Sarna del manzano : detección de la forma sexual (Venturia inaequalis ) en Mendoza (Argentina)

El aumento de los daños provocados por la sarna del manzano [Fusicladium dendritichum (Wallr.) Fuckl.] en montes frutales de Mendoza (Argentina), motivó estudios para aclarar la biología del hongo y buscar la presencia de la forma sexual, no detectada aún en la provincia. Al momento de la brotación se muestrearon hojas del año anterior que se encontraban en el suelo de dichos montes. En el laboratorio fueron observadas bajo estereomicroscopio. Los cuerpos negros inmersos en el tejido de la hoja fueron montados en preparados microscópicos y observados para establecer su identidad. Se confirmó la presencia en la zona de la forma teleomórfica del agente causal de la sarna del manzano. Este hecho tiene importancia en la epidemiología de la enfermedad y en las estrategias de control.

Duraznos O'Henry : impacto económico por pérdidas de calidad

El durazno O'Henry para consumo en fresco es una variedades apta para la exportación. Consecuentemente se propone un método sencillo para determinar las pérdidas de calidad que impiden obtener mejores precios en los mercados.. Las mismas se relacionan con una mala gestión en todos los niveles: producción, empaque y comercialización, y la deficiente capacitación del personal involucrado. En la temporada 1999/2000 el granizo fue la causa más frecuente en precosecha; sus marcas explican el 70 % de los frutos con falta de calidad. En cambio, el principal factor en cosecha fueron las picaduras de grafolita, dando 80 % de frutos sin calidad exportable. En galpón de empaque, el mal manejo poscosecha -revelado en machucones, marcas de uñas y rajaduras- originó el 30 % de las causas de no calidad; si se le agregan las picaduras de grafolita, el porcentaje se aproxima al 60 %. Ambas causas pueden minimizarse con buena planificación y capacitación. Con simulación económica se detectó -en la precitada temporada- 20 % de pérdida de calidad de una partida lo cual motiva una disminución superior al 13 % en los ingresos. Esta diferencia depende no sólo del peso relativo de las distintas calidades sino también del precio pactado.

Manejo de la carga frutal y productividad de duraznos conserveros

Para determinar la relación entre la carga frutal y la productividad, medida en términos de producción destinada a mitades (PDM) y producción para pulpa, se realizó un ensayo en un monte comercial de duraznero variedad Bowen, Andross y Ross. Se utilizaron árboles injertados sobre patrón Nemaguard, conducidos en vaso tipo californiano e implantados a 6 m entre hileras y a 5 m entre plantas (333 árboles/ha), los que fueron raleados 20 días después del inicio de endurecimiento de carozo. La carga frutal quedó establecida en el rango de 750 a 1350 frutos/planta en Bowen y Ross y entre 850 y 1400 frutos/planta en Andross. A la cosecha se evaluó: tamaño de frutos; producción de los árboles, a través de las variables producción total, producción destinada a mitades (PDM) y producción para pulpa; y valor de la producción. La carga frutal se vinculó, con un alto grado de correlación, con todas las variables analizadas y se observó una respuesta a la carga de frutos muy similar en las tres variedades en estudio. A medida que se incrementó la carga frutal, aumentó la producción total, aunque disminuyó el peso medio de frutos. La PDM aumentó en función de la carga frutal, hasta alcanzar valores máximos con 1200 frutos/planta; por encima de un valor crítico en la carga frutal, la PDM empezó a disminuir. Los rendimientos de pulpa, en fábrica, no se relacionaron linealmente con los rendimientos totales del monte frutal y respondieron a una función similar a la que relaciona carga frutal y PDM. Los resultados indicaron que, con independencia del destino de la fruta, el raleo de frutos aseguró los rendimientos y la calidad de frutos que permitieron maximizar la rentabilidad del productor, convirtiéndose en una tarea cultural imprescindible, desde el punto de vista técnico y económico.

Uso de agua en industrias de elaboración de conservas de tomate y de durazno de Mendoza, Argentina

El objetivo de este trabajo es conocer, a nivel de cuenca, el volumen de agua utilizado por las industrias de elaboración de conservas de tomate y de durazno de Mendoza. Para ello se estima la materia prima utilizada en la elaboración de estas conservas a partir de datos de superficie cultivada para tal destino y de rendimientos por superficie obtenidos en el Registro Permanente de Uso de la Tierra de Mendoza y el Instituto Nacional de Tecnología Agropecuaria. Se emplearon coeficientes de volumen de agua utilizada por unidad de materia prima procesada, que varían entre 5 y 25 L kg-1 de producto procesado, sin incluir el uso de agua para riego en fincas. Los resultados se analizaron para diferentes valores de coeficientes asociados a la eficiencia del uso del agua, en escenarios optimista y pesimista. Se concluye que las industrias elaboradoras de conservas de tomate y de durazno de Mendoza utilizan entre 0,66 y 6,15 hm3/año. El mayor consumo de agua de las conserveras de tomate ocurre en la cuenca Norte, alcanzando el 64,9% del total demandado por tales industrias. Para las conserveras de durazno, el mayor consumo se produce en la cuenca Sur con un 46% de total demandado.

Palynology of marine sediment cores from the West African coast

Seven sediment cores from the cruises of the "Meteor" and "Valdivia" were examined palynologically. The cores were retrieved from the lower continental slope in the area of between 33.5° N and 8° N, off the West African coast. Most of the cores contain sediments from the last Glacial and Interglacial period. In some cases, the Holocene sediments are missing. Some individual cores contain sediments also from earlier Glacial and Interglacial periods. The main reason for making this palynological study was to find out the differences between the vegetation of Glacial and Interglacial periods in those parts of West Africa which at present belong to the Mediterranean zone, the Sahara and the zones of the savannas and tropical forests. In today's Mediterranean vegetation zone at core 33.5° N, forests and deciduous forests in particular, are missing during Glacial conditions. Semi-deserts are found instead of these. In the early isotope stage 1, there is a very significant development of forests which contain evergreen oaks; this is the Mediterranean type of vegestation development. The Sahara type of vegetation development is shown in four cores from between 27° N and 19° N. The differences between Glacial and Interglacial periods are very small. It must be assumed therefore that in this latitudes, both Glacial and Interglacial conditions gave rise to desert generally. The results are in favour of a slightly more arid climate during Glacial and more humid one during Interglacial periods. The southern boundary of the Sahara and the adjacent savannas with grassland and tropical woods were situated more to the south during the Glacial periods than they were during the Interglacial ones. In front of today's savanna belt, it can be seen from the palynological results that there are considerable differences between the vegetation of Glacial and Interglacial periods. The woods are more important in Interglacial periods. During the Glacial periods these are replaced from north to south decreasingly by grassland (savanna and rainforest type of vegetation development). The southern limit of the Sahara during stage 2 was somewhat between 12° N and 8° N which is between 1.5 and 5 degrees in latitude further south than it i s today. Not only do these differences in climate and vegetation apply to the maximum of the last Glacial and for the Holocene, but they apparently apply also to the older Glacial and Interglacial periods, where they have been found in the profiles. The North African deset belt can be said to have expanded during Glacial times both towards the north and towards the south. All the available evidence of this study indicates that the grass land or the semi-desert of the Southern Europe cam einto connection with those of the N Africa; there could not have been any forest zone between them. The present study was also a good opportunity for investigating some of the basic marine palynological problems. The very well known overrepresentation of pollen grains of the genus Pinus in marine sediments can be traced as fa as 21° N. The present southern limit for the genus Pinus is on the Canaries and on the African continent as approximately 31° N. Highest values of Ephedra pollen grains even occur south of the main area of the present distribution of that genus. These does not seem to be any satisfactory explanation for this. In general, it would appear that the transport of pollen grains from the north is more important than transport from the south. The results so far, indicate strongly that further palynological studies are necessary. These should concentrate particularly on cores from between 33° N and 27° N as well as between 17° N and 10° N. It would also be useful to have a more detailed examination of sediments from the last Intergalcial period (substage 5 e). Absolute pollen counts and more general examination of surface samples would be desirable. Surface samples should be taken from the shelf down to the bottom of the continental slope in different latitudes.

Podredumbre parda del melocotonero (Monilinia spp.) : detección, identificación de especies y contribución a la epidemiología de la enfermedad

Monilinia spp. (M. laxa, M. fructigena y M. fructicola) causa marchitez en brotes y flores, chancros en ramas y podredumbre de la fruta de hueso provocando pérdidas económicas importantes en años con climatología favorable para el desarrollo de la enfermedad, particularmente en variedades tardías de melocotonero y nectarino. En estos huéspedes en España, hasta el momento, la especie predominante es M. laxa y, en menor proporción, M. fructigena. La reciente introducción en Europa de la especie de cuarentena M. fructicola hace necesaria una detección e identificación rápida de cada una de las especies. Además, hay diversos aspectos de la etiología y epidemiología de la enfermedad que no se conocen en las condiciones de cultivo españolas. En un primer objetivo de esta Tesis se ha abordado la detección e identificación de las especies de Monilinia spp. causantes de podredumbre parda. El estudio de las bases epidemiológicas para el control de la enfermedad constituye el fin del segundo objetivo. Para la detección del género Monilinia en material vegetal por PCR, diferenciándolo de otros hongos presentes en la superficie del melocotonero, se diseñaron una pareja de cebadores siguiendo un análisis del ADN ribosomal. La discriminación entre especies de Monilinia se consiguió utilizando marcadores SCAR (región amplificada de secuencia caracterizada), obtenidos después de un estudio de marcadores polimórficos de ADN amplificados al azar (RAPDs). También fue diseñado un control interno de amplificación (CI) basado en la utilización de un plásmido con secuencias de los cebadores diferenciadores del género, para ser utilizado en el protocolo de diagnóstico de la podredumbre parda con el fin de reconocer falsos negativos debidos a la inhibición de PCR por componentes celulares del material vegetal. Se disponía de un kit comercial que permitía distinguir Monilinia de otros géneros y M. fructicola del resto de especies mediante anticuerpos monoclonales utilizando la técnica DAS-ELISA. En esta Tesis se probaron diferentes fuentes de material como micelio ó conidias procedentes de cultivos en APD, o el micelio de la superficie de frutas o de momias frescas, como formas de antígeno. Los resultados obtenidos con ELISA se compararon con la identificación por métodos morfológico-culturales y por PCR con los cebadores desarrollados en esta Tesis. Los resultados demostraron la posibilidad de una detección temprana en frutas frescas por este método, realzando las posibilidades de una diagnosis temprana para una prevención más eficaz de M. fructicola en fruta de hueso. El estudio epidemiológico de la enfermedad comenzó con la determinación de las principales fuentes de inóculo primario y su importancia relativa en melocotoneros y nectarinos del valle del Ebro. Para ello se muestrearon 9 huertos durante los años 2003 a 2005 recogiendo todas las momias, frutos abortados, gomas, chancros, y brotes necróticos en los árboles. También se recogieron brotes aparentemente sanos y muestras de material vegetal situados en el suelo. En estas muestras se determinó la presencia de Monilinia spp. Los resultados mostraron que la fuente principal de inóculo son las momias que se quedan en los árboles en las que la supervivencia del hongo tras el invierno es muy alta. También son fuentes de inóculo las momias del suelo y los brotes necróticos. De aquí se deriva que una recomendación importante para los agricultores es que deben eliminar este material de los huertos. Un aspecto no estudiado en melocotonero o nectarino en España es la posible relación que puede darse entre la incidencia de infecciones latentes en los frutos inmaduros a lo largo del cultivo y la incidencia de podredumbre en los frutos en el momento de la recolección y en postcosecha. Esta relación se había observado previamente en otros frutales de hueso infectados con M. fructicola en diversos países del mundo. Para estudiar esta relación se realizaron ensayos en cinco huertos comerciales de melocotonero y nectarino situados en el Valle del Ebro en cuatro estados fenológicos durante los años 2000-2002. No se observaron infecciones latentes en botón rosa, dándose la máxima incidencia en precosecha, aunque en algunos huertos se daba otro pico en el endurecimiento del embrión. La especie prevaleciente fue M. laxa. Se obtuvo una correlación positiva significativa entre la incidencia de infecciones latentes y la incidencia de podredumbre en postcosecha. Se desarrolló también un modelo de predicción de las infecciones latentes en función de la temperatura (T) y el periodo de humectación (W). Este modelo indicaba que T y W explicaban el 83% de la variación en la incidencia de infecciones latentes causadas por Monilinia spp. Por debajo de 8ºC no se predecían latentes, necesitándose más de 22h de W para predecir la ocurrencia de latentes con T = 8ºC, mientras que solo se necesitaban 5h de W a 25ºC. Se hicieron también ensayos en condiciones controladas para determinar la relación entre la incidencia de las infecciones latentes, las condiciones ambientales (T y W), la concentración de inóculo del patógeno (I) y el estado de desarrollo del huésped (S) y para validar el modelo de predicción desarrollado con los experimentos de campo. Estos ensayos se llevaron cabo con flores y frutos de nectarino procedentes de un huerto comercial en seis estados fenológicos en los años 2004 y 2005, demostrándose que la incidencia de podredumbre en postcosecha y de infecciones latentes estaba afectada por T, W, I y S. En los frutos se producían infecciones latentes cuando la T no era adecuada para el desarrollo de podredumbre. Una vez desarrollado el embrión eran necesarias más de 4-5h de W al día y un inóculo superior a 104 conidias ml-1 para que se desarrollase o podredumbre o infección latente. La ecuación del modelo obtenido con los datos de campo era capaz de predecir los datos observados en estos experimentos. Para evaluar el efecto del inóculo de Monilinia spp. en la incidencia de infecciones latentes y de podredumbre de frutos en postcosecha se hicieron 11 experimentos en huertos comerciales de melocotonero y nectarino del Valle del Ebro durante 2002 a 2005. Se observó una correlación positiva entre los números de conidias de Monilinia spp. en la superficie de los frutos y la incidencia de infecciones latentes De los estudios anteriores se deducen otras dos recomendaciones importantes para los agricultores: las estrategias de control deben tener en cuenta las infecciones latentes y estimar el riesgo potencial de las mismas basándose en la T y W. Deben tener también en cuenta la concentración de esporas de Monilinia spp. en la superficie de los frutos para disminuir el riesgo de podredumbre parda. El conocimiento de la estructura poblacional de los patógenos sienta las bases para establecer métodos más eficaces de manejo de las enfermedades. Por ello en esta Tesis se ha estudiado el grado de diversidad genética entre distintas poblaciones de M. laxa en diferentes localidades españolas utilizando 144 marcadores RAPDs (59 polimórficos y 85 monomórficos) y 21 aislados. El análisis de la estructura de la población reveló que la diversidad genética dentro de las subpoblaciones (huertos) (HS) representaba el 97% de la diversidad genética (HT), mientras que la diversidad genética entre subpoblaciones (DST) sólo representaba un 3% del total de esta diversidad. La magnitud relativa de la diferenciación génica entre subpoblaciones (GST) alcanzaba 0,032 y el número estimado de migrantes por generación (Nm) fue de 15,1. Los resultados obtenidos en los dendrogramas estaban de acuerdo con el análisis de diversidad génica. Las agrupaciones obtenidas eran independientes del huerto de procedencia, año o huésped. En la Tesis se discute la importancia relativa de las diferentes fuentes evolutivas en las poblaciones de M. laxa. Finalmente se realizó un muestreo en distintos huertos de melocotonero y nectarino del Valle del Ebro para determinar la existencia o no de aislados resistentes a los fungicidas del grupo de los benzimidazoles y las dicarboximidas, fungicidas utilizados habitualmente para el control de la podredumbre parda y con alto riesgo de desarrollar resistencia en las poblaciones patógenas. El análisis de 114 aislados de M. laxa con los fungicidas Benomilo (bencimidazol) (1Bg m.a ml-1), e Iprodiona (dicarboximida) (5Bg m.a ml-1), mostró que ninguno era resistente en las dosis ensayadas. Monilinia spp. (M. laxa, M. fructigena and M. fructicola) cause bud and flower wilt, canker in branches and stone fruit rot giving rise important economic losses in years with appropriate environmental conditions, it is particularly important in late varieties of peach and nectarine. Right now, M. laxa is the major species for peach and nectarine in Spain followed by M. fructigena, in a smaller proportion. The recent introduction of the quarantine organism M. fructicola in Europe makes detection and identification of each one of the species necessary. In addition, there are different aspects of disease etiology and epidemiology that are not well known in Spain conditions. The first goal of this Thesis was the detection and identification of Monilinia spp. causing brown rot. Study of the epidemiology basis for disease control was the second objective. A pair of primers for PCR detection was designed based on the ribosomal DNA sequence in order to detect Monilinia spp. in plant material and to discriminate it from other fungi colonizing peach tree surface. Discrimination among Monilinia spp. was successful by SCAR markers (Sequence Characterized Amplified Region), obtained after a random amplified polymorphic DNA markers (RAPDs) study. An internal control for the PCR (CI) based on the use of a mimic plasmid designed on the primers specific for Monilinia was constructed to be used in diagnosis protocol for brown rot in order to avoid false negatives due to the inhibition of PCR as consequence of remained plant material. A commercial kit based on DAS-ELISA and monoclonals antibodies was successfully tested to distinguish Monilinia from other fungus genera and M. fructicola from other Monilinia species. Different materials such as micelium or conidias from APD cultures, or micelium from fresh fruit surfaces or mummies were tested in this Thesis, as antigens. Results obtained by ELISA were compared to classical identification by morphologic methods and PCR with the primers developed in this Thesis. Results demonstrated the possibility of an early detection in fresh fruits by this method for a more effective prevention of M. fructicola in stone fruit. The epidemiology study of the disease started with the determination of the main sources of primary inoculum and its relative importance in peach trees and nectarines in the Ebro valley. Nine orchards were evaluated during years 2003 to 2005 collecting all mummies, aborted fruits, rubbers, cankers, and necrotic buds from the trees. Apparently healthy buds and plant material located in the ground were also collected. In these samples presence of Monilinia spp. was determined. Results showed that the main inoculum sources are mummies that stay in the trees and where fungus survival after the winter is very high. Mummies on the ground and the necrotics buds are also sources of inoculum. As consequence of this an important recommendation for the growers is the removal of this material of the orchards. An important issue not well studied in peach or nectarine in Spain is the possible relationship between the incidence of latent infections in the immature fruits and the incidence of fruit rot at harvesting and postharvesting. This relationship had been previously shown in other stone fruit trees infected with M. fructicola in different countries over the world. In order to study this relationship experiments were run in five commercial peach and nectarine orchards located in the Ebro Valley in four phenologic states from 2000 to 2002. Latent infections were not observed in pink button, the maxima incidence arise in preharvest, although in some orchards another increase occurred in the embryo hardening. The most prevalence species was M. laxa. A significant positive correlation between the incidence of latent infections and the incidence of rot in postharvest was obtained. A prediction model of the latent infections based on the temperature (T) and the wetness duration (W) was also developed. This model showed that T and W explained 83% of the variation in latent infection incidence caused by Monilinia spp. Below 8ºC latent infection was not predicted, more than 22h of W with T = 8ºC were needed to predict latent infection occurrence of, whereas at 25ºC just 5h of W were enough. Tests under controlled conditions were also made to determine the relationship among latent infections incidence, environmental conditions (T and W), inoculum concentration of the pathogen (I) and development state of the host (S) to validate the prediction model developed on the field experiments. These tests were made with flowers and fruits of nectarine coming from a commercial orchard, in six phenologic states in 2004 and 2005, showing that incidence of rot in postharvest and latent infections were affected by T, W, I and S. In fruits latent infections took place when the T was not suitable for rot development. Once developed the embryo, more than 4-5h of W per day and higher inoculums (104 conidia ml-1) were necessary for rot or latent infection development. The equation of the model obtained with the field data was able to predict the data observed in these experiments. In order to evaluate the effect of inoculum of Monilinia spp. in the incidence of latent infections and of rot of fruits in postharvest, 11 experiments in commercial orchards of peach and nectarine of the Ebro Valley were performed from 2002 to 2005. A positive correlation between the conidial numbers of Monilinia spp. in the surface of the fruits and the incidence of latent infections was observed. Based on those studies other two important recommendations for the agriculturists are deduced: control strategies must consider the latent infections and potential risk based on the T and W. Spores concentration of Monilinia spp. in the surface of fruits must be also taken in concern to reduce the brown rot risk. The knowledge of the population structure of the pathogens determines the bases to establish more effective methods of diseases handling. For that reason in this Thesis the degree of genetic diversity among different M. laxa populations has been studied in different Spanish locations using 144 RAPDs markers (59 polymorphic and 85 monomorphics) on 21 fungal isolates. The analysis of the structure of the population revealed that the genetic diversity within the subpopulations (orchards) (HS) represented 97% of the genetic diversity (HT), whereas the genetic diversity between subpopulations (DST) only represented a 3% of the total of this diversity. The relative magnitude of the genic differentiation between subpopulations (GST) reached 0.032 and the considered number of migrantes by generation (Nm) was of 15.1. The results obtained in dendrograms were in agreement with the analysis of genic diversity. The obtained groupings were independent of the orchard of origin, year or host. In the Thesis the relative importance of the different evolutionary sources in the populations from M. laxa is discussed. Finally a sampling of resistant isolates in different orchards from peach and nectarine of Ebro Valley was made to determine the existence of fungicide resistance of the group of benzimidazoles and the dicarboximidas, fungicides used habitually for the control of rot brown and with high risk of resistance developing in the pathogenic populations. The analysis of 114 isolated ones of M. laxa with the fungicides Benomilo (bencimidazol) (1Bg m.a ml-1), and Iprodiona (dicarboximida) (5Bg m.a ml-1), showed no resistant in the doses evaluated.

The involvement of thaumatin-like proteins in plant food cross-reactivity: a multicenter study using a specific protein microarray.

Cross-reactivity of plant foods is an important phenomenon in allergy, with geographical variations with respect to the number and prevalence of the allergens involved in this process, whose complexity requires detailed studies. We have addressed the role of thaumatin-like proteins (TLPs) in cross-reactivity between fruit and pollen allergies. A representative panel of 16 purified TLPs was printed onto an allergen microarray. The proteins selected belonged to the sources most frequently associated with peach allergy in representative regions of Spain. Sera from two groups of well characterized patients, one with allergy to Rosaceae fruit (FAG) and another against pollens but tolerant to food-plant allergens (PAG), were obtained from seven geographical areas with different environmental pollen profiles. Cross-reactivity between members of this family was demonstrated by inhibition assays. Only 6 out of 16 purified TLPs showed noticeable allergenic activity in the studied populations. Pru p 2.0201, the peach TLP (41%), chestnut TLP (24%) and plane pollen TLP (22%) proved to be allergens of probable relevance to fruit allergy, being mainly associated with pollen sensitization, and strongly linked to specific geographical areas such as Barcelona, Bilbao, the Canary Islands and Madrid. The patients exhibited mayor que50% positive response to Pru p 2.0201 and to chestnut TLP in these specific areas. Therefore, their recognition patterns were associated with the geographical area, suggesting a role for pollen in the sensitization of these allergens. Finally, the co-sensitizations of patients considering pairs of TLP allergens were analyzed by using the co-sensitization graph associated with an allergen microarray immunoassay. Our data indicate that TLPs are significant allergens in plant food allergy and should be considered when diagnosing and treating pollen-food allergy.

Models for Internal Quality Fruit Sorting, Based on Time- Domain Laser Reflectance Spectroscopy (TDRS)

Time domain laser reflectance spectroscopy (TDRS) was applied for the first time to evaluate internal fruit quality. This technique, known in medicine-related knowledge areas, has not been used before in agricultural or food research. It allows the simultaneous non-destructive measuring of two optical characteristics of the tissues: light scattering and absorption. Models to measure firmness, sugar & acid contents in kiwifruit, tomato, apple, peach, nectarine and other fruits were built using sequential statistical techniques: principal component analysis, multiple stepwise linear regression, clustering and discriminant analysis. Consistent correlations were established between the two parameters measured with TDRS, i.e. absorption & transport scattering coefficients, with chemical constituents (sugars and acids) and firmness, respectively. Classification models were built to sort fruits into three quality grades, according to their firmness, soluble solids and acidity.

Woolliness assessment in peaches. Comparison between human and instrumental procedures and results

Woolliness, a negative attribute of sensory texture, is characterised by the lack of juiciness without variation of the tissue water content an incapacity of ripening although there is external ripe appearance. In this study, peaches cv Springcrest (early and soft flesh peaches) and Miraflores (late and hard flesh peaches) corresponding to three different maturity stages at harvest, stored 0, 1, 2, 3 and 4 weeks at 1 and 5°C have been tested by instrumental and sensory means. A Instrumental classification of woolliness has been compared to the sensory assessment. For Springcrest peaches the sensory results match with those found for the instrumental procedure. In this case , Woolliness appears after 2 weeks of storage at 5°C, changing abruptly from crispy to woolly. Miraflores peaches did not develop woolliness during storage. After comparing with sensory results, it is shown that a common instrumental scale may be appropriate to classify for woolliness all peach varieties.

Tri a 14, wheat Lipid Transfer Protein, a marker of wheat respiratory allergy

Over 30 wheat allergens have been associated to baker’s asthma and much of them have been also implied in food allergy. Few of them have rendered as major allergens. Tri a 14, wheat LTP, has been associated to baker’s asthma as major allergen in patients that can consume peach and wheat derived foodstuffs. In Spanish baker’s asthma patients, 60% showed positive response to Tri a 14 and 45% to Pru p 3. However, the cross-reactivity between peach and wheat has been unusual in allergic population (1,8). Moreover, wheat allergy is not so often as should be attending to the high consume.

Influence of climate change on the flowering of temperate fruit trees

It is well known that winter chilling is necessary for the flowering of temperate trees. The chilling requirement is a criterion for choosing a species or variety at a given location. Also chemistry products can be used for reducing the chilling-hours needs but make our production more expensive. This study first analysed the observed values of chilling hours for some representative agricultural locations in Spain for the last three decades and their projected changes under climate change scenarios. Usually the chilling is measured and calculated as chilling-hours, and different methods have been used to calculate them (e.g. Richarson et al., 1974 among others) according to the species considered. For our objective North Carolina method (Shaltout and Unrath, 1983) was applied for apples, Utah method (Richardson et al. 1974) for peach and grapevine and the approach used by De Melo-Abreu et al. (2004) for olive trees. The influence of climate change in temperate trees was studied by calculating projections of chilling-hours with climate data from Regional Climate Models (RCMs) at high resolution (25 km) from the European Project ENSEMBLES (http://www.ensembles-eu.org/). These projections will allow for analysing the modelled variations of chill-hours between 2nd half of 20C and 1st half of 21C at the study locations.

Sensitization profiles and cross-reactivity among plant allergens. Molecular mechanisms of food allergy development and the role of enhancers

La prevalencia de las alergias está aumentando desde mediados del siglo XX, y se estima que actualmente afectan a alrededor del 2-8 % de la población, pero las causas de este aumento aún no están claras. Encontrar el origen del mecanismo por el cual una proteína inofensiva se convierte en capaz de inducir una respuesta alérgica es de vital importancia para prevenir y tratar estas enfermedades. Aunque la caracterización de alérgenos relevantes ha ayudado a mejorar el manejo clínico y a aclarar los mecanismos básicos de las reacciones alérgicas, todavía queda un largo camino para establecer el origen de la alergenicidad y reactividad cruzada. El objetivo de esta tesis ha sido caracterizar las bases moleculares de la alergenicidad tomando como modelo dos familias de panalergenos (proteínas de transferencia de lípidos –LTPs- y taumatinas –TLPs-) y estudiando los mecanismos que median la sensibilización y la reactividad cruzada para mejorar tanto el diagnóstico como el tratamiento de la alergia. Para ello, se llevaron a cabo dos estrategias: estudiar la reactividad cruzada de miembros de familias de panalérgenos; y estudiar moléculas-co-adyuvantes que pudieran favorecer la capacidad alergénica de dichas proteínas. Para estudiar la reactividad cruzada entre miembros de la misma familia de proteínas, se seleccionaron LTPs y TLPs, descritas como alergenos, tomando como modelo la alergia a frutas. Por otra parte, se estudiaron los perfiles de sensibilización a alérgenos de trigo relacionados con el asma del panadero, la enfermedad ocupacional más relevante de origen alérgico. Estos estudios se llevaron a cabo estandarizando ensayos tipo microarrays con alérgenos y analizando los resultados por la teoría de grafos. En relación al estudiar moléculas-co-adyuvantes que pudieran favorecer la capacidad alergénica de dichas proteínas, se llevaron a cabo estudios sobre la interacción de los alérgenos alimentarios con células del sistema inmune humano y murino y el epitelio de las mucosas, analizando la importancia de moléculas co-transportadas con los alérgenos en el desarrollo de una respuesta Th2. Para ello, Pru p 3(LTP y alérgeno principal del melocotón) se selección como modelo para llevarlo a cabo. Por otra parte, se analizó el papel de moléculas activadoras del sistema inmune producidas por patógenos en la inducción de alergias alimentarias seleccionando el modelo kiwi-alternaria, y el papel de Alt a 1, alérgeno mayor de dicho hongo, en la sensibilización a Act d 2, alérgeno mayor de kiwi. En resumen, el presente trabajo presenta una investigación innovadora aportando resultados de gran utilidad tanto para la mejora del diagnóstico como para nuevas investigaciones sobre la alergia y el esclarecimiento final de los mecanismos que caracterizan esta enfermedad. ABSTRACT Allergies are increasing their prevalence from mid twentieth century, and they are currently estimated to affect around 2-8% of the population but the underlying causes of this increase remain still elusive. The understanding of the mechanism by which a harmless protein becomes capable of inducing an allergic response provides us the basis to prevent and treat these diseases. Although the characterization of relevant allergens has led to improved clinical management and has helped to clarify the basic mechanisms of allergic reactions, it seems justified in aspiring to molecularly dissecting these allergens to establish the structural basis of their allergenicity and cross-reactivity. The aim of this thesis was to characterize the molecular basis of the allergenicity of model proteins belonging to different families (Lipid Transfer Proteins –LTPs-, and Thaumatin-like Proteins –TLPs-) in order to identify mechanisms that mediate sensitization and cross reactivity for developing new strategies in the management of allergy, both diagnosis and treatment, in the near future. With this purpose, two strategies have been conducted: studies of cross-reactivity among panallergen families and molecular studies of the contribution of cofactors in the induction of the allergic response by these panallergens. Following the first strategy, we studied the cross-reactivity among members of two plant panallergens (LTPs , Lipid Transfer Proteins , and TLPs , Thaumatin-like Proteins) using the peach allergy as a model. Similarly, we characterized the sensitization profiles to wheat allergens in baker's asthma development, the most relevant occupational disease. These studies were performed using allergen microarrays and the graph theory for analyzing the results. Regarding the second approach, we analyzed the interaction of plant allergens with immune and epithelial cells. To perform these studies , we examined the importance of ligands and co-transported molecules of plant allergens in the development of Th2 responses. To this end, Pru p 3, nsLTP (non-specific Lipid Transfer Protein) and peach major allergen, was selected as a model to investigate its interaction with cells of the human and murine immune systems as well as with the intestinal epithelium and the contribution of its ligand in inducing an allergic response was studied. Moreover, we analyzed the role of pathogen associated molecules in the induction of food allergy. For that, we selected the kiwi- alternaria system as a model and the role of Alt a 1 , major allergen of the fungus, in the development of Act d 2-sensitization was studied. In summary, this work presents an innovative research providing useful results for improving diagnosis and leading to further research on allergy and the final clarification of the mechanisms that characterize this disease.

Quality assurance systems in the production and marketing of fresh peaches.

Due to the fast rate of peach post-harvest ripening, damage due to mechanical handling, externally appreciated as bruises and soft areas, is a real problem that leads to an early harvesting and poor quality of the fruits, as perceived by the consumers. More and more, the European consumer asks for good taste and freshness of fruits and vegetables, and these quality factors are not included in standards, nor in most of the producers' practices. Fruit processing and marketing centres (co-operatives) are increasingly interested in adopting quality controls in their processes. ISO 9000 procedures are being applied in some food areas, primarily milk and meat processors, but no generalised procedures have been developed until the present time to be applied to fresh product processes. All different peach and nectarine varieties that are harvested and handled in Murcia cooperatives and sold in a large supermarket in Madrid were analysed during the whole 1997 season (early May to late August). A total number of 78 samples of 25 fruits (co-operative) or 10 fruits (market), were tested in the laboratory for mechanical, optical, chemical and tasting quality. The variability and relationships between all these quality parameters are presented and discussed, and sampling unit sizes which would be advisable for quality control are calculated.

Fruit by fruit identification of mealy peaches. Extraction of storage features.

Mealiness is a negative attribute of sensory texture, characterised by the lack of juiciness decrease in the total amount of of water content of tissues. Peach mealy textures are known as \ and leatheriness. Besides the lack of juiciness and flavour, that characterises mealy fruits, in associated with internal browning near the stone and an incapacity of ripening although there i ripe appearance. It is considered as a physiological disorder that appears in stone fruits probably < unbalanced pectolitic enzyme activity during storage. Since January 1996, a wide EC Project entitled: "Mealiness in fruits. Consumer perception and i detection" is being carried out. Within it, the Physical Properties Laboratory (ETSIA-UPM) working to develop instrumental procedures to detect mealiness in different types of fruits (s contributions by Barreiro to AgEng). The results obtained have shown to correlate well with \ measurements in apples (Barreiro et al), also we have succeeded in identifying individual mealy j the basis of instrumental measurement in peaches. The definition of these texture categories will be used in further studies as a base for new individual classification.

Non-Destructive Identification of Woolly Peaches combining Mechanical Impact Response and NIR Spectroscopy.

Woolliness (mealiness in other fruits) is a negative attribute of peach sensory texture that is a physiological disorder associated with inadequate cold storage. It is characterised by lack of crispness and juiciness without variation in the tissue water content (Harker and Hallet, 1992). Many attempts have been made to develop destructive instrumental procedures to detect mealiness and woolliness. Non-destructive procedures attempted include using nuclear magnetic resonance (Sonego et al., 1995). However, this technique has economical limitations and is not practical at present. Non-destructive impact tests and NIR are non-destructive techniques which have been used to assess internal characteristics of fruits (Chen and Sun, 1991). The objective of this study was to develop a novel non-destructive procedure to identify woolly peaches by combining impact and NIR approaches.