Effects of elevated CO2 on fish behaviour undiminished by transgenerational acclimation

Behaviour and sensory performance of marine fishes are impaired at CO2 levels projected to occur in the ocean in the next 50-100 years, and there is limited potential for within-generation acclimation to elevated CO2. However, whether fish behaviour can acclimate or adapt to elevated CO2 over multiple generations remains unanswered. We tested for transgenerational acclimation of reef fish olfactory preferences and behavioural lateralization at moderate (656 µatm) and high (912 µatm) end-of-century CO2 projections. Juvenile spiny damselfish, Acanthochromis polyacanthus, from control parents (446 µatm) exhibited an innate avoidance to chemical alarm cue (CAC) when reared in control conditions. In contrast, juveniles lost their innate avoidance of CAC and even became strongly attracted to CAC when reared at elevated CO2 levels. Juveniles from parents maintained at mid-CO2 and high-CO2 levels also lost their innate avoidance of CAC when reared in elevated CO2, demonstrating no capacity for transgenerational acclimation of olfactory responses. Behavioural lateralization was also disrupted for juveniles reared under elevated CO2, regardless of parental conditioning. Our results show minimal potential for transgenerational acclimation in this fish, suggesting that genetic adaptation will be necessary to overcome the effects of ocean acidification on behaviour.

Independent component analysis for the identification of sources of variation on an industrial nirs application

A Near Infrared Spectroscopy (NIRS) industrial application was developed by the LPF-Tagralia team, and transferred to a Spanish dehydrator company (Agrotécnica Extremeña S.L.) for the classification of dehydrator onion bulbs for breeding purposes. The automated operation of the system has allowed the classification of more than one million onion bulbs during seasons 2004 to 2008 (Table 1). The performance achieved by the original model (R2=0,65; SEC=2,28ºBrix) was enough for qualitative classification thanks to the broad range of variation of the initial population (18ºBrix). Nevertheless, a reduction of the classification performance of the model has been observed with the passing of seasons. One of the reasons put forward is the reduction of the range of variation that naturally occurs during a breeding process, the other is the variations in other parameters than the variable of interest but whose effects would probably be affecting the measurements [1]. This study points to the application of Independent Component Analysis (ICA) on this highly variable dataset coming from a NIRS industrial application for the identification of the different sources of variation present through seasons.

Fusarium proliferatum isolated from garlic in Spain: Identification, toxigenic potential and pathogenicity on related Allium species

Fusarium proliferatum has been reported on garlic in the Northwest USA, Spain and Serbia, causing water-soaked tan-colored lesions on cloves. In this work, Fusarium proliferatum was isolated from 300 symptomatic garlic bulbs. Morphological identification of Fusarium was confirmed using species-specific PCR assays and EF-1α sequencing. Confirmation of pathogenicity was conducted with eighteen isolates. Six randomly selected F. proliferatum isolates from garlic were tested for specific pathogenicity and screened for fusaric acid production. Additionally, pathogenicity of each F. proliferatum isolate was tested on healthy seedlings of onion (Allium cepa), leek (A. porrum), scallions (A. fistulosum), chives (A. schoenoprasum) and garlic (A. sativum). A disease severity index (DSI) was calculated as the mean severity on three plants of each species with four test replicates. Symptoms on onion and garlic plants were observed three weeks after inoculation. All isolates tested produced symptoms on all varieties inoculated. Inoculation of F. proliferatum isolates from diseased garlic onto other Allium species provided new information on host range and pathogenicity. The results demonstrated differences in susceptibility with respect to host species and cultivar. The F. proliferatum isolates tested all produced fusaric acid (FA); correlations between FA production and isolate pathogenicity are discussed. Additionally, all isolates showed the presence of the FUM1 gene suggesting the ability of Spanish isolates to produce fumonisins.

The effects of storage duration, temperature and cultivar on the severity of garlic clove rot caused by Fusarium proliferatum.

Diseases that affect garlic during storage can lead to severe economic losses for farmers worldwide. One causal agent of clove rot is Fusarium proliferatum. Here, the progress of clove rot caused by F. proliferatum and its dependence on different storage conditions and cultivar type were studied. The effect of temperature on mycelial growth, conidial viability, and fungal survival during garlic commercial storage was documented. Samples of 50 bulbs from a randomized field trial with three different clonal generations for purple garlic (F3, F4 and F5) and the F4 clonal generation for white garlic were labeled and stored for two months (short-term storage). In addition, another sample of the F5 clonal generation of purple garlic was stored for 6 months after harvest (long-term storage). The presence of the pathogen and the percentage of symptomatic cloves were evaluated. A notable difference in the rot severity index (RSI) of different garlic varieties was observed. In all studied cases, clove rot increased with storage time at 20 ◦ C, and the white garlic variety had a higher index of rot severity after two months of storage. Additionally, there were clear differences between the growth rates of F. proliferatum isolates. Studies conducted on the temperature responses of the pathogen propagules showed that expo- sure for at least 20 min at 50 ◦ C was highly effective in significantly reducing the viability of fungal conidia. Pathogenicity studies showed that the fungus is pathogenic in all commercial varieties. However, there were significant differences in varietal susceptibility between Chinese and white garlic type cultivars (81.84 ± 16.44% and 87.5 ± 23.19% symptomatic cloves, respectively) and purple cultivars (49.06 ± 13.42% symptomatic cloves)

VLC oriented energy efficient driver techniques

LEDs are substituting fluorescent and incandescent bulbs as illumination sources due to their low power consumption and long lifetime. Visible Light Communications (VLC) makes use of the LEDs short switching times to transmit information. Although LEDs switching speed is around Mbps range, higher speeds (hundred of Mbps) can be reached by using high bandwidth-efficiency modulation techniques. However, the use of these techniques requires a more complex driver which elevates drastically its power consumption. In this work an energy efficiency analysis of the different VLC modulation techniques and drivers is presented. Besides, the design of new schemes of VLC drivers is described.

Caracterización de LEDs mediante el uso y programación de espectrómetro

En la búsqueda de dispositivos cada vez más eficientes, de larga duración y bajo coste de mantenimiento en el mundo de la iluminación, aparecen los LEDs. Estos pequeños dispositivos van poco a poco sustituyendo a las bombillas tradicionales de incandescencia, tomando un papel cada vez más importante entre las fuentes de iluminación. Las primeras funciones prácticas que tuvieron estos LEDs fueron como indicadores, y sus primeros usos fueron en pantallas de calculadoras, electrodomésticos, etcétera, y más adelante, con el desarrollo de nuevos materiales, se empezaron a utilizar como dispositivos de iluminación. Ha sido en estos últimos años cuando se ha producido un salto cuantitativo gracias a la aparición de los POWER LED (LEDs de potencia) o de alto brillo, que son los que han permitido ampliar el uso de estos dispositivos como fuentes de iluminación en, por ejemplo, hogares, alumbrado público, e incluso llegando a sustituir los faros halógenos de vehículos por iluminación LED en algunos modelos. Es por ello que mientras su potencia lumínica va aumentado, su rango de utilización también lo hace. Para caracterizar estas fuentes lumínicas y otras a las que se les pueden dar diferentes usos, se desarrolla este proyecto mediante el análisis de su espectro. Para ello, además, se hará un análisis del resto de instrumentación necesaria que forma parte del proyecto. Este análisis abarca el estudio del propio espectrómetro tanto a nivel de hardware como de software, que modificaremos según los intereses del proyecto. También se estudiará la fibra óptica y el driver para controlar los dispositivos LEDs de potencia, así como los propios LEDs. Para ello se medirán las características de estos LEDs y se compararán con las facilitadas por el fabricante. ABSTRACT. Searching for more efficient, long lasting an low-maintenance devices in lighting world, LEDs appear. These small devices are gradually replacing traditional incandescent bulbs. LEDs are taking an increasingly important role between the light sources. At the begining they were only used as indicators and their first use were in screens calculators, appliances, etc., and later, with the development of new materials, were progressively used as lighting devices. Nowadays a great development has happened in LED lighting with the apparition of the POWER LED or high bright. Power LEDs are allowed to extend the use of these devices as lighting sources for example for homes, street lighting, and even coming to replace halogen headlights LED lighting in vehicles of some models. That's the reason the more their lighting power increases the more their use increases too. The aim of this project is to characterize these light sources and others that can be given different uses by analyzing its spectrum. Moreover, necessary instruments will also be analysed. This study involves both hardware and software spectrometer analysis itself by modifying its software according to the interests of the project. Furthermore, optical fiber and the driver to control LED power devices will be studied by measuring LEDs characteristics and comparing with those provided by the manufacturer.

User experience with immersive and interactive media = Experiencia de usuario en entornos 3D inmersivos e interactivos

Estudios recientes promueven la integración de estímulos multisensoriales en activos multimedia con el fin de mejorar la experiencia de usuario mediante la estimulación de nuevos sentidos, más allá de la tradicional experiencia audiovisual. Del mismo modo, varios trabajos proponen la introducción de componentes de interacción capaces de complementar con nuevas características, funcionalidades y/o información la experiencia multimedia. Efectos sensoriales basados en el uso de nuevas técnicas de audio, olores, viento, vibraciones y control de la iluminación, han demostrado tener un impacto favorable en la sensación de Presencia, en el disfrute de la experiencia multimedia y en la calidad, relevancia y realismo de la misma percibidos por el usuario. Asimismo, los servicios basados en dos pantallas y la manipulación directa de (elementos en) la escena de video tienen el potencial de mejorar la comprensión, la concentración y la implicación proactiva del usuario en la experiencia multimedia. El deporte se encuentra entre los géneros con mayor potencial para integrar y explotar éstas soluciones tecnológicas. Trabajos previos han demostrado asimismo la viabilidad técnica de integrar éstas tecnologías con los estándares actualmente adoptados a lo largo de toda la cadena de transmisión de televisión. De este modo, los sistemas multimedia enriquecidos con efectos sensoriales, los servicios interactivos multiplataforma y un mayor control del usuario sobre la escena de vídeo emergen como nuevas formas de llevar la multimedia immersiva e interactiva al mercado de consumo de forma no disruptiva. Sin embargo, existen numerosas interrogantes relativas a los efectos sensoriales y/o soluciones interactivas más adecuadas para complementar un contenido audiovisual determinado o a la mejor manera de de integrar y combinar dichos componentes para mejorar la experiencia de usuario de un segmento de audiencia objetivo. Además, la evidencia científica sobre el impacto de factores humanos en la experiencia de usuario con estas nuevas formas de immersión e interacción en el contexto multimedia es aún insuficiente y en ocasiones, contradictoria. Así, el papel de éstos factores en el potencial de adopción de éstas tecnologías ha sido amplia-mente ignorado. La presente tesis analiza el impacto del audio binaural, efectos sensoriales (de iluminación y olfativos), interacción con objetos 3D integrados en la escena de vídeo e interacción con contenido adicional utilizando una segunda pantalla en la experiencia de usuario con contenidos de deporte. La posible influencia de dichos componentes en las variables dependientes se explora tanto a nivel global (efecto promedio) como en función de las características de los usuarios (efectos heterogéneos). Para ello, se ha llevado a cabo un experimento con usuarios orientado a explorar la influencia de éstos componentes immersivos e interactivos en dos grandes dimensiones de la experiencia multimedia: calidad y Presencia. La calidad de la experiencia multimedia se analiza en términos de las posibles variaciones asociadas a la calidad global y a la calidad del contenido, la imagen, el audio, los efectos sensoriales, la interacción con objetos 3D y la interacción con la segunda pantalla. El posible impacto en la Presencia considera dos de las dimensiones definidas por el cuestionario ITC-SOPI: Presencia Espacial (Spatial Presence) e Implicación (Engagement). Por último, los individuos son caracterizados teniendo en cuenta los siguientes atributos afectivos, cognitivos y conductuales: preferencias y hábitos en relación con el contenido, grado de conocimiento de las tecnologías integradas en el sistema, tendencia a involucrarse emocionalmente, tendencia a concentrarse en una actividad bloqueando estímulos externos y los cinco grandes rasgos de la personalidad: extroversión, amabilidad, responsabilidad, inestabilidad emocional y apertura a nuevas experiencias. A nivel global, nuestro estudio revela que los participantes prefieren el audio binaural frente al sistema estéreo y que los efectos sensoriales generan un aumento significativo del nivel de Presencia Espacial percibido por los usuarios. Además, las manipulaciones experimentales realizadas permitieron identificar una gran variedad de efectos heterogéneos. Un resultado interesante es que dichos efectos no se encuentran distribuidos de forma equitativa entre las medidas de calidad y Presencia. Nuestros datos revelan un impacto generalizado del audio binaural en la mayoría de las medidas de calidad y Presencia analizadas. En cambio, la influencia de los efectos sensoriales y de la interacción con la segunda pantalla se concentran en las medidas de Presencia y calidad, respectivamente. La magnitud de los efectos heterogéneos identificados está modulada por las siguientes características personales: preferencias en relación con el contenido, frecuencia con la que el usuario suele ver contenido similar, conocimiento de las tecnologías integradas en el demostrador, sexo, tendencia a involucrarse emocionalmente, tendencia a a concentrarse en una actividad bloqueando estímulos externos y niveles de amabilidad, responsabilidad y apertura a nuevas experiencias. Las características personales consideradas en nuestro experimento explicaron la mayor parte de la variación en las variables dependientes, confirmando así el importante (y frecuentemente ignorado) papel de las diferencias individuales en la experiencia multimedia. Entre las características de los usuarios con un impacto más generalizado se encuentran las preferencias en relación con el contenido, el grado de conocimiento de las tecnologías integradas en el sistema y la tendencia a involucrarse emocionalmente. En particular, los primeros dos factores parecen generar un conflicto de atención hacia el contenido versus las características/elementos técnicos del sistema, respectivamente. Asimismo, la experiencia multimedia de los fans del fútbol parece estar modulada por procesos emociona-les, mientras que para los no-fans predominan los procesos cognitivos, en particular aquellos directamente relacionados con la percepción de calidad. Abstract Recent studies encourage the integration of multi-sensorial stimuli into multimedia assets to enhance the user experience by stimulating other senses beyond sight and hearing. Similarly, the introduction of multi-modal interaction components complementing with new features, functionalities and/or information the multimedia experience is promoted. Sensory effects as odor, wind, vibration and light effects, as well as an enhanced audio quality, have been found to favour media enjoyment and to have a positive influence on the sense of Presence and on the perceived quality, relevance and reality of a multimedia experience. Two-screen services and a direct manipulation of (elements in) the video scene have the potential to enhance user comprehension, engagement and proactive involvement of/in the media experience. Sports is among the genres that could benefit the most from these solutions. Previous works have demonstrated the technical feasibility of implementing and deploying end-to-end solutions integrating these technologies into legacy systems. Thus, sensorially-enhanced media, two-screen services and an increased user control over the displayed scene emerge as means to deliver a new form of immersive and interactive media experiences to the mass market in a non-disruptive manner. However, many questions remain concerning issues as the specific interactive solutions or sensory effects that can better complement a given audiovisual content or the best way in which to integrate and combine them to enhance the user experience of a target audience segment. Furthermore, scientific evidence on the impact of human factors on the user experience with these new forms of immersive and interactive media is still insufficient and sometimes, contradictory. Thus, the role of these factors on the potential adoption of these technologies has been widely ignored. This thesis analyzes the impact of binaural audio, sensory (light and olfactory) effects, interaction with 3D objects integrated into the video scene and interaction with additional content using a second screen on the sports media experience. The potential influence of these components on the dependent variables is explored both at the overall level (average effect) and as a function of users’ characteristics (heterogeneous effects). To these aims, we conducted an experimental study exploring the influence of these immersive and interactive elements on the quality and Presence dimensions of the media experience. Along the quality dimension, we look for possible variations on the quality scores as-signed to the overall media experience and to the media components content, image, audio, sensory effects, interaction with 3D objects and interaction using the tablet device. The potential impact on Presence is analyzed by looking at two of the four dimensions defined by the ITC-SOPI questionnaire, namely Spatial Presence and Engagement. The users’ characteristics considered encompass the following personal affective, cognitive and behavioral attributes: preferences and habits in relation to the content, knowledge of the involved technologies, tendency to get emotionally involved and tendency to get absorbed in an activity and block out external distractors and the big five personality traits extraversion, agreeableness, conscientiousness, neuroticism and openness to experience. At the overall level, we found that participants preferred binaural audio than standard stereo audio and that sensory effects increase significantly the level of Spatial Presence. Several heterogeneous effects were also revealed as a result of our experimental manipulations. Interestingly, these effects were not equally distributed across the quality and Presence measures analyzed. Whereas binaural audio was foud to have an influence on the majority of the quality and Presence measures considered, the effects of sensory effects and of interaction with additional content through the tablet device concentrate mainly on the dimensions of Presence and on quality measures, respectively. The magnitude of these effects was modulated by individual’s characteristics, such as: preferences in relation to the content, frequency of viewing similar content, knowledge of involved technologies, gender, tendency to get emotionally involved, tendency to absorption and levels of agreeableness, conscientiousness and openness to experience. The personal characteristics collected in our experiment explained most of the variation in the dependent variables, confirming the frequently neglected role of individual differences on the media experience. Preferences in relation to the content, knowledge of involved technologies and tendency to get emotionally involved were among the user variables with the most generalized influence. In particular, the former two features seem to present a conflict in the allocation of attentional resources towards the media content versus the technical features of the system, respectively. Additionally, football fans’ experience seems to be modulated by emotional processes whereas for not fans, cognitive processes (and in particular those related to quality judgment) prevail.

Respuesta fisiológica, productiva y cualitativa del cultivar Syrah (Vitis vinifera L) con diferentes frecuencias de riego y densidad de goteros en suelos arcillosos

La disponibilidad hídrica es uno de los principales factores que determinan el rendimiento del viñedo en muchas regiones vitícolas, por lo que sus consecuencias han sido ampliamente estudiadas. Sin embargo, para una cantidad de agua de riego determinada, otros aspectos como la frecuencia de aplicación, o la combinación entre el caudal de los goteros y la distancia entre los mismos (es decir, el patrón de distribución de agua en el suelo), pueden jugar un papel relevante, pero estos factores han sido poco estudiados. El objetivo de este trabajo ha sido evaluar las implicaciones agronómicas y fisiológicas de dos frecuencias de riego (IrrF, cada 2 y 4 días) y dos patrones de distribución de agua (DisP, goteros de 2 L h-1 separados 0,6 m vs. goteros de 4 L h-1 separados 1,2 m). El experimento se llevó a cabo durante cuatro temporadas consecutivas en un viñedo cv. Syrah con un suelo arcilloso en el centro de España, y los dos factores fueron evaluados bajo dos condiciones de disponibilidad hídrica (Baja: 20% de ETo y Media: 40% de ETo). El efecto de la frecuencia de riego y el patrón de distribución de agua en la respuesta agronómica del cv. Syrah se ha estudiado en el capítulo 1. La frecuencia de riego y el patrón de distribución de agua en el suelo afectaron a algunos aspectos de los componentes de rendimiento y desarrollo vegetativo en las dos condiciones de disponibilidad hídrica, aunque los efectos observados no fueron los mismos todos los años. Los efectos fueron más evidentes para IrrF en condiciones de baja disponibilidad hídrica y para DisP en condiciones de disponibilidad hídrica media. Dos de los cuatro años del experimento, el pasar de frecuencia de riego de 2 días a 4 días causó un incremento medio de rendimiento del 20% para la situación de baja disponibilidad hídrica. La textura del suelo, sin duda ha condicionado los resultados obtenidos en los tratamientos regados con el 20% de la ETo, ya que regar cada dos días implicaba la aplicación de pequeñas cantidades de agua y se formaban bulbos de riego superficiales, probablemente favoreciendo las pérdidas por evaporación. En el capítulo 2, se ha analizado el efecto de la frecuencia de riego y del patrón de distribución de agua en el estado hídrico de la planta y el intercambio gaseoso a nivel de hoja con el fin de explicar las diferencias observadas en la respuesta agronómica. En lo que respecta a la frecuencia de riego, en condiciones de baja disponibilidad hídrica, las plantas regadas cada 4 días (plantas 4d), mostraron mayores tasas de asimilación neta y conductancia estomática que las plantas regadas cada 2 días (plantas 2d), lo que es consistente con la hipótesis de que con la frecuencia de riego de 2 días se produjo una pérdida de eficiencia del uso del agua, probablemente debido a una mayor evaporación como consecuencia del hecho de que el volumen de suelo mojado creado era pequeño y cerca de la superficie. En condiciones de disponibilidad hídrica media, las diferencias en el intercambio gaseoso a nivel de hoja fueron mucho más pequeñas. Al comienzo del verano cada frecuencia de riego se comportó mejor uno de los días de medida, compensando al final del ciclo de riego de 4 días. Sin embargo, a medida que avanzó el verano y el déficit de agua se hizo más alto, las diferencias significativas aparecieron sólo en el 'día 4' del ciclo de riego, cuando las plantas 2d se comportaron mejor que las plantas regadas 4d que llevaban tres días sin regarse. Estas diferencias fisiológicas fueron menores que en condiciones de baja disponibilidad hídrica y al parecer no suficientes para afectar el comportamiento agronómico. En cuanto al patrón de distribución de agua, el efecto fue poco significativo, pero la mayor densidad de goteros tendió a presentar un mayor intercambio gaseoso a nivel de hoja, especialmente a media mañana. El efecto fue más importante para las condiciones de disponibilidad hídrica media. En el capítulo 3, se han comparado las relaciones entre el intercambio gaseoso a nivel de hoja, el estado hídrico y la demanda atmosférica, con el fin de explicar los cambios en la intensidad de la respuesta fisiológica observados en el Capítulo 2. No se han encontrado diferencias en dichas relaciones para el patrón de distribución de agua, por lo que sólo se ha analizado el efecto de la frecuencia de riego. El estudio se ha centrado fundamentalmente en si las plantas mostraron una respuesta fisiológica diferente a los cambios en el estado hídrico y en la demanda atmosférica según el tiempo transcurrido desde el último riego. Las diferencias observadas explican los resultados obtenidos en los capítulos anteriores, y sugieren la existencia de procesos de aclimatación vinculados a la frecuencia de riego y a la disponibilidad hídrica. Las plantas bajo condiciones de baja disponibilidad hídrica se mostraron más aclimatadas al estrés hídrico que aquellas en condiciones de disponibilidad hídrica media. La frecuencia de riego afectó claramente la relación entre los parámetros de intercambio gaseoso a nivel de hoja, el estado hídrico de la planta y las condiciones atmosféricas, y junto con la cantidad de agua aplicada tuvo implicaciones en el desarrollo de mecanismos de aclimatación que afectaron a la respuesta fisiológica de la planta, afectando a la eficiencia del riego. ABSTRACT Water availability is one of the major factors that determine vineyard performance in many grape growing regions, so its implications have been widely studied before. However, for a given irrigation water amount, other aspects such as application frequency, or emitter spacing and flow rate (i.e., distribution pattern), may play a relevant role, but these factors have been scarcely studied. The aim of this work was to evaluate the agronomic and physiological implications of two irrigation frequencies (IrrF, every 2 and 4 days) and two water distribution patterns (DisP, 2 L h−1 emitters every 0.6 m vs. 4 L h−1 emitters every 1.2 m). The experiment was carried out during four consecutive seasons in a cv. Syrah vineyard with a clay soil in central Spain, and the two factors were evaluated under two water availability conditions (LOW WA: 20% of ETo and MEDIUM WA: 40% of ETo). The effect of irrigation frequency and water distribution pattern on the agronomical response of cv. Syrah was studied in Chapter 1. IrrF and DisP affected some aspects of vegetative development and yield components under both water availability conditions, although the effects observed were not the same every year. The effects were more evident for IrrF under low water availability and for DisP under medium water availability. Two out of the four years of the experiment, the change of irrigation frequency from 2 days to 4 days promoted an average yield increase of 20% for the LOW WA situation. Soil texture certainly conditioned the results obtained under LOW WA conditions, since high frequency irrigation implied applying small amounts of water that resulted in limited superficial water bulbs, which probably favored water evaporation. In Chapter 2, the effect of irrigation frequency and water distribution pattern on plant water status and leaf gas exchange was analyzed to explain the differences observed in the agronomical response. Concerning irrigation frequency, under LOW WA conditions, applying irrigation every 4 days, resulted in higher net assimilation rates and stomatal conductance than doing it every 2 days, supporting the hypothesis that the latter frequency resulted in a water use efficiency loss, probably due to higher evaporation as a consequence of the fact the wetted soil volume created was small and close to the surface. Under MEDIUM WA conditions, differences in leaf gas exchange were much smaller. At the beginning of the summer each irrigation frequency behaved better one of the measurements days, compensating at the end of the 4-day irrigation cycle. However, as the summer progressed and water deficit became higher, significant differences appeared only on ‘day 4’ of the irrigation cycle, when 2d plants behaved better than 4d plants. These physiological differences were smaller than under LOW WA conditions and apparently not sufficient to affect agronomical performance. Regarding water distribution pattern, the effect was less significant but the closest emitter spacing resulted in general terms in a higher leaf gas exchange, especially at midmorning. The effect was more noticeable for MEDIUM WA conditions. In Chapter 3, the relationships between leaf gas exchange and leaf water status and atmospheric demand were compared to explain the changes in the intensity of the physiological response observed in Chapter 2. No differences were found in the relationships for water distribution pattern, so only the effect of irrigation frequency was analyzed focusing on whether the plants have a different physiological response to changes in water status and atmospheric demand according to the time elapsed since the last irrigation. Differences observed in the relationships explained the results obtained in the previous chapters, and point at the occurrence of acclimation processes linked to irrigation frequency and to water availability. Plants under LOW WATER AVAILABILITY conditions seemed to be more acclimated to water stress than those under MEDIUM WATER AVAILABILITY conditions. Irrigation frequency clearly affected the relationship between leaf gas exchange parameters, plant water status and atmospheric conditions, and together with the amount of water applied had implications in the development of acclimation mechanisms that affected plant physiological response, thus affecting irrigation efficiency.

Region-dependent dynamics of cAMP response element-binding protein phosphorylation in the basal ganglia

The cAMP response element-binding protein (CREB) is an activity-dependent transcription factor that is involved in neural plasticity. The kinetics of CREB phosphorylation have been suggested to be important for gene activation, with sustained phosphorylation being associated with downstream gene expression. If so, the duration of CREB phosphorylation might serve as an indicator for time-sensitive plastic changes in neurons. To screen for regions potentially involved in dopamine-mediated plasticity in the basal ganglia, we used organotypic slice cultures to study the patterns of dopamine- and calcium-mediated CREB phosphorylation in the major subdivisions of the striatum. Different durations of CREB phosphorylation were evoked in the dorsal and ventral striatum by activation of dopamine D1-class receptors. The same D1 stimulus elicited (i) transient phosphorylation (≤15 min) in the matrix of the dorsal striatum; (ii) sustained phosphorylation (≤2 hr) in limbic-related structures including striosomes, the nucleus accumbens, the fundus striati, and the bed nucleus of the stria terminalis; and (iii) prolonged phosphorylation (up to 4 hr or more) in cellular islands in the olfactory tubercle. Elevation of Ca2+ influx by stimulation of L-type Ca2+ channels, NMDA, or KCl induced strong CREB phosphorylation in the dorsal striatum but not in the olfactory tubercle. These findings differentiate the response of CREB to dopamine and calcium signals in different striatal regions and suggest that dopamine-mediated CREB phosphorylation is persistent in limbic-related regions of the neonatal basal ganglia. The downstream effects activated by persistent CREB phosphorylation may include time-sensitive neuroplasticity modulated by dopamine.

The type 2 iodothyronine deiodinase is expressed primarily in glial cells in the neonatal rat brain

Thyroid hormone plays an essential role in mammalian brain maturation and function, in large part by regulating the expression of specific neuronal genes. In this tissue, the type 2 deiodinase (D2) appears to be essential for providing adequate levels of the active thyroid hormone 3,5,3′-triiodothyronine (T3) during the developmental period. We have studied the regional and cellular localization of D2 mRNA in the brain of 15-day-old neonatal rats. D2 is expressed in the cerebral cortex, olfactory bulb, hippocampus, caudate, thalamus, hypothalamus, and cerebellum and was absent from the white matter. At the cellular level, D2 is expressed predominantly, if not exclusively, in astrocytes and in the tanycytes lining the third ventricle and present in the median eminence. These results suggest a close metabolic coupling between subsets of glial cells and neurons, whereby thyroxine is taken up from the blood and/or cerebrospinal fluid by astrocytes and tanycytes, is deiodinated to T3, and then is released for utilization by neurons.

Gestational exposure to ethanol suppresses msx2 expression in developing mouse embryos

Ethanol acts as a teratogen in developing fetuses causing abnormalities of the brain, heart, craniofacial bones, and limb skeletal elements. To assess whether some teratogenic actions of ethanol might occur via dysregulation of msx2 expression, we examined msx2 expression in developing mouse embryos exposed to ethanol on embryonic day (E) 8 of gestation and subjected to whole mount in situ hybridization on E11–11.5 using a riboprobe for mouse msx2. Control mice exhibited expression of msx2 in developing brain, the developing limb buds and apical ectodermal ridge, the lateral and nasal processes, olfactory pit, palatal shelf of the maxilla, the eye, the lens of the eye, otic vesicle, prevertebral bodies (notochord), and endocardial cushion. Embryos exposed to ethanol in utero were significantly smaller than their normal counterparts and did not exhibit expression of msx2 in any structures. Similarly, msx2 expression, as determined by reverse transcription–PCR and Northern blot hybridization, was reduced ≈40–50% in fetal mouse calvarial osteoblastic cells exposed to 1% ethanol for 48 hr while alkaline phosphatase was increased by 2-fold and bone morphogenetic protein showed essentially no change. Transcriptional activity of the msx2 promoter was specifically suppressed by alcohol in MC3T3-E1 osteoblasts. Taken together, these data demonstrate that fetal alcohol exposure decreases msx2 expression, a known regulator of osteoblast and myoblast differentiation, and suggest that one of the “putative” mechanisms for fetal alcohol syndrome is the inhibition of msx2 expression during key developmental periods leading to developmental retardation, altered craniofacial morphogenesis, and cardiac defects.

Molecular determinants of the modulation of cyclic nucleotide-activated channels by calmodulin

The action of calmodulin (CaM) on target proteins is important for a variety of cellular functions. We demonstrate here, however, that the presence of a CaM-binding site on a protein does not necessarily imply a functional effect. The α-subunit of the cGMP-gated cation channel of human retinal cones has a CaM-binding site on its cytoplasmic N-terminal region, but the homomeric channel that it forms is not functionally modulated by CaM. Mutational analysis based on comparison to the highly homologous olfactory cyclic nucleotide-gated channel α-subunit, which does form a CaM-modulated channel, indicates that residues downstream of the CaM-binding domain on these channels are also important for CaM to have an effect. These findings suggest that a CaM-binding site and complementary structural features in a protein probably evolve independently, and an effect caused by CaM occurs only in the presence of both elements. More generally, the same may be true for other recognized binding sites on proteins for modulators or activators, so that a demonstrated physical interaction does not necessarily imply functional consequence.

Postnatal mouse subventricular zone neuronal precursors can migrate and differentiate within multiple levels of the developing neuraxis

The mammalian subventricular zone (SVZ) of the lateral wall of the forebrain ventricle retains a population of proliferating neuronal precursors throughout life. Neuronal precursors born in the postnatal and adult SVZ migrate to the olfactory bulb where they differentiate into interneurons. Here we tested the potential of mouse postnatal SVZ precursors in the environment of the embryonic brain: (i) a ubiquitous genetic marker, (ii) a neuron-specific transgene, and (iii) a lipophilic-dye were used to follow the fate of postnatal day 5–10 SVZ cells grafted into embryonic mouse brain ventricles at day 15 of gestation. Graft-derived cells were found at multiple levels of the neuraxis, including septum, thalamus, hypothalamus, and in large numbers in the midbrain inferior colliculus. We observed no integration into the cortex. Neuronal differentiation of graft derived cells was demonstrated by double-staining with neuron-specific β-tubulin antibodies, expression of the neuron-specific transgene, and the dendritic arbors revealed by the lipophilic dye. We conclude that postnatal SVZ cells can migrate through and differentiate into neurons within multiple embryonic brain regions other than the olfactory bulb.

A genetic screen for mutations that disrupt an auditory response in Drosophila melanogaster

Hearing is one of the last sensory modalities to be subjected to genetic analysis in Drosophila melanogaster. We describe a behavioral assay for auditory function involving courtship among groups of males triggered by the pulse component of the courtship song. In a mutagenesis screen for mutations that disrupt the auditory response, we have recovered 15 mutations that either reduce or abolish this response. Mutant audiograms indicate that seven mutants reduced the amplitude of the response at all intensities. Another seven abolished the response altogether. The other mutant, 5L3, responded only at high sound intensities, indicating that the threshold was shifted in this mutant. Six mutants were characterized in greater detail. 5L3 had a general courtship defect; courtship of females by 5L3 males also was affected strongly. 5P1 males courted females normally but had reduced success at copulation. 5P1 and 5N18 showed a significant decrement in olfactory response, indicating that the defects in these mutations are not specific to the auditory pathway. Two other mutants, 5M8 and 5N30, produced amotile sperm although in 5N30 this phenotype was genetically separable from the auditory phenotype. Finally, a new adult circling behavior phenotype, the pirouette phenotype, associated with massive neurodegeneration in the brain, was discovered in two mutants, 5G10 and 5N18. This study provides the basis for a genetic and molecular dissection of auditory mechanosensation and auditory behavior.

Network of tangential pathways for neuronal migration in adult mammalian brain

Cells in the brains of adult mammals continue to proliferate in the subventricular zone (SVZ) throughout the lateral wall of the lateral ventricle. Here we show, using whole mount dissections of this wall from adult mice, that the SVZ is organized as an extensive network of chains of neuronal precursors. These chains are immunopositive to the polysialylated form of NCAM, a molecule present at sites of plasticity, and TuJ1, an early neuronal marker. The majority of the chains are oriented along the rostrocaudal axis and many join the rostral migratory stream that terminates in the olfactory bulb. Using focal microinjections of DiI and transplantation of SVZ cells carrying a neuron-specific reporter gene, we demonstrate that cells originating at different rostrocaudal levels of the SVZ migrate rostrally and reach the olfactory bulb where they differentiate into neurons. Our results reveal an extensive network of pathways for the tangential chain migration of neuronal precursors throughout the lateral wall of the lateral ventricle in the adult mammalian brain.