The manipulation of apoptosis-related genes to generate resistance to Fusarium wilt and water stress in banana

Bananas are susceptible to a diverse range of biotic and abiotic stresses, many of which cause serious production constraints worldwide. One of the most destructive banana diseases is Fusarium wilt caused by the soil-borne fungus, Fusarium oxysporum f. sp. cubense (Foc). No effective control strategy currently exists for this disease which threatens global banana production. Although disease resistance exists in some wild bananas, attempts to introduce resistance into commercially acceptable bananas by conventional breeding have been hampered by low fertility, long generation times and association of poor agronomical traits with resistance genes. With the advent of reliable banana transformation protocols, molecular breeding is now regarded as a viable alternative strategy to generate disease-resistant banana plants. Recently, a novel strategy involving the expression of anti-apoptosis genes in plants was shown to result in resistance against several necrotrophic fungi. Further, the transgenic plants showed increased resistance to a range of abiotic stresses. In this thesis, the use of anti-apoptosis genes to generate transgenic banana plants with resistance to Fusarium wilt was investigated. Since water stress is an important abiotic constraint to banana production, the resistance of the transgenic plants to water stress was also examined. Embryogenic cell suspensions (ECS) of two commercially important banana cultivars, Grand Naine (GN) and Lady Finger (LF), were transformed using Agrobacterium with the anti-apoptosis genes, Bcl-xL, Bcl-xL G138A, Ced-9 and Bcl- 2 3’ UTR. An interesting, and potentially important, outcome was that the use of anti-apoptosis genes resulted in up to a 50-fold increase in Agrobacterium-mediated transformation efficiency of both LF and GN cells over vector controls. Regenerated plants were subjected to a complete molecular characterisation in order to detect the presence of the transgene (PCR), transcript (RT-PCR) and gene product (Western blot) and to determine the gene copy number (Southern blot). A total of 36 independently-transformed GN lines (8 x Bcl-xL, 5 x Bcl-xL G138A, 15 x Ced-9 and 8 x Bcl-2 3’ UTR) and 41 independently-transformed LF lines (8 x Bcl-xL, 7 x BclxL G138A, 13 x Ced-9 and 13 x Bcl-2 3’ UTR) were identified. The 41 transgenic LF lines were multiplied and clones from each line were acclimatised and grown under glasshouse conditions for 8 weeks to allow monitoring for phenotypic abnormalities. Plants derived from 3 x Bcl-xL, 2 x Ced-9 and 5 x Bcl-2 3’ UTR lines displayed a variety of aberrant phenotypes. However, all but one of these abnormalities were off-types commonly observed in tissue-cultured, non-transgenic banana plants and were therefore unlikely to be transgene-related. Prior to determining the resistance of the transgenic plants to Foc race 1, the apoptotic effects of the fungus on both wild-type and Bcl-2 3’ UTR-transgenic LF banana cells were investigated using rapid in vitro root assays. The results from these assays showed that apoptotic-like cell death was elicited in wild-type banana root cells as early as 6 hours post-exposure to fungal spores. In contrast, these effects were attenuated in the root cells of Bcl-2 3’ UTR-transgenic lines that were exposed to fungal spores. Thirty eight of the 41 transgenic LF lines were subsequently assessed for resistance to Foc race 1 in small-plant glasshouse bioassays. To overcome inconsistencies in rating the internal (vascular discolouration) disease symptoms, a MatLab-based computer program was developed to accurately and reliably assess the level of vascular discolouration in banana corms. Of the transgenic LF banana lines challenged with Foc race 1, 2 x Bcl-xL, 3 x Ced-9, 2 x Bcl-2 3’ UTR and 1 x Bcl-xL G138A-transgenic line were found to show significantly less external and internal symptoms than wild-type LF banana plants used as susceptible controls at 12 weeks post-inoculation. Of these lines, Bcl-2 3’ UTR-transgenic line #6 appeared most resistant, displaying very mild symptoms similar to the wild-type Cavendish banana plants that were included as resistant controls. This line remained resistant for up to 23 weeks post-inoculation. Since anti-apoptosis genes have been shown to confer resistance to various abiotic stresses in other crops, the ability of these genes to confer resistance against water stress in banana was also investigated. Clonal plants derived from each of the 38 transgenic LF banana plants were subjected to water stress for a total of 32 days. Several different lines of transgenic plants transformed with either Bcl-xL, Bcl-xL G138A, Ced-9 or Bcl-2 3’ UTR showed a delay in visual water stress symptoms compared with the wild-type control plants. These plants all began producing new growth from the pseudostem following daily rewatering for one month. In an attempt to determine whether the protective effect of anti-apoptosis genes in transgenic banana plants was linked with reactive oxygen species (ROS)-associated programmed cell death (PCD), the effect of the chloroplast-targeting, ROS-inducing herbicide, Paraquat, on wild-type and transgenic LF was investigated. When leaf discs from wild-type LF banana plants were exposed to 10 ìM Paraquat, complete decolourisation occurred after 48 hours which was confirmed to be associated with cell death and ROS production by trypan blue and 3,3-diaminobenzidine (DAB) staining, respectively. When leaf discs from the transgenic lines were exposed to Paraquat, those derived from some lines showed a delay in decolourisation, suggesting only a weak protective effect from the transgenes. Finally, the protective effect of anti-apoptosis genes against juglone, a ROS-inducing phytotoxin produced by the causal agent of black Sigatoka, Mycosphaerella fijiensis, was investigated. When leaf discs from wild-type LF banana plants were exposed to 25 ppm juglone, complete decolourisation occurred after 48 hours which was again confirmed to be associated with cell death and ROS production by trypan blue and DAB staining, respectively. Further, TdT-mediated dUTP nick-end labelling (TUNEL) assays on these discs suggested that the cell death was apoptotic. When leaf discs from the transgenic lines were exposed to juglone, discs from some lines showed a clear delay in decolourisation, suggesting a protective effect. Whether these plants are resistant to black Sigatoka is unknown and will require future glasshouse and field trials. The work presented in this thesis provides the first report of the use of anti-apoptosis genes as a strategy to confer resistance to Fusarium wilt and water stress in a nongraminaceous monocot, banana. Such a strategy may be exploited to generate resistance to necrotrophic pathogens and abiotic stresses in other economically important crop plants.

Public accountability : the perceived usefulness of School Annual reports

Annual reports are an important component of New Zealand schools’ public accountability. Through the annual report the governance body informs stakeholders about school aims, objectives, achievements, use of resources, and financial performance. We examine the perceived usefulness of the annual report in the discharge of public accountability. We find that 15% of the recipients (mainly parents/caregivers) do not read the annual report because they trust in the school to do the right thing or rely on others to monitor school activities. We find that the annual report is used for a variety of purposes including to determine if the school has conducted its activities effectively and achieved stated objectives and goals; to examine student achievements; to assess financial accountability and performance; and to make decisions about the school as a suitable environment for their child/children. We find that other forms of communication such as school newsletters, parent-teacher interviews, children and other parents are more important sources of information about the school than the annual report.

Molecular architecture of the human sinus node: insights into the function of the cardiac pacemaker.

BACKGROUND: Although we know much about the molecular makeup of the sinus node (SN) in small mammals, little is known about it in humans. The aims of the present study were to investigate the expression of ion channels in the human SN and to use the data to predict electrical activity. METHODS AND RESULTS: Quantitative polymerase chain reaction, in situ hybridization, and immunofluorescence were used to analyze 6 human tissue samples. Messenger RNA (mRNA) for 120 ion channels (and some related proteins) was measured in the SN, a novel paranodal area, and the right atrium (RA). The results showed, for example, that in the SN compared with the RA, there was a lower expression of Na(v)1.5, K(v)4.3, K(v)1.5, ERG, K(ir)2.1, K(ir)6.2, RyR2, SERCA2a, Cx40, and Cx43 mRNAs but a higher expression of Ca(v)1.3, Ca(v)3.1, HCN1, and HCN4 mRNAs. The expression pattern of many ion channels in the paranodal area was intermediate between that of the SN and RA; however, compared with the SN and RA, the paranodal area showed greater expression of K(v)4.2, K(ir)6.1, TASK1, SK2, and MiRP2. Expression of ion channel proteins was in agreement with expression of the corresponding mRNAs. The levels of mRNA in the SN, as a percentage of those in the RA, were used to estimate conductances of key ionic currents as a percentage of those in a mathematical model of human atrial action potential. The resulting SN model successfully produced pacemaking. CONCLUSIONS: Ion channels show a complex and heterogeneous pattern of expression in the SN, paranodal area, and RA in humans, and the expression pattern is appropriate to explain pacemaking.

60Sox Report Volume 1: from education to work in Australia's digital content industries. The opinions and practices of aspiring creatives in the creative industries

This report presents findings from the largest survey of aspiring creatives who work or intend to work in the digital content industries ever undertaken in Australia. Survey respondents included those with aspirations to work in the publicly-supported, less commercial end of the Creative Industries spectrum as well as those with aspirations to work in the digital content industries. The survey gathered rich data on their characteristics, skills and attributes, barriers to employment, workforce mobility, career intentions, professional development, mentors and industry supports, and participation in communities of practice. The survey sought to determine if aspiring creatives have the necessary skills and attributes to work effectively in the digital content industries. This task also involved finding out how they develop their skills and attributes, and what they need to develop them further.

From education to work in Australia’s creative digital industries : comparing the opinions and practices of employers and aspiring creatives

This report represents the second of two reports that aim to explore views about the employability of aspiring creatives. The first report, released in June 2009, presented findings from a survey of 507 aspiring creatives, defined as recent graduates and/or people with less than two years industry experience. It presents findings from a project that administered an survey to employers in Australia’s Creative Digital Industries. The survey included questions on employer characteristics, recruitment and training practices, employers’ views of the capabilities of aspiring creatives, and participation in communities of interest/networks, mentoring and internships. The main purpose of the project was to identify capability gaps of aspiring creatives as well as those factors that enhance or inhibit employers’ views of the capabilities of aspiring creatives – both of which impact on the ability of aspiring creatives to find work in their preferred occupations in Australia’s Creative Digital Industries.

Online scheduling in the operating theatre

Online scheduling is considered in this paper for the Operating Theatre. Robust elective schedules are determined in the offline environment prior to the day of surgery for the online environment. Changes to the offline schedule during project implementation are minimized using an online scheduling model that operates in real-time. The model aims to minimise cancellations of pre-scheduled elective patients whilst also allowing for additional scheduling of emergency cases, time permitting, which may arise during the schedules implementation. Surgical durations are modelled with a lognormal distribution. The single theatre case is solved and the computationally complex multiple theatre case, which is left for future work, is discussed.

User-Created Content and Online Social Networks

Traditional media are under assault from digital technologies. Online advertising is eroding the financial basis of newspapers and television, demarcations between different forms of media are fading, and audiences are fragmenting. We can podcast our favourite radio show, data accompanies television programs, and we catch up with newspaper stories on our laptops. Yet mainstream media remain enormously powerful. The Media and Communications in Australia offers a systematic introduction to this dynamic field. Fully updated and revised to take account of recent developments, this third edition outlines the key media industries and explains how communications technologies are impacting on them. It provides a thorough overview of the main approaches taken in studying the media, and includes new chapters on social media, gaming, telecommunications, sport and cultural diversity. With contributions from some of Australia's best researchers and teachers in the field, The Media and Communications in Australia is the most comprehensive and reliable introduction to media and communications available. It is an ideal student text, and a reference for teachers of media and anyone interested in this influential industry.

Working sheep at 'Glen Shiel'

Working Sheep on 'Glen Shiel' is a key work of the author's exhibition Lightsite, which toured Western Australian galleries from February 2006 to November 2007. It is a five-minute-long exposure photographic image captured inside a purpose-built, room-sized pinhole camera which is demountable and does not have a floor. The work depicts octogenarian Ian Mangan who is both one of the first and last soldier settler farmers in the Gairdner-Jerramungup district in the Great Southern Region of Western Australia. Ian, his son, Stuart and Grandson Jacob, are preparing the last mob of sheep for sale before they move off their farm. The pinhole camera-room is sited amongst the sheep in the farm's sheep yards. Stuart and Jacob are depicted here standing amongst the sheep. The light from this exterior landscape is 'projected' inside the camera-room and illuminates the interior scene which includes that part of the sheep yards upon which the floorless room is erected, along with Ian who is standing motionless inside. The image evokes the temporality of light. Here, light itself is portrayed as the primary medium through which we both perceive and describe landscape. In this way it is through the agency of light that we construct our connectivity to landscape.

Canopy dweller

This article discusses the renovation of a house located in western Brisbane, Queensland, originally designed by architects Lindsay and Kerry Clare and now renovated by architect Stuart Vokes of Owen and Vokes. Features of the renovation include keeping the Queenslander style while updating with new timber finishing, white painted surfaces, opening up living areas and designing a new kitchen.

Market access of Papua New Guinea bananas (Musa spp.) with particular respect to banana fly (Bactrocera musae (Tryon)) (Diptera: Tephritidae)

International market access for fresh commodities is regulated by international accepted phytosanitary guidelines, the objectives of which are to reduce the biosecurity risk of plant pest and disease movement. Papua New Guinea (PNG) has identified banana as a potential export crop and to help meet international market access requirements, this thesis provides information for the development of a pest risk analysis (PRA) for PNG banana fruit. The PRA is a three step process which first identifies the pests associated with a particular commodity or pathway, then assesses the risk associated with those pests, and finally identifies risk management options for those pests if required. As the first step of the PRA process, I collated a definitive list on the organisms associated with the banana plant in PNG using formal literature, structured interviews with local experts, grey literature and unpublished file material held in PNG field research stations. I identified 112 organisms (invertebrates, vertebrate, pathogens and weeds) associated with banana in PNG, but only 14 of these were reported as commonly requiring management. For these 14 I present detailed information summaries on their known biology and pest impact. A major finding of the review was that of the 14 identified key pests, some research information occurs for 13. The single exception for which information was found to be lacking was Bactrocera musae (Tryon), the banana fly. The lack of information for this widely reported ‘major pest on PNG bananas’ would hinder the development of a PNG banana fruit PRA. For this reason the remainder of the thesis focused on this organism, particularly with respect to generation of information required by the PRA process. Utilising an existing, but previously unanalysed fruit fly trapping database for PNG, I carried out a Geographic Information System analysis of the distribution and abundance of banana in four major regions of PNG. This information is required for a PRA to determine if banana fruit grown in different parts of the country are at different risks from the fly. Results showed that the fly was widespread in all cropping regions and that temperature and rainfall were not significantly correlated with banana fly abundance. Abundance of the fly was significantly correlated (albeit weakly) with host availability. The same analysis was done with four other PNG pest fruit flies and their responses to the environmental factors differed to banana fly and each other. This implies that subsequent PRA analyses for other PNG fresh commodities will need to investigate the risk of each of these flies independently. To quantify the damage to banana fruit caused by banana fly in PNG, local surveys and one national survey of banana fruit infestation were carried out. Contrary to expectations, infestation was found to be very low, particularly in the widely grown commercial cultivar, Cavendish. Infestation of Cavendish fingers was only 0.41% in a structured, national survey of over 2 700 banana fingers. Follow up laboratory studies showed that fingers of Cavendish, and another commercial variety Lady-finger, are very poor hosts for B. musae, with very low host selection rates by female flies and very poor immature survival. An analysis of a recent (within last decade) incursion of B. musae into the Gazelle Peninsula of East New Britain Province, PNG, provided the final set of B. musae data. Surveys of the fly on the peninsular showed that establishment and spread of the fly in the novel environment was very rapid and thus the fly should be regarded as being of high biosecurity concern, at least in tropical areas. Supporting the earlier impact studies, however, banana fly has not become a significant banana fruit problem on the Gazelle, despite bananas being the primary starch staple of the region. The results of the research chapters are combined in the final Discussion in the form of a B. musae focused PRA for PNG banana fruit. Putting the thesis in a broader context, the Discussion also deals with the apparent discrepancy between high local abundance of banana fly and very low infestation rates. This discussion focuses on host utilisation patterns of specialist herbivores and suggests that local pest abundance, as determined by trapping or monitoring, need not be good surrogate for crop damage, despite this linkage being implicit in a number of international phytosanitary protocols.

What's your other job : a census analysis of arts employment in Australia

This report applies CCI’s creative trident methodology with the definition of the arts as established by the Australia Council for the Arts to data sourced from Australia’s national census data (from 1996, 2001 and the most recent one in 2006). Analysis has been conducted on employment, income, gender, age and the nature of employment for artists and arts related workers within and beyond the arts industries, as well as other support workers in the arts industries.