Intra-annual patterns of tracheid size in the Mediterranean tree Juniperus thurifera as an indicator of seasonal water stress

Because climate can affect xylem cell anatomy, series of intra-annual cell anatomical features have the potential to retrospectively supply seasonal climatic information. In this study, we explored the ability to extract information about water stress conditions from tracheid features of the Mediterranean conifer Juniperus thurifera L. Tracheidograms of four climatic years from two drought-sensitive sites in Spain were compared to evaluate whether it is possible to link intra-annual cell size patterns to seasonal climatic conditions. Results indicated site-specific anatomical adjustment such as smaller and thicker tracheids at the dryer site but also showed a strong climatic imprint on the intra-annual pattern of tracheid size. Site differences in cell size reflected expected structural adjustments against cavitation failures. Differences between intra-annual patterns, however, indicated a response to seasonal changes in water availability whereby cells formed under drought conditions were smaller and thicker, and vice versa. This relationship was more manifest and stable at the dryer site

Metabolomics demonstrates divergent responses of two Eucalyptus species to water stress

Past studies of water stress in Eucalyptus spp. generally highlighted the role of fewer than five “important” metabolites, whereas recent metabolomic studies on other genera have shown tens of compounds are affected. There are currently no metabolite profiling data for responses of stress-tolerant species to water stress. We used GC–MS metabolite profiling to examine the response of leaf metabolites to a long (2 month) and severe (Ψpredawn < −2 MPa) water stress in two species of the perennial tree genus Eucalyptus (the mesic Eucalyptus pauciflora and the semi-arid Eucalyptus dumosa). Polar metabolites in leaves were analysed by GC–MS and inorganic ions by capillary electrophoresis. Pressure–volume curves and metabolite measurements showed that water stress led to more negative osmotic potential and increased total osmotically active solutes in leaves of both species. Water stress affected around 30–40% of measured metabolites in E. dumosa and 10–15% in E. pauciflora. There were many metabolites that were affected in E. dumosa but not E. pauciflora, and some that had opposite responses in the two species. For example, in E. dumosa there were increases in five acyclic sugar alcohols and four low-abundance carbohydrates that were unaffected by water stress in E. pauciflora. Re-watering increased osmotic potential and decreased total osmotically active solutes in E. pauciflora, whereas in E. dumosa re-watering led to further decreases in osmotic potential and increases in total osmotically active solutes. This experiment has added several extra dimensions to previous targeted analyses of water stress responses in Eucalyptus, and highlights that even species that are closely related (e.g. congeners) may respond differently to water stress and re-watering

Implications of the mesophyll conductance to CO2 for photosynthesis and water-use efficiency during long-term water stress and recovery in two contrasting Eucalyptu species

Water stress (WS) slows growth and photosynthesis (An), but most knowledge comes from short-time studies that do not account for longer term acclimation processes that are especially relevant in tree species. Using two Eucalyptus species that contrast in drought tolerance, we induced moderate and severe water deficits by withholding water until stomatal conductance (gsw) decreased to two pre-defined values for 24 d, WS was maintained at the target gsw for 29 d and then plants were re-watered. Additionally, we developed new equations to simulate the effect on mesophyll conductance (gm) of accounting for the resistance to refixation of CO2. The diffusive limitations to CO2, dominated by the stomata, were the most important constraints to An. Full recovery of An was reached after re-watering, characterized by quick recovery of gm and even higher biochemical capacity, in contrast to the slower recovery of gsw. The acclimation to long-term WS led to decreased mesophyll and biochemical limitations, in contrast to studies in which stress was imposed more rapidly. Finally, we provide evidence that higher gm under WS contributes to higher intrinsic water-use efficiency (iWUE) and reduces the leaf oxidative stress, highlighting the importance of gm as a target for breeding/genetic engineering.

Water stress thresholds for regulated deficit irrigation in pistachio trees

The response of "Kerman" pistachio trees budded on three different rootstocks (Pistacia terebinthus, Pista-cia atlantica and Pistacia integerrima) to regulated deficit irrigation (RDI) in shallow soils was studied for3 years. The trees were either fully irrigated (C treatment) or subjected to deficit irrigation during Stage IIof fruit growth with two water stress thresholds (T1 and T2). The irrigation scheduling for fully-irrigatedtrees and water-stressed trees was managed by means of midday stem water potential (?stem) measure-ments. The use of direct measurements of the water status allowed estimating accurately the irrigationrequirements for pistachio trees, with water reductions ranging from 46 to 205 mm in fully-irrigatedtrees. The combination of the ?stemuse and the RDI regime saved 43?70% in T1 and 48?73% in T2 ofwater compared to the calculated crop evapotranspiration (ETc) for fully irrigated treatment (C).Deficit irrigation during Stage II significantly reduced the vegetative growth of the trees. Yield and fruitquality were not affected by any irrigation regime, except during the first year of the study. Thus, theresults indicate that full irrigation scheduling and RDI can be achieved successfully using ?stemtool onpistachio trees growing in shallow soils. A ?stemthreshold of ?1.5 MPa during stage II (T1) was suggestedfor RDI scheduling, as it did not reduce the yield or the production value. However a ?stemthresholdof ?2.0 MPa (T2) resulted in a significant reduction and an extensive delay in the recovery of stomatalconductance (gl),with negative effects on long-term pistachio production.P. integerrima showed a weaker capacity of adaptation to the study conditions compared to P. atlanticaand P. terebinthus, having a tendency to get more stressed and to produce a lower quality crop.

Midday measurements of leaf water potential and stomatal conductance are highly correlated with daily water use of Thompson Seedless grapevines

A study was conducted to determine the relationship between midday measurements of vine water status and daily water use of grapevines measured with a weighing lysimeter. Water applications to the vines were terminated on August 24th for 9 days and again on September 14th for 22 days. Daily water use of the vines in the lysimeter (ETLYS) was approximately 40 L vine−1 (5.3 mm) prior to turning the pump off, and it decreased to 22.3 L vine−1 by September 2nd. Pre-dawn leaf water potential (ΨPD) and midday Ψl on August 24th were −0.075 and −0.76 MPa, respectively, with midday Ψl decreasing to −1.28 MPa on September 2nd. Leaf g s decreased from ~500 to ~200 mmol m−2 s−1 during the two dry-down periods. Midday measurements of g s and Ψl were significantly correlated with one another (r = 0.96) and both with ETLYS/ETo (r = ~0.9). The decreases in Ψl, g s, and ETLYS/ETo in this study were also a linear function of the decrease in volumetric soil water content. The results indicate that even modest water stress can greatly reduce grapevine water use and that short-term measures of vine water status taken at midday are a reflection of daily grapevine water use

Effect of row orientation on soil water content and vine water status on a Cabernet franc vineyard in Madrid, spain

The aim of this study was to evaluate the effects of row orien¬tation on vine and soil water status in an irrigated vineyard. The trial was developed during 2006, 2007 and 2008, in the South East region of Madrid (Spain) on 5-year old Cabernet franc grapevines (Vitis vinifera L.) grafted onto 140Ru. Plant spacing was 2.5 m x 1.5 m and vines were trained to a VSP. Four orientations were stu¬died: North-South (N-S), East-West (E-W), Northeast-Southwest (N+45) and North-South +20o (N+20). Irrigation (0.4•ET0) started when shoot growth stopped. Soil water availability was measured using a TDR technique with forty buried probes. Row orientation did not have any effect on water consumption in the vineyard. At maturity, leaf water potential was measured at predawn, early mor¬ning, midday and 14:00 solar time, on both canopy sides - sun and shade – ; the early morning measurement was the one that better differentiated treatments. Leaf water potential was a good indica¬tor of plant water status. Differences between (N-S and E-W) and (N+20 and N+45) treatments were obtained both on sun and shade canopy sides, N+20 and N+45 having lower leaf water potentials then drier leaves. The water stress integral shows that N-S and E-W reach the end of maturation with a greater level of hydration than N+45 and N+20. As a whole, N+45 and N+20 orientations, without affecting too much the soil available water content, induce regularly more water stress to the vine at some periods, probably due to an higher sunlight interception in early morning which makes water limitation for the vine more early and thus more severe during the day.

Latin American Agricultural Trade: The Role of the WTO in Sustainable Virtual Water Flows, European Association of Agricultural Economists

International agricultural trade has been growing significantly during the last decade. Many countries rely on imports to ensure adequate food supplies to the people. A few are becoming food baskets of the world. This process raises issues about the food security in depending countries and potentially unsustainable land and water use in exporting countries. In this paper, we analyse the impacts of amplified farm trade on natural resources, especially water. Farm exports and imports of five Latin America countries (Brazil, Argentina, Mexico, Peru and Chile) are examined carefully. A preliminary analysis indicates that virtual water imports can save valuable water resources in water-short countries, such as Mexico and Chile. Major exporting countries, including Brazil and Argentina, have become big exporters due to abundant natural resource endowments. The opportunity costs of agricultural production in those countries are identified as being low, because of the predominant green water use. It is concluded that virtual water trade can be a powerful tool to alleviate water stress in semi-arid countries. However, for exporting nations a sustainable water use can only be guaranteed if environmental production costs are fully reflected in the commodity prices. There is no basis for erecting environmental trade tariffs on exporters though. Setting up legal foundations for them in full compliance with WTOs processes would be a daunting task.

Balancing groundwater conservation and rural livelihoods under water and climate uncertainties: An integrated hydro-economic modeling framework

In arid countries worldwide, social conflicts between irrigation-based human development and the conservation of aquatic ecosystems are widespread and attract many public debates. This research focuses on the analysis of water and agricultural policies aimed at conserving groundwater resources and maintaining rurallivelihoods in a basin in Spain's central arid region. Intensive groundwater mining for irrigation has caused overexploitation of the basin's large aquifer, the degradation of reputed wetlands and has given rise to notable social conflicts over the years. With the aim of tackling the multifaceted socio-ecological interactions of complex water systems, the methodology used in this study consists in a novel integration into a common platform of an economic optimization model and a hydrology model WEAP (Water Evaluation And Planning system). This robust tool is used to analyze the spatial and temporal effects of different water and agricultural policies under different climate scenarios. It permits the prediction of different climate and policy outcomes across farm types (water stress impacts and adaptation), at basin's level (aquifer recovery), and along the policies’ implementation horizon (short and long run). Results show that the region's current quota-based water policies may contribute to reduce water consumption in the farms but will not be able to recover the aquifer and will inflict income losses to the rural communities. This situation would worsen in case of drought. Economies of scale and technology are evidenced as larger farms with cropping diversification and those equipped with modern irrigation will better adapt to water stress conditions. However, the long-term sustainability of the aquifer and the maintenance of rurallivelihoods will be attained only if additional policy measures are put in place such as the control of illegal abstractions and the establishing of a water bank. Within the policy domain, the research contributes to the new sustainable development strategy of the EU by concluding that, in water-scarce regions, effective integration of water and agricultural policies is essential for achieving the water protection objectives of the EU policies. Therefore, the design and enforcement of well-balanced region-specific polices is a major task faced by policy makers for achieving successful water management that will ensure nature protection and human development at tolerable social costs. From a methodological perspective, this research initiative contributes to better address hydrological questions as well as economic and social issues in complex water and human systems. Its integrated vision provides a valuable illustration to inform water policy and management decisions within contexts of water-related conflicts worldwide.

Analyzing climate change adaptation in the agriculture and water sectors: screening risks and opportunities.

As part of the Mediterranean area, the Guadiana basin in Spain is particularly exposed to increasing water stress due to climate change. Future warmer and drier climate will have negative implications for the sustainability of water resources and irrigation agriculture, the main socio- economic sector in the region. This paper illustrates a systematic analysis of climate change impacts and adaptation in the Guadiana basin based on a two-stage modeling approach. First, an integrated hydro-economic modeling framework was used to simulate the potential effects of regional climate change scenarios for the period 2000-2069. Second, a participatory multi-criteria technique, namely the Analytic Hierarchy Process (AHP), was applied to rank potential adaptation measures based on agreed criteria. Results show that, in the middle-long run and under severe climate change, reduced water availability, lower crop yields and increased irrigation demands might lead to water shortages, crop failure, and up to ten percent of income losses to irrigators. AHP results show how private farming adaptation measures, including improving irrigation efficiency and adjusting crop varieties, are preferred to public adaptation measures, such as building new dams. The integrated quantitative and qualitative methodology used in this research can be considered a socially-based valuable tool to support adaptation decision-making.

Responses of riparian guilds to flowalterations in a Mediterranean stream

Questions: Do Mediterranean riparian guilds show distinct responses to stream water declines? If observed,which are the most sensitive and resilient guilds and theirmost affected attributes? Location: Tie¿tar river below the Rosarito dam, central-western Spain. Methods: We identified riparian guilds based on key woody species features and species distribution within this Mediterranean river corridor, and evaluated similarity of their responses to long-term flow alteration (i.e. stream water declines since dam construction in 1959). Hierarchical cluster analysis was used to group surveyed vegetation bands according to species composition. The groups were designated as riparian guilds where each vegetation group comprising a guild: (1) contains species sharing similar features (using PCA); and (2) shares a similar environment (using DCA). Changes in several guild attributes (i.e. dominance and species composition, diversity and establishment patterns) during the regulated period were compared statistically. We used pre- and post-dam established vegetation bands identified based on old (1956) and modern (2006) aerial photographs and field measurements of woody species diameter. Results: Responses to flow alterations varied between guilds according to ecological requirements of their species. The ability to survive water stress (i.e. ?Xeric? guilds) and drag forces caused by floods (?Torrential? guilds) allowed certain pioneer shrub-dominated guilds (e.g. Flueggea tinctoria and Salix salviifolia) to spread on newly emerged surfaces downward to the main channel after flow alterations, although new shrubland had less species diversity than pre-dam shrubland. In contrast, new hydromorphological conditions following damming limited recruitment of native late-successional tree guilds sensitive to floods (to drag forces, inundation and anoxia; i.e. ?Slow-water? and ?Flood-sensitive?, respectively) and those with greater water requirements (i.e. ?Hydric?) (e.g. Alnus glutinosa and Celtis australis), although species diversity increased in this mature forest through co-existence of remaining riparian species and new arrival of upland species. Conclusions: Changes in several riparian attributes after flow alterations differed between guilds. Stream water declines after damming caused shifts in species-poor pioneer shrubland downwards to the watered channel, resulting in severe declines ofmaturenative forest.Understanding vegetation guild responses provides information about general trends in plant populations and assemblage structures expected to occur during river development and flow regulation, increasing our capacity to detect and synthesize complex flowalteration?riparian ecosystem response relationships, and anticipate irreversible impacts.

Optimization of the irrigation time and irrigation frequency by using Hydrus-2D and a capacitance FDR sensor

The evolution of water content on a sandy soil during the sprinkler irrigation campaign, in the summer of 2010, of a field of sugar beet crop located at Valladolid (Spain) is assessed by a capacitive FDR (Frequency Domain Reflectometry) EnviroScan. This field is one of the experimental sites of the Spanish research center for the sugar beet development (AIMCRA). The objective of the work focus on monitoring the soil water content evolution of consecutive irrigations during the second two weeks of July (from the 12th to the 28th). These measurements will be used to simulate water movement by means of Hydrus-2D. The water probe logged water content readings (m3/m3) at 10, 20, 40 and 60 cm depth every 30 minutes. The probe was placed between two rows in one of the typical 12 x 15 m sprinkler irrigation framework. Furthermore, a texture analysis at the soil profile was also conducted. The irrigation frequency in this farm was set by the own personal farmer 0 s criteria that aiming to minimizing electricity pumping costs, used to irrigate at night and during the weekend i.e. longer irrigation frequency than expected. However, the high evapotranspiration rates and the weekly sugar beet water consumption—up to 50mm/week—clearly determined the need for lower this frequency. Moreover, farmer used to irrigate for six or five hours whilst results from the EnviroScan probe showed the soil profile reaching saturation point after the first three hours. It must be noted that AIMCRA provides to his members with a SMS service regarding weekly sugar beet water requirement; from the use of different meteorological stations and evapotranspiration pans, farmers have an idea of the weekly irrigation needs. Nevertheless, it is the farmer 0 s decision to decide how to irrigate. Thus, in order to minimize water stress and pumping costs, a suitable irrigation time and irrigation frequency was modeled with Hydrus-2D. Results for the period above mentioned showed values of water content ranging from 35 and 30 (m3/m3) for the first 10 and 20cm profile depth (two hours after irrigation) to the minimum 14 and 13 (m3/m3) ( two hours before irrigation). For the 40 and 60 cm profile depth, water content moves steadily across the dates: The greater the root activity the greater the water content variation. According to the results in the EnviroScan probe and the modeling in Hydrus-2D, shorter frequencies and irrigation times are suggested.

Seasonal evolution of soil respiration in a mixed forestry plantation of walnuts (Juglans × intermedia Carr.) and alders (Alnus cordata (Loisel.) Duby) in Domaine de Restinclières (France)

The ecological intensification of crops is proposed as a solution to the growing demand of agricultural and forest resources, in opposition to intensive monocultures. The introduction of mixed cultures as mixtures between nitrogen fixing species and non nitrogen fixing species intended to increase crop yield as a result of an improvement of the available nitrogen and phosphorus in soil. Relationship between crops have received little attention despite the wide range of advantages that confers species diversity to these systems, such as increased productivity, resilience to disruption and ecological sustainability. Forests and forestry plantations can develop an important role in storing carbon in their tissues, especially in wood which become into durable product. A simplifying parameter to analyze the amount allocated carbon by plantation is the TBCA (total belowground carbon allocation), whereby, for short periods and mature plantations, is admitted as the subtraction between soil carbon efflux and litterfall. Soil respiration depends on a wide range of factors, such as soil temperature and soil water content, soil fertility, presence and type of vegetation, among others. The studied orchard is a mixed forestry plantation of hybrid walnuts(Juglans × intermedia Carr.) for wood and alders (Alnus cordata (Loisel.) Duby.), a nitrogen fixing specie through the actinomycete Frankia alni ((Woronin, 1866) Von Tubeuf 1895). The study area is sited at Restinclières, a green area near Montpellier (South of France). In the present work, soil respiration varied greatly throughout the year, mainly influenced by soil temperature. Soil water content did not significantly influence the response of soil respiration as it was constant during the measurement period and under no water stress conditions. Distance between nearest walnut and measurement was also a highly influential factor in soil respiration. Generally there was a decreasing trend in soil respiration when the distance to the nearest tree increased. It was also analyzed the response of soil respiration according to alder presence and fertilizer management (50 kg N·ha-1·año-1 from 1999 to 2010). None of these treatments significantly influenced soil respiration, although previous studies noticed an inhibition in rates of soil respiration under fertilized conditions and high rates of available nitrogen. However, treatments without fertilization and without alder presence obtained higher respiration rates in those cases with significant differences. The lack of significant differences between treatments may be due to the high coefficient of variation experienced by soil respiration measurements. Finally an asynchronous fluctuation was observed between soil respiration and litterfall during senescence period. This is possibly due to the slowdown in the emission of exudates by roots during senescence period, which are largely related to microbial activity.

Genes implicados en la respuesta molecular al estrés hídrico en Pinus pinaster Ait

La presente tesis doctoral se centra en el estudio de la respuesta molecular de las coníferas mediterráneas al estrés hídrico. Para ello se ha escogido como especie modelo Pinus pinaster Ait., la conífera más abundante en España, y que habita un amplio rango de situaciones ecológicas, especialmente en lo relativo a la disponibilidad de agua. En primer lugar, se ha aplicado un estrés hídrico controlado en cultivo hidropónico y se ha generando una genoteca sustractiva con objeto de identificar los genes inducidos por el estrés, analizando su expresión en raíces, tallos y acículas. A continuación, se ha analizado, la expresión de los genes anteriormente obtenidos así como de otros seleccionados de las bases de datos disponibles, durante una sequía prolongada en tierra, similar a las que las plantas deben afrontar en la naturaleza. Se ha utilizado en este caso, además de P. pinaster, P. pinea, otra conífera mediterránea adaptada a las sequías recurrentes. Este trabajo ha permitido identificar genes candidato expresionales, presumiblemente comunes en la respuesta molecular de las coníferas al déficit hídrico. Se han detectado diferencias notables en la expresión de determinados genes, que podrían ser los responsables de las diferencias exhibidas por ambas especies en el comportamiento frente a la sequía. Entre los genes identificados como inducidos por el estrés hídrico se encuentran varios miembros de la familia de las deshidrinas. Trabajos previos han utilizado deshidrinas como genes candidato; no obstante, la falta de especificidad de ciertos fragmentos y marcadores utilizados, debido a la complejidad estructural de esta familia, resta fiabilidad a algunos de los resultados publicados. Por este motivo, se ha estudiado en detalle esta familia en P. pinaster, se han identificado y caracterizado 8 miembros y se ha analizado su patrón de expresión frente a sequía. Este estudio ha permitido describir por primera vez unos segmentos conservados en la secuencia de aminoácidos de las deshidrinas de pináceas, cuya presencia y número de repeticiones parece estar relacionado con su especificidad. Por último, se han escogido tres genes implicados en distintas fases de la respuesta al estrés hídrico para su análisis exhaustivo: una deshidrina, una nodulina y un factor de transcripción tipo AP2. Se ha caracterizado su estructura exón/intrón y secuenciado su región promotora. Además, se han obtenido líneas transformadas que sobreexpresan estos genes tanto de forma heteróloga, en la especie modelo Arabidopsis thaliana, como en el propio P. pinaster. Este material facilitará la realización de futuros estudios sobre la función y el mecanismo de actuación de estos genes en la respuesta al estrés hídrico. ABSTRACT This thesis focuses in the study of the molecular response to water stress in Mediterranean conifers. For this purpose, P. pinaster was selected as model species. It’s the most abundant conifer in Spain, living in a wide range of ecological conditions, especially regarding water availability. First, we have applied a controlled polyethylene glycol-induced water stress in hydroponic culture and obtained a suppression subtractive hybridization (SSH) library, with the aim of identifying genes induced by water stress, analysing their expression in roots, stems and needles. We have then analysed the expression patterns of the identified genes, together with other genes selected from public databases. This study was conducted throughout a prolonged drought stress in soil, similar to the ones plants have to face in nature. In this case not only P. pinaster was analysed but also P. pinea, another Mediterranean conifer well adapted to recurrent droughts. This work has enabled us to identify of reliable candidate genes, presumably shared with other conifers in the response to water stress. We observed remarkable differences in the expression of some genes, which could be involved in the differential behaviour that these species show in the water stress response. Within the genes induced by water stress, several members of the dehydrin gene family were identified. Due to the structural complexity of the family, certain ambiguities and inconsistencies have been detected in previous works that have used dehydrins as candidate genes. For this reason, we have analysed thoroughly this gene family in P. pinaster, and have identified and characterized eight different members, whose expression patterns during drought have also been assessed. This study has allowed us to identify for the first time novel conserved segments in the amino acids sequences of Pinaceae. The presence and number of repetitions of these segments could be associated with the functional specificity of these proteins. Finally, three genes involved in different steps of the water stress response were selected for an exhaustive analysis: a dehydrin, a nodulin and an AP2 transcription factor. For all of them, the exon/intron structure was established and their promoter region was sequenced. Also, transformed lines were obtained both in Arabidopsis thaliana and in P. pinaster for the constitutive overexpression of these genes. This material will facilitate the development of further studies to investigate the function of these genes during the water stress response

Caracterización molecular del mutante riso 1508 de cebada : modificaciones transcripcionales y posttranscripcionales en el desarrollo de la semilla

La semilla es el principal órgano reproductivo de las plantas espermatofitas, permitiendo la dispersión de las poblaciones y asegurando su supervivencia gracias a su tolerancia a la desecación y a su capacidad para germinar bajo condiciones ambientales óptimas. El rendimiento y valor económico de los cereales, que constituyen la primera cosecha mundial, depende, en buena medida, de la eficacia con que se acumulan en la semilla sustancias de reserva: proteínas, carbohidratos y lípidos. El principal carbohidrato acumulado en la semilla de cebada es el almidón y la fracción mayoritaria de proteínas es la de las prolaminas (solubles en etanol al 70%); estas proteínas tienen muy bajo contenido en lisina, un aminoácido esencial en la dieta de animales monogástricos. Con el fin de mejorar el valor nutricional de la semilla de cebada, se han obtenido diferentes mutantes con un mayor contenido en este aminoácido. Riso 1508 es un mutante de cebada rico en lisina cuya mutación lys3a, de efectos pleiotrópicos, segrega como un único gen mendeliano. Entre otros, presenta una reducción drástica de la expresión de algunos genes que codifican proteínas de reserva de tipo prolamina, en concreto, presenta reducida la expresión de los genes que codifican B-, C- y ϒ-Hordeínas y del inhibidor de tripsina CMe, pero no tiene alterada la expresión del gen que codifica las D-Hordeínas. Este último gen carece en su promotor del motivo GLM (5’‐(G/A)TGA(G/C)TCA(T/C)‐3’), que es reconocido por factores transcripcionales bZIP. En este trabajo, el mutante de cebada Riso 1508 se ha utilizado como herramienta para profundizar en el conocimiento de la regulación génica en semillas durante las fases de la maduración y la germinación. Para ello, en una primera aproximación, se llevó a cabo un análisis transcriptómico comparando el genotipo mutante con el silvestre durante la maduración de la semilla. Además de confirmar variaciones en los genes que codifican proteínas de reserva, este análisis indicó que también estaban afectados los genes relacionados con metabolismo de carbohidratos. Por ello se decidió caracterizar la familia multigénica de sacarosas sintasa (SUSy) en cebada. Se anotaron dos nuevos genes, HvSs3 y HvSs4, cuya expresión se comparó con la de los genes HvSs1 y HvSs2, previamente descritos en el laboratorio. La expresión de los cuatro genes en tejidos diferentes y su respuesta a estreses abióticos se analizó mediante RT-qPCR. HvSs1 y HvSs2 se expresaron preferencialmente durante el desarrollo del endospermo, y HvSs1 también fue un tránscrito abundante durante la germinación. HvSs1 se indujo en hojas en condiciones de anoxia y HvSs3 por estrés hídrico, y ambos genes se indujeron por tratamientos de frío. La localización subcelular de las cuatro isoformas no fue sólo citoplásmica, sino que también se localizaron en zonas próximas a retículo endoplásmico y en la cara interna de la membrana plasmática; además, se observó una co-localización de HvSS1 con el marcador de mitocondrias. Estos datos sugieren un papel distinto aunque parcialmente solapante de las cuatro Sacarosa Sintasas de cebada, descritas hasta la fecha. Las cinéticas de expresión de los genes que codifican los TFs más importantes implicados en la regulación génica durante el desarrollo del endospermo de cebada, se analizaron por RT-qPCR en ambos genotipos, demostrando que los TFs de la clase DOF aparecieron desregulados durante todo el proceso en Riso 1508 comparado con el cv. Bomi, aunque también se observaron diferencias significativas en algunos de los que codifican bZIPs. Estudios previos indicaban que el ortólogo de BLZ2 en maíz, O2, se regula post-traduccionalmente mediante un mecanismo de fosforilación/defosforilación reversible, y que la forma defosforilada es la fisiológicamente activa. En este trabajo se demostró que BLZ2 está sujeto a este tipo de regulación y que la proteín-fosfatasa HvPP2C2 está implicada en el proceso. La interacción de HvPP2C2 y BLZ2 tiene lugar en el núcleo celular únicamente en presencia de 100 μM ABA. En el mutante Riso 1508, BLZ2 se encuentra en un estado hiperfosforilado tanto durante la maduración como durante la germinación de la semilla, lo que dificultaría la unión de BLZ2 a las secuencias GLM en los promotores de los genes que codifican B-, C-,y ϒ- Hordeínas y CMe. Summary The seed is the main reproductive organ of spermatophyte plants allowing the spread of populations and ensuring their survival through its desiccation tolerance and because of their ability to germinate under optimum environmental conditions. Yield and economic value of cereal crops, that constitute the first world crop, depend largely on the efficiency with which they accumulate in the seed reserve substances: proteins, carbohydrates and lipids. The main carbohydrate accumulated in the barley seed is starch and the major protein fraction is that of prolamins (soluble in 70% ethanol); these proteins have a very low lysine content, an essential amino-acid for the diet of monogastric animals. In order to improve the nutritional value of the barley seed, different mutants have been obtained with a higher content of this amino-acid. Riso 1508 is one lysine-rich mutant whose mutation (lys3a) segregates as a single Mendelian gene with pleiotropic effects, such as a drastic reduction of genes encoding the trypsin inhibitor CMe and the B-, C-and ϒ-hordeins, but has not altered the expression of the gene encoding the D-hordeins. This latter gene lacks in its promotor the GLM motif (5’‐(G/A)TGA(G/C)TCA(T/C)‐3’), that is recognised by bZIP transcription factors In this work we have used the barley mutant Riso 1508 as a tool for better understanding gene regulation in seeds during the maturation and germination phases. To this aim, a transcriptomic analysis was performed comparing wild and mutant genotypes during seed maturation. Besides confirming variations in the expression of genes encoding reserve proteins, this analysis indicated that some genes related with carbohydrate metabolism were also affected. It was therefore decided to characterize the multigene family of sucrose synthases (SUSy) in barley. Two new genes were annotated, HvSs3 and HvSs4, and its expression was compared with that of genes HvSs1 and HvSs2, previously described in our laboratory. The expression of the four genes in different tissues and in response to abiotic stresses was analyzed by RTqPCR. HvSs1 and HvSs2 were preferentially expressed during the development of the endosperm, and the HvSs1 transcript was also abundant upon germination. HvSs1 was induced in leaves by anoxic conditions, HvSs3 by water stress, and both genes were induced by cold treatments. The subcellular localization of all four isoforms was not only cytoplasmic, but they could be found along the endoplasmic reticulum and at the inner side of the cell membrane; HvSS1, was also associated with the mitochondrial marker. These data suggest a distinct but partially overlapping roles for the barley sucrose synthases, described so far. The expression kinetics of the genes encoding the most important TFs involved in gene regulation during barley endosperm development was analyzed by RT-qPCR in both genotypes. These data show that the genes encoding DOF TFs were mis-regulated throughout the process in Riso 1508, although significant differences were also found among some of those encoding bZIPs. Previous studies indicated that the BLZ2 orthologue in maize, O2, was post-translationally regulated by reversible phosphorylation/dephosphorylation and that the dephosphorylated protein is the physiologically active form. In this work we demostrate that BLZ2 is under a similar regulation and that the proteinphosphatase HvPP2C2 is implicated in the process. The interaction between HvPP2C2 and BLZ2 takes place in the cell nucleus only in the presence of 100 μM ABA. In the Riso 1508 mutant, BLZ2 is found in a hyperphosphorylated state in the maturation phase and upon seed germination; because of this, the BLZ2 binding to the GLM promoter sequences of genes encoding B-, C- y ϒ- Hordeins and CMe would be decreased in the mutant.

Non invasive moisture measurement in agricultural fiel using a rolling spherical robot

Irrigation management in large crop fields is a very important practice. Since the farm management costs and the crop results are directly connected with the environmental moisture, water control optimization is a critical factor for agricultural practices, as well as for the planet sustainability. Usually, the crop humidity is measured through the water stress index (WSI), using imagery acquired from satellites or airplanes. Nevertheless, these tools have a significant cost, lack from availability, and dependability from the weather. Other alternative is to recover to ground tools, such as ground vehicles and even static base stations. However, they have an outstanding impact in the farming process, since they can damage the cultivation and require more human effort. As a possible solution to these issues, a rolling ground robot have been designed and developed, enabling non-invasive measurements within crop fields. This paper addresses the spherical robot system applied to intra-crop moisture measurements. Furthermore, some experiments were carried out in an early stage corn field in order to build a geo-referenced WSI map.