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Linear mixed models (LMMs) provide a still underused methodological perspective on combining experimental and individual-differences research. Here we illustrate this approach with two-rectangle cueing in visual attention (Egly et al., 1994). We replicated previous experimental cue-validity effects relating to a spatial shift of attention within an object (spatial effect), to attention switch between objects (object effect), and to the attraction of attention toward the display centroid (attraction effect), also taking into account the design-inherent imbalance of valid and other trials. We simultaneously estimated variance/covariance components of subject-related random effects for these spatial, object, and attraction effects in addition to their mean reaction times (RTs). The spatial effect showed a strong positive correlation with mean RT and a strong negative correlation with the attraction effect. The analysis of individual differences suggests that slow subjects engage attention more strongly at the cued location than fast subjects. We compare this joint LMM analysis of experimental effects and associated subject-related variances and correlations with two frequently used alternative statistical procedures
Within this work, three physicochemical methods for the hydrophobization of initially hydrophilic solid particles are investigated. The modified particles are then used for the stabilization of oil-in-water (o/w) emulsions. For all introduced methods electrostatic interactions between strongly or weakly charged groups in the system are es-sential. (i) Short chain alkylammonium bromides (C4 – C12) adsorb on oppositely charged solid particles. Macroscopic contact angle measurements of water droplets under air and hexane on flat silica surfaces in dependency of the surface charge density and alkylchain-length allow the calculation of the surface energy and give insights into the emulsification properties of solid particles modified with alkyltrimethylammonium bromides. The measure-ments show an increase of the contact angle with increasing surface charge density, due to the enhanced adsorp-tion of the oppositely charged alkylammonium bromides. Contact angles are higher for longer alkylchain lengths. The surface energy calculations show that in particular the surface-hexane or surface-air interfacial en-ergy is being lowered upon alkylammonium adsorption, while a significant increase of the surface-water interfa-cial energy occurs only at long alkyl chain lengths and high surface charge densities. (ii) The thickness and the charge density of an adsorbed weak polyelectrolyte layer (e.g. PMAA, PAH) influence the wettability of nanoparticles (e.g. alumina, silica, see Scheme 1(b)). Furthermore, the isoelectric point and the pH range of colloidal stability of particle-polyelectrolyte composites depend on the thickness of the weak polye-lectrolyte layer. Silica nanoparticles with adsorbed PAH and alumina nanoparticles with adsorbed PMAA be-come interfacially active and thus able to stabilize o/w emulsions when the degree of dissociation of the polye-lectrolyte layer is below 80 %. The average droplet size after emulsification of dodecane in water depends on the thickness and the degree of dissociation of the adsorbed PE-layer. The visualization of the particle-stabilized o/w emulsions by cryogenic SEM shows that for colloidally stable alumina-PMAA composites the oil-water interface is covered with a closely packed monolayer of particles, while for the colloidally unstable case closely packed aggregated particles deposit on the interface. (iii) By emulsifying a mixture of the corrosion inhibitor 8-hydroxyquinoline (8-HQ) and styrene with silica nanoparticles a highly stable o/w emulsion can be obtained in a narrow pH window. The amphoteric character of 8-HQ enables a pH dependent electrostatic interaction with silica nanoparticles, which can render them interfa-cially active. Depending on the concentration and the degree of dissociation of 8-HQ the adsorption onto silica results from electrostatic or aromatic interactions between 8-HQ and the particle-surface. At intermediate amounts of adsorbed 8-HQ the oil wettability of the particles becomes sufficient for stabilizing o/w emulsions. Cryogenic SEM visualization shows that the particles arrange then in a closely packed shell consisting of partly of aggregated domains on the droplet interface. For further increasing amounts of adsorbed 8-HQ the oil wet-tability is reduced again and the particles ability to stabilize emulsions decreases. By the addition of hexadecane to the oil phase the size of the droplets can be reduced down to 200 nm by in-creasing the silica mass fraction. Subsequent polymerization produces corrosion inhibitor filled (20 wt-%) poly-styrene-silica composite particles. The measurement of the release of 8-hydroxyquinoline shows a rapid increase of 8-hydroxyquinoline in a stirred aqueous solution indicating the release of the total content in less than 5 min-utes. The method is extended for the encapsulation of other organic corrosion inhibitors. The silica-polymer-inhibitor composite particles are then dispersed in a water based alkyd emulsion, and the dispersion is used to coat flat aluminium substrates. After drying and cross-linking the polmer-film Confocal Laser Scanning Micros-copy is employed revealing a homogeneous distribution of the particles in the film. Electrochemical Impedance Spectroscopy in aqueous electrolyte solutions shows that films with aggregated particle domains degrade with time and don’t provide long-term corrosion protection of the substrate. However, films with highly dispersed particles have high barrier properties for corrosive species. The comparison of films containing silica-polystyrene composite particles with and without 8-hydroxyquinoline shows higher electrochemical impedances when the inhibitor is present in the film. By applying the Scanning Vibrating Electrode Technique the localized corrosion rate in the fractured area of scratched polymer films containing the silica-polymer-inhibitor composite particles is studied. Electrochemical corrosion cannot be suppressed but the rate is lowered when inhibitor filled composite particles are present in the film. By depositing six polyelectrolyte layers on particle stabilized emulsion droplets their surface morphology changes significantly as shown by SEM visualization. When the oil wettability of the outer polyelectrolyte layer increases, the polyelectrolyte coated droplets can act as emulsion stabilizers themselves by attaching onto bigger oil droplets in a closely packed arrangement. In the presence of 3 mM LaCl3 8-HQ hydrophobized silica particles aggregate strongly on the oil-water inter-face. The application of an ultrasonic field can remove two dimensional shell-compartments from the droplet surface, which are then found in the aqueous bulk phase. Their size ranges up to 1/4th of the spherical particle shell.
Business Process Management (BPM) emerged as a means to control, analyse, and optimise business operations. Conceptual models are of central importance for BPM. Most prominently, process models define the behaviour that is performed to achieve a business value. In essence, a process model is a mapping of properties of the original business process to the model, created for a purpose. Different modelling purposes, therefore, result in different models of a business process. Against this background, the misalignment of process models often observed in the field of BPM is no surprise. Even if the same business scenario is considered, models created for strategic decision making differ in content significantly from models created for process automation. Despite their differences, process models that refer to the same business process should be consistent, i.e., free of contradictions. Apparently, there is a trade-off between strictness of a notion of consistency and appropriateness of process models serving different purposes. Existing work on consistency analysis builds upon behaviour equivalences and hierarchical refinements between process models. Hence, these approaches are computationally hard and do not offer the flexibility to gradually relax consistency requirements towards a certain setting. This thesis presents a framework for the analysis of behaviour consistency that takes a fundamentally different approach. As a first step, an alignment between corresponding elements of related process models is constructed. Then, this thesis conducts behavioural analysis grounded on a relational abstraction of the behaviour of a process model, its behavioural profile. Different variants of these profiles are proposed, along with efficient computation techniques for a broad class of process models. Using behavioural profiles, consistency of an alignment between process models is judged by different notions and measures. The consistency measures are also adjusted to assess conformance of process logs that capture the observed execution of a process. Further, this thesis proposes various complementary techniques to support consistency management. It elaborates on how to implement consistent change propagation between process models, addresses the exploration of behavioural commonalities and differences, and proposes a model synthesis for behavioural profiles.
Motivation | Societal and economic needs of East Africa rely entirely on the availability of water, which is governed by the regular onset and retreat of the rainy seasons. Fluctuations in the amounts of rainfall has tremendous impact causing widespread famine, disease outbreaks and human migrations. Efforts towards high resolution forecasting of seasonal precipitation and hydrological systems are therefore needed, which requires high frequency short to long-term analyses of available climate data that I am going to present in this doctoral thesis by three different studies. 15,000 years - Suguta Valley | The main study of this thesis concentrated on the understanding of humidity changes within the last African Humid Period (AHP, 14.8-5.5 ka BP). The nature and causes of intensity variations of the West-African (WAM) and Indian Summer monsoons (ISM) during the AHP, especially their exact influence on regional climate relative to each other, is currently intensely debated. Here, I present a high-resolution multiproxy lake-level record spanning the AHP from the remote Suguta Valley in the northern Kenya Rift, located between the WAM and ISM domains. The presently desiccated valley was during the AHP filled by a 300 m deep and 2200 km2 large palaeo-lake due to an increase in precipitation of only 26%. The record explains the synchronous onset of large lakes in the East African Rift System (EARS) with the longitudinal shift of the Congo Air Boundary (CAB) over the East African and Ethiopian Plateaus, as the direct consequence of an enhanced atmospheric pressure gradient between East-Africa and India due to a precessional-forced northern hemisphere insolation maximum. Pronounced, and abrupt lake level fluctuations during the generally wet AHP are explained by small-scale solar irradiation changes weakening this pressure gradient atmospheric moisture availability preventing the CAB from reaching the study area. Instead, the termination of the AHP occurred, in a non-linear manner due to a change towards an equatorial insolation maximum ca. 6.5 ka ago extending the AHP over Ethiopia and West-Africa. 200 years - Lake Naivasha | The second part of the thesis focused on the analysis of a 200 year-old sediment core from Lake Naivasha in the Central Kenya Rift, one of the very few present freshwater lakes in East Africa. The results revealed and confirmed, that the appliance of proxy records for palaeo-climate reconstruction for the last 100 years within a time of increasing industrialisation and therefore human impact to the proxy-record containing sites are broadly limited. Since the middle of the 20th century, intense anthropogenic activity around Lake Naivasha has led to cultural eutrophication, which has overprinted the influence of natural climate variation to the lake usually inferred from proxy records such as diatoms, transfer-functions, geochemical and sedimentological analysis as used in this study. The results clarify the need for proxy records from remote unsettled areas to contribute with pristine data sets to current debates about anthropologic induced global warming since the past 100 years. 14 years - East African Rift | In order to avoid human influenced data sets and validate spatial and temporal heterogeneities of proxy-records from East Africa, the third part of the thesis therefore concentrated on the most recent past 14 years (1996-2010) detecting climate variability by using remotely sensed rainfall data. The advancement in the spatial coverage and temporal resolutions of rainfall data allow a better understanding of influencing climate mechanisms and help to better interpret proxy-records from the EARS in order to reconstruct past climate conditions. The study focuses on the dynamics of intraseasonal rainfall distribution within catchments of eleven lake basins in the EARS that are often used for palaeo-climate studies. We discovered that rainfall in adjacent basins exhibits high complexities in the magnitudes of intraseasonal variability, biennial to triennial precipitation patterns and even are not necessarily correlated often showing opposite trends. The variability among the watersheds is driven by the complex interaction of topography, in particular the shape, length and elevation of the catchment and its relative location to the East African Rift System and predominant influence of the ITCZ or CAB, whose locations and intensities are dependent on the strength of low pressure cells over India, SST variations in the Atlantic, Pacific or Indian Ocean, QBO phases and the 11-year solar cycle. Among all seasons we observed, January-September is the season of highest and most complex rainfall variability, especially for the East African Plateau basins, most likely due to the irregular penetration and sensitivity of the CAB.
The heterogeneity in species assemblages of epigeal spiders was studied in a natural forest and in a managed forest. Additionally the effects of small-scale microhabitat heterogeneity of managed and unmanaged forests were determined by analysing the spider assemblages of three different microhabitat structures (i. vegetation, ii. dead wood. iii. litter cover). The spider were collected in a block design by pitfall traps (n=72) in a 4-week interval. To reveal key environmental factors affecting the spider distribution abiotic and biotic habitat parameters (e.g. vegetation parameters, climate parameters, soil moisture) were assessed around each pitfall trap. A TWINSPAN analyses separated pitfall traps from the natural forest from traps of the managed forest. A subsequent discriminant analyses revealed that the temperature, the visible sky, the plant diversity and the mean diameter at breast height as key discriminant factors between the microhabitat groupings designated by the TWINSPAN analyses. Finally a Redundant analysis (RDA) was done revealing similar environmental factors responsible for the spider species distribution, as a good separation of the different forest types as well as the separation of the microhabitat groupings from the TWINSPAN. Overall the study revealed that the spider communities differed between the forest types as well as between the microhabitat structures and thus species distribution changed within a forest stand on a fine spatial scale. It was documented that the structure of managed forests affects the composition of spider assemblages compared to natural forests significantly and even small scale-heterogeneity seems to influence the spider species composition.
Regulation of gene transcription plays a major role in mediating cellular responses and physiological behavior in all known organisms. The finding that similar genes are often regulated in a similar manner (co-regulated or "co-expressed") has directed several "guilt-by-association" approaches in order to reverse-engineer the cellular transcriptional networks using gene expression data as a compass. This kind of studies has been considerably assisted in the recent years by the development of high-throughput transcript measurement platforms, specifically gene microarrays and next-generation sequencing. In this thesis, I describe several approaches for improving the extraction and interpretation of the information contained in microarray based gene expression data, through four steps: (1) microarray platform design, (2) microarray data normalization, (3) gene network reverse engineering based on expression data and (4) experimental validation of expression-based guilt-by-association inferences. In the first part test case is shown aimed at the generation of a microarray for Thellungiella salsuginea, a salt and drought resistant close relative to the model plant Arabidopsis thaliana; the transcripts of this organism are generated on the combination of publicly available ESTs and newly generated ad-hoc next-generation sequencing data. Since the design of a microarray platform requires the availability of highly reliable and non-redundant transcript models, these issues are addressed consecutively, proposing several different technical solutions. In the second part I describe how inter-array correlation artifacts are generated by the common microarray normalization methods RMA and GCRMA, together with the technical and mathematical characteristics underlying the problem. A solution is proposed in the form of a novel normalization method, called tRMA. The third part of the thesis deals with the field of expression-based gene network reverse engineering. It is shown how different centrality measures in reverse engineered gene networks can be used to distinguish specific classes of genes, in particular essential genes in Arabidopsis thaliana, and how the use of conditional correlation can add a layer of understanding over the information flow processes underlying transcript regulation. Furthermore, several network reverse engineering approaches are compared, with a particular focus on the LASSO, a linear regression derivative rarely applied before in global gene network reconstruction, despite its theoretical advantages in robustness and interpretability over more standard methods. The performance of LASSO is assessed through several in silico analyses dealing with the reliability of the inferred gene networks. In the final part, LASSO and other reverse engineering methods are used to experimentally identify novel genes involved in two independent scenarios: the seed coat mucilage pathway in Arabidopsis thaliana and the hypoxic tuber development in Solanum tuberosum. In both cases an interesting method complementarity is shown, which strongly suggests a general use of hybrid approaches for transcript expression-based inferences. In conclusion, this work has helped to improve our understanding of gene transcription regulation through a better interpretation of high-throughput expression data. Part of the network reverse engineering methods described in this thesis have been included in a tool (CorTo) for gene network reverse engineering and annotated visualization from custom transcription datasets.
Answer Set Programming (ASP) is an emerging paradigm for declarative programming, in which a computational problem is specified by a logic program such that particular models, called answer sets, match solutions. ASP faces a growing range of applications, demanding for high-performance tools able to solve complex problems. ASP integrates ideas from a variety of neighboring fields. In particular, automated techniques to search for answer sets are inspired by Boolean Satisfiability (SAT) solving approaches. While the latter have firm proof-theoretic foundations, ASP lacks formal frameworks for characterizing and comparing solving methods. Furthermore, sophisticated search patterns of modern SAT solvers, successfully applied in areas like, e.g., model checking and verification, are not yet established in ASP solving. We address these deficiencies by, for one, providing proof-theoretic frameworks that allow for characterizing, comparing, and analyzing approaches to answer set computation. For another, we devise modern ASP solving algorithms that integrate and extend state-of-the-art techniques for Boolean constraint solving. We thus contribute to the understanding of existing ASP solving approaches and their interconnections as well as to their enhancement by incorporating sophisticated search patterns. The central idea of our approach is to identify atomic as well as composite constituents of a propositional logic program with Boolean variables. This enables us to describe fundamental inference steps, and to selectively combine them in proof-theoretic characterizations of various ASP solving methods. In particular, we show that different concepts of case analyses applied by existing ASP solvers implicate mutual exponential separations regarding their best-case complexities. We also develop a generic proof-theoretic framework amenable to language extensions, and we point out that exponential separations can likewise be obtained due to case analyses on them. We further exploit fundamental inference steps to derive Boolean constraints characterizing answer sets. They enable the conception of ASP solving algorithms including search patterns of modern SAT solvers, while also allowing for direct technology transfers between the areas of ASP and SAT solving. Beyond the search for one answer set of a logic program, we address the enumeration of answer sets and their projections to a subvocabulary, respectively. The algorithms we develop enable repetition-free enumeration in polynomial space without being intrusive, i.e., they do not necessitate any modifications of computations before an answer set is found. Our approach to ASP solving is implemented in clasp, a state-of-the-art Boolean constraint solver that has successfully participated in recent solver competitions. Although we do here not address the implementation techniques of clasp or all of its features, we present the principles of its success in the context of ASP solving.
Genetic variation is crucial for the long-term survival of the species as it provides the potential for adaptive responses to environmental changes such as emerging diseases. The Major Histocompatibility Complex (MHC) is a gene family that plays a central role in the vertebrate’s immune system by triggering the adaptive immune response after exposure to pathogens. MHC genes have become highly suitable molecular markers of adaptive significance. They synthesize two primary cell surface molecules namely MHC class I and class II that recognize short fragments of proteins derived respectively from intracellular (e.g. viruses) and extracellular (e.g. bacteria, protozoa, arthropods) origins and present them to immune cells. High levels of MHC polymorphism frequently observed in natural populations are interpreted as an adaptation to detect and present a wide array of rapidly evolving pathogens. This variation appears to be largely maintained by positive selection driven mainly by pathogenic selective pressures. For my doctoral research I focused on MHC I and II variation in free-ranging cheetahs (Acinonyx jubatus) and leopards (Panthera pardus) on Namibian farmlands. Both felid species are sympatric thus subject to similar pathogenic pressures but differ in their evolutionary and demographic histories. The main aims were to investigate 1) the extent and patterns of MHC variation at the population level in both felids, 2) the association between levels of MHC variation and disease resistance in free-ranging cheetahs, and 3) the role of selection at different time scales in shaping MHC variation in both felids. Cheetahs and leopards represent the largest free-ranging carnivores in Namibia. They concentrate in unprotected areas on privately owned farmlands where domestic and other wild animals also occur and the risk of pathogen transmission is increased. Thus, knowledge on adaptive genetic variation involved in disease resistance may be pertinent to both felid species’ conservation. The cheetah has been used as a classic example in conservation genetics textbooks due to overall low levels of genetic variation. Reduced variation at MHC genes has been associated with high susceptibility to infectious diseases in cheetahs. However, increased disease susceptibility has only been observed in captive cheetahs whereas recent studies in free-ranging Namibian cheetahs revealed a good health status. This raised the question whether the diversity at MHC I and II genes in free-ranging cheetahs is higher than previously reported. In this study, a total of 10 MHC I alleles and four MHC II alleles were observed in 149 individuals throughout Namibia. All alleles but one likely belong to functional MHC genes as their expression was confirmed. The observed alleles belong to four MHC I and three MHC II genes in the species as revealed by phylogenetic analyses. Signatures of historical positive selection acting on specific sites that interact directly with pathogen-derived proteins were detected in both MHC classes. Furthermore, a high genetic differentiation at MHC I was observed between Namibian cheetahs from east-central and north-central regions known to differ substantially in exposure to feline-specific viral pathogens. This suggests that the patterns of MHC I variation in the current population mirrors different pathogenic selective pressure imposed by viruses. Cheetahs showed low levels of MHC diversity compared with other mammalian species including felids, but this does not seem to influence the current immunocompetence of free-ranging cheetahs in Namibia and contradicts the previous conclusion that the cheetah is a paradigm species of disease susceptibility. However, it cannot be ruled out that the low MHC variation might limit a prosperous immunocompetence in the case of an emerging disease scenario because none of the remaining alleles might be able to recognize a novel pathogen. In contrast to cheetahs, leopards occur in most parts of Africa being perhaps the most abundant big cat in the continent. Leopards seem to have escaped from large-scale declines due to epizootics in the past in contrast to some free-ranging large carnivore populations in Africa that have been afflicted by epizootics. Currently, no information about the MHC sequence variation and constitution in African leopards exists. In this study, I characterized genetic variation at MHC I and MHC II genes in free-ranging leopards from Namibia. A total of six MHC I and six MHC II sequences were detected in 25 individuals from the east-central region. The maximum number of sequences observed per individual suggests that they likely correspond to at least three MHC I and three MHC II genes. Hallmarks of MHC evolution were confirmed such as historical positive selection, recombination and trans-species polymorphism. The low MHC variation detected in Namibian leopards is not conclusive and further research is required to assess the extent of MHC variation in different areas of its geographic range. Results from this thesis will contribute to better understanding the evolutionary significance of MHC and conservation implications in free-ranging felids. Translocation of wildlife is an increasingly used management tool for conservation purposes that should be conducted carefully as it may affect the ability of the translocated animals to cope with different pathogenic selective pressures.
Subcellular compartmentation of primary carbon metabolism in mesophyll cells of Arabidopsis thaliana
(2011)
Metabolism in plant cells is highly compartmented, with many pathways involving reactions in more than one compartment. For example, during photosynthesis in leaf mesophyll cells, primary carbon fixation and starch synthesis take place in the chloroplast, whereas sucrose is synthesized in the cytosol and stored in the vacuole. These reactions are tightly regulated to keep a fine balance between the carbon pools of the different compartments and to fulfil the energy needs of the organelles. I applied a technique which fractionates the cells under non-aqueous conditions, whereby the metabolic state is frozen at the time of harvest and held in stasis throughout the fractionation procedure. With the combination of non-aqueous fractionation and mass spectrometry based metabolite measurements (LC-MS/MS, GC-MS) it was possible to investigate the intracellular distributions of the intermediates of photosynthetic carbon metabolism and its products in subsequent metabolic reactions. With the knowledge about the in vivo concentrations of these metabolites under steady state photosynthesis conditions it was possible to calculate the mass action ratio and change in Gibbs free energy in vivo for each reaction in the pathway, to determine which reactions are near equilibrium and which are far removed from equilibrium. The Km value and concentration of each enzyme were compared with the concentrations of its substrates in vivo to assess which reactions are substrate limited and so sensitive to changes in substrate concentration. Several intermediates of the Calvin-Benson cycle are substrates for other pathways, including dihydroxyacetone-phosphate (DHAP,sucrose synthesis), fructose 6-phosphate (Fru6P, starch synthesis), erythrose 4-phosphate (E4P,shikimate pathway) and ribose 5-phosphate (R5P, nucleotide synthesis). Several of the enzymes that metabolise these intermediates, and so lie at branch points in the pathway, are triose-phosphate isomerase (DHAP), transketolase (E4P, Fru6P), sedoheptulose-1,7-bisphosphate aldolase (E4P) and ribose-5-phosphate isomerase (R5P) are not saturated with their respective substrate as the metabolite concentration is lower than the respective Km value. In terms of metabolic control these are the steps that are most sensitive to changes in substrate availability, while the regulated irreversible reactions of fructose-1,6-bisphosphatase and sedoheptulose-1,7-bisphosphatase are relatively insensitive to changes in the concentrations of their substrates. In the pathway of sucrose synthesis it was shown that the concentration of the catalytic binding site of the cytosolic aldolase is lower than the substrate concentration of DHAP, and that the concentration of Suc6P is lower than the Km of sucrose-phosphatase for this substrate. Both the sucrose-phosphate synthase and sucrose-phosphatase reactions are far removed from equilibrium in vivo. In wild type A. thaliana Columbia-0 leaves, all of the ADPGlc was found to be localised in the chloroplasts. ADPglucose pyrophosphorylase is localised to the chloroplast and synthesises ADPGlc from ATP and Glc1P. This distribution argues strongly against the hypothesis proposed by Pozueta-Romero and colleagues that ADPGlc for starch synthesis is produced in the cytosol via ADP-mediated cleavage of sucrose by sucrose synthase. Based on this observation and other published data it was concluded that the generally accepted pathway of starch synthesis from ADPGlc produced by ADPglucose pyrophosphorylase in the chloroplasts is correct, and that the alternative pathway is untenable. Within the pathway of starch synthesis the concentration of ADPGlc was found to be well below the Km value of starch synthase for ADPGlc, indicating that the enzyme is substrate limited. A general finding in the comparison of the Calvin-Benson cycle with the synthesis pathways of sucrose and starch is that many enzymes in the Calvin Benson cycle have active binding site concentrations that are close to the metabolite concentrations, while for nearly all enzymes in the synthesis pathways the active binding site concentrations are much lower than the metabolite concentrations.
AM symbiosis has a positive influence on plant P-nutrition and growth, but little is known about the molecular mechanism of the symbiosis adaptation to different phosphate conditions. The recently described induction of several pri-miR399 transcripts in mycorrhizal shoots and subsequent accumulation of mature miR399 in mycorrhizal roots indicates that local PHO2 expression must be controlled during symbiosis, presumably in order to sustain AM symbiosis development, in spite of locally increased Pi-concentration. A reverse genetic approach used in this study demonstrated that PHO2 and thus the PHR1-miR399-PHO2 signaling pathway, is involved in certain stages of progressive root colonization. In addition, a transcriptomic approach using a split-root system provided a comprehensive insight into the systemic transcriptional changes in mycorrhizal roots and shoots of M. truncatula in response to high phosphate conditions. With regard to the transcriptional responses of the root system, the results indicate that, although the colonization is drastically reduced, AM symbiosis is still functional at high Pi concentrations and might still be beneficial to the plant. Additionally, the data suggest that a specific root-borne mycorrhizal signal systemically induces protein synthesis, amino acid metabolism and photosynthesis at low Pi conditions, which is abolished at high Pi conditions. MiRNAs, such as miR399, are involved in long-distance signaling and are therefore potential systemic signals involved in AM symbiosis. A deep-sequencing approach identified 243 novel miRNAs in the root tissue of M. truncatula. Read-count analysis, qRT-PCR measurements and in situ hybridizations clearly indicated a regulation of miR5229a/b, miR5204, miR160f*, miR160c, miR169 and miR169d*/l*/m*/e.2* during arbuscular mycorrhizal symbiosis. Moreover, miR5204* represses a GRAS TF, which is specifically transcribed in mycorrhizal roots. Since miR5204* is induced by high Pi it might represent a further Pi status-mediating signal beside miR399. This study provides additional evidence that MtNsp2, a key regulator of symbiosis-signaling, is regulated and presumably spatially restricted by miR171h cleavage. In summary, a repression of mycorrhizal root colonization at high phosphate status is most likely due to a repression of the phosphate starvation responses and the loss of beneficial responses in mycorrhizal shoots. These findings provide a new basis for investigating the regulatory network leading to cellular reprogramming during interaction between plants, arbuscular mycorrhizal fungi and different phosphate conditions.