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.

A review of system safety failure probability objectives for Unmanned Aircraft Systems

Unmanned Aircraft Systems (UAS) are one of a number of emerging aviation sectors. Such new aviation concepts present a significant challenge to National Aviation Authorities (NAAs) charged with ensuring the safety of their operation within the existing airspace system. There is significant heritage in the existing body of aviation safety regulations for Conventionally Piloted Aircraft (CPA). It can be argued that the promulgation of these regulations has delivered a level of safety tolerable to society, thus justifying the “default position” of applying these same standards, regulations and regulatory structures to emerging aviation concepts such as UAS. An example of this is the proposed “1309” regulation for UAS, which is based on the 1309 regulation for CPA. However, the absence of a pilot on-board an unmanned aircraft creates a fundamentally different risk paradigm to that of CPA. An appreciation of these differences is essential to the justification of the “default position” and in turn, to ensure the development of effective safety standards and regulations for UAS. This paper explores the suitability of the proposed “1309” regulation for UAS. A detailed review of the proposed regulation is provided and a number of key assumptions are identified and discussed. A high-level model characterising the expected number of third party fatalities on the ground is then used to determine the impact of these assumptions. The results clearly show that the “one size fits all” approach to the definition of 1309 regulations for UAS, which mandates equipment design and installation requirements independent of where the UAS is to be operated, will not lead to an effective management of the risks.

Sustainable, smart, safe : a 3'S' approach towards a modern transportation system

Sustainability, smartness and safety are three sole components of a modern transportation system. The objective of this study is to introduce a modern transportation system in the light of a 3‘S’ approach: sustainable, smart and safe. In particular this paper studies the transportation system of Singapore to address how this system is progressing in this three-pronged approach towards a modern transportation system. While sustainability targets environmental justice and social equity without compromising economical efficiency, smartness incorporates qualities like automated sensing, processing and decision making, and action-taking into the transportation system. Since a system cannot be viable without being safe, the safety of the modern transportation system aims minimizing crash risks of all users including motorists, motorcyclists, pedestrians, and bicyclists. Various policy implications and technology applications inside the transportation system of Singapore are discussed to illustrate a modern transportation system within the framework of the 3‘S’ model.

Relative entropy rate based model selection for linear hybrid system filters of uncertain nonlinear systems

Hybrid system representations have been exploited in a number of challenging modelling situations, including situations where the original nonlinear dynamics are too complex (or too imprecisely known) to be directly filtered. Unfortunately, the question of how to best design suitable hybrid system models has not yet been fully addressed, particularly in the situations involving model uncertainty. This paper proposes a novel joint state-measurement relative entropy rate based approach for design of hybrid system filters in the presence of (parameterised) model uncertainty. We also present a design approach suitable for suboptimal hybrid system filters. The benefits of our proposed approaches are illustrated through design examples and simulation studies.

System identification, estimation and control for a cost effective open-source quadcopter

This paper describes system identification, estimation and control of translational motion and heading angle for a cost effective open-source quadcopter — the MikroKopter. The dynamics of its built-in sensors, roll and pitch attitude controller, and system latencies are determined and used to design a computationally inexpensive multi-rate velocity estimator that fuses data from the built-in inertial sensors and a low-rate onboard laser range finder. Control is performed using a nested loop structure that is also computationally inexpensive and incorporates different sensors. Experimental results for the estimator and closed-loop positioning are presented and compared with ground truth from a motion capture system.

Strategy taxonomy and classification system development : study of two state governments

There is limited understanding about business strategies related to parliamentary government's departments. This study focuses on the strategies of departments of two state governments in Australia. The strategies are derived from department strategic plans available in public domain and collected from respective websites. The results of this research indicate that strategies fall into seven categories: internal, development, political, partnership, environment, reorientation and status quo. The strategies of the departments are mainly internal or development where development strategy is mainly the focus of departments such as transport, and infrastructure. Political strategy is prevalent for departments related to communities, and education and training. Further three layers of strategies are identified as kernel, cluster and individual, which are mapped to the developed taxonomy.

An improved model of tumour-immune system interactions

The immune system plays an important role in defending the body against tumours and other threats. Currently, mechanisms involved in immune system interactions with tumour cells are not fully understood. Here we develop a mathematical tool that can be used in aiding to address this shortfall in understanding. This paper de- scribes a hybrid cellular automata model of the interaction between a growing tumour and cells of the innate and specific immune system including the effects of chemokines that builds on previous models of tumour-immune system interactions. In particular, the model is focused on the response of immune cells to tumour cells and how the dynamics of the tumour cells change due to the immune system of the host. We present results and predictions of in silico experiments including simulations of Kaplan-Meier survival-like curves.

The relationship between listening and other language skills in International English Language Testing System

Listening comprehension is the primary channel of learning a language. Yet of the four dominant macro-skills (listening, speaking, reading and writing), it is often difficult and inaccessible for second and foreign language learners due to its implicit process. The secondary skill, speaking, proceeds listening cognitively. Aural/oral skills precede the graphic skills, such as reading and writing, as they form the circle of language learning process. However, despite the significant relationship with other language skills, listening comprehension is treated lightly in the applied linguistics research. Half of our daily conversation and three quarters of classroom interaction are virtually devoted to listening comprehension. To examine the relationship of listening skill with other language skills, the outcome of 1800 Iranian participants undertaking International English Language Testing System (IELTS) in Tehran indicates the close correlation between listening comprehension and the overall language proficiency.

Child Victims of Sexual Abuse: Children's Experience of the Investigative Process in the Criminal Justice System

This paper is a selected review of research on issues surrounding the investigation of intra-familial child sexual abuse for children aged eight and above, in the criminal justice system. Particular attention is paid to features of the investigative interview in relation to the child's level of understanding, ability to report and likely emotional response when the proceedings take place. Best practice by police and social care agencies involves establishing valid and reliable information from children while attending to their developmental level and emotional state. The review aims to distil principles optimising this process from both the investigative judicial perspective and the child's focus, as well as from the inter-agency perspective and information sharing. Recommendations are made for improving the interview process based on research and methods from a range of disciplines and to optimise information recording in a format easily shared between agencies. Updated and ongoing training procedures are key to successful practice with training shared across police and social work agencies. The focus of this review is informed by preliminary findings from pilot research in progress on behalf of the Metropolitan Police Child Abuse Investigation Command.