Recent trends in Inner Asian forest dynamics to temperature and precipitation indicate high sensitivity to climate change

Semi-arid ecosystems play an important role in regulating global climate with the fate of these ecosystems in the Anthropocene depending upon interactions among temperature, precipitation, and CO2. However, in cool-arid environments, precipitation is not the only limitation to forest productivity. Interactions between changes in precipitation and air temperature may enhance soil moisture stress while simultaneously extending growing season length, with unclear consequences for net carbon uptake. This study evaluates recent trends in productivity and phenology of Inner Asian forests (in Mongolia and Northern China) using satellite remote sensing, dendrochronology, and dynamic global vegetation model (DGVM) simulations to quantify the sensitivity of forest dynamics to decadal climate variability and trends. Trends in photosynthetically active radiation fraction (FPAR) between 1982 and 2010 show a greening of about 7% of the region in spring (March, April, May), and 3% of the area ‘browning’ during summertime (June, July, August). These satellite observations of FPAR are corroborated by trends in NPP simulated by the LPJ DGVM. Spring greening trends in FPAR are mainly explained by long-term trends in precipitation whereas summer browning trends are correlated with decreasing precipitation. Tree ring data from 25 sites confirm annual growth increments are mainly limited by summer precipitation (June, July, August) in Mongolia, and spring precipitation in northern China (March, April, May), with relatively weak prior-year lag effects. An ensemble of climate projections from the IPCC CMIP3 models indicates that warming temperatures (spring, summer) are expected to be associated with higher summer precipitation, which combined with CO2 causes large increases in NPP and possibly even greater forest cover in the Mongolian steppe. In the absence of a strong direct CO2 fertilization effect on plant growth (e.g., due to nutrient limitation), water stress or decreased carbon gain from higher autotrophic respiration results in decreased productivity and loss of forest cover. The fate of these semi-arid ecosystems thus appears to hinge upon the magnitude and subtleties of CO2 fertilization effects, for which experimental observations in arid systems are needed to test and refine vegetation models.

Resistance of a lowland rain forest to increasing drought intensity in Sabah, Borneo

Occasional strong droughts are an important feature of the climatic environment of tropical rain forest in much of Borneo. This paper compares the response of a lowland dipterocarp forest at Danum, Sabah, in a period of low (LDI) and a period of high (HDI) drought intensity (1986-96, 9.98 y;1996-99, 2.62 y). Mean annual drought intensity was two-fold higher in the HDI than LDI period (1997 v. 976 mm), and each period had one moderately strong main drought (viz. 1992, 1998). Mortality of `all' trees greater than or equal to 10 cm gbh (girth at breast height) and stem growth rates of `small' trees 10less than or equal to50 cm gbh were measured in sixteen 0.16-ha subplots (half on ridge, half on lower slope sites) within two 4-ha plots. These 10-50-cm trees were composed largely of true understorey species. A new procedure was developed to correct for the effect of differences in length of census interval when comparing tree mortality rates. Mortality rates of small trees declined slightly but not significantly between the LDI and HDI periods (1.53 to 1.48% y(-1)): mortality of all trees showed a similar pattern. Relative growth rates declined significantly by 23% from LDI to HDI periods (11.1 to 8.6 mm m(-1) y(-1)): for absolute growth rates the decrease was 28% (2.45 to 1.77 mm y(-1)). Neither mortality nor growth rates were significantly influenced by topography. For small trees, across subplots, absolute growth rate was positively correlated in the LDI period, but negatively correlated in the HDI period, with mortality rate. There was no consistent pattern in the responses among the 19 most abundant species (n greater than or equal to 50 trees) which included a proposed drought-tolerant guild. In terms of tree survival, the forest at Danum was resistant to increasing drought intensity, but showed decreased stem growth attributable to increasing water stress.

Insect oral secretions suppress wound-induced responses in Arabidopsis

The induction of plant defences and their subsequent suppression by insects is thought to be an important factor in the evolutionary arms race between plants and herbivores. Although insect oral secretions (OS) contain elicitors that trigger plant immunity, little is known about the suppressors of plant defences. The Arabidopsis thaliana transcriptome was analysed in response to wounding and OS treatment. The expression of several wound-inducible genes was suppressed after the application of OS from two lepidopteran herbivores, Pieris brassicae and Spodoptera littoralis. This inhibition was correlated with enhanced S. littoralis larval growth, pointing to an effective role of insect OS in suppressing plant defences. Two genes, an ERF/AP2 transcription factor and a proteinase inhibitor, were then studied in more detail. OS-induced suppression lasted for at least 48 h, was independent of the jasmonate or salicylate pathways, and was not due to known elicitors. Interestingly, insect OS attenuated leaf water loss, suggesting that insects have evolved mechanisms to interfere with the induction of water-stress-related defences.

Flor y fruto de vid [Vitis vinifera L.] : : micrografía aplicada a viticultura y enología

Un conocimiento apropiado de la anatomía de los diferentes tipos florales de la vid permite interpretar procesos de floración y fertilidad. Se ha observado durante cinco años las características de distintas variedades de vid en Mendoza (Argentina). Debido a las diferencias de variedades y viñedos, las observaciones se han tratado desde un punto de vista general. El estudio de las características anatómicas de la baya permite comprender fenómenos de maduración, estrés hídrico, deficiencias y productividad. En cuanto a los procedimientos tecnológicos, la observación de los componentes celulares del fruto: pared celular, polifenoles vacuolares, plástidos y ráfides de cristales de tartrato de calcio permite evaluar métodos enológicos, procesos de maceración y extracción de compuestos fenólicos y los tratamientos que deberían aplicarse para mejorar la calidad del vino.

Relaciones entre cavitación y cierre estomático en vides sometidas a déficit hídrico

La cavitación es una disfunción fisiológica que ocurre en el xilema de las plantas cuando estas están bajo déficit hídrico, y que entraña una pérdida de su conductancia hidráulica (kL), cuando algunos vasos se llenan de aire. Esto incide negativamente sobre la oferta de agua y afecta el potencial hídrico foliar (ΨL) y la hidratación de la canopia. El cierre estomático es una respuesta efectiva ante la disminución del contenido hídrico. Dependiendo de la especie vegetal, los estomas suelen cerrase para evitar la cavitación catastrófica. Una planta poco vulnerable a cavitar puede mantenerlos abiertos por más tiempo y por lo tanto seguir fotosintetizando. Por el contrario, plantas vulnerables a la cavitación deben cerrar sus estomas con anterioridad para poder evitar cavitaciones graves. En este trabajo se estudió, el ajuste estomático como mecanismo para evitar la cavitación en cuatro variedades contrastantes de vid (Grenache, Syrah, Malbec y Chardonnay). Se hipotetizó que las vides sometidas a déficit hídrico disminuyen gs para evitar sufrir cavitaciones catastróficas y que algunas variedades de vid, cuando crecen bajo restricción hídrica, se aclimatan mejor desarrollando un ajuste estomático más preciso, ó un sistema conductor menos vulnerable a cavitar, o ambas. Se diseñó un experimento aleatorizado en invernáculo donde se probaron dos situaciones hídricas edáficas (100% y 50% de FTSW). Luego de dos meses, se midieron, a lo largo del día, las variables gs, transpiración y potencial hídrico. Luego se construyeron las curvas de cavitación y se calculó la conduactancia hidráulica de la planta (kL) y el embolismo a lo largo del día. Finalmente se obtuvo la relación entre gs, kL y el embolismo. Mediante un modelo mecanístico, que se construyó teniendo en cuenta los flujos de agua y vapor, las kL y gs, y la vulnerabilidad del xilema a cavitar; se probó que gs no es la única variable responsable de frenar la embolia. Se determinó que gs y kL están íntimamente asociadas y que este acople entre ambas conductancias es lo que frena la embolia. Se concluyó que, en la vid y bajo niveles de estrés hídrico moderado, no es necesario un cierre estomático para controlar la cavitación, sino una disminución de la diferencia entre gs y kL (Δgs). Por esto, el mecanismo de control de la cavitación en la vid no conlleva un costo en términos de intercambio gaseoso. También se descubrió que bajo niveles moderados de déficit hídrico la vulnerabilidad xilemática no aumenta con respecto a las plantas que no sufren déficit, sin importar de qué variedad se trate.

Regulación de la hidratación y la turgencia foliares por mecanismos evitadores del estrés, y resistencia a déficit hídrico en vid

El transporte del agua en las plantas es impulsado por diferencias de energía libre entre el suelo y la atmósfera, y está regulado por mecanismos biológicos evitadores, como el cierre estomático. La hidratación y la turgencia foliares resultan del equilibrio entre ΨL del apoplasto, el potencial osmótico del simplasto y la elasticidad de los tejidos. Sobre esta base se conjeturó que las interacciones de los mecanismos evitadores del estrés hídrico de la planta tienen un rol clave en la definición de su resistencia a déficit hídrico. Para probar esta hipótesis se construyó un modelo mecanístico basado en las leyes del flujo de savia de Van de Honert, de difusión de Fick, de elasticidad de Hooke, la ecuación de Gardner para el flujo del agua en la rizósfera y el modelo de conductancia estomática (gs) de Buckley. Mediante el modelo se demostró teóricamente que la hidratación y la turgencia foliares dependen de la oferta de agua edáfica (representada por el potencial hídrico del suelo) y de la demanda evaporativa de la atmósfera (representada por la radiación absorbida, la temperatura del aire, la velocidad del viento y el déficit de presión de vapor de la atmósfera). También que los mecanismos evitadores del estrés hídrico -i.e., conductancia hidráulica de la planta, conductancia estomática, elasticidad del tejido y potencial osmótico a turgencia máxima- son todos necesarios para determinar la hidratación y la turgencia foliares. El modelo también demostró que la conductancia hidráulica suelo-hoja (kL) depende de la fracción de agua edáfica transpirable (FTSW) con un patrón de decaimiento sigmoide, a medida que el suelo se seca. Esto implica que las variables que dependen en parte de kL (i.e., gs, transpiración, fotosíntesis y superficie foliar) también dependen de FTSW con el mismo patrón. El modelo se probó experimentalmente a distintos niveles de humedad edáfica (desde déficit hídrico nulo, hasta severo) en cinco variedades de vid y mostró un poder predictivo superior al 90%. En todas las variedades las gs se asociaron linealmente con las kL observadas, al considerar todas las situaciones de déficit hídrico en conjunto, si bien la pendiente de estas relaciones fueron distintas en cada variedad. La contrastación experimental mostró que, en una escala de tiempo de varios meses, las variedades más evitadoras -i.e., Grenache y Cereza- mantuvieron mayor kL, ajuste osmótico y rigidez de los tejidos y una menor pendiente de la relación de gs vs. kL, que las variedades menos evitadoras -i.e., Malbec y Syrah-. La menor pendiente de la relación entre gs y kL, en las variedades más evitadoras, estuvo asociada a una mayor cantidad de estomas, en relación con la cantidad de células epidérmicas. Los variedades más evitadoras bajo déficit hídrico moderado -i.e., con una fracción de agua edáfica transpirable entre 0,6 y 0,4- tuvieron mayor superficie foliar y produjeron más biomasa, favoreciendo raíces profundas y densas, y ahorrando agua. Chardonnay mantuvo una alta hidratación y turgencia a expensas de un alto gasto de agua debido a que privilegiaba una alta kL por sobre el ajuste estomático, por lo que no podría considerarse en forma estricta como muy evitadora.

Respuesta al déficit hídrico en el crecimiento de forestales del bosque urbano de Mendoza : análisis comparativo en árboles jóvenes

El objetivo del trabajo fue evaluar la respuesta de plantas jóvenes de cuatro especies forestales urbanas a diferentes niveles de déficit hídrico. El ensayo se realizó en vivero, durante tres ciclos vegetativos, con plantas de Platanus x hispanica cv. acerifolia (plátano), Morus alba (morera), Fraxinus americana (fresno americano) y Acacia visco (visco). Los tratamientos de riego fueron: control (reposición del 100% transpirado); riego deficitario moderado (reposición del 66% transpirado) y riego deficitario severo (reposición del 33% transpirado). Las variables de respuesta evaluadas fueron altura, diámetro de tallos, área foliar y ancho de anillos de crecimiento. Las plantas bajo riego deficitario severo presentaron disminuciones en todos los parámetros de crecimiento considerados. Acacia visco resultó con crecimientos comparables al control, bajo riego deficitario equivalente al 66% del agua transpirada, con lo cual puede ser considerada especie de interés para forestaciones en condiciones de estrés hídrico moderado.

Plant-wax, alkenone and major element analyses of ODP Site 659 for the Last Glacial cycle and associated coretop data

We present a hydrologic reconstruction of the Sahara-Sahel transition, covering the complete last glacial cycle (130 ka), based on a combination of plant-wax-specific hydrogen (dD) and carbon isotopes (d13C). The dD and d13C signatures of long-chain n-alkanes from ODP Site 659 off NW Africa reveal a significant anti-correlation. Complementary to published pollen data, we infer that this plant-wax signal reflects sensitive responses of the vegetation cover to precipitation changes in the Sahel region, as well as varying contributions from biomes north of the Sahara (C3 domain) by North-East Trade Winds (NETW). During arid phases, especially the northern parts of the Sahel likely experienced crucial water stress, which resulted in a pronounced contraction of the vegetation cover, thus reducing the amount of C4 plant waxes from the region. The increase in NETW strength during dry periods further promoted a more pronounced C3-plant-wax signal derived from the North African C3 plant domain. During humid periods, the C4-dominated Sahelian environments spread northward into the Saharan realm, in association with lower NETW inputs of C3 plant waxes. Arid-humid cycles deduced from plant-wax dD are in accordance with concomitant changes in weathering intensity reflected in varying major element distributions. Environmental shifts are generally linked to periods with large fluctuations in Northern Hemisphere summer insolation. During Marine Isotope Stages 2 and 3, when insolation variability was low, coupling of the hydrologic regime to alkenone-based estimates of NE Atlantic sea-surface temperatures becomes apparent.

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.

Effect of solute and matric potential on growth rate of fungal species isolated from cheese

The effect of water potential ( J w ) on the growth of 15 fungal species isolated from cheeses was analysed. The species, identified mainly by analysis of DNA sequences, belonged to genera Penicillium , Geotrichum , Mucor , Aspergillus , Microascus and Talaromyces . Particularly, the effect of matric potential ( J m ), and ionic (NaCl) and non-ionic (glycerol) solute potentials ( J s ) on growth rate was studied. The response of strains was highly dependent on the type of J w . For J s , clear profiles for optimal, permissive and marginal conditions for growth were obtained, and differences in growth rate were achieved comparing NaCl and glycerol for most of the species. Conversely, a sustained growth was obtained for J m in all the strains, with the exception of Aspergillus pseudoglaucus , whose growth increased proportionally to the level of water stress. Our results might help to understand the impact of environmental factors on the ecophysiology and dynamics of fungal populations associated to cheeses.