Many environmental stresses result in increased generation of active oxygen species (AOS) in plant cells, leading to the induction of protective mechanisms. In this study, signalling components linking AOS perception to downstream responses were examined, with particular emphasis on H₂O₂ signalling. All AOS investigated had an early [Ca²⁺]_cyt peak in common, but differed in other aspects of their Ca²⁺ signatures, indicating that the plant is able to discriminate between different types of AOS. An early event in AOS signal transduction may involve changes in the cellular redox balance as reduction of glutathione levels prior to stress application increased the height of the first [Ca²⁺]_cyt peak. Inhibiting or enhancing the height of the H₂O₂-triggered Ca²⁺ signature lead to inhibition or enhancement of GST1 and APX1 induction, respectively, demonstrating that the Ca²⁺ signature is required for induction of genes encoding antioxidant enzymes. OX1, encoding a putative ser/thr kinase, was shown to be involved in signal transduction in response to H₂O₂-generating stresses. Transcript levels of OX1 were increased upon treatment with H₂O₂ and a range of abiotic and biotic stresses as well as ABA, all of which have been shown to result in H₂O₂ accumulation. Inhibition of stress-induced [Ca²⁺]_cyt elevations inhibited OX1 induction, placing the OX1 kinase downstream of Ca²⁺ in the signalling chain. OX1 is required for full activation of AtMPKS and AtMPK6 in response to ozone fumigation, indicating that OX1 functions upstream of these MAP kinases. An ox1 null-mutant displayed enhanced susceptibility to infection with a virulent Peronospora parasitica isolate as well as reduced induction of several defence genes. In addition, the ox1 mutant exhibited shorter root hairs and an early flowering phenotype. AOS treatment induced several genes encoding AtERF transcription factors, but did not have an effect on other members of this family. Induction occurred in an ethylene-independent but Ca²⁺-dependent manner.

The sequencing of eukaryotic model organisms has provided us an unprecedented opportunity for a genome-wide characterization of Transposable Elements (TEs) and the study of TE-host relationships. By developing methodologies on database mining, we explored the existence of MUtator (Mu)-Like Elements (MULEs) in Arabidopsis thaliana and Oryza sativa. Mu elements were first discovered in Zea mays; so far, a dozen of the elements have been identified in the genome. We identified a total of 1392 MULEs from the sequenced Arabidopsis genome. They represent one of the most abundant, diversified, yet still mobile DNA transposon families in eukaryotes. The Arabidopsis MULEs are composed of not only the elements showing the typical Mu-family-specific terminal structure (that is the long Terminal Inverted Repeat, TIR), but also a novel type of non-TIR MULEs. Some of this latter type of elements was found to be active both transcriptionally and transpositionally. To understand host-mediated genome-wide regulation(s) on the MULE system in Arabidopsis, we characterized 235 MULE mobility-specific genes (or mudrA-like genes) by mapping them on the sequenced Arabidopsis chromosomes and performing a genome-wide expression assay utilizing Arabidopsis METHYLTRANSFERSE1 (MET1) mutant (met2) plants, we showed that MET1-mediated global CpG methylation can only repress a portion of the gene family; its efficiency depends largely on the gene locations within the context of Arabidopsis chromatin remodeling: stronger in heterochromatic regions but weaker in euchromatic ones. This finding suggests that the Arabidopsis heterochromatic regions are not just a graveyard for the accumulation of defective elements; rather, they may have been playing an important role on the repression of TE activity via, at least in part, exerting MET1-mediated silencing effect. Our expression analysis also suggested that a TIR structure is not necessarily required for the MET1-mediated si

The brevipedicellus (bp) mutant, caused by a loss-of-function mutation in the KNAT1 homeobox gene, is known to affect the stem morphogenesis. BP and ERECTA (ER) genes are required to promote internode and pedicel development and delineate nodal boundaries to maintain the radial symmetry of stems and pedicels. My research aims to identify genes acting on the BP pathway by utilizing a forward genetics approach. The suppressor4 mutant, identified from the bp er mutant screen, exhibits moderate length and perpendicularly-oriented pedicels with partially formed distal pedicel bulges, absent in the bp mutant. The kinky mutant, identified from the bp mutant screen, develops severe bends at the floral nodes and enhanced achlorophyllous stripes. These results suggest the SUPPRESSOR4 gene contribution in inhibiting the development of distal pedicel bulge and influencing both pedicel angle and length whereas the KINKY gene might act with BP in regulating proper inflorescence development.

Auxins are plant hormones involved in virtually all aspects of plant life. Despite long-term commercial and horticultural use of the auxin Indole-3-Butyric Acid (IBA), a full recognition of its natural occurrence in plants was made only recently. I have used multiple approaches to dissect the role of IBA in Arabidopsis thaliana. This thesis includes the first characterization of a mutant with an altered response to IBA that retains wild-type sensitivity to Indole-3-Acetic Acid (IAA), the most studied endogenous auxin. This mutant, named resistant to IBA ( rib1), has modified root architecture and gravitropism and is resistant to auxin transport inhibitors. As these phenotypes are reminiscent of those of characterized auxin transport mutants, movement of IAA and IBA was studied in wild-type and mutant plants. IBA is transported in seedlings in three distinct flows, like IAA, and this transport is saturable, indicating it is carrier mediated. However, unlike IAA, IBA is not polarly transported in inflorescence axes, and IBA transport is not sensitive to IAA transport inhibitors. These results suggest IAA and IBA transport could be mediated or regulated by different mechanisms. In rib1 seedlings, all flows of IBA transport are modified, while IAA transport levels are unchanged. Modifications in IBA transport match phenotypic differences between rib1 and wild-type, and analyses of the physiological effects of IBA also suggest IBA has a role in defining wild-type seedling morphology in Arabidopsis. Though IAA transport levels are not changed in rib1, one flow of IAA transport is rendered insensitive to IAA transport inhibitors, perhaps revealing cross-talk between IAA and IBA transport regulation. Additionally, double mutant analyses reveal that IAA transport and response mutants can suppress some phenotypes of rib1, and some mutant combinations produce novel phenotypes, further suggesting cross-talk between IBA and IAA transport and response p

Interactions between plant and virus proteins play pivotal roles in many processes during the viral infection cycle. Analysis of protein-protein interactions is crucial for understanding virus and host protein functions and the molecular mechanisms underlying viral infection. Several interactions between virus-encoded proteins have been reported. However, few interactions between viral and plant proteins have been identified so far. To examine interactions between Turnip mosaic potyvirus (TuMV) proteins and plant proteins, recombinant proteins were produced and used in ELISA-type assays and in in vitro co-immunoprecipitation experiments. An interaction between TuMV P1 proteinase and wheat poly(A)-binding protein (PABP) was identified. An interaction between P1 protein and the plant Arabidopsis thaliana eukaryotic initiation factor (iso)4E [eIF(iso)4E] was also found. Finally, potential interactions between both TuMV CI and P1 proteins and between TuMV CI protein and eIF(iso)4E were identified.

Recent preliminary data have suggested that microbe-to-plant signals, and plant internal signals elicited by microbial signals, affect aspects of plant physiology, development and growth. The reported research investigated the responses of plants to signal compounds of microbial and plant origin, such as lipo-chitooligosaccharides (LCOs - signal molecules in rhizobia-legume associations), chitin and chitosan (present in fungal cell walls), and phenolic compounds (salicylic acid, acetylsalicylic acid and gentisic acid - internal signals in plants, often affected by signals from microbes). Phenylalanine ammonia-lyase (PAL) and tyrosine ammonia-lyase (TAL) are key enzymes of the phenylpropanoid pathway. Oligomers of chitin and chitosan increased the activities of both PAL and TAL in soybean leaves. The degree of increase was dependent on oligomer chain length and time after treatment. LCO [Nod Bj V (C18:1 , MeFuc)] was isolated from Bradyrhizobium japonicum strain 532C. When Arabidopsis thaliana plants were grown for two weeks on agar containing this LCO (10-8M) or chitin pentamer (10-4 M), they had greater root length, root diameter, root surface area and number of root tips than control plants. Chitosan (tetramer and pentamer) did not have this effect. Chitin and chitosan were also tested for effects on corn and soybean photosynthetic rates and growth. High molecular weight chitosan generally reduced photosynthetic rates, but did not reduce the growth of corn or soybean. However, foliar application of 10-6 M LCO to corn leaves increased photosynthetic rates (up to 36%). Foliar application of lumichrome (10-5 and 10-6 M), a breakdown product of riboflavin produced by some rhizosphere bacteria, to corn (C4 plant) and soybean (C3 plant) increased photosynthetic rates (up to 6%). Foliar application of lumichrome (10-5 M) increased soybean leaf area and shoot dry weight. Foliar application of SA, acetyl salicylic acid (ASA) and gentisic acid (GT

Successful reproduction in plants is a highly-regulated process reliant on the integration of both endogenous and external cues. Different accessions of the model plant Arabidopsis thaliana have been collected, including those with a winter annual or rapid-cycling flowering habit. Natural variation and mutant screens have enabled many flowering time genes to be identified. A key regulator of flowering is FLOWERING LOCUS C (FLC). FLC is a repressor of flowering and is regulated by a number of genes, including those in the autonomous and FRIGIDA-mediated pathways. Of particular interest are FRIGIDA (FRI) and FRIGIDA-LIKE 1 (FRL1) and the autonomous pathway members, FCA and FY. FRI and FRL 1 promote FLC expression making them dominant repressors of flowering. FRI is proposed to initiate chromatin remodelling at the FLC locus leading to increased FLC expression. Once elevated, FLC levels are maintained until plants undergo an extended period of cold, therefore flowering occurs in spring. In contrast, FCA and FY promote flowering by repressing FLC expression. FCA has also been identified as a receptor of the plant hormone abscisic acid (ABA). Upon binding to FCA, ABA is proposed to disrupt/inhibit the FCA:FY interaction which results in delayed flowering. To characterise the FCA:ABA interaction and identify the ABA binding site, a number of truncated FCA proteins were utilised. Initially a FCA:FY GST-pulldown was used to identify the ABA binding site. However, when ABA failed to inhibit the FCA:FY interaction a direct binding assay using [�H]-ABA was employed. Another Arabidopsis ABA receptor, G-protein coupled receptor 2 (GCR2), was used as a positive control in these binding assays. Both FCA and GCR2 failed to bind [�H]-ABA suggesting a broader issue with the binding assay. The identification of FCA and GCR2 as ABA receptors can be attributed to the quality of the protein assayed, the sensitivity of the binding assay and the subsequent data analysis. This study resulted in the retraction of the original paper (Razem et at, 2006) reporting FCA as an ABA receptor. To investigate the molecular mechanism by which FRI and FRL1 act as positive regulators of FLC expresion, a biochemical approach was taken. FRI and FRL1 have no known homology to any other protein or domain and the only method for assessing protein function is through plant complementation experiments. In the absence of sequence homology, or a timely functional assay, a classical approach was taken to produce soluble protein for analysis. Truncation of predicted regions of disorder and expression, solubility and stability screens produced soluble protein of reasonable purity. This allowed characterisation of the biochemical properties of FRI and FRL1. Interaction studies between FRI and FRL1, and the zinc finger protein SUPRESSOR OF FRIGIDA 4 (SUF4), were also carried out. Polyclonal antibodies against FRI and FRL1, made during this study, were useful for protein detection in these experiments. The interaction studies, together with plant complementation experiments, suggest that the C-terminus of FRI is essential for protein function, while the N-terminus improves FRI activity. These findings provide a better understanding of how the components of the proposed "FRI-complex" may interact to promote FLC expression.

Bibliography: leaves 175-186. / 186 leaves : ill. (some col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Aims to examine the AP in A. thaliana and to produce an inducible antisense plant to assist future studies of the role of AOX. / Thesis (Ph.D.)--University of Adelaide, Dept. of Botany, 1999?

Bibliography: leaves 89-104. / vi, 104 leaves, [15] leaves of plates : ill. (chiefly col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / In this study an EMS-mutated Arabidopsis mutant pho2, which accumulates Pi in leaves, was used to study Pi uptake and transport by comparing it to wild-type seedlings. The study aimed to define the physiological lesions in pho2 mutant and to obtain evidence regarding the function of the PHO2 gene in P nutrition in higher plants. Accumulation of Pi in leaves of pho2 was found to reside in the symplast and was not related to Zn-deficiency. The physiology of the pho2 mutant is consistent with either a block in Pi transport in phloem from shoots to roots or an inability of shoot cells to regulate internal Pi concentration. Southern block analysis revealed that the two transporter genes, APT1 and APT2 were not responsible for the pho2 mutant. Data from the mapping of the PHO2 gene along with information from the Arabidopsis genome sequencing will form the basis for cloning the PHO2 gene in the future. / Thesis (Ph.D.)--University of Adelaide, Dept. of Plant Science, 1999

Thesis (M.S.)--Worcester Polytechnic Institute. / Keywords: Turnip crinkle virus; Hypersensitive response. Includes bibliographical references (leaves 52-61).