TaNF-YB3 is involved in the regulation of photosynthesis genes in Triticum aestivum

Nuclear Factor Y (NF-Y) transcription factor is a heterotrimer comprised of three subunits: NF-YA, NF-YB and NF-YC. Each of the three subunits in plants is encoded by multiple genes with differential expression profiles, implying the functional specialisation of NF-Y subunit members in plants. In this study, we investigated the roles of NF-YB members in the light-mediated regulation of photosynthesis genes. We identified two NF-YB members from Triticum aestivum (TaNF-YB3 & 7) which were markedly upregulated by light in the leaves and seedling shoots using quantitative RT-PCR. A genome-wide coexpression analysis of multiple Affymetrix Wheat Genome Array datasets revealed that TaNF-YB3-coexpressed transcripts were highly enriched with the Gene Ontology term photosynthesis. Transgenic wheat lines constitutively overexpressing TaNF-YB3 had a significant increase in the leaf chlorophyll content, photosynthesis rate and early growth rate. Quantitative RT-PCR analysis showed that the expression levels of a number of TaNF-YB3-coexpressed transcripts were elevated in the transgenic wheat lines. The mRNA level of TaGluTR encoding glutamyl-tRNA reductase, which catalyses the rate limiting step of the chlorophyll biosynthesis pathway, was significantly increased in the leaves of the transgenic wheat. Significant increases in the expression level in the transgenic plant leaves were also observed for four photosynthetic apparatus genes encoding chlorophyll a/b-binding proteins (Lhca4 and Lhcb4) and photosystem I reaction center subunits (subunit K and subunit N), as well as for a gene coding for chloroplast ATP synthase  subunit. These results indicate that TaNF-YB3 is involved in the positive regulation of a number of photosynthesis genes in wheat.

Characterisation of the TaNF-Y family of transcription factors in wheat

Light plays a unique role for plants as it is both a source of energy for growth and a signal for development. Light captured by the pigments in the light harvesting complexes is used to drive the synthesis of the chemical energy required for carbon assimilation. The light perceived by photoreceptors activates effectors, such as transcription factors (TFs), which modulate the expression of light-responsive genes. Recently, it has been speculated that increasing the photosynthetic rate could further improve the yield potential of three carbon (C3) crops such as wheat. However, little is currently known about the transcriptional regulation of photosynthesis genes, particularly in crop species. Nuclear factor Y (NF-Y) TF is a functionally diverse regulator of growth and development in the model plant species, with demonstrated roles in embryo development, stress response, flowering time and chloroplast biogenesis. Furthermore, a light-responsive NF-Y binding site (CCAAT-box) is present in the promoter of a spinach photosynthesis gene. As photosynthesis genes are co-regulated by light and co-regulated genes typically have similar regulatory elements in their promoters, it seems likely that other photosynthesis genes would also have light-responsive CCAAT-boxes. This provided the impetus to investigate the NF-Y TF in bread wheat. This thesis is focussed on wheat NF-Y members that have roles in light-mediated gene regulation with an emphasis on their involvement in the regulation of photosynthesis genes. NF-Y is a heterotrimeric complex, comprised of the three subunits NF-YA, NF-YB and NF-YC. Unlike the mammalian and yeast counterparts, each of the three subunits is encoded by multiple genes in Arabidopsis. The initial step taken in this study was the identification of the wheat NF-Y family (Chapter 3). A search of the current wheat nucleotide sequence databases identified 37 NF-Y genes (10 NF-YA, 11 NF-YB, 14 NF-YC & 2 Dr1). Phylogenetic analysis revealed that each of the three wheat NF-Y (TaNF-Y) subunit families could be divided into 4-5 clades based on their conserved core regions. Outside of the core regions, eleven motifs were identified to be conserved between Arabidopsis, rice and wheat NF-Y subunit members. The expression profiles of TaNF-Y genes were constructed using quantitative real-time polymerase chain reaction (RT-PCR). Some TaNF-Y subunit members had little variation in their transcript levels among the organs, while others displayed organ-predominant expression profiles, including those expressed mainly in the photosynthetic organs. To investigate their potential role in light-mediated gene regulation, the light responsiveness of the TaNF-Y genes were examined (Chapters 4 and 5). Two TaNF-YB and five TaNF-YC members were markedly upregulated by light in both the wheat leaves and seedling shoots. To identify the potential target genes of the light-upregulated NF-Y subunit members, a gene expression correlation analysis was conducted using publically available Affymetrix Wheat Genome Array datasets. This analysis revealed that the transcript expression levels of TaNF-YB3 and TaNF-YC11 were significantly correlated with those of photosynthesis genes. These correlated express profiles were also observed in the quantitative RT-PCR dataset from wheat plants grown under light and dark conditions. Sequence analysis of the promoters of these wheat photosynthesis genes revealed that they were enriched with potential NF-Y binding sites (CCAAT-box). The potential role of TaNF-YB3 in the regulation of photosynthetic genes was further investigated using a transgenic approach (Chapter 5). Transgenic wheat lines constitutively expressing TaNF-YB3 were found to have significantly increased expression levels of photosynthesis genes, including those encoding light harvesting chlorophyll a/b-binding proteins, photosystem I reaction centre subunits, a chloroplast ATP synthase subunit and glutamyl-tRNA reductase (GluTR). GluTR is a rate-limiting enzyme in the chlorophyll biosynthesis pathway. In association with the increased expression of the photosynthesis genes, the transgenic lines had a higher leaf chlorophyll content, increased photosynthetic rate and had a more rapid early growth rate compared to the wild-type wheat. In addition to its role in the regulation of photosynthesis genes, TaNF-YB3 overexpression lines flower on average 2-days earlier than the wild-type (Chapter 6). Quantitative RT-PCR analysis showed that there was a 13-fold increase in the expression level of the floral integrator, TaFT. The transcript levels of other downstream genes (TaFT2 and TaVRN1) were also increased in the transgenic lines. Furthermore, the transcript levels of TaNF-YB3 were significantly correlated with those of constans (CO), constans-like (COL) and timing of chlorophyll a/b-binding (CAB) expression 1 [TOC1; (CCT)] domain-containing proteins known to be involved in the regulation of flowering time. To summarise the key findings of this study, 37 NF-Y genes were identified in the crop species wheat. An in depth analysis of TaNF-Y gene expression profiles revealed that the potential role of some light-upregulated members was in the regulation of photosynthetic genes. The involvement of TaNF-YB3 in the regulation of photosynthesis genes was supported by data obtained from transgenic wheat lines with increased constitutive expression of TaNF-YB3. The overexpression of TaNF-YB3 in the transgenic lines revealed this NF-YB member is also involved in the fine-tuning of flowering time. These data suggest that the NF-Y TF plays an important role in light-mediated gene regulation in wheat.

From the consciousness industry to the creative industries : consumer-created content, social network markets, & the growth of knowledge

In studies of media industries, too much attention has been paid to providers and firms, too little to consumers and markets. But with user-created content, the question first posed more than a generation ago by the uses & gratifications method and taken up by semiotics and the active audience tradition (‘what do audiences do with media?’), has resurfaced with renewed force. What’s new is that where this question (of what the media industries and audiences did with each other) used to be individualist and functionalist, now, with the advent of social networks using Web 2.0 affordances, it can be re-posed at the level of systems and populations as well.

Search engine content analysis

Search engines have forever changed the way people access and discover knowledge, allowing information about almost any subject to be quickly and easily retrieved within seconds. As increasingly more material becomes available electronically the influence of search engines on our lives will continue to grow. This presents the problem of how to find what information is contained in each search engine, what bias a search engine may have, and how to select the best search engine for a particular information need. This research introduces a new method, search engine content analysis, in order to solve the above problem. Search engine content analysis is a new development of traditional information retrieval field called collection selection, which deals with general information repositories. Current research in collection selection relies on full access to the collection or estimations of the size of the collections. Also collection descriptions are often represented as term occurrence statistics. An automatic ontology learning method is developed for the search engine content analysis, which trains an ontology with world knowledge of hundreds of different subjects in a multilevel taxonomy. This ontology is then mined to find important classification rules, and these rules are used to perform an extensive analysis of the content of the largest general purpose Internet search engines in use today. Instead of representing collections as a set of terms, which commonly occurs in collection selection, they are represented as a set of subjects, leading to a more robust representation of information and a decrease of synonymy. The ontology based method was compared with ReDDE (Relevant Document Distribution Estimation method for resource selection) using the standard R-value metric, with encouraging results. ReDDE is the current state of the art collection selection method which relies on collection size estimation. The method was also used to analyse the content of the most popular search engines in use today, including Google and Yahoo. In addition several specialist search engines such as Pubmed and the U.S. Department of Agriculture were analysed. In conclusion, this research shows that the ontology based method mitigates the need for collection size estimation.

Exploring the influence of a science content course incorporating explicit nature of science and argumentation instruction on preservice primary teachers' views of nature of science

There exists a general consensus in the science education literature around the goal of enhancing students. and teachers. views of nature of science (NOS). An emerging area of research in science education explores NOS and argumentation, and the aim of this study was to explore the effectiveness of a science content course incorporating explicit NOS and argumentation instruction on preservice primary teachers. views of NOS. A constructivist perspective guided the study, and the research strategy employed was case study research. Five preservice primary teachers were selected for intensive investigation in the study, which incorporated explicit NOS and argumentation instruction, and utilised scientific and socioscientific contexts for argumentation to provide opportunities for participants to apply their NOS understandings to their arguments. Four primary sources of data were used to provide evidence for the interpretations, recommendations, and implications that emerged from the study. These data sources included questionnaires and surveys, interviews, audio- and video-taped class sessions, and written artefacts. Data analysis involved the formation of various assertions that informed the major findings of the study, and a variety of validity and ethical protocols were considered during the analysis to ensure the findings and interpretations emerging from the data were valid. Results indicated that the science content course was effective in enabling four of the five participants. views of NOS to be changed. All of the participants expressed predominantly limited views of the majority of the examined NOS aspects at the commencement of the study. Many positive changes were evident at the end of the study with four of the five participants expressing partially informed and/or informed views of the majority of the examined NOS aspects. A critical analysis of the effectiveness of the various course components designed to facilitate the development of participants‟ views of NOS in the study, led to the identification of three factors that mediated the development of participants‟ NOS views: (a) contextual factors (including context of argumentation, and mode of argumentation), (b) task-specific factors (including argumentation scaffolds, epistemological probes, and consideration of alternative data and explanations), and (c) personal factors (including perceived previous knowledge about NOS, appreciation of the importance and utility value of NOS, and durability and persistence of pre-existing beliefs). A consideration of the above factors informs recommendations for future studies that seek to incorporate explicit NOS and argumentation instruction as a context for learning about NOS.

Encounters and content sharing in an urban village : reading texts through an archaeological lens

Archaeology provides a framework of analysis and interpretation that is useful for disentangling the textual layers of a contemporary lived-in urban space. The producers and readers of texts may include those who planned and developed the site and those who now live, visit and work there. Some of the social encounters and content sharing between these people may be artificially produced or manufactured in the hope that certain social situations will occur. Others may be serendipitous. With archaeology’s original focus on places that are no longer inhabited it is often only the remaining artefacts and features of the built environment that form the basis for interpreting the social relationships of past people. Our analysis however, is framed within a contemporary notion of archaeological artefacts in an urban setting. Unlike an excavation, where the past is revealed through digging into the landscape, the application of landscape archaeology within a present day urban context is necessarily more experiential, visual and based on recording and analysing the physical traces of social encounters and relationships between residents and visitors. These physical traces are present within the creative content, and the built and natural elements of the environment. This chapter explores notions of social encounters and content sharing in an urban village by analysing three different types of texts: the design of the built environment; content produced by residents through a geospatial web application; and, print and online media produced in digital storytelling workshops.

Women's participation in the Australian Digital Content Industry : initial case study findings

An exploratory case study which seeks to understand better the problem of low participation rates of women in Information Communication Technology (ICT) is currently being conducted in Queensland, Australia. Contextualised within the Digital Content Industry (DCI) multimedia and games production sectors, the emphasis is on women employed as interactive content creators rather than as users of the technologies. Initial findings provide rich descriptive insights into the perceptions and experiences of female DCI professionals. Influences on participation such as: existing gender ratios, gender and occupational stereotypes, access into the industry and future parental responsibilities have emerged from the data. Bandura’s (1999) Social Cognitive Theory (SCT) is used as a “scaffold” (Walsham, 1995:76) to guide data analysis and assist analytic generalisation of the case study findings. We propose that the lens of human agency and theories such as SCT assist in explaining how influences are manifested and affect women’s agency and ultimately participation in the DCI. The Sphere of Influence conceptual model (Geneve et al, 2008), which emerges from the data and underpinning theory, is proposed as a heuristic framework to further explore influences on women’s participation in the DCI industry context.