results: Four of the 13 participants achieved the target of 1500 kcal wk−1 of ExEE and all achieved 1000 kcal wk−1. The average ExEE achieved by the group across the 8 weeks was 1434 ± 237 kcal wk−1. Vigorous physical activity, as assessed by accelerometry, increased during the intervention by an average of 30 min per day. Non-exercise activity thermogenesis (NEAT) decreased, on average, by 175 kcal d−1 (−22%) from baseline to the intervention and baseline fitness was correlated with change in NEAT. Conclusions: Potential alterations in non-exercise activity should be considered when exercise is prescribed. The provision of appropriate education on how to self-monitor daily activity levels may improve intervention outcomes in groups who are new to exercise. Practice implications: Strategies to sustain incidental and light physical activity should be offered to help empower individuals as they develop and maintain healthy and long-lasting lifestyle habits.

Efficacy of a progressive walking program and glucosamine sulphate supplementation on osteoarthritic symptoms of the hip and knee: a feasibility trial

Introduction: Management of osteoarthritis (OA) includes the use of non-pharmacological and pharmacological therapies. Although walking is commonly recommended for reducing pain and increasing physical function in people with OA, glucosamine sulphate has also been used to alleviate pain and slow the progression of OA. This study evaluated the effects of a progressive walking program and glucosamine sulphate intake on OA symptoms and physical activity participation in people with mild to moderate hip or knee OA. Methods: Thirty-six low active participants (aged 42 to 73 years) were provided with 1500 mg glucosamine sulphate per day for 6 weeks, after which they began a 12-week progressive walking program, while continuing to take glucosamine. They were randomized to walk 3 or 5 days per week and given a pedometer to monitor step counts. For both groups, step level of walking was gradually increased to 3000 steps/day during the first 6 weeks of walking, and to 6000 steps/day for the next 6 weeks. Primary outcomes included physical activity levels, physical function (self-paced step test), and the WOMAC Osteoarthritis Index for pain, stiffness and physical function. Assessments were conducted at baseline and at 6-, 12-, 18-, and 24-week follow-ups. The Mann Whitney Test was used to examine differences in outcome measures between groups at each assessment, and the Wilcoxon Signed Ranks Test was used to examine differences in outcome measures between assessments. Results: During the first 6 weeks of the study (glucosamine supplementation only), physical activity levels, physical function, and total WOMAC scores improved (P<0.05). Between the start of the walking program (Week 6) and the final follow-up (Week 24), further improvements were seen in these outcomes (P<0.05) although most improvements were seen between Weeks 6 and 12. No significant differences were found between walking groups. Conclusions: In people with hip or knee OA, walking a minimum of 3000 steps (~30 minutes), at least 3 days/week, in combination with glucosamine sulphate, may reduce OA symptoms. A more robust study with a larger sample is needed to support these preliminary findings. Trial Registration: Australian Clinical Trials Registry ACTRN012607000159459.

The old face of 'new' social networks : the republic of letters as a virtual community

This paper explores the idea of virtual participation through the historical example of the republic of letters in early modern Europe (circa 1500-1800). By reflecting on the construction of virtuality in a historical context, and more specifically in a pre-digital environment, this paper calls attention to accusations of technological determinism in ongoing research concerning the affordances of the Internet and related media of communication. It argues that ‘the virtual’ is not synonymous with ‘the digital’ and suggests that, in order to articulate what is novel about modern technologies, we must first understand the social interactions underpinning the relationships which are facilitated through those technologies. By analysing the construction of virtuality in a pre-digital environment, this paper thus offers a baseline from which scholars might consider what is different about the modes of interaction and communication being engaged in via modern media.

Numerical evaluation of pollutant dispersion at a toll plaza based on system dynamics and Computational Fluid Dynamics models

The health of tollbooth workers is seriously threatened by long-term exposure to polluted air from vehicle exhausts. Using traffic data collected at a toll plaza, vehicle movements were simulated by a system dynamics model with different traffic volumes and toll collection procedures. This allowed the average travel time of vehicles to be calculated. A three-dimension Computational Fluid Dynamics (CFD) model was used with a k–ε turbulence model to simulate pollutant dispersion at the toll plaza for different traffic volumes and toll collection procedures. It was shown that pollutant concentration around tollbooths increases as traffic volume increases. Whether traffic volume is low or high (1500 vehicles/h or 2500 vehicles/h), pollutant concentration decreases if electronic toll collection (ETC) is adopted. In addition, pollutant concentration around tollbooths decreases as the proportion of ETC-equipped vehicles increases. However, if the proportion of ETC-equipped vehicles is very low and the traffic volume is not heavy, then pollutant concentration increases as the number of ETC lanes increases.

Teaching sustainability : vehicle or endpoint?

In 2006, the Faculty of Built Environment and Engineering introduced the first faculty wide unit dedicated to sustainability at any Australian University. BEB200 Introducing Sustainability has semester enrolments of up to 1500 students. Instruments such as lectures, readings, field visits, group projects and structured tutorial activities are used and have evolved over the last five years in response to student and staff feedback and attempts to better engage students. More than seventy staff have taught in the unit, which is in its final offering in this form in 2010. This paper reflects on the experiences of five academics who have played key roles in the development and teaching of this unit over the last five years. They argue that sustainability is a paradigm that allows students to explore other ways of knowing as they engage with issues in a complex world, not an end in itself. From the students’ perspective, grappling with such issues enables them to move towards a context in which they can understand their own discipline and its role in the contradictory and rapidly changing professional world. Insights are offered into how sustainability units may be developed in the future.

Endochondral ossification in vitro is influenced by mechanical bending

Bone development is influenced by the local mechanical environment. Experimental evidence suggests that altered loading can change cell proliferation and differentiation in chondro- and osteogenesis during endochondral ossification. This study investigated the effects of three-point bending of murine fetal metatarsal bone anlagen in vitro on cartilage differentiation, matrix mineralization and bone collar formation. This is of special interest because endochondral ossification is also an important process in bone healing and regeneration. Metatarsal preparations of 15 mouse fetuses stage 17.5 dpc were dissected en bloc and cultured for 7 days. After 3 days in culture to allow adherence they were stimulated 4 days for 20 min twice daily by a controlled bending of approximately 1000-1500 microstrain at 1 Hz. The paraffin-embedded bone sections were analyzed using histological and histomorphometrical techniques. The stimulated group showed an elongated periosteal bone collar while the total bone length was not different from controls. The region of interest (ROI), comprising the two hypertrophic zones and the intermediate calcifying diaphyseal zone, was greater in the stimulated group. The mineralized fraction of the ROI was smaller in the stimulated group, while the absolute amount of mineralized area was not different. These results demonstrate that a new device developed to apply three-point bending to a mouse metatarsal bone culture model caused an elongation of the periosteal bone collar, but did not lead to a modification in cartilage differentiation and matrix mineralization. The results corroborate the influence of biophysical stimulation during endochondral bone development in vitro. Further experiments with an altered loading regime may lead to more pronounced effects on the process of endochondral ossification and may provide further insights into the underlying mechanisms of mechanoregulation which also play a role in bone regeneration.