Phosphate (Pi) deficiency is a global problem for food production. Plants have evolved complex mechanisms to adapt to low Pi. We focused on the initial aspects of adaptation to low Pi - perception and immediate-early responses to changes in external Pi. To examine whether a labile repressor controls expression of the high affinity Pi transporter, Pht1;1, we performed electrophoretic mobility shift assays (EMSA) but observed only weak protein-DNA binding activity using extracts from Arabidopsis suspension cultures or seedlings. The regulatory role of different regions in Pht1;1 promoter was dissected by promoter deletion analysis, using uidA as a reporter. We identified two domains important for regulation: sequences between -1898 bp and - 932 bp are important for induction of Pht1;1 in low Pi; the intron in the 5’UTR impacts Pht1;1 expression in the young part of both primary and lateral root apices. A complementary approach to identify repressors of Pi starvation responses was pursued: We identified ZAT18, a putative transcription factor, as a candidate repressor. ZAT18 contains an EAR motif, a repressor domain in plants; the expression of ZAT18 responds to Pi starvation. Using transgenic lines with promoter::ZAT18-VENUS constructs, we studied its expression, localization and abundance in different levels of Pi availability: ZAT18 is mainly expressed in the nucleus of Arabidopsis root hair cells. Its accumulation was induced by 4 day Pi starvation. We also performed a microarray analysis to examine global gene expression levels during Pi starvation and rapid recovery. Our data indicated that 258 genes were induced and 188 genes were suppressed during Pi starvation. For most of these genes, responses were reversed after 4 hour Pi recovery. Further study of these genes will help to define targets of the early Pi starvation-signalling pathway.

The worldwide occurrence of sites contaminated with toxic metals and the associated high costs of remediating them using chemical and mechanical methods have led to calls to develop inexpensive and sustainable approaches based on the use of plants that naturally accumulate large amounts of metals in their tissues. The ability of plants to remediate metals has been rigorously studied and some species have been identified as excellent phytoremediators. However, the growth of phytoremediators is often retarded under high soil metal concentrations, rendering them ineffective. Meanwhile, some plants do not have remediating abilities but are capable of growing in contaminated environments with little or no sign of stress. Despite the volume of research dedicated to the screening and evaluation of phytoremediators, major questions remain about why some plants survive but do not remediate while the growth of phytoremediators is mostly hindered. The growth and metal-remediating efficiency of plants exposed to toxic concentrations of metals can be enhanced by inoculating phytoremediating plants with certain bacteria but the mechanisms behind this process remain unclear. Furthermore, the use of leguminous plants to improve the growth of a target plant under a mixed planting system has long been recognised as an effective yield-enhancing cropping system. However, the possibility of a non-remediating but tolerant leguminous plant conferring metal tolerance to a phytoremediator has not been explored. This thesis reports results from repeated glasshouse and lab-based growth experiments on the phytoremediating plant Brassica juncea exposed to 400 – 600 mg Zn kg-1. The aim was to investigate the abilities of two plant growth promoting bacteria (PGPB) species Pseudomonas brassicacearum and Rhizobium leguminosarum, and a leguminous plant Vicia sativa to promote B. juncea growth and enhance remediation of Zn-contaminated soil. B. juncea plant roots were analysed using synchrotron based micro-focus X-ray Fluorescence (μXRF) imaging and X-ray Absorption Near Edge Structure (μXANES) analysis to probe Zn speciation. P. brassicacearum exhibited the poorest plant growth promoting ability, while R. leguminosarum alone and in combination with P. brassicacearum significantly enhanced B. juncea growth and Zn bioaccumulation. X-ray Absorption Spectroscopy (XAS) analysis showed that reduced plant growth was due to root accumulation of Zn as Zn sulphate, Zn oxalate and Zn polygalacturonic acids. The better growth and increased metal accumulation observed in plants inoculated with R. leguminosarum and its combination with P. brassicacearum was attributed to root storage of Zn in the chelated forms of Zn phytate and Zn cysteine. A subcellular analysis of plant root also showed that the PGPB enhanced tolerance to Zn contamination by enhancing epidermal Zn compartmentalisation depending on the nature of root colonization, and induced changes in Zn speciation to less toxic Zn species in the epidermis and endodermis of plant root. The thesis therefore identifies enhanced Zn compartmentalization at the root epidermis and bacterial mediated changes in Zn toxicity through changes in Zn speciation as key complimentary mechanisms of plant growth promotion and enhanced Zn accumulation in plants by PGPB. Further experiments investigating alternative phytoremediation strategies showed that the use of the leguminous plant V. sativa in a mixed planting system with B. juncea plants completely out performed the effects of bacteria in promoting the growth and remediation potential of B. juncea under Zn contamination. By combining PGPB with mixed planting, B. juncea recovered full growth while also achieving maximum phytoremediation efficiency. The novel legume assistedmicrobial phytoremediation method that is reported in this thesis is the first to demonstrate complete plant growth recovery in plants exposed to 400 – 450 mg kg-1 soil Zn contamination for 5 weeks. Survival of V. sativa was attributed to its root storage of Zn in the chelated forms of Zn histidine and cysteine whereas in the roots of stunted B. juncea plants the majority of Zn was present as Zn oxalate and toxic Zn sulphate. Although the use of natural and synthetic chelates has been reported to enhance phytoremediation, this thesis recommends a legume-assisted-microbialphytoremediation system as a more sustainable method for Zn bioremediation.

Begonia L. is one of the largest plant genera, comprising over 1500 species. Weak species cohesion, and the rapid evolution of reproductive barriers in allopatry, are two processes that have been postulated to explain the generation of such hyper-diversity of taxa within a single genus of plants. The aim of this thesis is to investigate whether these factors are likely to have been important contributors to the diversity of species found in Central American Begonia. Species cohesion was analysed in the widespread Central American species Begonia heracleifolia and B. nelumbiifolia. Interpopulation seed flow was estimated with seven plastid microsatellites. Breeding system estimates and measures of genetic differentiation at nine nuclear microsatellites were used to infer levels of interpopulation pollen flow. Controlled crosses were employed to assess the strength of reproductive barriers both between populations within species, and between species differing in ecology. The potential for gene flow between species in the wild was assessed in natural hybrid zones using molecular markers. Finally a quantitative trait locus (QTL) approach was employed to investigate the genetic basis of reproductive traits that differ between species. No plastid polymorphisms were found in B. nelumbiifolia, suggesting it has been through a recent population bottleneck. In contrast, B. heracleifolia possessed many plastid haplotypes that were strongly differentiated between populations (G’ST = 0.829). Nuclear microsatellites showed high genetic differentiation within species, and both species were self-compatible and self-fertilize at a moderate rate (B. heracleifolia F’ST = 0.506, FIS = 0.249; B. nelumbiifolia F’ST = 0.439, FIS = 0.380). F1s between ecologically similar B. heracleifolia and B. sericoneura were partly fertile (2-5% seed set), and F1s and early generation backcrosses were found in a hybrid swarm. F1s between B. heracleifolia and the ecologically contrasting B. nelumbiifolia were pollen sterile, and 3 hybrid swarms showed no evidence of hybrids beyond the F1 generation. Seven QTL were found for reproductive traits, including: sex ratio, pollen sterility and stamen number. The population biology of Begonia, with limited seed and pollen dispersal, small population sizes and frequent self-fertilization predisposes them to genetic isolation, increasing the chances that reproductive barriers evolve. These characteristics may underlie the large number of endemics in Begonia.

The use of Arabidopsis thaliana populations to understand the genetic basis for natural variation has been highlighted in recent years. The role of adaptation in natural variation remains of key interest. Here, natural variation in growth rate, flowering time and seed production were examined in local populations of A. thaliana from the Edinburgh area using a common garden approach. Growth rate and seed production were found to be highly genetically determined and sometimes correlated, and some genotypes were found to perform consistently better as winter annuals and others as summer annuals, suggesting that adaptation to different seasons might maintain natural variation locally. In order to dissect the environmental factors that could affect growth, these genotypes were also grown under controlled conditions. Photoperiod and temperature were identified as two of the seasonal variables to which different genotypes may be adapted. The relationship between growth rate and competition was also examined. In general, competition exaggerated the differences in performance between genotypes, although the identity of neighbours was observed to have an effect on both growth rate and fitness of A. thaliana in competition. To understand the genetic basis of growth rate variation, the genetic relationships between local populations was examined. Local accessions were usually found to be more closely related to each other than to world-wide accessions, suggesting that their variation did not reflect recent immigration. To examine the genetic architecture of growth rate variation, hybrids between local genotypes with different growth rates were used in QTL analysis. Four chromosomal regions were detected; these regions represent potential growth-rate associated QTL.

In the UK, nearly half of the plants short listed for high conservation priority in the UK Biodiversity Action Plan are found in taxonomically complex groups. It is thought that a shift from species- to process-based conservation strategies, aimed at conserving the processes that generate diversity as opposed to simply the end product of these dynamic interactions, may benefit these groups. One group for which this strategy has been proposed is tetraploid Euphrasia. The underlying taxonomic complexity in this group is hypothesised to arise via breeding systems, hybridisation and local ecotypic adaptation. The goal of this thesis is to use morphological, ecological and molecular marker data to examine taxon limits and evolutionary processes in order to further understand the mechanisms involved in maintaining species boundaries and generating taxonomic complexity in tetraploid Euphrasia. This will not only make conservation in this group more effective, but will also provide a broader insight into some of the processes involved in plant speciation. A detailed study of two widespread, small flowered, tetraploid taxa, E. micrantha and E. scottica, showed that offspring are almost exclusively the result of self-fertilization. These taxa maintain distinctive morphologies, habitat preferences and chloroplast DNA variation throughout their range, suggesting that they represent coherent lineages within Scotland. As in other widespread inbreeding species, there are high levels of microsatellite differentiation among different populations of the same species. In northwest Scotland three complex populations of tetraploid Euphrasia were identified which comprised an array of many different morphs (recognised species, and putative hybrids). Analysis of chloroplast and microsatellite markers suggests that these different morphs represent distinct genetic groups. Within each site there is evidence both for habitat heterogeneity, and for association of morphs with this habitat variation. Intermediate morphs were not simple F1 hybrids, but are likely to have originated via hybridisation and subsequent selfing, surviving as independent recombinant lines, perhaps specialised for habitat types different from that of their progenitor parents. These stable morphs of hybrid origin could represent groups with adaptive potential that may result in the origin of a novel Euphrasia species. It will be important to further examine the processes involved in generating novel diversity in Euphrasia. For the time being, these complex populations must be recognised as sites requiring special protection within the context of a process-based conservation strategy.

Bei der arbuskulären Myorrhiza-Symbiose (AM) und der Wurzelknöllchen-Symbiose (RNS) handelt es sich um symbiotische Interaktionen, die einen großen Vorteil für Pflanzenwachstum und kultivierung mit sich bringen. Während bei der AM Pilze die Pflanze mit verschiedenen Nährstoffen aus dem Boden versorgen, stellen die in den Wurzelknöllchen lokalisierten Rhizobien der Pflanze fixierte Stickstoffverbindungen zur Verfügung. Folglich ist es von großem Interesse, die Entwicklung dieser Symbiosen im Detail zu verstehen. Für die Erkennung der arbuskulären Mykorrhiza-Pilze und der Stickstoff-fixierenden Rhizobien durch die Pflanze sind lösliche symbiotische Signalmoleküle essentiell, die zu der Gruppe der Lipochitinoligosaccharide (LCOs) gehören. Während der Entwicklung der AM und der RNS erkennen die Pflanzenwurzeln diese LCOs über Lysin-Motiv-Rezeptor-ähnliche Kinasen der Plasmamembran. Eine der ersten Antworten der Wurzelzellen auf Nod-LCOs ist eine Depolarisierung des Membranpotentials. An dieser Antwort sind mit großer Wahrscheinlichkeit Anionenkanäle der Plasmamembran beteiligt, da sie auch bei Depolarisierungen als Antwort auf andere Stimuli bzw. Stressantworten involviert sind. In Arabidopsis stellt die S-Typ-Familie eine bedeutende Gruppe von Anionenkanälen dar, die von Calcium-abhängigen Kinasen (CPKs) aktiviert werden. Da Nod-LCOs repetitive Veränderungen des zytosolischen Calcium-Levels induzieren, wurde in dieser Arbeit die Hypothese aufgestellt, dass Calcium-Signale CPKs aktivieren. CPKs sorgen im Gegenzug für die Stimulation von S-Typ-Anionenkanälen in Wurzelzellen. Die Änderungen des Membranpotentials in M. truncatula-Wurzelhaarzellen als Antwort auf Nod- und Myc-LCOs wurden mittels intrazellulärer Mikroelektroden analysiert. Es wurde gezeigt, dass Nod-LCOs in M. truncatula-Wurzelhaarzellen eine Depolarisierung des Membranpotentials induzieren. Doch Wurzelhaarzellen reagieren nicht nur auf Nod-LCOs. So konnte in dieser Studie zum ersten Mal eine Depolarisierung als Antwort auf sulfatisierte Myc-LCOs nachgewiesen werden. Eine zweite Gruppe von Myc-LCOs, denen die Sulfatgruppe fehlt, löste keine Reaktion des Membranpotentials aus. Diese Daten deuten darauf hin, dass Wurzelhaarzellen für die Erkennung von sulfatisierten LCOs von symbiotischen Pilzen und Bakterien dasselbe Perzeptionssystem nutzen. Diese Schlussfolgerung wird von Experimenten unterstützt, in denen vor der Stimulation durch Nod-LCOs ein sulfatisierter Myc-LCO hinzugegeben wurde. Diese sukzessive Zugabe von zwei Stimuli führte zu einer einzigen Depolarisierung. Die sulfatisierten Myc-LCOs unterdrückten die Antwort des Membranpotentials auf Nod-LCOs. Die Beziehung zwischen Nod-LCO-induzierten zytosolischen Calcium-Signalen und Änderungen des Membranpotentials wurde mit einer Kombination aus intrazellulären Mikroelektroden und Imaging eines Calcium-sensitiven Fluoreszenzfarbstoffs analysiert. In Messungen der zytosolischen Calcium-Konzentration wurde keine transiente Zunahme innerhalb der ersten vier Minuten nach der Applikation der Nod-LCOs beobachtet. Die durch Nod-LCOs induzierten Depolarisierungen traten früher auf und erreichten ihr Maximum normalerweise nach drei Minuten. Demnach geht die Depolarisierung des Membranpotentials den zytosolischen Calcium-Signalen voraus. Diese Beobachtung wurde von simultanen Messungen beider Antworten bestätigt. Um der Möglichkeit einer Beteiligung von S-Typ-Anionenkanälen an der LCO-abhängigen Depolarisierung nachzugehen, wurden zwei in den Wurzeln exprimierte M. truncatula-Orthologe der AtSLAC1-Anionenkanal-Familie identifiziert. Die klonierten Anionenkanäle, MtSLAC1, MtSLAH2-3A und MtSLAH2-3B zeigten bei der Untersuchung in Xenopus-Oozyten die typischen Charakteristika von S-Typ-Anionenkanälen. So konnte gezeigt werden, dass MtSLAH2-3A und MtSLAH2-3B eine Proteinkinase sowie externes Nitrat zur Aktivierung benötigen. Außerdem zeichnen sie sich durch eine sehr viel höhere Permeabilität für Nitrat im Vergleich zu Chlorid aus. Ähnlich wie bei AtSLAH3 macht eine Koexpression mit AtSLAH1 genau wie eine intrazelluläre Azidifikation MtSLAH2-3A und MtSLAH2-3B zu Anionenkanälen, die unabhängig von externem Nitrat und einer Phosphorylierung durch eine Proteinkinase aktiv sind. Weil S-Typ-Anionenkanäle eine hohe Permeabilität für Nitrat aufweisen, wurde der Einfluss von Änderungen der extrazellulären Anionenkonzentration auf die Nod-LCO-induzierte Depolarisierung analysiert. Es stellte sich heraus, dass eine Verringerung der extrazellulären Nitratkonzentration die Antwort beschleunigt. Eine Erhöhung der extrazellulären Chlorid- und Sulfatkonzentration hingegen führte zu einer Verstärkung der Depolarisierung. Diese Beobachtung spricht dafür, dass andere Anionenkanal-Typen wie ALMT-Kanäle an der Depolarisierung des Membranpotentials durch LCOs beteiligt sind. Die Daten dieser Arbeit zeigen eine Abhängigkeit der Nod-LCO-induzierten Änderungen des Membranpotentials vom M. truncatula-Genotyp. Neben Nod-LCOs lösen auch sulfatisierte Myc-LCOs eine Depolarisierung des Membranpotentials aus. Vermutlich werden sulfatisierte Nod- und Myc-LCOs von demselben Rezeptorsystem erkannt. Die Nod-LCO-induzierte Depolarisierung ist unabhängig von Änderungen des zytosolischen Calcium-Levels. Folglich sind in die Depolarisierung keine S-Typ-Anionenkanäle involviert, die ausschließlich durch Calcium-abhängige Protein-Kinasen aktiviert werden. Interessanterweise lassen sich die MtSLAH2-3-Anionenkanäle aus M. truncatula im Gegensatz zu AtSLAH3 von Calcium-unabhängigen SnRK2/OST1-Proteinkinasen aktivieren. Dies ermöglicht die Aktivierung der MtSLAH2-3-Anionenkanäle in Abwesenheit eines Calcium-Signals. In weiterführenden Studien sollten die Genexpressionsprofile von Calcium-unabhängigen Proteinkinasen wie SnRK2 und S-Typ-Anionenkanälen aus M. truncatula sowie deren Interaktionen untersucht werden. So könnte eine Aussage darüber getroffen werden, ob diese Proteinkinasen die Anionenkanäle MtSLAH2-3 Nod-LCO-spezifisch aktivieren. Außerdem wäre es von großem Interesse, verschiedene M. truncatula-Mutanten zu untersuchen, denen Gene für MtSLAH2-3A, MtSLAH2-3B und R-Typ-Anionenkanäle fehlen. Diese Experimente könnten zur Identifizierung von Genen führen, die an der frühen Entwicklung der Symbiose beteiligt sind und erklären, warum nur eine kleine Gruppe von Pflanzen dazu in der Lage ist, eine RNS einzugehen, während die AM im Pflanzenreich weit verbreitet ist. / Arbuscular Mycorrhiza (AM) and Root Nodule Symbiosis (RNS) are symbiotic interactions with a high benefit for plant growth and crop production. In the soil, AM fungi supply the plant with a broad range of nutrients, whereas the rhizobium bacteria in the root nodules provide fixed nitrogen sources. Thus, it is of great interest to understand the developmental process of these symbiotic interactions. For recognition of AM fungi and nitrogen-fixing bacteria by plants, diffusible symbiotic signals are essential, which belong to the group of lipochitinoligosaccharides (LCOs). During the development of AM and RNS, plant roots sense these LCOs with pairs of lysin motiv domain receptor-like kinases that are located in the plasma membrane. One of the earliest Nod-LCO-triggered responses of root cells represents the depolarization of the plasma membrane. It is likely that plasma membrane anion channels are essential for this reaction, as these channels are required for depolarization in response to a number of other stimuli/stress responses. In Arabidopsis, the S-type family is a prominent group of anion channels that are activated by calcium-dependent Protein Kinases (CPKs). As Nod-LCOs can trigger repetitive elevations of the cytosolic calcium level, we hypothesized that calcium signals activate CPKs, which in turn stimulate S-type anion channels in root cells. The membrane potential changes of M. truncatula root hair cells in response to Nod- and Myc-LCOs were analyzed by using intracellular micro electrodes. In accordance with previous studies in M. sativa, Nod-LCOs evoked a membrane depolarization in root hairs cells of M. truncatula. Root hair cells not only were sensitive to Nod-LCOs, but for the first time a depolarization response was also shown in response to sulphated Myc-LCOs. However, a second group of Myc-LCO-signals, which lack the sulfate group, did not initiate any reaction of the membrane potential. These data thus suggest that root hair cells use the same perception system to sense sulfated LCOs of symbiotic fungi and bacteria. This conclusion was supported by experiments in which a sulfated Myc-LCO was applied, prior to stimulation with Nod LCOs. This successive application of two stimuli resulted only in a single transient depolarization, as sulfated Myc-LCOs repressed plasma membrane responses to Nod-LCOs. The relations between Nod-LCO-induced cytosolic calcium signals and membrane potential changes were studied with a combination of intracellular micro electrodes and calcium sensitive reporter dye imaging. In measurements of the cytosolic calcium concentration the first transient increase was not observed within four minutes after application of Nod-LCOs. Nod-LCO-induced depolarizations occurred earlier and normally peaked after three minutes. In contrast to current models as well as the initial hypothesis of this project, the membrane depolarization thus precedes the cytosolic calcium signals, which was confirmed by simultaneous measurement of both responses. As S-type anion channels are good candidates for the induction of the LCO-dependent depolarization, we indentified two root-expressed M. truncatula orthologues of AtSLAC1-family. The cloned S-type anion channels, MtSLAC1, MtSLAH2-3A and MtSLAH2-3B showed typical characteristics of S-type anion channels, when studied in Xenopus oocytes. Thereby we could show that both M. truncatula anion channels, MtSLAH2-3A and MtSLAH2-3B, need a protein kinase and external nitrate for activation. They are characterized by a much higher permeability for nitrate compared to chloride. Similarly, to AtSLAH3 coexpression with AtSLAH1 or intracellular acidification rendered MtSLAH2-3A/B independent from phosphorylation via protein kinases and external nitrate. Because S-type anion channels show a high permeability for nitrate, we tested the influence of changes in the extracellular anion concentration on the Nod-LCO induced depolarization. It turned out that the response was accelerated when the concentration gradient for nitrate was decreased. However, increasing the extracellular chloride and sulfate concentrations also enhanced the magnitude of the depolarization, which indicates that other types of anion channels, such as ALMT channels may contribute to the LCO-triggered depolarization of root hairs. The data generated in this project show that the Nod-LCO induced membrane potential change is strongly dependent on the genotype of M. truncatula. This early response in the recognition of symbiotic microorganisms is also induced by sulfated Myc-LCOs, which seem to be perceived via the same receptor system as Nod-LCOs. In contrast to our expectations, the depolarization response to Nod-LCOs is independent of changes in the cytosolic calcium level. Consequently, S-type anion channels, activated solely by calcium-dependent protein kinases are not involved in this response. Interestingly, in contrast to the Arabidopsis SLAH3, the SLAH2-3s from M. truncatula are activated via calcium-independent SnRK2/OST1-like kinases which would allow the activation of the channels even in the absence of calcium transients. Thus, in future studies the expression profile and interaction of calcium-independent protein kinases like SnRK2s and S-type anion channels in M. truncatula should be determined to investigate whether these proteins are capable of activating MtSLAH2-3A/B in a Nod-LCO-specific manner. Moreover, the further analysis of M. truncatula mutants that lack MtSLAH2-3A/B as well as M. truncatula R-type anion channels will be of great interest. These experiments can thus lead to the identification of genes that are involved in early symbiosis-related events, which may explain why only a small group of plants is able to develop root nodules, whereas the interaction with mycorrhiza is found for a large variety of plant species.

Plant performance under reduced water availability has traditionally been assessed as drought resistance and more recently as water use efficiency (WUE). An extensive body of work has been established over the past 15 years where the natural variation of water use efficiency has been studied in the model species Arabidopsis thaliana (Arabidopsis). At the same time, a substantial degree of criticism has arisen with respect to the use of drought resistance and WUE as measures of plant performance, due to the lack of relatedness of these parameters to reproductive performance, i.e. yield. The work in this thesis is centered on understanding the physiological and genetic basis of water use and water productivity as alternative measures of plant performance under the context of reduced water availability. The first part of this study describes an extensive assessment of the natural variation of water use and water productivity in Arabidopsis in relation to numerous key physiological, phenological, and developmental parameters. Furthermore, this work concisely relates plasticity of key traits to historical climatic variation. A fundamental aspect of this work was the clarification that it is possible to estimate long term water use to a high degree of accuracy based on short term water use, i.e. soil drying rate, and flowering time. Flowering time was demonstrated to be the predominant driver of vegetative performance and water use, however it appeared to be genetically uncoupled from reproductive performance. This is in contrast to previous work that suggests WUE, measured as the ratio of C12 to C13 isotopes (δ13C), is positively associated with flowering time. Additionally, it was demonstrated that multiple commonly employed proxies of reproductive performance including total biomass, WUE, and flowering time, were not sufficient at predicting seed yield in Arabidopsis across multiple environments. The second part of this study involved the genetic dissection of water use and productivity related traits in Arabidopsis through a quantitative trait loci (QTL) mapping study and a genome wide association study (GWAS). QTL mapping using a recombinant inbred line (RIL) population developed from the ecotypes Col-0 and C24 revealed two key flowering time genes, FLOWERING LOCUS C (FLC) and FRIGIDA (FRI), as key regulators of water use. It was demonstrated that a combination of non-functional alleles of both FLC and FRI reduced long term water use via a shorted life cycle, which is again in contrast to previous work relating to the genetic dissection of WUE in Arabidopsis. Crucially, it was observed that reduced water use mediated in this fashion did not detrimentally impact upon reproductive performance. GWAS was employed subsequent to the QTL mapping in order to identify candidate genes underlying the variation for productivity as a unique trait and also as a factor of water use, i.e. water productivity. GWAS identified multiple promising candidate genes that potentially underlie the heritable genetic variation for flowering time, water use, and water productivity.

The genus Clematis L. (Ranunculaceae) was used as a new model group to assess the role of the Himalayan orogeny on generation of biodiversity through investigations of its phylogeny, phylogeography and taxonomy. Although existing checklists include 28 species of Clematis from Nepal, a comprehensive taxonomic revision of available material in herbaria and additional sampling from fieldwork during this study has led to the recognition of 21 species of Clematis in Nepal, including one species (C. kilungensis) not previously recorded from Nepal. Exisiting phylogenetic and taxonomic concepts were tested with the addition of new samples from Nepal. The results highlight the shortcomings of the previous studies which were poorly resolved and indicate the need for a thorough revision of the sectional classification. Despite the increased sampling the results are still equivocal due to poor statistical support along the backbone of the phylogeny. Groups of species in well supported terminal clades are broadly comparable with results from previous studies although there are fewer clearly recognisable and well supported clades. The published dates for the evolution of Clematis were tested and the methodology of the previous study critically reappraised. The results indicate that the genus Clematis is approximately twice as old as previously reported and evolved in the middle Miocene. The phylogeny also demonstrates that, even allowing for poor support for the relationships between groups of species within Clematis, the extant Nepalese species must have multiple independent origins from at least 6 different colonisations. With their occurrence in the Pliocene and Pleistocene, these events are relatively recent in relation to the Himalayan orogeny, and may be linked more to the dispersal ability of Clematis than to the direct effects of the orogeny. Additional Nepalese samples of Koenigia and Meconopsis were added to exisiting datasets and these were reanalysed. The result from Clematis, Koenigia and Meconopsis were appraised in light of the the geocientific literature and previously published phylogeographic studies to create an overview of the drivers behind speciation in the Himalaya.

Plants are sessile photo-autotrophic organisms and need to adapt constantly to a dynamic environment. Light is of utmost importance for plants to be able to monitor their surroundings. Ultraviolet-B radiation (UV-B; 280-315 nm) is an intrinsic part of sunlight and, depending on the wavelength and the fluence rate, it may be a stressful signal or an “informational” one. The so called photomorphogenic responses of plants to UV-B are largely mediated by the UV-B specific photoreceptor UV RESISTANCE LOCUS 8 (UVR8), which “senses” UV-B via a tryptophan based mechanism. UVR8 is localised in the cytoplasm and the nucleus mainly as a homodimer. Upon UV-B irradiation it splits to its monomers and accumulates in the nucleus where it has been found to interact with the E3 Ubiquitin ligase COP1. In the nucleus UVR8 has been shown to associate with chromatin on loci of UV-B responsive genes, including that encoding for the bZIP transcription factor (TF) ELONGATED HYPOCOTYL 5 (HY5), a key effector of UVR8-dependent signalling pathways. The binding of UVR8 to chromatin appears to take place via interaction with histones (H2B in particular) rather than DNA itself. However, this association with chromatin seems not to be UV-B specific. The above data suggest a mechanistic basis for an assumed function of UVR8 in the regulation transcription. It seems likely that UVR8 interacts with other proteins associated with chromatin to promote remodelling and/or recruits/activates TFs which in turn stimulate transcription of its target genes. The main objective of this study was to address the above working hypothesis.

The aquatic macrophyte Crassula helmsii is a non-native species and classed as 'invasive' in the UK. This study was carried out in an attempt to provide empirical evidence as to whether the establishment and growth of Crassula helmsii has a measurable impact on the ecology of invaded sites. Dispersal and distribution patterns across England were explored using databases and GIS interpretation. Ecological impact was measured by comparing invaded sites with uninvaded control sites on a range of habitats and waterbody types across Kent and East Sussex. The analysis of spread patterns provided evidence that the plant is likely to have been dispersed due to the horticultural trade, as well as natural vectors into neighbouring habitats. Macrophyte analysis showed that species losses did not occur when C. helmsii was present. Changes to species composition did occur, with rarer plant species being associated with the presence of C. helmsii. Freshwater macroinvertebrates showed no change in either species number or species rarity. The seed banks of invaded and control sites showed no difference, but active management was shown to reduce the total number of seeds in the soil significantly. The water chemistry of invaded and uninvaded sites showed a relationship between the presence of C. helmsii and reduced total organic nitrogen. The results of this study show that the expected species loss associated with non-native species may not be occurring with C. helmsii. Active management may be impacting the ability of native species to recolonise. Further work on other habitats and waterbodies across a wider geographic range are required to explore whether this is a localised effect.