Search

As society's demands for goods and services from forests increase, there is a corresponding increase in demand for high quality science which can help shape forestry policies for the sustainable management of natural resources. Forestry research is now undertaken in a very diverse range of institutions. The increased outputs from researchers in domains other than traditional forestry schools have not been matched by systematic methods of utilising new or existing information for policy or to identify future research needs that reflect the priorities of people in forestry's extended networks. An analysis of academic papers and informal articles indicated that the term 'evidence-based' is widely used in environmental literature, matching the rates of popularity of the term 'evidence-based medicine', but without the rigorous methodological processes that medicine has adopted to justify the phrase. Two strands of participatory research were undertaken to examine the potential for adopting rigorous evidence-based frameworks for forestry. The first focussed on a process for identifying important questions for policy-focussed forestry research. Using a combination of internet-based surveys and a workshop, some 480 people representing broad interests in the UK forestry sector set out a priority list of research questions. In the second, a group of international researchers and policy makers explored the possibility of using systematic reviews for validating the evidence base for carbon stock assessment under the REDD (Reducing Emissions from Deforestation and forest Degradation) process. The results indicate that there is capacity to collaborate to frame questions and critically appraise evidence for important policy issues. The longer-term benefits of adopting gold-standard systematic reviews are clear, however, there are also short-term advantages to be gained from moving incrementally towards this goal. In the aftermath of society's concerns about possible manipulation of science by experts (in the climate science arena), it would be folly for the forestry sector to ignore the benefits that would accrue from adopting genuinely evidence-based procedures for matters of global significance.

Phylogenetic studies are providing powerful new insights into the evolution of complex traits. Metal hyperaccumulation is an unusual and complex physiological trait found in about 500 plant species and is associated with an exceptionally high degree of tolerance of metalliferous soils. Alyssum L. (Brassicaceae) is the largest known hyperaccumulator genus, comprising approximately 188 species distributed throughout the Mediterranean region and south-west Asia. Approximately one-quarter of these are largely restricted to areas of serpentine soils and have the ability to accumulate nickel to high concentrations in shoot tissue. This genus provides a good example in which to study the origins of a complex physiological trait, but its phylogeny is currently poorly understood. To produce a well-resolved phylogenetic tree to investigate the number and timing of origins of nickel hyperaccumulation within Alyssum, DNA sequences were generated for four chloroplast regions (matK, rps16–trnK, trnD–T and trnL–F) from 170 of 255 species in the tribe Alysseae. Additional sequencing was carried out for the chloroplast genes ndhF and rbcL and the nuclear gene PHYA. A Bayesian analysis employing a relaxed uncorrelated lognormal molecular clock and multiple fossil-age calibration points was carried out to reconstruct a time-calibrated phylogeny of this tribe using appropriate outgroups. Optimization of the nickel hyperaccumulation trait onto the resulting phylogenetic tree suggests that nickel hyperaccumulation arose twice in the Alysseae in the late Miocene/early Pliocene: 3.3–8.3 Mya in Alyssum and 6.3–8.8 Mya in Bornmuellera. The single origin in Alyssum is strongly associated with a significant acceleration in net species diversification rate, suggesting the ability to hyperaccumulate nickel could have provided a key evolutionary innovation facilitating rapid range expansion and subsequent species diversification. The scattered distribution of nickel hyperaccumulators across small island-like patches of serpentine soil suggests that allopatric speciation may have driven rapid diversification in this clade.

Tomatoes are the fourth most valuable commodity in agriculture after rice, wheat and soybeans globally with 151 million tonnes of fruit being produced in 2012. The tomato fruit is also a model system for fleshy fruit development. During ethylene-regulated fruit ripening there are complex changes in fruit chemical composition due to degradation and synthesis of a number of soluble and volatile metabolites. Ultimately, these changes control the composition of the ripe fruit and dictate its flavour and texture. It is known that ripening can proceed when mature green fruit are removed from the plant (and indeed this is standard commercial practice) but the extent to which metabolic changes are sustained when fruit are ripened in this way has yet to be established. A modelling approach such as constraints-based modelling can provide system-level insights into the workings of the complex tomato metabolic network during ripening. The first aim of this thesis was therefore to construct a genome-scale metabolic network model for tomato and to use this model to explore metabolic network flux distributions during the transitions between the stages of fruit ripening. The flux distributions predicted provided insight into the production and usage of energy and reductants, into routes for climacteric CO2 release, and the metabolic routes underlying metabolite conversions during ripening. The second aim of this thesis was to use the model to explore metabolic engineering strategies for increased production of lycopene in tomato fruit. The model predictions showed that rearrangement of dominant metabolic fluxes were required to cope with the increased demand for reductants at high lycopene accumulation, which came at a cost of a lower accumulation of other secondary metabolites. Overall the thesis provides an approach to connect underlying metabolic mechanisms to the known metabolic processes that happen during ripening.

Among the most compelling and least well-understood tree species growing in the temperate forests of southern South America is Fitzroya cupressoides, a high biomass species and the second longest-lived tree species in the world. This thesis quantified the main components of the carbon cycle in Fitzroya forests (i.e. net primary productivity (NPP) and soil respiration) and evaluated the environmental variables that are most related to them. The study was focused on medium-age and old-growth forests growing in the Coastal Range (Alerce Costero National Park, AC) and the Andean Cordillera (Alerce Andino National Park, AA) of southern Chile, respectively. The specific objectives of this thesis were to: 1) assess the forest structure, species composition and characterise the environmental conditions of these forests; 2) assess biomass, aboveground NPP, carbon allocation and mean wood residence time in these forests; 3) assess soil respiration and relate it to soil environmental conditions. Additionally, to use a mass balance approach to estimate fine root productivity; 4) estimate total NPP using biometric and indirect estimates of productivity; 5) evaluate the climatic factors mainly related to Fitzroya stem radial change on an intra-annual basis; and 6) evaluate changes in Fitzroya's tree growth and carbon isotopes during recent decades, and determine which environmental factors are more related to them. The last two objectives focus on Fitzroya as the dominant species and the subject of this study. Two 0.6 ha plots were installed within each national park; NPP was estimated for a year and soil respiration and high resolution stem growth measurements were monitored over almost two years. Aboveground biomass estimates for the Andean site are among the most massive reported in the world and carbon fluxes in Fitzroya forests are among the lowest reported for temperate wet forests worldwide. The longevity as well as the particularly rainy and nutrient poor soil conditions where these ecosystems grow may influence their exceptionally slow carbon dynamics. Differences in carbon fluxes between sites seem most probably driven by different environmental conditions rather than by developmental stage. Moreover, carbon fluxes were more sensitive to interannual climate variability in AC than AA. Warmer and drier summer conditions, likely to become more common under future climate change, more significantly affected stem growth and soil respiration in the Coastal Range than in the Andes. Regarding long-term changes, tree growth has been decreasing in the coastal site in the last 40 years and increasing in the Andes since the 1900s. These trends have been accompanied by an increase in intrinsic water use efficiency which is likely caused by rises in CO2 and changes in climate conditions in both sites. Although Fitzroya grows in particularly wet and cool areas, projected drier and warmer conditions may have a negative effect on Fitzroya stem growth and carbon sequestration in both study sites. This effect would be more critical in the Coastal Range though, because of its more Mediterranean climate influence and more restrictive soil conditions in this area. Adequate resources are needed for the monitoring and conservation of these slow growth and massive forests especially in the Coastal Range, in order to avoid ongoing illegal cuttings and threatening forest fires.

Tropical forests have existed for up to one hundred million years, and today provide many ecosystem services vital for human well-being. They also harbour great biodiversity, which, in addition to its intrinsic value, plays a key role in the functioning of these ecosystems. Despite their local to global significance, there are still many knowledge gaps concerning the dynamic processes that govern the functioning of tropical forests. Rapid rates of deforestation and landscape conversion, predominantly for logging and industrial agriculture, are limiting the time and opportunity available to collect the information needed to fill these gaps. This research aims to shed light on the long-term ecological functioning of tropical forests, specifically investigating the history of disturbance in these ecosystems and the response of forest vegetation to past perturbations. The carbon-rich tropical peat swamp forests found along the coast of Sarawak, Malaysian Borneo, are a central focus of this study. For these forests in particular, a large deficit of knowledge surrounding their history and unique ecological functioning is coupled with some of the highest conversion rates of all tropical forest ecosystems across the world. In this thesis, palaeoecological data has been used to reconstruct temporal variability in forest vegetation coincident with external perturbations in order to identify changes in the resilience of these ecosystems through time, via indicators such as slowing rates of recovery and reduced regeneration of forest vegetation. Results suggest that tropical forest ecosystems have, for the most part, shown resilience to natural disturbances in the past, ranging from instantaneous localised tree-fall to longer-term regional climatic change; but that recent anthropogenic disturbances, of novel forms and greater intensities, are jeopardizing the potential for forest recovery and thus compromising ecosystem resilience. These findings enhance our understanding of the ecology of tropical peat swamp forests, and tropical forests more broadly. They also provide a context for contemporary tropical forest management, allowing for predictions on future responses to disturbance and enabling more ecologically sustainable landscape planning.

The presence of exotic plants both in Africa and in Asia has long attracted the attention of scholars who have attempted to understand the human activities linked to them. Archaeological and ethnographic evidence for the reconstruction of these activities is often very limited, but indirect methods such as the study of DNA have become useful tools in building models of early human dispersal. Among the plants that were carried across the Indian Ocean, sometimes known as the “tropical food kit”, the staple crop taro (Colocasia esculenta (L.) Schott) has continued to be the subject of ongoing research. The use of this crop in antiquity is well documented by discoveries of ancient taro starch granules found on archaeological artefacts from sites in Southeast Asia and the Pacific islands, making it one of the oldest plants consumed by people. However, less is known about the use of taro in Africa and the Mediterranean region, where it is found both in the wild and under cultivation - often representing a staple crop in Sub-Saharan Africa. In this doctoral thesis, genetic analysis was performed on modern samples of taro collected from Africa and other regions of the Indian Ocean, using four molecular markers. Two main clusters have been identified, and within this main sub-division four populations of taro have been detected in Africa. By integrating the genetic results with historical and linguistic research, and extensive ethnobotanical fieldwork in Africa, two of these populations are proposed to represent early translocations, with modern distribution patterns suggesting diverse dispersal routes at different times. These results open up a new scenario in which the “tropical food kit” is finally unpacked, with important historical implications for each of the crops contained within it.

It is commonly assumed that transpiration does not occur at night because leaf stomata are closed in the dark. We tested this assumption across a diversity of ecosystems and woody plant species by various methods to explore the circumstances when this assumption is false. Our primary goals were: (1) to evaluate the nature and magnitude of nighttime transpiration, E(n), or stomatal conductance, g(n); and (2) to seek potential generalizations about where and when it occurs. Sap-flow, porometry and stable isotope tracer measurements were made on 18 tree and eight shrub species from seven ecosystem types. Coupled with environmental data, our findings revealed that most of these species transpired at night. For some species and circumstances, nighttime leaf water loss constituted a significant fraction of total daily water use. Our evidence shows that E(n) or g(n) can occur in all but one shrub species across the systems we investigated. However, under conditions of high nighttime evaporative demand or low soil water availability, stomata were closed and E(n) or g(n) approached zero in eleven tree and seven shrub species. When soil water was available, E(n) or g(n) was measurable in these same species demonstrating plasticity for E(n) or g(n). We detected E(n) or g(n) in both trees and shrubs, and values were highest in plants from sites with higher soil water contents and in plants from ecosystems that were less prone to atmospheric or soil water deficits. Irrespective of plant or ecosystem type, many species showed E(n) or g(n) when soil water deficits were slight or non-existent, or immediately after rainfall events that followed a period of soil water deficit. The strongest relationship was between E(n) or g(n) and warm, low humidity and (or) windy (> 0.8 m s(-1)) nights when the vapor pressure deficit remained high (> 0.2 kPa in wet sites, > 0.7 kPa in dry sites). Why E(n) or g(n) occurs likely varies with species and ecosystem type; however, our data support four plausible explanations: (1) it may facilitate carbon fixation earlier in the day because stomata are already open; (2) it may enhance nutrient supply to distal parts of the crown when these nutrients are most available (in wet soils) and transport is rapid; (3) it may allow for the delivery of dissolved O(2) via the parenchyma to woody tissue sinks; or (4) it may occur simply because of leaky cuticles in older leaves or when stomata cannot close fully because of obstructions from stomatal (waxy) plugs, leaf endophytes or asymmetrical guard cells (all non-adaptive reasons). We discuss the methodological, ecophysiological, and theoretical implications of the occurrence of E(n) or g(n) for investigations at a variety of scales.

Japan is one of the world’s 34 biodiversity hotspots, according to Conservation International (CI). The methods used by various organisations to define priorities differ, however, and all have weaknesses when trying to identify hotspots at finer resolutions. The goal of this thesis is to investigate how biodiversity hotspots in Japan could be revealed and mapped in order to encapsulate conservation elements of biodiversity in practical ways and at various scales. Bioquality is a term that emphasises the concentration within a community of elements of biodiversity with high conservation value. It evaluates the global rarity and taxonomic distinctiveness of plant species or infra-specific taxa using four Star categories. At a plant community level, the Genetic Heat Index (GHI), which is a standardised global range size rarity score, is calculated using weighted Star statuses of species in the community. Bioquality hotspots are assessed here for the first time for the flora and vegetation in Japan – and for temperate Asia – by categorising the Japanese flora into Stars and by applying GHI to survey data and literature-based sources. Keys to Stars are developed for the Japanese flora, with adjustments for variability in species geographic range size information and for taxonomic relatedness. A Flora of Japan (FOJ) database was compiled as a BRAHMS database, containing 8,262 accepted names (30,656 taxon names in total, including synonyms) in 258 families – the first full database of Japanese vascular plants. A total of 7,145 taxa are assigned Stars; from the rarest to the widespread class, there are 884 Black, 756 Gold, 833 Blue, and 4,672 Green Star taxa, confirming that Japan as a whole contains a high proportion of globally rare taxa (23% taxa in Black or Gold). A protocol for calibrating the weight of Stars based on species geographic range is developed based on fine-resolution distribution maps within Japan and coarse–resolution Taxonomic Database Working Group (TDWG) code information. The protocol optimises calculation for temperate regions. The first ever bioquality hotspot maps of Japan are produced using two independent data sources on species distribution at national level: 1) 50 botanical prefectures using 4,830 species from the FOJ database; 2) 1,418 Horikawa ‘geoquadrats’ (0.1° latitude by 0.15° longitude grid) maps covering 829 species. The Ryukyu Islands and Ogasawara Islands are identified as bioquality hotspots, and high mountain ranges in mainland Japan are predicted to contain areas potentially high in GHI; the spatial patterns of GHI are generally concordant between maps of different resolutions. These findings highlight that bioquality assessment can be applied meaningfully at various spatial resolutions. Using field sampling data and existing literature, three study sites are further investigated on a local level: 1) the satochi-satoyama landscape, the current national priority area for biodiversity conservation; 2) various vegetation types of Okinawa-jima Island, the Ryukyu Islands; and 3) the Utaki sacred groves within the predicted hotspot of the Ryukyu Islands. The Ryukyu Islands are confirmed to contain bioquality hotspots within many individual sites, while there was generally low GHI across the satochi-satoyama landscape. The field study outcomes, together with a gap analysis of the existing coverage of protected areas, highlight three important points that are directly relevant to national biodiversity conservation planning: 1) the Ryukyu Islands urgently need newly designated protected areas; 2) the satochi-satoyama landscape conservation should redirect its focus on cultural benefits to the public; 3) the existing protected areas, particularly on mountain areas, need re-evaluation in terms of upgrading their status in light of the bioquality assessment.

Little is known about the composition of tropical epiphytic communities and the influence of environmental variables on community composition. In this thesis I quantify the diversity and biomass of bromeliads, and other vascular epiphytes along an altitudinal transect on the eastern slope of the southeast Peruvian Andes and then look for species’ adaptations related to patterns of diversity and biomass. I compare patterns with those of woody species. Bromeliad species, like tree species, were found to form ecological zones related to climate. The lowest altitude ecological zone (below 1250 m) is the lowland rainforest (LRF), which has the warmest climate and highest evapotranspiration. In LRF vascular epiphytes are less prominent than other ecological zones, with the lowest bromeliad species richness and lowest vascular epiphyte biomass. However, low water-availability gives rise to most variable shoot morphology of bromeliads. The tropical montane forest (TMF), between 1250 m and 2250 m, is intermediate in climate between the LRF and the tropical montane cloud forest (TCF). The TMF has the highest α-diversity, but species richness is lower than the TCF. The shoot morphology of bromeliads is intermediate between TCF and LRF. The highest altitude ecological zone with forest is the TCF (above 2250 m). The TCF has the highest bromeliad species richness, and lowest diversity of shoot forms. The low diversity of shoot forms represents the need for a large phytotelm (water-impounding shoot) to intercept and store precipitation. The TCF has the highest vascular epiphyte biomass, although the biomass is variable as a consequence of the natural disturbance caused by landslides. Along the transect bromeliad species with CAM photosynthesis are only present in the LRF. Terrestrial bromeliad distribution records covering the Neotropics show CAM photosynthesis is more prevalent in drier environments showing that CAM photosynthesis is primarily an adaptation to drought. Epiphytic bromeliads, pre-adapted to a water-stressed environment show no differences in presence along rainfall gradients, but species with CAM photosynthesis occupy warmer environments.

This thesis addresses the methodological challenges of determining the variability of large herbivore populations through time and their impact on European vegetation. Large herbivores are at the heart of conservation policy. However, opinions widely diverge on whether we should aim for fewer herbivores and managed populations or, on the contrary as advocated by the rewilding movement, more herbivores and self-regulating populations acting as ecosystem engineers. This controversy has roots in a debate regarding the nature of ecosystems before the prevalence of human activities. Baseline ecosystems are either described as continuous forest cover with passive large herbivores, or, in contrast, as mosaics with patchy forest cover driven inter alia by bison, aurochs and horses, now rare or extinct in Europe. The main obstacle in moving this debate forward is a poor understanding of large-herbivore densities in the past. I analysed modern pollen and spore assemblages from known environmental settings to improve palaeoecological interpretation of fossil assemblages dating from the pre-human (baseline) period. The sites investigated are the rewilded grasslands of the Oostvaardersplassen (The Netherlands), the mosaic habitats of The New Forest (UK) and the old-growth closedcanopy forest of Białowieża (Poland). I demonstrate that the common practice of interpreting pollen percentages fails to estimate past forest cover in situations with natural grazing. As an explanation, I suggest that pollen productivity fluctuates with biotic factors such as herbivory and canopy shading. As a result, new insights into the baseline debate require additional lines of evidence. In this thesis, I develop an existing methodology to reconstruct past herbivore presence using fossil dung fungal spores. I synthesise current knowledge of this method with an emphasis on spore identification and, finally, I demonstrate that dung fungal spore abundance in lake sediments can be translated into large herbivore numbers. The evidence presented in this thesis contributes to the debate on re-wilding and addresses a fundamental challenge of nature conservation in the human-dominated landscapes of Europe.

Tomato is an important food crop and a model for fleshy fruit development. The process of fruit ripening involves changes in chemical composition and in particular the accumulation of sugars, organic, amino acids and carotenes. The research described in this thesis aimed to identify key regulatory aspects associated with the accumulation of the major acids in tomato fruit by analysis of introgression lines resulting from a cross between a cultivated variety, Solanum lycopersicum, and a wild progenitor species, Solanum pennellii. Line 2-5 showed increases in citrate, malate, aspartate and glutamate in fruit grown under greenhouse conditions.The genetic differences between line 2-5, its overlapping lines, sub-introgression lines and the recurrent parent were used to link the metabolite phenotypes to smaller chromosomal regions. This analysis suggested multiple epistatic loci control fruit metabolite accumulation. Investigation of the biochemical differences between line 2-5 and the recurrent parent revealed that organic and amino acid accumulation did not dependent upon increased TCA cycle capacity. Regulation at the metabolic level was identified for citrate accumulation with changes in cytosolic aconitase in line 2-5. As these metabolites accumulate in the vacuole, tonoplast transport was investigated. Correlation of ATPase-dependent malate influx with altered malate content suggested malate tonoplast transport plays a role in malate accumulation and highlights the importance of vacuolar storage and transport in the regulation of organic and amino acid accumulation.

Rab GTPases are major regulatory proteins of vesicle traffic and thus responsible for membrane identity, vesicle targeting and vesicle fusion. The angiosperm Rab GTPase family is grouped into eight clades (Rab-A to Rab-H) that are broadly conserved in animals and yeasts. It has been proposed that the Rab-A clade has diversified in land plants giving rise to six plant- specific structural subclasses, Rab-A1 to Rab-A6. Previous work suggests that the Arabidopsis Rab-A2 and Rab-A3 proteins define a novel endosomal compartment that lies on a pathway between the Golgi and the plasma membrane. In dividing cells, the Rab-A2/A3 compartment is implicated in biosynthetic traffic to the cell plate but it is unclear what traffics through this compartment in non-dividing cells. In this project, I investigated a range of membrane trafficking pathways in Arabidopsis thaliana. These were probed for dependency on RAB-A2a function, using the dominant negative approach combined with fluorescent marker technology. The data presented in this thesis suggests that RAB-A2a acts on a protein recycling pathway that is used by PIN2:GFP.

Small Island Developing States (SIDS) of the Caribbean are one of the world’s ‘hottest’ ‘biodiversity hotspots’. However, this biodiversity continues to be threatened by habitat loss, and now, by climate change. The research reported here investigated the potential of species distribution modelling (SDM) as a plant conservation tool within Caribbean SIDS, using Trinidad as a case study. Prior to the application of SDM, ancillary analyses including: (i) quantification and mapping of forest cover change (1969 to 2007) and deforestation rates, and (ii) assessment of the island’s vegetation community distribution and associated drivers were carried out.Community distribution and commercial importance and global/regional rarity were used to generate a list of species for assessing the potential of SDM within Trinidad. Species occurrence data were used to generate species distribution models for present climate conditions within the SDM algorithm, MaxEnt. These results were assessed through expert appraisal and concurrence with results of ecological analyses. These models were used to forecast suitable species climate space forty years into an SRES A2 future. Present and future models were then combined to produce a ‘collective change map’ which showed projected areas of species’ range expansion, contraction or stability for this group of species with respect to Trinidad’s Protected Areas (PAs) network.Despite the models being indicative rather than accurate, it was concluded that species’ climate space is likely to decrease or disappear across Trinidad. Extended beyond Trinidad into the remainder of the Caribbean region, SDM may be a crucial tool in identifying which PAs within the region (and not individual islands) will facilitate future survival of given target species. Consideration of species conservation from a regional, rather than an individual island perspective, is strongly recommended for aiding the Caribbean SIDS to adapt in response to climate change.

A key question in developmental biology is how form is generated. The model species Arabidopsis thaliana produces simple leaves with marginal outgrowths termed serrations. Serration development in A. thaliana requires both the transcription factor CUP-SHAPED COTYLEDON2 (CUC2) and the auxin efflux facilitator PIN-FORMED1 (PIN1), which regulates polar auxin transport by forming convergence points (Hay et al., 2006; Nikovics et al., 2006; Scarpella et al., 2006). In Chapter 3, I investigate how CUC2, PIN1 and auxin interact to control serration development. I demonstrate that CUC2 promotes PIN1 convergence point and auxin activity foci formation along the margin of the leaf, whilst high auxin activity represses CUC2 expression. Furthermore, interspersed peaks of CUC2 and auxin activity pattern serration development along the proximo-distal axis of the leaf. Thus, auxin, PIN1 and CUC2 form a negative feedback loop that patterns serration development.CUC genes and PIN1 are required for leaflet development in Cardamine hirsuta (Barkoulas et al., 2008; Blein et al., 2008), a close relative of A. thaliana that produces compound leaves subdivided into units termed leaflets. However, it is unclear how CUC and PIN1 interact to control leaflet development. In Chapter 4, I demonstrate that similar to A. thaliana, CUC genes promote PIN1 convergence point and auxin activity foci formation at the C. hirsuta leaf margin, whilst high auxin activity represses CUC2 expression. These genetic interactions likely create interspersed peaks of CUC2 and auxin activity that pattern leaflet development. Thus, the same negative feedback loop between CUC, PIN1 and auxin patterns both leaflet development in C. hirsuta and serration development in A. thaliana.KNOTTED1-LIKE HOMEOBOX (KNOX) genes are expressed in C. hirsuta leaves, and interact with ChCUC and PIN1 in positive and negative feedback loops, respectively, to control leaflet development (Barkoulas et al., 2008; Blein et al., 2008). KNOX genes are not expressed in A. thaliana leaves, but deeply lobed margins reminiscent of leaflets develop in association with ectopic KNOX expression in leaves (Chuck et al., 1996; Hay et al., 2006). However, it is unclear whether regulatory interactions of PIN1, CUC and KNOX which occur in C. hirsuta leaflets are employed during KNOX-induced lobe development in A. thaliana. In Chapter 5, I demonstrate that CUC2 and polar auxin transport are required for ectopic KNOX expression. Conversely, I show that KNOX misexpression up-regulates CUC2 expression in A. thaliana leaves. Thus, interactions between KNOX, CUC and PIN1 that occur in leaflet development in C. hirsuta also occur in association with KNOX-induced lobe development in A. thaliana.In addition to investigating the regulatory interactions between known components of leaf development pathways, I sought to identify novel genes that mediate CUC2-dependent serration development in A. thaliana. In Chapter 6, I identify a suppressor of the smooth margin phenotype of cuc2 leaves that partially restores PIN1 localisation in the absence of functional CUC2. Finally, in the General Discussion I evaluate how interlinking feedback loops between CUC, KNOX and auxin pattern serration and leaflet development. I then discuss why interlinking feedback loops may have been deployed to control outgrowths in both plant and animal systems.

Africa's miombo woodlands constitute one of the most important dry tropical forests on earth, yet the hydraulic function of these woodlands remains poorly researched. Given the current predictions of increased aridity by the end of this century in the miombo ecoregion, understanding the likely response of miombo woodlands tree species to water stress is crucial in planning adaptation strategies. Predicting the response of miombo woodlands to future climate trends is hampered by a lack of knowledge on the physiology of the common miombo woodlands tree species. In particular, plant-water relations for this woodlands type are not well understood.An understanding of plant-water relations for this woodlands type will provide insights into how water limits tree species distribution in this ecosystem. This will also improve our prediction model on the likely response of this ecosystem to predicted climate change. For this reason, the overall objective of this research was to evaluate the hydraulic architecture and xylem vulnerability to cavitation for nine principal miombo woodlands tree species differing in drought tolerance ability and habitat preference. This was achieved by; examining the hydraulic properties and evaluating the extent to which each hydraulic design was vulnerable to water stress-induced xylem cavitation; investigating how seasonal changes in plant-water relations influences seasonal patterns of leaf display and; analyzing the relationship between stem hydraulic supply and leaf functional traits related to drought tolerance ability.This research has found that drought-intolerant tree species with mesic specialization have more efficient stem hydraulic systems than co-occurring habitat broad ranging species. Broad ranging tree species attain wider habitat distribution by adjusting their hydraulic supply in response to changing ecosystem water availability. The finding that hydraulic properties differ significantly between tree species with contrasting habitat preference suggests that tree hydraulic design may have some adaptive ecological role in influencing species habitat preferences in miombo woodlands.The evaluation of xylem vulnerability to cavitation revealed that mesic specialized tree species were more vulnerable to water stress-induced cavitation than habitat broad ranging tree species. Vulnerability to cavitation in individuals from the same broad-ranging species growing in contrasting habitats showed only marginal and statistically insignificant (P > 0.05) differences between wet and dry sites.In the investigation of the influence of seasonal changes in stem water relations on seasonal leaf display, seasonal rhythms in stem water status were found to exert significant controls on leaf phenology. Mesic specialists had strong stem water controls throughout the year in comparison to broad ranging tree species.An analysis of the relationship between stem hydraulic supply and leaf functional traits suggests that stem hydraulic supply constrains leaf biomass allocation patterns among miombo tree species. Mesic specialists tend to invest more in leaf longevity than broad ranging tree species.This thesis has uncovered some interesting relationships between plant-water-relations and the distribution of miombo woodlands tree species. These results lead to the conclusion that in an event of increased ecosystem drying under future climate trends, tree species with mesic specialisation are at a greater risk of experiencing cavitation related species mortality than broad ranging ones.

Background Archaeobotany, the study of plant remains from sites of ancient human activity, provides data for studying the initial evolution of domesticated plants. An important background to this is defining the domestication syndrome, those traits by which domesticated plants differ from wild relatives. These traits include features that have been selected under the conditions of cultivation. From archaeological remains the easiest traits to study are seed size and in cereal crops the loss of natural seed dispersal. Scope The rate at which these features evolved and the ordering in which they evolved can now be documented for a few crops of Asia and Africa. This paper explores this in einkorn wheat (Triticum monococcum) and barley (Hordeum vulgare) from the Near East, rice (Oryza sativa) from China, mung (Vigna radiata) and urd (Vigna mungo) beans from India, and pearl millet (Pennisetum glaucum) from west Africa. Brief reference is made to similar data on lentils (Lens culinaris), peas (Pisum sativum), soybean (Glycine max) and adzuki bean (Vigna angularis). Available quantitative data from archaeological finds are compiled to explore changes with domestication. The disjunction in cereals between seed size increase and dispersal is explored, and rates at which these features evolved are estimated from archaeobotanical data. Contrasts between crops, especially between cereals and pulses, are examined. Conclusions These data suggest that in domesticated grasses, changes in grain size and shape evolved prior to non-shattering ears or panicles. Initial grain size increases may have evolved during the first centuries of cultivation, within perhaps 500–1000 years. Non-shattering infructescences were much slower, becoming fixed about 1000–2000 years later. This suggests a need to reconsider the role of sickle harvesting in domestication. Pulses, by contrast, do not show evidence for seed size increase in relation to the earliest cultivation, and seed size increase may be delayed by 2000–4000 years. This implies that conditions that were sufficient to select for larger seed size in Poaceae were not sufficient in Fabaceae. It is proposed that animal-drawn ploughs (or ards) provided the selection pressure for larger seeds in legumes. This implies different thresholds of selective pressure, for example in relation to differing seed ontogenetics and underlying genetic architecture in these families. Pearl millet (Pennisetum glaucum) may show some similarities to the pulses in terms of a lag-time before truly larger-grained forms evolved.

The relative contributions of biome history and geological setting to historical assembly of species richness in biodiversity hotspots remain poorly understood. The tropical Andes is one of the world’s top biodiversity hotspots, and with its diverse biomes and the relatively recent but dramatic uplift, the Andes provides an ideal study system to address these questions. To gain insights into the historical species assembly of the tropical Andes, this study focuses on investigating patterns of plant species diversification in the Andean seasonally dry tropical forest (SDTF) biome. Three plant genera are used as study groups: Amicia (Leguminosae, Papilionoideae), Tecoma (Bignoniaceae), and Mimosa (Leguminosae, Mimosoideae). Species limits are re-evaluated to enable dense sampling of species and intraspecific diversity for phylogeny reconstruction for each group. Time-calibrated phylogenies for Amicia and Mimosa are presented and used to determine patterns of species diversification in time and space. For Tecoma, incongruence between nuclear and chloroplast gene trees precludes straightforward estimation of a species tree and this incongruence is attributed to possible reticulation caused by hybridization. Divergence time estimates and patterns of diversification for Amicia and Mimosa are compared with other Andean SDTF groups (Cyathostegia, Coursetia, Poissonia; Leguminosae) using isolation by distance and phylogenetic geographic structure analyses. Consistently deep divergences between sister species and high geographic structure across all five groups suggest that Andean SDTF lineages have persisted over the past 10 million years (My) with high endemism driven by dispersal limitation, caused by geographic isolation, following the most recent episode of rapid mountain uplift 5-10 My ago. This prolonged stasis of the Andean SDTF biome is in line with Miocene fossil and paleoclimate evidence. Finally, wider analyses of the contrasting evolutionary timescales of older SDTF and more recent high-altitude grassland diversity suggest that the exceptional plant species diversity in the Andes is the outcome of highly heterogeneous evolutionary histories reflecting the physiographical heterogeneity of the Andean biodiversity hotspot.

Among the most compelling and least well-understood tree species growing in the temperate forests of southern South America is Fitzroya cupressoides, a high biomass species and the second longest-lived tree species in the world. This thesis quantified the main components of the carbon cycle in Fitzroya forests (i.e. net primary productivity (NPP) and soil respiration) and evaluated the environmental variables that are most related to them. The study was focused on medium-age and old-growth forests growing in the Coastal Range (Alerce Costero National Park, AC) and the Andean Cordillera (Alerce Andino National Park, AA) of southern Chile, respectively. The specific objectives of this thesis were to: 1) assess the forest structure, species composition and characterise the environmental conditions of these forests; 2) assess biomass, aboveground NPP, carbon allocation and mean wood residence time in these forests; 3) assess soil respiration and relate it to soil environmental conditions. Additionally, to use a mass balance approach to estimate fine root productivity; 4) estimate total NPP using biometric and indirect estimates of productivity; 5) evaluate the climatic factors mainly related to Fitzroya stem radial change on an intra-annual basis; and 6) evaluate changes in Fitzroya's tree growth and carbon isotopes during recent decades, and determine which environmental factors are more related to them. The last two objectives focus on Fitzroya as the dominant species and the subject of this study. Two 0.6 ha plots were installed within each national park; NPP was estimated for a year and soil respiration and high resolution stem growth measurements were monitored over almost two years. Aboveground biomass estimates for the Andean site are among the most massive reported in the world and carbon fluxes in Fitzroya forests are among the lowest reported for temperate wet forests worldwide. The longevity as well as the particularly rainy and nutrient poor soil conditions where these ecosystems grow may influence their exceptionally slow carbon dynamics. Differences in carbon fluxes between sites seem most probably driven by different environmental conditions rather than by developmental stage. Moreover, carbon fluxes were more sensitive to interannual climate variability in AC than AA. Warmer and drier summer conditions, likely to become more common under future climate change, more significantly affected stem growth and soil respiration in the Coastal Range than in the Andes. Regarding long-term changes, tree growth has been decreasing in the coastal site in the last 40 years and increasing in the Andes since the 1900s. These trends have been accompanied by an increase in intrinsic water use efficiency which is likely caused by rises in CO2 and changes in climate conditions in both sites. Although Fitzroya grows in particularly wet and cool areas, projected drier and warmer conditions may have a negative effect on Fitzroya stem growth and carbon sequestration in both study sites. This effect would be more critical in the Coastal Range though, because of its more Mediterranean climate influence and more restrictive soil conditions in this area. Adequate resources are needed for the monitoring and conservation of these slow growth and massive forests especially in the Coastal Range, in order to avoid ongoing illegal cuttings and threatening forest fires.

A major goal in biology is to understand the genetic basis of morphological variation at different evolutionary scales, for example between and within species. Here I investigate this issue by using plant leaves as an example. Previously comparative studies between the simple leaf model plant Arabidopsis thaliana and its dissected leaf relative Cardamine hirsuta have shown that inter-specific differences in leaf shape mostly result from variation in local tissue growth and patterning (Vlad et al., 2014; Hay et al., 2006; Barkoulas et al., 2008). Here, I aim to elucidate the genetic basis of natural variation in leaf form within species, by using divergent strains of C. hirsuta. I present evidence that variation in six strains collected from geographically diverse locations results from different rates of progression of an age-dependent leaf development programme in a phenomenon known as heteroblasty. By using Quantitative trait loci (QTL) mapping with a recombinant inbred line (RIL) population derived from a cross between the Oxford and Azores strains, I detected six QTL that influence leaflet production on multiple leaves. A QTL located on the 4th linkage group was validated and selected for further analysis. Characterisation of QTL effect indicated that the QTL influences leaf form by altering the rate of heteroblastic development. Subsequently I fine mapped this QTL to a DNA segment of 48 kb containing the gene SQUAMOSA PROMOTER PROTEIN BINDING LIKE 9 (ChSPL9), a previously characterised regulator of age dependent development. The parental alleles of ChSPL9 show variation in their sequence and were transformed into A. thaliana to evaluate whether they contribute to the QTL effect. Resultant phenotypes mirrored the QTL effect suggesting that ChSPL9 does indeed contribute to this QTL effect. These results indicate that age-dependent leaf shape progression underlies variation in leaflet number within species and more broadly suggest that in the case of plant leaves different processes might underlie morphological variation between and within species.

The presence of exotic plants both in Africa and in Asia has long attracted the attention of scholars who have attempted to understand the human activities linked to them. Archaeological and ethnographic evidence for the reconstruction of these activities is often very limited, but indirect methods such as the study of DNA have become useful tools in building models of early human dispersal. Among the plants that were carried across the Indian Ocean, sometimes known as the “tropical food kit”, the staple crop taro (Colocasia esculenta (L.) Schott) has continued to be the subject of ongoing research. The use of this crop in antiquity is well documented by discoveries of ancient taro starch granules found on archaeological artefacts from sites in Southeast Asia and the Pacific islands, making it one of the oldest plants consumed by people. However, less is known about the use of taro in Africa and the Mediterranean region, where it is found both in the wild and under cultivation - often representing a staple crop in Sub-Saharan Africa. In this doctoral thesis, genetic analysis was performed on modern samples of taro collected from Africa and other regions of the Indian Ocean, using four molecular markers. Two main clusters have been identified, and within this main sub-division four populations of taro have been detected in Africa. By integrating the genetic results with historical and linguistic research, and extensive ethnobotanical fieldwork in Africa, two of these populations are proposed to represent early translocations, with modern distribution patterns suggesting diverse dispersal routes at different times. These results open up a new scenario in which the “tropical food kit” is finally unpacked, with important historical implications for each of the crops contained within it.

This thesis addresses the methodological challenges of determining the variability of large herbivore populations through time and their impact on European vegetation. Large herbivores are at the heart of conservation policy. However, opinions widely diverge on whether we should aim for fewer herbivores and managed populations or, on the contrary as advocated by the rewilding movement, more herbivores and self-regulating populations acting as ecosystem engineers. This controversy has roots in a debate regarding the nature of ecosystems before the prevalence of human activities. Baseline ecosystems are either described as continuous forest cover with passive large herbivores, or, in contrast, as mosaics with patchy forest cover driven inter alia by bison, aurochs and horses, now rare or extinct in Europe. The main obstacle in moving this debate forward is a poor understanding of large-herbivore densities in the past. I analysed modern pollen and spore assemblages from known environmental settings to improve palaeoecological interpretation of fossil assemblages dating from the pre-human (baseline) period. The sites investigated are the rewilded grasslands of the Oostvaardersplassen (The Netherlands), the mosaic habitats of The New Forest (UK) and the old-growth closedcanopy forest of Białowieża (Poland). I demonstrate that the common practice of interpreting pollen percentages fails to estimate past forest cover in situations with natural grazing. As an explanation, I suggest that pollen productivity fluctuates with biotic factors such as herbivory and canopy shading. As a result, new insights into the baseline debate require additional lines of evidence. In this thesis, I develop an existing methodology to reconstruct past herbivore presence using fossil dung fungal spores. I synthesise current knowledge of this method with an emphasis on spore identification and, finally, I demonstrate that dung fungal spore abundance in lake sediments can be translated into large herbivore numbers. The evidence presented in this thesis contributes to the debate on re-wilding and addresses a fundamental challenge of nature conservation in the human-dominated landscapes of Europe.

Tomatoes are the fourth most valuable commodity in agriculture after rice, wheat and soybeans globally with 151 million tonnes of fruit being produced in 2012. The tomato fruit is also a model system for fleshy fruit development. During ethylene-regulated fruit ripening there are complex changes in fruit chemical composition due to degradation and synthesis of a number of soluble and volatile metabolites. Ultimately, these changes control the composition of the ripe fruit and dictate its flavour and texture. It is known that ripening can proceed when mature green fruit are removed from the plant (and indeed this is standard commercial practice) but the extent to which metabolic changes are sustained when fruit are ripened in this way has yet to be established. A modelling approach such as constraints-based modelling can provide system-level insights into the workings of the complex tomato metabolic network during ripening. The first aim of this thesis was therefore to construct a genome-scale metabolic network model for tomato and to use this model to explore metabolic network flux distributions during the transitions between the stages of fruit ripening. The flux distributions predicted provided insight into the production and usage of energy and reductants, into routes for climacteric CO2 release, and the metabolic routes underlying metabolite conversions during ripening. The second aim of this thesis was to use the model to explore metabolic engineering strategies for increased production of lycopene in tomato fruit. The model predictions showed that rearrangement of dominant metabolic fluxes were required to cope with the increased demand for reductants at high lycopene accumulation, which came at a cost of a lower accumulation of other secondary metabolites. Overall the thesis provides an approach to connect underlying metabolic mechanisms to the known metabolic processes that happen during ripening.

Japan is one of the world’s 34 biodiversity hotspots, according to Conservation International (CI). The methods used by various organisations to define priorities differ, however, and all have weaknesses when trying to identify hotspots at finer resolutions. The goal of this thesis is to investigate how biodiversity hotspots in Japan could be revealed and mapped in order to encapsulate conservation elements of biodiversity in practical ways and at various scales. Bioquality is a term that emphasises the concentration within a community of elements of biodiversity with high conservation value. It evaluates the global rarity and taxonomic distinctiveness of plant species or infra-specific taxa using four Star categories. At a plant community level, the Genetic Heat Index (GHI), which is a standardised global range size rarity score, is calculated using weighted Star statuses of species in the community. Bioquality hotspots are assessed here for the first time for the flora and vegetation in Japan – and for temperate Asia – by categorising the Japanese flora into Stars and by applying GHI to survey data and literature-based sources. Keys to Stars are developed for the Japanese flora, with adjustments for variability in species geographic range size information and for taxonomic relatedness. A Flora of Japan (FOJ) database was compiled as a BRAHMS database, containing 8,262 accepted names (30,656 taxon names in total, including synonyms) in 258 families – the first full database of Japanese vascular plants. A total of 7,145 taxa are assigned Stars; from the rarest to the widespread class, there are 884 Black, 756 Gold, 833 Blue, and 4,672 Green Star taxa, confirming that Japan as a whole contains a high proportion of globally rare taxa (23% taxa in Black or Gold). A protocol for calibrating the weight of Stars based on species geographic range is developed based on fine-resolution distribution maps within Japan and coarse–resolution Taxonomic Database Working Group (TDWG) code information. The protocol optimises calculation for temperate regions. The first ever bioquality hotspot maps of Japan are produced using two independent data sources on species distribution at national level: 1) 50 botanical prefectures using 4,830 species from the FOJ database; 2) 1,418 Horikawa ‘geoquadrats’ (0.1° latitude by 0.15° longitude grid) maps covering 829 species. The Ryukyu Islands and Ogasawara Islands are identified as bioquality hotspots, and high mountain ranges in mainland Japan are predicted to contain areas potentially high in GHI; the spatial patterns of GHI are generally concordant between maps of different resolutions. These findings highlight that bioquality assessment can be applied meaningfully at various spatial resolutions. Using field sampling data and existing literature, three study sites are further investigated on a local level: 1) the satochi-satoyama landscape, the current national priority area for biodiversity conservation; 2) various vegetation types of Okinawa-jima Island, the Ryukyu Islands; and 3) the Utaki sacred groves within the predicted hotspot of the Ryukyu Islands. The Ryukyu Islands are confirmed to contain bioquality hotspots within many individual sites, while there was generally low GHI across the satochi-satoyama landscape. The field study outcomes, together with a gap analysis of the existing coverage of protected areas, highlight three important points that are directly relevant to national biodiversity conservation planning: 1) the Ryukyu Islands urgently need newly designated protected areas; 2) the satochi-satoyama landscape conservation should redirect its focus on cultural benefits to the public; 3) the existing protected areas, particularly on mountain areas, need re-evaluation in terms of upgrading their status in light of the bioquality assessment.

Tropical forests have existed for up to one hundred million years, and today provide many ecosystem services vital for human well-being. They also harbour great biodiversity, which, in addition to its intrinsic value, plays a key role in the functioning of these ecosystems. Despite their local to global significance, there are still many knowledge gaps concerning the dynamic processes that govern the functioning of tropical forests. Rapid rates of deforestation and landscape conversion, predominantly for logging and industrial agriculture, are limiting the time and opportunity available to collect the information needed to fill these gaps. This research aims to shed light on the long-term ecological functioning of tropical forests, specifically investigating the history of disturbance in these ecosystems and the response of forest vegetation to past perturbations. The carbon-rich tropical peat swamp forests found along the coast of Sarawak, Malaysian Borneo, are a central focus of this study. For these forests in particular, a large deficit of knowledge surrounding their history and unique ecological functioning is coupled with some of the highest conversion rates of all tropical forest ecosystems across the world. In this thesis, palaeoecological data has been used to reconstruct temporal variability in forest vegetation coincident with external perturbations in order to identify changes in the resilience of these ecosystems through time, via indicators such as slowing rates of recovery and reduced regeneration of forest vegetation. Results suggest that tropical forest ecosystems have, for the most part, shown resilience to natural disturbances in the past, ranging from instantaneous localised tree-fall to longer-term regional climatic change; but that recent anthropogenic disturbances, of novel forms and greater intensities, are jeopardizing the potential for forest recovery and thus compromising ecosystem resilience. These findings enhance our understanding of the ecology of tropical peat swamp forests, and tropical forests more broadly. They also provide a context for contemporary tropical forest management, allowing for predictions on future responses to disturbance and enabling more ecologically sustainable landscape planning.

Rab GTPases are major regulatory proteins of vesicle traffic and thus responsible for membrane identity, vesicle targeting and vesicle fusion. The angiosperm Rab GTPase family is grouped into eight clades (Rab-A to Rab-H) that are broadly conserved in animals and yeasts. It has been proposed that the Rab-A clade has diversified in land plants giving rise to six plant- specific structural subclasses, Rab-A1 to Rab-A6. Previous work suggests that the Arabidopsis Rab-A2 and Rab-A3 proteins define a novel endosomal compartment that lies on a pathway between the Golgi and the plasma membrane. In dividing cells, the Rab-A2/A3 compartment is implicated in biosynthetic traffic to the cell plate but it is unclear what traffics through this compartment in non-dividing cells. In this project, I investigated a range of membrane trafficking pathways in Arabidopsis thaliana. These were probed for dependency on RAB-A2a function, using the dominant negative approach combined with fluorescent marker technology. The data presented in this thesis suggests that RAB-A2a acts on a protein recycling pathway that is used by PIN2:GFP.

Phylogenetic studies are providing powerful new insights into the evolution of complex traits. Metal hyperaccumulation is an unusual and complex physiological trait found in about 500 plant species and is associated with an exceptionally high degree of tolerance of metalliferous soils. Alyssum L. (Brassicaceae) is the largest known hyperaccumulator genus, comprising approximately 188 species distributed throughout the Mediterranean region and south-west Asia. Approximately one-quarter of these are largely restricted to areas of serpentine soils and have the ability to accumulate nickel to high concentrations in shoot tissue. This genus provides a good example in which to study the origins of a complex physiological trait, but its phylogeny is currently poorly understood. To produce a well-resolved phylogenetic tree to investigate the number and timing of origins of nickel hyperaccumulation within Alyssum, DNA sequences were generated for four chloroplast regions (matK, rps16–trnK, trnD–T and trnL–F) from 170 of 255 species in the tribe Alysseae. Additional sequencing was carried out for the chloroplast genes ndhF and rbcL and the nuclear gene PHYA. A Bayesian analysis employing a relaxed uncorrelated lognormal molecular clock and multiple fossil-age calibration points was carried out to reconstruct a time-calibrated phylogeny of this tribe using appropriate outgroups. Optimization of the nickel hyperaccumulation trait onto the resulting phylogenetic tree suggests that nickel hyperaccumulation arose twice in the Alysseae in the late Miocene/early Pliocene: 3.3–8.3 Mya in Alyssum and 6.3–8.8 Mya in Bornmuellera. The single origin in Alyssum is strongly associated with a significant acceleration in net species diversification rate, suggesting the ability to hyperaccumulate nickel could have provided a key evolutionary innovation facilitating rapid range expansion and subsequent species diversification. The scattered distribution of nickel hyperaccumulators across small island-like patches of serpentine soil suggests that allopatric speciation may have driven rapid diversification in this clade.

Little is known about the composition of tropical epiphytic communities and the influence of environmental variables on community composition. In this thesis I quantify the diversity and biomass of bromeliads, and other vascular epiphytes along an altitudinal transect on the eastern slope of the southeast Peruvian Andes and then look for species’ adaptations related to patterns of diversity and biomass. I compare patterns with those of woody species. Bromeliad species, like tree species, were found to form ecological zones related to climate. The lowest altitude ecological zone (below 1250 m) is the lowland rainforest (LRF), which has the warmest climate and highest evapotranspiration. In LRF vascular epiphytes are less prominent than other ecological zones, with the lowest bromeliad species richness and lowest vascular epiphyte biomass. However, low water-availability gives rise to most variable shoot morphology of bromeliads. The tropical montane forest (TMF), between 1250 m and 2250 m, is intermediate in climate between the LRF and the tropical montane cloud forest (TCF). The TMF has the highest α-diversity, but species richness is lower than the TCF. The shoot morphology of bromeliads is intermediate between TCF and LRF. The highest altitude ecological zone with forest is the TCF (above 2250 m). The TCF has the highest bromeliad species richness, and lowest diversity of shoot forms. The low diversity of shoot forms represents the need for a large phytotelm (water-impounding shoot) to intercept and store precipitation. The TCF has the highest vascular epiphyte biomass, although the biomass is variable as a consequence of the natural disturbance caused by landslides. Along the transect bromeliad species with CAM photosynthesis are only present in the LRF. Terrestrial bromeliad distribution records covering the Neotropics show CAM photosynthesis is more prevalent in drier environments showing that CAM photosynthesis is primarily an adaptation to drought. Epiphytic bromeliads, pre-adapted to a water-stressed environment show no differences in presence along rainfall gradients, but species with CAM photosynthesis occupy warmer environments.

Japan is one of the world’s 34 biodiversity hotspots, according to Conservation International (CI). The methods used by various organisations to define priorities differ, however, and all have weaknesses when trying to identify hotspots at finer resolutions. The goal of this thesis is to investigate how biodiversity hotspots in Japan could be revealed and mapped in order to encapsulate conservation elements of biodiversity in practical ways and at various scales. Bioquality is a term that emphasises the concentration within a community of elements of biodiversity with high conservation value. It evaluates the global rarity and taxonomic distinctiveness of plant species or infra-specific taxa using four Star categories. At a plant community level, the Genetic Heat Index (GHI), which is a standardised global range size rarity score, is calculated using weighted Star statuses of species in the community. Bioquality hotspots are assessed here for the first time for the flora and vegetation in Japan – and for temperate Asia – by categorising the Japanese flora into Stars and by applying GHI to survey data and literature-based sources. Keys to Stars are developed for the Japanese flora, with adjustments for variability in species geographic range size information and for taxonomic relatedness. A Flora of Japan (FOJ) database was compiled as a BRAHMS database, containing 8,262 accepted names (30,656 taxon names in total, including synonyms) in 258 families – the first full database of Japanese vascular plants. A total of 7,145 taxa are assigned Stars; from the rarest to the widespread class, there are 884 Black, 756 Gold, 833 Blue, and 4,672 Green Star taxa, confirming that Japan as a whole contains a high proportion of globally rare taxa (23% taxa in Black or Gold). A protocol for calibrating the weight of Stars based on species geographic range is developed based on fine-resolution distribution maps within Japan and coarse–resolution Taxonomic Database Working Group (TDWG) code information. The protocol optimises calculation for temperate regions. The first ever bioquality hotspot maps of Japan are produced using two independent data sources on species distribution at national level: 1) 50 botanical prefectures using 4,830 species from the FOJ database; 2) 1,418 Horikawa ‘geoquadrats’ (0.1° latitude by 0.15° longitude grid) maps covering 829 species. The Ryukyu Islands and Ogasawara Islands are identified as bioquality hotspots, and high mountain ranges in mainland Japan are predicted to contain areas potentially high in GHI; the spatial patterns of GHI are generally concordant between maps of different resolutions. These findings highlight that bioquality assessment can be applied meaningfully at various spatial resolutions. Using field sampling data and existing literature, three study sites are further investigated on a local level: 1) the satochi-satoyama landscape, the current national priority area for biodiversity conservation; 2) various vegetation types of Okinawa-jima Island, the Ryukyu Islands; and 3) the Utaki sacred groves within the predicted hotspot of the Ryukyu Islands. The Ryukyu Islands are confirmed to contain bioquality hotspots within many individual sites, while there was generally low GHI across the satochi-satoyama landscape. The field study outcomes, together with a gap analysis of the existing coverage of protected areas, highlight three important points that are directly relevant to national biodiversity conservation planning: 1) the Ryukyu Islands urgently need newly designated protected areas; 2) the satochi-satoyama landscape conservation should redirect its focus on cultural benefits to the public; 3) the existing protected areas, particularly on mountain areas, need re-evaluation in terms of upgrading their status in light of the bioquality assessment.

A key question in developmental biology is how form is generated. The model species Arabidopsis thaliana produces simple leaves with marginal outgrowths termed serrations. Serration development in A. thaliana requires both the transcription factor CUP-SHAPED COTYLEDON2 (CUC2) and the auxin efflux facilitator PIN-FORMED1 (PIN1), which regulates polar auxin transport by forming convergence points (Hay et al., 2006; Nikovics et al., 2006; Scarpella et al., 2006). In Chapter 3, I investigate how CUC2, PIN1 and auxin interact to control serration development. I demonstrate that CUC2 promotes PIN1 convergence point and auxin activity foci formation along the margin of the leaf, whilst high auxin activity represses CUC2 expression. Furthermore, interspersed peaks of CUC2 and auxin activity pattern serration development along the proximo-distal axis of the leaf. Thus, auxin, PIN1 and CUC2 form a negative feedback loop that patterns serration development.CUC genes and PIN1 are required for leaflet development in Cardamine hirsuta (Barkoulas et al., 2008; Blein et al., 2008), a close relative of A. thaliana that produces compound leaves subdivided into units termed leaflets. However, it is unclear how CUC and PIN1 interact to control leaflet development. In Chapter 4, I demonstrate that similar to A. thaliana, CUC genes promote PIN1 convergence point and auxin activity foci formation at the C. hirsuta leaf margin, whilst high auxin activity represses CUC2 expression. These genetic interactions likely create interspersed peaks of CUC2 and auxin activity that pattern leaflet development. Thus, the same negative feedback loop between CUC, PIN1 and auxin patterns both leaflet development in C. hirsuta and serration development in A. thaliana.KNOTTED1-LIKE HOMEOBOX (KNOX) genes are expressed in C. hirsuta leaves, and interact with ChCUC and PIN1 in positive and negative feedback loops, respectively, to control leaflet development (Barkoulas et al., 2008; Blein et al., 2008). KNOX genes are not expressed in A. thaliana leaves, but deeply lobed margins reminiscent of leaflets develop in association with ectopic KNOX expression in leaves (Chuck et al., 1996; Hay et al., 2006). However, it is unclear whether regulatory interactions of PIN1, CUC and KNOX which occur in C. hirsuta leaflets are employed during KNOX-induced lobe development in A. thaliana. In Chapter 5, I demonstrate that CUC2 and polar auxin transport are required for ectopic KNOX expression. Conversely, I show that KNOX misexpression up-regulates CUC2 expression in A. thaliana leaves. Thus, interactions between KNOX, CUC and PIN1 that occur in leaflet development in C. hirsuta also occur in association with KNOX-induced lobe development in A. thaliana.In addition to investigating the regulatory interactions between known components of leaf development pathways, I sought to identify novel genes that mediate CUC2-dependent serration development in A. thaliana. In Chapter 6, I identify a suppressor of the smooth margin phenotype of cuc2 leaves that partially restores PIN1 localisation in the absence of functional CUC2. Finally, in the General Discussion I evaluate how interlinking feedback loops between CUC, KNOX and auxin pattern serration and leaflet development. I then discuss why interlinking feedback loops may have been deployed to control outgrowths in both plant and animal systems.

Small Island Developing States (SIDS) of the Caribbean are one of the world’s ‘hottest’ ‘biodiversity hotspots’. However, this biodiversity continues to be threatened by habitat loss, and now, by climate change. The research reported here investigated the potential of species distribution modelling (SDM) as a plant conservation tool within Caribbean SIDS, using Trinidad as a case study. Prior to the application of SDM, ancillary analyses including: (i) quantification and mapping of forest cover change (1969 to 2007) and deforestation rates, and (ii) assessment of the island’s vegetation community distribution and associated drivers were carried out.Community distribution and commercial importance and global/regional rarity were used to generate a list of species for assessing the potential of SDM within Trinidad. Species occurrence data were used to generate species distribution models for present climate conditions within the SDM algorithm, MaxEnt. These results were assessed through expert appraisal and concurrence with results of ecological analyses. These models were used to forecast suitable species climate space forty years into an SRES A2 future. Present and future models were then combined to produce a ‘collective change map’ which showed projected areas of species’ range expansion, contraction or stability for this group of species with respect to Trinidad’s Protected Areas (PAs) network.Despite the models being indicative rather than accurate, it was concluded that species’ climate space is likely to decrease or disappear across Trinidad. Extended beyond Trinidad into the remainder of the Caribbean region, SDM may be a crucial tool in identifying which PAs within the region (and not individual islands) will facilitate future survival of given target species. Consideration of species conservation from a regional, rather than an individual island perspective, is strongly recommended for aiding the Caribbean SIDS to adapt in response to climate change.

Tomato is an important food crop and a model for fleshy fruit development. The process of fruit ripening involves changes in chemical composition and in particular the accumulation of sugars, organic, amino acids and carotenes. The research described in this thesis aimed to identify key regulatory aspects associated with the accumulation of the major acids in tomato fruit by analysis of introgression lines resulting from a cross between a cultivated variety, Solanum lycopersicum, and a wild progenitor species, Solanum pennellii. Line 2-5 showed increases in citrate, malate, aspartate and glutamate in fruit grown under greenhouse conditions.The genetic differences between line 2-5, its overlapping lines, sub-introgression lines and the recurrent parent were used to link the metabolite phenotypes to smaller chromosomal regions. This analysis suggested multiple epistatic loci control fruit metabolite accumulation. Investigation of the biochemical differences between line 2-5 and the recurrent parent revealed that organic and amino acid accumulation did not dependent upon increased TCA cycle capacity. Regulation at the metabolic level was identified for citrate accumulation with changes in cytosolic aconitase in line 2-5. As these metabolites accumulate in the vacuole, tonoplast transport was investigated. Correlation of ATPase-dependent malate influx with altered malate content suggested malate tonoplast transport plays a role in malate accumulation and highlights the importance of vacuolar storage and transport in the regulation of organic and amino acid accumulation.

The relative contributions of biome history and geological setting to historical assembly of species richness in biodiversity hotspots remain poorly understood. The tropical Andes is one of the world’s top biodiversity hotspots, and with its diverse biomes and the relatively recent but dramatic uplift, the Andes provides an ideal study system to address these questions. To gain insights into the historical species assembly of the tropical Andes, this study focuses on investigating patterns of plant species diversification in the Andean seasonally dry tropical forest (SDTF) biome. Three plant genera are used as study groups: Amicia (Leguminosae, Papilionoideae), Tecoma (Bignoniaceae), and Mimosa (Leguminosae, Mimosoideae). Species limits are re-evaluated to enable dense sampling of species and intraspecific diversity for phylogeny reconstruction for each group. Time-calibrated phylogenies for Amicia and Mimosa are presented and used to determine patterns of species diversification in time and space. For Tecoma, incongruence between nuclear and chloroplast gene trees precludes straightforward estimation of a species tree and this incongruence is attributed to possible reticulation caused by hybridization. Divergence time estimates and patterns of diversification for Amicia and Mimosa are compared with other Andean SDTF groups (Cyathostegia, Coursetia, Poissonia; Leguminosae) using isolation by distance and phylogenetic geographic structure analyses. Consistently deep divergences between sister species and high geographic structure across all five groups suggest that Andean SDTF lineages have persisted over the past 10 million years (My) with high endemism driven by dispersal limitation, caused by geographic isolation, following the most recent episode of rapid mountain uplift 5-10 My ago. This prolonged stasis of the Andean SDTF biome is in line with Miocene fossil and paleoclimate evidence. Finally, wider analyses of the contrasting evolutionary timescales of older SDTF and more recent high-altitude grassland diversity suggest that the exceptional plant species diversity in the Andes is the outcome of highly heterogeneous evolutionary histories reflecting the physiographical heterogeneity of the Andean biodiversity hotspot.

Africa's miombo woodlands constitute one of the most important dry tropical forests on earth, yet the hydraulic function of these woodlands remains poorly researched. Given the current predictions of increased aridity by the end of this century in the miombo ecoregion, understanding the likely response of miombo woodlands tree species to water stress is crucial in planning adaptation strategies. Predicting the response of miombo woodlands to future climate trends is hampered by a lack of knowledge on the physiology of the common miombo woodlands tree species. In particular, plant-water relations for this woodlands type are not well understood.An understanding of plant-water relations for this woodlands type will provide insights into how water limits tree species distribution in this ecosystem. This will also improve our prediction model on the likely response of this ecosystem to predicted climate change. For this reason, the overall objective of this research was to evaluate the hydraulic architecture and xylem vulnerability to cavitation for nine principal miombo woodlands tree species differing in drought tolerance ability and habitat preference. This was achieved by; examining the hydraulic properties and evaluating the extent to which each hydraulic design was vulnerable to water stress-induced xylem cavitation; investigating how seasonal changes in plant-water relations influences seasonal patterns of leaf display and; analyzing the relationship between stem hydraulic supply and leaf functional traits related to drought tolerance ability.This research has found that drought-intolerant tree species with mesic specialization have more efficient stem hydraulic systems than co-occurring habitat broad ranging species. Broad ranging tree species attain wider habitat distribution by adjusting their hydraulic supply in response to changing ecosystem water availability. The finding that hydraulic properties differ significantly between tree species with contrasting habitat preference suggests that tree hydraulic design may have some adaptive ecological role in influencing species habitat preferences in miombo woodlands.The evaluation of xylem vulnerability to cavitation revealed that mesic specialized tree species were more vulnerable to water stress-induced cavitation than habitat broad ranging tree species. Vulnerability to cavitation in individuals from the same broad-ranging species growing in contrasting habitats showed only marginal and statistically insignificant (P > 0.05) differences between wet and dry sites.In the investigation of the influence of seasonal changes in stem water relations on seasonal leaf display, seasonal rhythms in stem water status were found to exert significant controls on leaf phenology. Mesic specialists had strong stem water controls throughout the year in comparison to broad ranging tree species.An analysis of the relationship between stem hydraulic supply and leaf functional traits suggests that stem hydraulic supply constrains leaf biomass allocation patterns among miombo tree species. Mesic specialists tend to invest more in leaf longevity than broad ranging tree species.This thesis has uncovered some interesting relationships between plant-water-relations and the distribution of miombo woodlands tree species. These results lead to the conclusion that in an event of increased ecosystem drying under future climate trends, tree species with mesic specialisation are at a greater risk of experiencing cavitation related species mortality than broad ranging ones.