Climatic factors control rodent seed predation in Pinus pinea L. stands in Central Spain

- Context: Pinus pinea L. presents serious problems of natural regeneration in managed forest of Central Spain. The species exhibits specific traits linked to frugivore activity. Therefore, information on plant–animal interactions may be crucial to understand regeneration failure. - Aims: Determining the spatio-temporal pattern of P. pinea seed predation by Apodemus sylvaticus L. and the factors involved. Exploring the importance of A. sylvaticus L. as a disperser of P. pinea. Identifying other frugivores and their seasonal patterns. - Methods: An intensive 24-month seed predation trial was carried out. The probability of seeds escaping predation was modelled through a zero-inflated binomial mixed model. Experiments on seed dispersal by A. sylvaticus were conducted. Cameras were set up to identify other potential frugivores. - Results: Decreasing rodent population in summer and masting enhances seed survival. Seeds were exploited more rapidly nearby parent trees and shelters. A. sylvaticus dispersal activity was found to be scarce. Corvids marginally preyed upon P. pinea seeds. - Conclusions: Survival of P. pinea seeds is climate-controlled through the timing of the dry period together with masting occurrence. Should germination not take place during the survival period, establishment may be limited. A. sylvaticus mediated dispersal does not modify the seed shadow. Seasonality of corvid activity points to a role of corvids in dispersal.

Can seed production and restricted dispersal limit recruitment in Pinus pinaster Aiton from the Spanish Northern Plateau?

Natural regeneration faces increasing difficulties in dry forests from the Mediterranean basin, including for normally well-regenerating species such as maritime pine (Pinus pinaster Aiton). In this paper, we studied female fertility, seed dispersal and spread rates in P. pinaster from the Spanish Northern Plateau, where natural regeneration failure is a main concern for forest managers. For this purpose we periodically collected data from seed traps and trees located at two core locations across several years. We found significant variation in interannual cone production, with the best seed trees being the same across years. In addition, we found highly skewed distributions of female reproductive effort and large fertility differences across stands located few kilometres away. Annual seed dispersal kernels fitted lognormal or 2Dt models depending on the stand analysed, with median dispersal distances between 14 and 25 m. Kernels fitted for maximum dispersal periods showed an outstanding intraseasonal variation of median dispersal distances, from 10 to 54 m, in association to variable patterns of rainfall and maximum wind speed. The amount of seed produced appeared to be enough to guarantee the natural regeneration of the stands during the typical 20-year regeneration period. Colonisation simulations concluded that Mediterranean maritime pine has a notable dispersion capacity, which is strongly influenced by levels of fecundity and, especially, by the number and frequency of long-distance dispersal events. The latter play a key role in tree dispersion processes through enlarging the occupied area and fostering the invasion of abandoned crop land.

Estudio de los efectos en el viñedo de la modificación del déficit hídrico continuo en los periodos anterior y posterior al envero (cv. Cabernet Sauvignon; Vitis vinifera L.)

En regiones semiáridas, con veranos cálidos, evapotranspiraciones elevadas, humedades relativas bajas, además de precipitaciones intermitentes y escasas (400 mm al año) que reducen la recarga del perfil en invierno, se puede ver afectada la productividad y sostenibilidad de los viñedos. Por eso en estas zonas puede ser necesario utilizar el riego suplementario para mantener la calidad y aumentar la productividad. En este marco de circunstancias es en el que se desarrolla la mayoría de la viticultura española, y en concreto en la zona centro donde se realizó el estudio. Con este trabajo se pretende estudiar la influencia del agua del riego, con distintas dosis y aplicada en diferentes fases de crecimiento y maduración de la baya, en la producción y calidad de la uva. Se estudió la influencia del déficit moderado continuo aplicado antes y después de envero, en el aprovechamiento de los recursos ambientales y en la respuesta del cv. Cabernet Sauvignon. El ensayo se desarrolló en 2010 y 2011 en la finca “La Alcoholera”, perteneciente a bodegas LICINIA S.L., ubicada en la localidad de Chinchón, Comunidad de Madrid, España. Durante 2010 y 2011 se realizaron estudios en cepas del cv. Cabernet Sauvignon (clon 15), injertadas sobre 41B y plantadas en 2005. Con poda corta en Cordón Royat unilateral de 80 cm de altura, y cuya vegetación alcanzó una altura de 97 cm. Las cepas del ensayo fueron conducidas verticalmente en espaldera (VSP), con 10 yemas por metro lineal. Las plantas dentro de la fila fueron separadas 1 m y la distancia de la calle fue de 3 m. La orientación de las filas norte – sur. Se plantearon cuatro tratamientos experimentales con diferentes dosis y momento de aplicación del riego. - T: Testigo. Déficit ligero, con un aporte continuado de agua desde floración (420 mm). - DMc: Déficit moderado continuo. Manejo del riego convencional, empezando en pre-envero (154 mm) - DM1: Déficit moderado a partir de pre-envero. Aporte de riego continuado desde floración reduciéndose la dosis de riego en pre-envero (312 mm). - DM2: Déficit moderado hasta pre-envero. Se empezó a regar a partir de preenvero (230 mm). Se observó que bajo condiciones de estrés hídrico leve, el crecimiento se reduce disminuyendo la migración de fotoasimilados hacia los órganos vegetativos. Generando menor área foliar en las plantas sometidas a déficit hídrico moderado entre floración y cuajado. Disminuyendo su consumo. En maduración la humedad del suelo dependió principalmente del riego aplicado y del consumo de la planta; dicho consumo dependió a la vez del desarrollo foliar del canopy y de la necesidad hídrica de la vid. El aumento del déficit hídrico disminuyó el contenido de agua en el suelo, lo que provocó diferencias en el estado hídrico y en el intercambio gaseoso de las hojas. En condiciones de déficit moderado, la fotosíntesis se encontró altamente correlacionada con el potencial hídrico foliar medido a mediodía solar. Además en condiciones de déficit moderado continuo la disponibilidad hídrica, la demanda atmosférica y el nivel de hidratación de las hojas, interaccionaron de forma compleja en la regulación estomática de las hojas, condicionando el intercambio gaseoso y la eficiencia en el uso del agua. Al aumentar el volumen de agua aplicado el peso de madera de poda por metro de fila fue superior. Las diferencias encontradas en el rendimiento fueron debidas a las diferencias en el peso de baya. El cual estuvo condicionado por la estrategia de riego, ya que, el déficit hídrico moderado antes de envero fue más crítico para el rendimiento que el déficit impuesto durante la madurez. Por otro lado, la biomasa dependió de la actividad fisiológica de la planta, la cual fue altamente dependiente de la disponibilidad hídrica. Déficit hídrico moderado aplicado de cuajado a envero, generó bayas más pequeñas y aumentó la relación hollejo:pulpa. El déficit moderado aplicado después de envero favoreció la acumulación de IPT y antocianos extraíbles, mejorando la calidad de la uva, pero disminuyó la acidez de la baya. El riego aplicado de envero a vendimia desaceleró la concentración de azúcares en bayas que fueron sometidas a déficit antes de pre-envero. ABSTRACT Mediterranean climate is characterized by hot summers, high evapotranspiration rates, and scarce precipitations (400 mm per year) during grapevine cycle. These extremely dry conditions affect vineyard productivity and sustainability. Supplementary irrigation is needed practice in order to maintain yield and quality. Almost all Spanish grape growing regions are characterized by these conditions, especially in the center region, where this trial was performed. The main objective of this work is to study the influence of water irrigation on yield and quality. For this aim, different levels of irrigation (mm of water applied) were applied during different stages of growth and berry maturity of Caberent Sauvignon grapevines. The work was conducted from 2010 to 2011 and located in Licinia (40º 12´ N, 3º 28´ W), Madrid, Spain. The cultivar utilized was Cabernet Sauvignon, clone 15 grafted onto rootstock 41B, planted in 2005. The vineyard was oriented north-south, with spacing on 3 meters between rows and 1 meter between plants. Vines were spurpruned to 10 buds per meter and trained in unilateral cordon with a height of 80 cm. Shoots were positioned vertically (VSP). Considering the amount of water and the moment of the application, four experimental treatments were applied: - T: Control sample. Slight deficit (420 mm) applied from bloom to maturity. - DMc: Continuous moderate deficit. Traditional irrigation: application of 154 mm of water from pre-veraison to maturity. - DM1: Deficit moderate from pre-veraison. Irrigation of 312 mm of water from bloom to pre-veraison. - DM2: Deficit moderate to pre-veraison. Irrigation of 230 mm of water from preveraison to maturity Under moderated water stress conditions it was seen than leaf growth decreases due to the reduction of migration of photo-assimilates to vegetative organs. Vines with moderate water deficit between flowering and ripening develop less leaf area and decrease its water consumption. During maturation of berries, soil moisture depends on irrigation and plant consumption. This consumption further depends on vegetation development and on the vine needs for water. By increasing water deficit, the water content in the soil decreases. This causes differences in leaf water status and in the gases exchange. Under moderate deficit conditions, photosynthesis was found highly correlated with midday leaf water potential. Further, atmospheric demand and the level of leaves hydration interact in complex ways in the stomatal regulation, which affects leaf gas exchange and the efficiency of water use. The amount of water applied is directly proportional to pruning weight. Changes in berry weight cause differences in yield ratios. The differences in berry weights are conditioned by the irrigation strategy, the moderate water deficit before veraison influences more than the deficit applied from veraison. Biomass generated for the plant depends on its physiological activity, which is highly related to the water availability. Moderate water deficit applied from fruit set to ripening generates smaller berries, increasing the pulp/skin ratio. Moderate deficit applied after veraison promotes the accumulation of extractables anthocyans and TPI. Despite this treatment improves color parameters of the grapes, it decreases its total acidity. Irrigation applied from veraison to harvest slows down sugar accumulation in berries compared to those under deficit before veraison conditions.

bZIP transcription factors in the regulatory networks controlling seed maturation and germination of Arabidopsis thaliana

Se describe la expresión por RTqPCR de los genes que codifican los factores transcripcionales bZIP44 y bZIP9. Asimismo se establece la interacción entre ambas proteínas en el sistema de 2 híbridos de levadura y in planta por complementación bimolecular fluorescente.

Combinatorial interaction between AtbZIP44 and other bZIP transcription factors in the regulation of Arabidopsis thaliana seed maturation and germination

El gen AtbZIP44 se expresa en el endospermo micropilar y en la radícula de semillas en germinación. El mRNA del gen AtbZIP9 se localiza también en la radícula. Se realizan experimentos de interacción proteína-proteína entre los correspondientes factores transcripcionales.

A partial study of vertical distribution of conventional no-till seeders and spatial variability of seed depth placement of maize in the Alentejo region, Portugal

The requirements for a good stand in a no-till field are the same as those for conventional planting as well as added field and machinery management. Among the various factors that contribute towards producing a successful maize crop, seed depth placement is a key determinant. Although most no-till planters on the market work well under good soil and residue conditions, adjustments and even modifications are frequently needed when working with compacted or wet soils or with heavy residues. The main objective of this study, carried out in 2010, 2011 and 2012, was to evaluate the vertical distribution and spatial variability of seed depth placement in a maize crop under no-till conditions, using precision farming technologies and conventional no-till seeders. The results obtained indicate that the seed depth placement was affected by soil moisture content and forward speed. The seed depth placement was negatively correlated with soil resistance and seeding depth had a significant impact on mean emergence time and the percentage of emerged plants. Shallow average depth values and high coefficients of variation suggest a need for improvements in controlling the seeders’ sowing depth mechanism or more accurate calibration by operators in the field.

Spatial variability of seed depth placement of maize under no tillage in Alentejo, Portugal

Among the various factors that contribute towards producing a successful maize crop, seed depth placement is a key determinant, especially in a no-tillage system. The main objective of this work was to evaluate the spatial variability of seed depth placement and crop establishment in a maize crop under no-tillage conditions, using precision farming technologies. The obtained results indicate that seed depth placement was significantly affected by soil moisture content, while a very high coefficient of variation of 39% was found for seed depth. Seeding depth had a significant impact on mean emergence time and percentage of emerged plants. Shallow average depth values and the high coefficient of variation suggest a need for improvement in controlling the seeders sowing depth.

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.

Contribute to the Study of Vertical Distribution of Conventional No-Till Seeders and Spatial Variability of Seed Depth Placement of Maize in Alentejo, Portugal

Entre os vários fatores que contribuem para a produção de uma cultura de milho, a distribuição vertical dos semeadores avaliada através da localização da semente em profundidade é um fator-chave, especialmente na técnica de sementeira direta. Simultaneamente, dada a complexidade dos ecossistemas naturais e agrícolas em sistemas de agricultura de conservação, a gestão diferenciada e localizada das parcelas assume um importante papel na análise e gestão da variabilidade das propriedades do solo e estabelecimento das culturas, nomeadamente utilizando informação geo referenciada e tecnologia expedita. Assim, o principal objetivo desta Tese foi a avaliação em culturas de milho da variabilidade espacial da localização de semente em profundidade e estabelecimento da cultura em sementeira direta usando sistemas convencionais de controlo de profundidade, tendo-se comparado com diferentes sistemas de mobilização e recorrendo a tecnologias de agricultura de precisão. Os ensaios decorreram na região Mediterrânea do Alentejo, em propriedades agrícolas no decorrer das campanhas de 2010, 2011, 2012 e 2015 em 6 diferentes campos experimentais. O trabalho experimental consistiu em ensaios com avaliações in loco do solo e cultura, consumo de combustível das operações e deteção remota. Os resultados obtidos indicam que não só o sistema de mobilização afetou a localização da semente em profundidade, como em sementeira direta a profundidade de sementeira foi afetada pelo teor de humidade do solo, resistência do solo à profundidade e velocidade da operação de sementeira. Adicionalmente observaram-se condições heterogéneas de emergência e estabelecimento da cultura afetadas por condições físicas de compactação do solo. Comparando os diferentes sistemas de mobilização, obteve-se uma significativa redução de combustível para a técnica de sementeira direta, apesar de se terem observado diferenças estatísticas significativas considerando diferentes calibrações de profundidade de sementeira Do trabalho realizado nesta Tese ressalva-se a importância que as tecnologias de agricultura de precisão podem ter no acompanhamento e avaliação de culturas em sementeira direta, bem como a necessidade de melhores procedimentos no controlo de profundidade dos semeadores pelo respetivos operadores ou ao invés, a adoção de semeadores com mecanismos ativos de controlo de profundidade. ABSTRACT Among the various factors that contribute towards producing a successful maize crop, seeders vertical distribution evaluated through seed depth placement is a key determinant, especially under a no-tillage technique. At the same time in conservation agriculture systems due to the complexity of natural and agricultural ecosystems site specific management became an important approach to understand and manage the variability of soil properties and crop establishment, especially when using geo spatial information and affording readily technology Thus, the main objective of this Thesis was to evaluate the spatial variability of seed depth placement and crop establishment in maize crops under no-tillage conditions compared to different tillage systems, using conventional seed depth control no till seeders and precision farming technologies. Trials were carried out in the Mediterranean region of Alentejo, in private farms along the sowing operations season over the years 2010, 2011, 2012 and 2015 in 6 different experimental fields. Experimental work covered field tests with in loco soil and crop evaluations, fuel operation evaluations and aerial sensing. The results obtained indicate that not only tillage system affected seed depth placement but under no till conditions seed depth was affected by soil moisture content, soil resistance to penetration and seeders forward speed. In addition uneven crop seedling and establishment depended on seed depth placement and could be affected by physical problems of compaction layers. Significant reduction in fuel consumption was observed for no till operations although significant differences observed according to different setting calibrations of seed depth control. According to the results, precision agriculture is an important tool to evaluate crops under no till conditions and seed depth mechanisms should be more accurate by the operators or is determinant the adoption of new active depth control technology to improve seeders performance.

Seed germination characteristics of Phillyrea angustifolia L. and P. latifolia L. (Oleaceae), two Mediterranean shrub species having lignified endocarp

The aim of this study was to determine the germination characteristics of Phillyrea angustifolia L. and P. latifolia L. seeds in order to develop an optimized propagation protocol for Phillyrea species. Seeds of P. angustifolia and P. latifolia were collected from wild plants growing in Cáceres province (CW Spain) and Andalucía (S Spain), respectively. Percentage of water uptake for P. latifolia seeds was calculated. Seeds with and without endocarp were germinated at different constant and alternating temperatures. Seeds without endocarp were soaked in distilled water or gibberellic acid, and then set to germinate. Seeds with endocarp of both species were stratified at 5 ºC for 30 or 90 days and then the endocarp was completely removed from the seeds before they were sowed. Chemical scarification with sulfuric acid and mechanical scarification were tested on P. angustifolia seeds with endocarp. Phillyrea endocarp was permeable to water, since Phillyrea seeds with endocarp imbibed water, but water uptake was faster when the endocarp was removed. Moreover, the encodarp could interfere mechanically in the emergence of the radicle, since seed germination of Phillyrea species was promoted by the complete removal of the lignified endocarp surrounding each seed. Optimal germination temperature for both species was 15 ºC, and lower temperatures produced secondary dormancy. Soaking in distilled water or gibberellic acid did not significantly enhance seed germination. Cold stratification and chemical scarification treatments were detrimental for seed germination. Keywords cold stratification, Phillyrea species, treatments before sowing, seed germination, seed scarification, lignified endocarp.

Mannans and endo-beta-mannanases (MAN) in Brachypodium distachyon: expression profiling and possible role of the BdMAN genes during coleorhiza-limited seed germination

Immunolocalization of mannans in the seeds of Brachypodium distachyon reveals the presence of these polysaccharides in the root embryo and in the coleorhiza in the early stages of germination (12h), decreasing thereafter to the point of being hardly detected at 27h. Concurrently, the activity of endo-β-mannanases (MANs; EC 3.2.1.78) that catalyse the hydrolysis of β-1,4 bonds in mannan polymers, increases as germination progresses. The MAN gene family is represented by six members in the Brachypodium genome, and their expression has been explored in different organs and especially in germinating seeds. Transcripts of BdMAN2, BdMAN4 and BdMAN6 accumulate in embryos, with a maximum at 24–30h, and are detected in the coleorhiza and in the root by in situ hybridization analyses, before root protrusion (germination sensu stricto). BdMAN4 is not only present in the embryo root and coleorhiza, but is abundant in the de-embryonated (endosperm) imbibed seeds, while BdMAN2 and BdMAN6 are faintly expressed in endosperm during post-germination (36–42h). BdMAN4 and BdMAN6 transcripts are detected in the aleurone layer. These data indicate that BdMAN2, BdMAN4 and BdMAN6 are important for germination sensu stricto and that BdMAN4 and BdMAN6 may also influence reserve mobilization. Whether the coleorhiza in monocots and the micropylar endosperm in eudicots have similar functions, is discussed.

Enhanced methionine levels and increased nutritive value of seeds of transgenic lupins (Lupinus angustifolius L.) expressing a sunflower seed albumin gene

With the aim of improving the nutritive value of an important grain legume crop, a chimeric gene specifying seed-specific expression of a sulfur-rich, sunflower seed albumin was stably transformed into narrow-leafed lupin (Lupinus angustifolius L.). Sunflower seed albumin accounted for 5% of extractable seed protein in a line containing a single tandem insertion of the transferred DNA. The transgenic seeds contained less sulfate and more total amino acid sulfur than the nontransgenic parent line. This was associated with a 94% increase in methionine content and a 12% reduction in cysteine content. There was no statistically significant change in other amino acids or in total nitrogen or total sulfur contents of the seeds. In feeding trials with rats, the transgenic seeds gave statistically significant increases in live weight gain, true protein digestibility, biological value, and net protein utilization, compared with wild-type seeds. These findings demonstrate the feasibility of using genetic engineering to improve the nutritive value of grain crops.

Directed seed dispersal by bellbirds in a tropical cloud forest

A fundamental goal of plant population ecology is to understand the consequences for plant fitness of seed dispersal by animals. Theories of seed dispersal and tropical forest regeneration suggest that the advantages of seed dispersal for most plants are escape from seed predation near the parent tree and colonization of vacant sites, the locations of which are unpredictable in space and time. Some plants may gain in fitness as a fortuitous consequence of disperser behavior if certain species of dispersers nonrandomly place seeds in sites predictably favorable for seedling establishment. Such patterns of directed dispersal by vertebrates long have been suggested but never demonstrated for tropical forest trees. Here we report the pattern of seed distribution and 1-year seedling survival generated by five species of birds for a neotropical, shade-tolerant tree. Four of the species dispersed seeds to sites near the parent trees with microhabitat characteristics similar to those at random locations, whereas the fifth species, a bellbird, predictably dispersed seeds under song perches in canopy gaps. The pattern of seedling recruitment was bimodal, with a peak near parent trees and a second peak, corresponding to bellbird song perches, far (>40 m) from parent trees. Seedling survival was higher for seeds dispersed by bellbirds than by the other species, because of a reduction in seedling mortality by fungal pathogens in gaps. Thus, bellbirds play a significant role in seed dispersal by providing directed dispersal to favorable sites and therefore may influence plant recruitment patterns and species diversity in Neotropical forests.

Cloning and Characterization of TPE4A, a Thiol-Protease Gene Induced during Ovary Senescence and Seed Germination in Pea1

A cDNA clone encoding a thiol-protease (TPE4A) was isolated from senescent ovaries of pea (Pisum sativum) by reverse transcriptase-polymerase chain reaction. The deduced amino acid sequence of TPE4A has the conserved catalytic amino acids of papain. It is very similar to VSCYSPROA, a thiol-protease induced during seed germination in common vetch. TPE4A mRNA levels increase during the senescence of unpollinated pea ovaries and are totally suppressed by treatment with gibberellic acid. In situ hybridization indicated that TPE4A mRNA distribution in senescent pea ovaries is different from that of previously reported thiol-proteases induced during senescence, suggesting the involvement of different proteases in the mobilization of proteins from senescent pea ovaries. TPE4A is also induced during the germination of pea seeds, indicating that a single protease gene can be induced during two different physiological processes, senescence and germination, both of which require protein mobilization.

Temporal and Spatial Patterns of Accumulation of the Transcript of Myo-Inositol-1-Phosphate Synthase and Phytin-Containing Particles during Seed Development in Rice1

Myo-inositol-1-phosphate (I[1]P) synthase (EC 5.5.1.4) catalyzes the reaction from glucose 6-phosphate to I(1)P, the first step of myo-inositol biosynthesis. Among the metabolites of I(1)P is inositol hexakisphosphate, which forms a mixed salt called phytin or phytate, a storage form of phosphate and cations in seeds. We have isolated a rice (Oryza sativa L.) cDNA clone, pRINO1, that is highly homologous to the I(1)P synthase from yeast and plants. Northern analysis of total RNA showed that the transcript accumulated to high levels in embryos but was undetectable in shoots, roots, and flowers. In situ hybridization of developing seeds showed that the transcript first appeared in the apical region of globular-stage embryos 2 d after anthesis (DAA). Strong signals were detected in the scutellum and aleurone layer after 4 DAA. The level of the transcript in these cells increased until 7 DAA, after which time it gradually decreased. Phytin-containing particles called globoids appeared 4 DAA in the scutellum and aleurone layer, coinciding with the localization of the RINO1 transcript. The temporal and spatial patterns of accumulation of the RINO1 transcript and globoids suggest that I(1)P synthase directs phytin biosynthesis in rice seeds.