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La tasa de cuajado es un factor determinante en la producción de un cultivar. Este proceso, regulado en gran medida por señales hormonales, representa la primera etapa del desarrollo de un fruto y depende del éxito de la polinización y de la fecundación. A pesar de su importancia agronómica, los mecanismos moleculares que regulan el proceso del cuajado del fruto son aún desconocidos. La identificación y caracterización de mutantes afectados en la tasa de cuajado es una de las mejores estrategias para lograr este propósito. En este trabajo de investigación hemos abordado la caracterización de tres mutantes insercionales de tomate afectados en la tasa de cuajado de fruto. Dos de estos mutantes, Lower fruit setting 2084 y 2448 (Lfs2084 y Lfs2448), identificados en el contexto de esta Tesis Doctoral, exhiben menor tasa de cuajado de fruto y presentan un patrón de herencia autosómica dominante. La caracterización de ambas mutaciones ha permitido descubrir que están asociadas a un fenómeno de letalidad del gameto masculino. La mutación Lfs2084 promueve la degeneración de los granos de polen que portan el inserto, pero también afecta al desarrollo reproductivo del ovario, lo que reduce de forma dramática la tasa de cuajado de fruto. El análisis del contenido endógeno de hormonas de este mutante parece indicar una alteración de la ruta del 2C-metil-Deritrol 4-fosfato (MEP), responsable de la síntesis de fitohormonas, pigmentos fotosintéticos y moléculas de defensa. Los menores contenidos de GA1 en Lfs2084, la giberelina activa más importante en el inicio del desarrollo del fruto de tomate, podrían estar asociados con una mayor actividad del enzima catabólico SlGA2ox2. Estos resultados, junto con los cambios en los perfiles de expresión de SlARF7 y TAG1, son coherentes con el fenotipo de menor tasa de cuajado observado en el mutante. La mutación Lfs2448 también promueve la degeneración de los granos de polen portadores del inserto, reduciendo en gran medida la cantidad de tubos polínicos que progresan a lo largo del estilo durante el proceso de polinización. Su caracterización molecular ha indicado que el fenotipo de Lfs2448 podría estar ocasionado por la alteración de la función de uno o dos genes que codifican proteínas CYP450 y que se expresan mayoritariamente en botones florales y frutos de 1 cm. Los resultados sugieren que la mutación desempeña un relevante papel en el desarrollo reproductivo de tomate, afectando al proceso de polinización y al cuajado de fruto. Además, la caracterización del mutante Arlequín (Alq) ha revelado alteraciones en su programa de desarrollo vegetativo y reproductivo. Respecto al desarrollo vegetativo, Alq promueve un patrón de crecimiento más vigoroso que el que exhiben las líneas hermanas que carecen de la mutación, así como cambios en la configuración espacial del desarrollo radicular. Con relación al desarrollo reproductivo, Alq conduce a un mayor cuajado de fruto como consecuencia de eventos de división celular en el pericarpio del ovario previos al proceso de polinización. Un aspecto esencial desde un punto de vista agronómico es que la mayor tasa de cuajado permite que Alq mantenga la producción de fruto en condiciones de moderada salinidad. / Fruit setting rate is one of the most determining factors in the yield of a cultivar. This process, largely regulated by hormonal signals, is the first stage of fruit development and depends on the successful completion of pollination and fertilization. Despite their agronomic importance, the molecular mechanisms regulating the fruit setting process are still unknown. The identification and characterization of mutations affecting fruit set rate is one of the best strategies to achieve this purpose. In this research work the characterization of three tomato insertional mutants altered in the fruit set rate has been addressed. Two of these mutants, Lower fruit setting 2084 y 2448 (Lfs2084 and Lfs2448), identified in the context of this Doctoral Thesis, exhibit decreased fruit set rate and present an autosomal dominant inheritance pattern. The characterization of both mutations allowed us to discover that they are associated with a phenomenon of lethality of the male gametes carrying the T-DNA insert. The Lfs2084 mutation not only promotes degeneration of pollen grains bearing the T-DNA, but also affects the reproductive development of the ovary, dramatically reducing the fruit set rate. Analysis of the endogenous hormone content of this mutant suggests an alteration of the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway, responsible for the synthesis of phytohormones, photosynthetic pigments and plant defence molecules. The lower GA1 content in Lfs2084 mutant, the main active form to induce fruit development, may be associated with increased activity of the SlGA2ox2 catabolic enzyme. These results, together with the changes in the expression profiles of both SlARF7 and TAG1 genes, are consistent with the phenotype of lower fruit set rate. The Lfs2448 mutation also promotes degeneration of the pollen grains carrying the T-DNA, largely reducing the number of pollen tubes that progress along the style during the pollination process. Its molecular characterization indicated that the Lfs2448 mutant phenotype could be caused by the functional alteration of one or two genes that encode CYP450 proteins mostly expressed in floral buds and young fruits. Results suggest that the tagged gene plays an important role in tomato reproductive development, affecting both normal pollination and fruit set process. Furthermore, the characterization of the Arlequín (Alq) mutant revealed alterations in its vegetative and reproductive developmental program. As regards vegetative development, Alq mutation promotes a more vigorous growth pattern, as well as changes in the spatial configuration of root system. Regarding reproductive development, Alq mutation leads to increased fruit set as a consequence of cell division events in the ovary pericarp prior to the pollination process. Finally, it should be noted that, thanks to its higher fruit set rate, the Arlequin mutant is capable of maintaining fruit production in moderate salinity conditions, which is of great importance from an agronomic point of view. / La taxa de quallat és un factor determinant en la producció d`un cultivar. Aquest procés, regulat en gran mesura per senyals hormonals, representa la primera etapa del desenvolupament d¿un fruit i depén de l¿èxit de la pol·linització i de la fecundació. A pesar de la seua importància agronòmica, els mecanismes moleculars que regulen el procés de quallat són encara desconeguts. La identificació i caracterització de mutants afectats en la taxa de quallat és una de les millors estratègies per aconseguir aquest propòsit. En aquest treball d¿investigació hem abordat la caracterització de tres mutants insercionals de tomaca afectats en la taxa de quallat de fruit. Dos d¿aquests mutants, Lower fruit setting 2084 i 2448 (Lfs2084 i Lfs2448), identificats en el context d¿aquesta Tesi Doctoral, exhibixen una menor taxa de quallat de fruit i presenten un patró d¿herència autosòmica dominant. La caracterització d¿ambdós mutacions ha permés descobrir que es troben associades a un fenomen de letalitat del gàmeta masculí. La mutació Lfs2084 promou la degeneració dels grans de pol·len que porten l¿insert, però també afecta el desenvolupament reproductiu de l¿ovari, la qual cosa reduïx de forma dramàtica la taxa de quallat de fruit. L¿anàl·lisi del contingut endogen d¿hormones d¿aquest mutant pareix indicar una alteració de la ruta 2C-metil-Deritrol 4-fosfato (MEP), responsable de la síntesi de fitohormones, pigments fotosintètics i molècules de defensa. Els menors continguts de GA1 en Lfs2084, la gibberel·lina activa més important a l'inici del desenvolupament del fruit de tomaca, podrien estar associats amb una major activitat de l¿enzim catabòlic SlGA2ox2. Aquests resultats, juntament amb els canvis en els perfils d¿expressió de SlARF7 i TAG1, són coherents amb el fenotip de menor taxa de quallat observat en el mutant. La mutació Lfs2448 també promou la degeneració dels grans de pol·len portadors de l¿insert, reduint en gran mesura la quantitat de tubs pol·línics que progressen al llarg de l¿estil durant el procés de pol·linització. La seua caracterització molecular ha indicat que el fenotip de Lfs2448 podria estar ocasionat per una alteració de la funció d¿un o dos gens que codifiquen proteïnes CYP450 i que s'expressen majoritàriament en botons florals i fruits d¿un centímetre. Els resultats suggerixen que la mutació té una funció important en el desenvolupament reproductiu de la tomaca, afectant el procés normal de pol·linització i el quallat de fruit. A més, la caracterització del mutant Arlequín (Alq) ha revelat alteracions en el seu programa de desenvolupament vegetatiu i reproductiu. Pel que fa al desenvolupament vegetatiu, Alq promou un patró d¿expressió més vigorós que el que exhibixen les línies germanes que no porten la mutació, així com canvis en la configuració espacial del desenvolupament radicular. Quant al desenvolupament reproductiu, Alq conduïx a un major quallat de fruit com a conseqüència d¿events de divisió cel·lulars en el pericarp de l¿ovari previs al procés de pol·linització. Un aspecte essencial des d¿un punt de vista agronòmic és que la major taxa de quallat pemet que Alq mantinga la producció de fruit en condicions de moderada salinitat. / Ribelles Alfonso, C. (2018). IDENTIFICACIÓN Y CARACTERIZACIÓN DE MUTANTES DE TOMATE RELACIONADOS CON EL CUAJADO DEPENDIENTE E INDEPENDIENTE DE POLINIZACIÓN [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/111833 / TESIS

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Harmful substances containing acid or neutral rock drainages (ARD and NRD) are a major challenge related to the management of extractive industry wastes. This issue is particularly related to deposits containing sulphide minerals, which are prone to oxidization under the influence of atmospheric oxygen and water. The drainage quality depends mainly on the mineralogical and chemical composition of the extractive wastes, and especially on the ratio of acid-producing and neutralizing minerals, combined with reactions catalysed by microbes. Since harmful drainages play a major role in the generation of environmental issues for extractive industry, the accurate prediction of the drainage quality is of utmost importance. To design appropriate extractive waste facilities and drainage management, the characterisation of extractive wastes and assessment of the behaviour of the waste material is essential already before the actual mining activities start. Several methods have been developed to characterize extractive waste materials and to predict their short and long term behaviour, including e.g. geochemical laboratory tests, static tests and longer term kinetic tests, and geochemical modelling. The characterisation methods for assessing the ARD risk can be divided into static and kinetic tests. Static tests are short term laboratory analyses, usually used for preliminary investigation and screening. Kinetic tests are longer term tests, revealing information on the time scale of drainage events. Commonly used static tests for ARD prediction include acid–base accounting (ABA) tests and the net acid generation (NAG) test. Since acid and neutralisation potential largely depend on the ratio and quality of acid-producing and neutralizing minerals, mineralogical calculations could also be used for ARD prediction. The mobility of potentially harmful substances from extractive waste can be preliminary assessed using different geochemical laboratory tests, including selective extraction and leaching methods. The most commonly used selective extraction method in Finland is the aqua regia (AR) extraction. In addition to some silicates and secondary precipitate minerals, it is intended to dissolve elements bound especially to sulphide phases. A less commonly used method for element mobility prediction is the analysis of the single addition NAG test leachate. In this study, several Finnish waste rock sites were investigated and the performances of different preliminary drainage quality test methods evaluated and compared. The assessed acid production potential methods included the ABA test as presented in the standard EN 15875, the single addition NAG test as presented in the AMIRA guidebook, and a SEM mineralogy-based calculation. The assessed methods for element mobility prediction included the single addition NAG test leachate analysis and the AR extraction. According to the results, pyrrhotite seems to be the main mineral contributing to acid production, and the silicate minerals the main contributors to the neutralisation potential at the most Finnish waste rock sites. Since silicate minerals appear to have a significant role in ARD prevention, the behaviour of these minerals in mining environment should be more thoroughly investigated. In the investigated Finnish waste rocks, Co, Cr, Cu and Ni often occurred as elevated concentrations, and the most widely abundant harmful elements in the waste rock drainages were Co, Cu, Ni and Zn. The results suggest that an acid production prediction based on SEM mineralogical calculation is at least as accurate as the commonly used static laboratory methods. The AR extraction indicates well which elements might occur as elevated concentrations in the drainage. Also the NAG test leachate analysis performed well in element mobility assessment, but only when the NAG test leachate was sufficiently acidic, the leachate pH being below of 3-6, depending on the element of interest.

Biotic and abiotic stresses are signifi cant factors limiting the production of food and other supporting materials required to sustain increasing world population. Plant health is directly related to human health and is increasingly becoming signifi cant and demands more attention towards limiting the damages caused by biotic and abiotic stresses. Signifi cant progress has been made towards our understanding of the processes, which mediate both biotic and abiotic stress signaling in plants. Signifi cant role is played by various plant hormones, e.g., salicylic acid (SA) and jasmonic acid (JA) in biotic stress and abscisic acid (ABA) in abiotic stress (Annu Rev Cell Dev Biol 28:489-521, 2012). Other hormones with minor role include the cytokinins (CK), auxins (indole 3 acetic acid. IAA), and the brassinosteroids (BR) (Annu Rev Cell Dev Biol 28:489-521, 2012). Cross talk between these plant hormones is signifi cant and may result in either synergistic or antagonistic effect on stress responses (Annu Rev Cell Dev Biol 28:489-521, 2012). In recent years, extensive research carried out in various laboratories has implicated cross talk between the ABA and the SA in abiotic stress response. This is signifi cant in light of SA being key player in biotic stress responses in plants. This review will discuss the role of SA in biotic and abiotic stress signaling and its cross talk with other hormones in mediating abiotic stress signaling in plants.

Ett cirkulärt samhälle strävar efter att efterlikna naturens kretslopp där material som blir över från en verksamhet används i en annan, för att minska uttag av jungfruliga resurser. Från verksamheter kopplade till vägbyggen är det främst geologiska material som hanteras. Bergmaterial kan innehålla sulfidmineral som vid oxidation kan producera syra. För att säkerställa att bergmaterialet kan användas på ett säkert sätt används olika predikteringsmetoder för att identifiera risker med materialet, projektet har studerat några av dessa predikteringsmetoder och resultatet visar att de ger mycket osäkra bedömningar av det studerade materialet. I detta examensarbete har 10 bergmaterial studerats med syfte att undersöka och diskutera predikteringsmetoder och hur dessa kan tillämpas och tolkas för sulfidförande berg.  Bergmaterialet har en kemiskt analyserad svavelhalt mellan 0,3 och 1,6 vikts-%. Undersökningen inkluderar statiska och kinetiska tester. De statiska tester som använts är ABA och NAGpH, mineralogiska studier, både kvalitativt och i optiskt mikroskop och kvantitativt i SEM, XRF-mätningar, även ett enkelt laktest har utförts på materialet. Kinetiskt fuktkammartest (40 veckor) har inom ramen för projektet påbörjats på tre av bergmaterialen. Resultaten av predikteringstesterna visar på en stor variation mellan materialen utifrån använda riktvärden. Bedömningen av syrapotentialen och neutralisationspotentialen visar också stora variationer utifrån nyttjade riktlinjer. Det finns behov att vidare undersöka vad det är som gör att predikteringsmetoderna ger det resultat de ger och vidare hur detta kan tolkas. Statiska predikteringstester visar inte heller någon tidsaspekt som är viktigt att betrakta vid prediktering av material. Kinetiska test, som fuktkammartest visar hur utlakningen varierar med tiden.  Hur provtagningen av materialet utförs har också en stor påverkan på resultatet, det är därför viktigt att fortsatt utreda hur en representativ provtagning kan utföras, och hur olika provtagningsmetoder påverkar analysresultaten / In a circular society that strives to close the cycle of resources, usage of rest material from one operation area to another plays a natural role. In operations connected to roadworks geological material are mainly handled. Geological material can contain sulfide minerals which can through oxidation produce acid. To ensure that the material can be used in constructions different prediction methods are used to estimate risks with the material. In this master thesis prediction methods has been studied on 10 different materials, the results shows that the studied prediction methods gives a big variation in results. 10 rock samples has been studied with the aim to investigate and discuss prediction methods for sulfidic rock.  The rock samples had a sulfur content of 0,3 to 1,6 weight- %. The studied prediction methods included static test as Acid Base Accounting (ABA) and Net Acid Generation (NAGpH), mineralogical studies on the material. A kinetic test, humidity cell test (HCT) was started up in this project but not completed. Full rock chemistry and a simple leaching test was also performed on the material. The results from the prediction test on the material shows a large variation between the materials and when compared to different guideline values interpreted by different authorities. Determination of the neutralization and acid potential of the material also present a difference. There is a need to further investigate how the results from the studied prediction methods should be interpreted, and which tests are appropriate to use depending on where the material are planned to be used. How the sampling and sample preparation is preformed will also affect the result from prediction test. Therefore it´s important to investigate how representative sampling can be done to fulfill the purpose and how different sampling and preparation methods affects the results from analysis.

Having a child with autism results in a unique set of challenges for both parents and siblings. Applied behavior analysis (ABA) has significant empirical support demonstrating its effectiveness as a treatment for children with autism. However, effective behavioral interventions should also consider how the family is affected. The purpose of this phenomenological study was to explore the lived experiences of 8 parents of children with autism who have received ABA treatment and their perceptions of how the family system is impacted by ABA outcomes, specifically focusing on parental well-being and sibling relationship quality. A family systems theoretical framework and phenomenological research method were used. Data were analyzed using content analysis to find the common themes that emerged, which were: parents of children with autism face a high level of stress, but ABA treatment results in a decrease of some sources of stress. Having a child with autism produces strains in family relationships; however, ABA treatment outcomes can help with improved communication between spouses and interactions between siblings. Parents acknowledge challenges associated with ABA treatment but believe that the benefits are worth it. Insights gained from this study were meaningful and practical for professionals who provide ABA treatment to children with autism, as well as for parents of children with autism who may be interested in seeking ABA treatment. Positive social change resulted from the identification of perceived benefits of ABA treatment, such as reduced parental stress and improved relationships amongst all members of the family.

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Abscisic acid is an important plant hormone in the responses to biotic and abiotic stresses, which also regulates various growth and developmental processes in plants. Three major components-receptors (PYRs), the PP2C type phosphatases and the SnRK2 subtype kinases form a double negative regulatory system: PYR/PYL/RCARs inhibit the activity of PP2Cs while PP2Cs inhibit that of SnRK2s in ABA signaling pathway. The results of my studies showed that ABA would directly affect the interaction between SnRK2.2 and ABI1 in absence of PYRs. Furthermore, ABA can inhibit the catalytic activity of the SnRK2.2 kinase. These findings indicated that ABA may directly interact with SnRK2.2. Posttranslational modifications play a key role in signal transduction. Phosphorylation is the most important posttranscriptional modification in ABA signal transduction. To dissect new components in ABA signaling network in plants, proteomics studies were carried out in Arabidopsis for identifying ABA- responsive phosphoproteins. Ten phosphoproteins, ATPB, ATPC1, FBA1, CTIMC, GGAT1, GAPC1, GAPC2, GAPA1 and ALDH11A3, were identified by 2-DE proteomic approach and LC-MS/MS analysis. These proteins are likely to be the potential phosphorylated targets of SnRK2s in ABA signaling network. Lysine acetylation (LysAc) also emerges as one of the important posttranslational modifications for protein regulation in plants. Eleven lysine acetylated proteins with altered acetylation in response to ABA were identified in Arabidopsis using proteomic approach. The increased LysAc of Rubisco and the decreased Rubisco activity by ABA treatment indicates the acetylation of Rubisco caused by ABA resulted in the inhibition of Rubisco activity. ABA has also been shown to exist and function in both lower animals and mammalians. The medical application of ABA has become a new area of investigation. To explore the role of protein phosphorylation in ABA-mediated function in mammalians, phosphoproteomic study was conducted in mouse 3T3-L1 cells. Ten phosphoproteins with significant changes in serine/threonine phosphorylation in response to ABA were identified. These results suggest these phosphoproteins are involved in ABA signaling network in mouse cells. The significance of the function of SFRS1, ANXA1 and Galectin-3 on human diseases indicated that ABA could be a potential treatment for some human diseases, such as cancer.

One goal of this study was to evaluate the emergence of mands and intraverbals following tact acquisition for individuals with aphasia due to acquired brain injury. A second goal was to evaluate the transfer of shortened latencies as a function of tact training across untrained operants. In Study 1, the dependent measure was accuracy of responding and in Study 2, the dependent measures were rate and latency of responding. Participants for Study 1 were two uninjured adults (pilot) and two adults with brain injury (ABI). Both sets of participants were directly taught to tact up to 6 stimuli. Once tacts were acquired, the response forms were assessed under mand and intraverbal conditions. All pilot participants and one ABI participant showed mand transfer for all stimuli. Tact to intraverbal transfer varied across participants. One adult with brain injury served as a participant for Study 2. Fluency training was used to teach tacts for 15 stimuli. Response latencies were gathered for all operants before and after training. The participant met the designated aim (rate of responding) and showed a decrease in latencies for tacts and untrained intraverbals. Changes in mand latencies varied. Fluency gains showed partial retention. Results from Study 1 provide further evidence for interdependence across operants during rehabilitative training. Results from Study 2 provide preliminary support for the utility of fluency training for verbal behavior after brain injury. Future research should explore the components of fluency training and their effects on the verbal behavior of individuals with ABI.