Automatic blood glucose classification for gestational diabetes with feature selection: decision trees vs neural networks

Automatic blood glucose classification may help specialists to provide a better interpretation of blood glucose data, downloaded directly from patients glucose meter and will contribute in the development of decision support systems for gestational diabetes. This paper presents an automatic blood glucose classifier for gestational diabetes that compares 6 different feature selection methods for two machine learning algorithms: neural networks and decision trees. Three searching algorithms, Greedy, Best First and Genetic, were combined with two different evaluators, CSF and Wrapper, for the feature selection. The study has been made with 6080 blood glucose measurements from 25 patients. Decision trees with a feature set selected with the Wrapper evaluator and the Best first search algorithm obtained the best accuracy: 95.92%.

A model of economic management of cultural heritage: the rehabilitation of fort christmas as an interpretation center of defensive architecture of the Mediterranean in Cartagena (Spain)

The Cultural Heritage constitutes a way to generate social identities and play an important role in the development of the Spanish Mediterranean cities that opt to sustainable quality tourism. The reflection on the necessity of intervention on this heritage, in addition to establishing what should be done, brings up the need to define the reasons for taking action, why and what-for. These decisions are essential to establish if its maintenance and recovery are economically sustainable. The Project "Cartagena Port of Cultures", with support from the European Union, is an example of effective instrument for ensuring the sustainability of our built heritage conservation. Its main objective was to enable sustainable development of tourism in Cartagena based on sustainability and seasonality. This was achieved through a process of recovery of heritage resources and their optimum promotion and marketing.

Urban trees and atmospheric pollutants in big cities: Effects in Madrid

Una investigación sobre la mejora de la contaminación del aire (CA) por medio de arbolado urbano se realizó en Madrid, una ciudad con casi 4 M de habitantes, 2,8 M de vehículos y casi 3 M de árboles de mantenimiento público. La mayoría de los árboles estaban en dos bosques periurbanos. Los 650.000 restantes era pies de alineación y parques. Los taxones estudiados fueron Platanus orientalis (97.205 árboles), Ulmus sp. (70.557), Pinus pinea (49.038), Aesculus hippocastanum (22.266), Cedrus sp. (13.678) y Quercus ilex (1.650), de calles y parques. Muestras foliares se analizaron en diferentes épocas del año, así como datos de contaminación por PM10 de 28 estaciones de medición de la contaminación durante 30 años, y también la intensidad del tráfico (IMD) en 2.660 calles. La acumulación de metales pesados (MP) sobre hojas y dentro de estas se estimó en relación con la CA y del suelo y la IMD del tráfico. La concentración media de Ba, Cd, Cr, Cu, Mn, Ni, Pb y Zn en suelo (materia seca) alcanzó: 489,5, 0,7, 49,4, 60,9, 460,9, 12,8, 155,9 y 190,3 mg kg-1 respectivamente. Los árboles urbanos, particularmente coníferas (debido a la mayor CA en invierno) contribuyen significativamente a mejorar la CA sobre todo en calles con alta IMD. La capacidad de las seis sp. para capturar partículas de polvo en su superficies foliares está relacionada con la IMD del tráfico y se estimó en 16,8 kg/año de MP tóxicos. Pb y Zn resultaron ser buenos marcadores antrópicos en la ciudad en relación con el tráfico, que fue la principal fuente de contaminación en los árboles y suelos de Madrid. Las especies de árboles variaron en función de su capacidad para capturar partículas (dependiendo de las propiedades de sus superficies foliares) y acumular los MP absorbidos de los suelos. Las concentraciones foliares de Pb y Zn estuvieron por encima de los límites establecidos en diferentes sitios de la ciudad. La microlocalización de Zn mediante microscópico mostró la translocación al xilema y floema. Se detectaron puntos de contaminación puntual de Cu and Cr en antiguos polígonos industriales y la distribución espacial de los MP en los suelos de Madrid mostró que en incluso en zonas interiores del El Retiro había ciertos niveles elevados de [Pb] en suelo, tal vez por el emplazamiento la Real Fábrica de Porcelana en la misma zona hace 200 años. Distintas áreas del centro de la ciudad también alcanzaron niveles altos de [Pb] en suelo. Según los resultados, el empleo de una combinación de Pinus pinea con un estrato intermedio de Ulmus sp. y Cedrus sp. puede ser la mejor recomendación como filtro verde eficiente. El efecto del ozono (O3) sobre el arbolado en Madrid fue también objeto de este estudio. A pesar de la reducción de precursores aplicada en muchos países industrializados, O3 sigue siendo la principal causa de CA en el hemisferio norte, con el aumento de [O3] de fondo. Las mayores [O3] se alcanzaron en regiones mediterráneas, donde el efecto sobre la vegetación natural es compensado por el xeromorfismo y la baja conductancia estomática en respuesta los episodios de sequía estival característicos de este clima. Durante una campaña de monitoreo, se identificaron daños abióticos en hojas de encina parecidos a los de O3 que estaban plantadas en una franja de césped con riego del centro de Madrid. Dada la poca evidencia disponible de los síntomas de O3 en frondosas perennifolias, se hizo un estudio que trató de 1) confirman el diagnóstico de daño de O3, 2) investigar el grado de los síntomas en encinas y 3) analizar los factores ambientales que contribuyeron a los daños por O3, en particular en lo relacionado con el riego. Se analizaron los marcadores macro y micromorfológicos de estrés por O3, utilizando las mencionadas encinas a modo de parcela experimental. Los síntomas consistieron en punteado intercostal del haz, que aumentó con la edad. Además de un punteado subyacente, donde las células superiores del mesófilo mostraron reacciones características de daños por O3. Las células próximas a las zonas dañadas, presentaron marcadores adicionales de estrés oxidativo. Estos marcadores morfológicos y micromorfológicos de estrés por O3 fueron similares a otras frondosas caducifolias con daños por O3. Sin embargo, en nuestro caso el punteado fue evidente con AOT40 de 21 ppm•h, asociada a riego. Análisis posteriores mostraron que los árboles con riego aumentaron su conductancia estomática, con aumento de senescencia, manteniéndose sin cambios sus características xeromórficas foliares. Estos hallazgos ponen de relieve el papel primordial de la disponibilidad de agua frente a las características xeromórficas a la hora de manifestarse los síntomas en las células por daños de O3 en encina. ABSTRACT Research about air pollution mitigation by urban trees was conducted in Madrid (Spain), a southern European city with almost 4 M inhabitants, 2.8 M daily vehicles and 3 M trees under public maintenance. Most trees were located in two urban forests, while 650'000 trees along urban streets and in parks. The urban taxa included Platanus orientalis (97'205 trees), Ulmus sp. (70’557), Pinus pinea (49'038), Aesculus hippocastanum (22’266), Cedrus sp. (13'678 and Quercus ilex (1'650) along streets and parks. Leave samples were analysed sequentially in different seasons, PM10 data from 28 air monitoring stations during 30 years and traffic density estimated from 2’660 streets. Heavy metal (HM) accumulation on the leaf surface and within leaves was estimated per tree related to air and soil pollution, and traffic intensity. Mean concentration of Ba, Cd, Cr, Cu, Mn, Ni, Pb and Zn in topsoil samples (dry mass) amounted in Madrid: 489.5, 0.7, 49.4, 60.9, 460.9, 12.8, 155.9 and 190.3 mg kg-1 respectively. Urban trees, particularly conifers (due to higher pollution in winter) contributed significantly to alleviate air pollution especially near to high ADT roads. The capacity of the six urban street trees species to capture air-born dust on the foliage surface as related to traffic intensity was estimated to 16.8 kg of noxious metals from exhausts per year. Pb and Zn pointed to be tracers of anthropic activity in the city with vehicle traffic as the main source of diffuse pollution on trees and soils. Tree species differed by their capacity to capture air-borne dust (by different leaf surface properties) and to allocate HM from soils. Pb and Zn concentrations in the foliage were above limits in different urban sites and microscopic Zn revelation showed translocation in xylem and phloem tissue. Punctual contamination in soils by Cu and Cr was identified in former industrial areas and spatial trace element mapping showed for central Retiro Park certain high values of [Pb] in soils even related to a Royal pottery 200 years ago. Different areas in the city centre also reached high levels [Pb] in soils. According to the results, a combination of Pinus pinea with understorey Ulmus sp. and Cedrus sp. layers can be recommended for the best air filter efficiency. The effects of ozone (O3) on trees in different areas of Madrid were also part of this study. Despite abatement programs of precursors implemented in many industrialized countries, ozone remained the main air pollutant throughout the northern hemisphere with background [O3] increasing. Some of the highest ozone concentrations were measured in regions with a Mediterranean climate but the effect on the natural vegetation is alleviated by low stomatal uptake and frequent leaf xeromorphy in response to summer drought episodes characteristic of this climate. During a bioindication survey, abiotic O3-like injury was identified in foliage. Trees were growing on an irrigated lawn strip in the centre of Madrid. Given the little structural evidence available for O3 symptoms in broadleaved evergreen species, a study was undertaken in 2007 with the following objectives 1) confirm the diagnosis, 2) investigate the extent of symptoms in holm oaks growing in Madrid and 3) analyse the environmental factors contributing to O3 injury, particularly, the site water supply. Therefore, macro- and micromorphological markers of O3 stress were analysed, using the aforementioned lawn strip as an intensive study site. Symptoms consisted of adaxial and intercostal stippling increasing with leaf age. Underlying stippling, cells in the upper mesophyll showed HR-like reactions typical of ozone stress. The surrounding cells showed further oxidative stress markers. These morphological and micromorphological markers of ozone stress were similar to those recorded in deciduous broadleaved species. However, stippling became obvious already at an AOT40 of 21 ppm•h and was primarily found at irrigated sites. Subsequent analyses showed that irrigated trees had their stomatal conductance increased and leaf life-span reduced whereas their leaf xeromorphy remained unchanged. These findings suggest a central role of water availability versus leaf xeromorphy for ozone symptom expression by cell injury in holm oak.

Evolutionary computation of forests with Degree- and Role-Constrained Minimum Spanning Trees

Finding the degree-constrained minimum spanning tree (DCMST) of a graph is a widely studied NP-hard problem. One of its most important applications is network design. Here we deal with a new variant of the DCMST problem, which consists of finding not only the degree- but also the role-constrained minimum spanning tree (DRCMST), i.e., we add constraints to restrict the role of the nodes in the tree to root, intermediate or leaf node. Furthermore, we do not limit the number of root nodes to one, thereby, generally, building a forest of DRCMSTs. The modeling of network design problems can benefit from the possibility of generating more than one tree and determining the role of the nodes in the network. We propose a novel permutation-based representation to encode these forests. In this new representation, one permutation simultaneously encodes all the trees to be built. We simulate a wide variety of DRCMST problems which we optimize using eight different evolutionary computation algorithms encoding individuals of the population using the proposed representation. The algorithms we use are: estimation of distribution algorithm, generational genetic algorithm, steady-state genetic algorithm, covariance matrix adaptation evolution strategy, differential evolution, elitist evolution strategy, non-elitist evolution strategy and particle swarm optimization. The best results are for the estimation of distribution algorithms and both types of genetic algorithms, although the genetic algorithms are significantly faster.

Development of somatic embryogenesis for cloning and conservation of mature holm oak trees (Quercus ilex L.)

La encina (Quercus ilex L.) es una de las especies forestales mediterráneas más importantes. Constituye gran parte del estrato arbóreo de dehesas o montados, produce bellota como alimento del ganado y establece simbiosis con hongos micorrizógenos de gran valor económico. La encina está considerada como una especie recalcitrante en términos de conservación de semillas y capacidad morfogénica, lo que dificulta los programas de conservación de recursos genéticos y la mejora de la especie. La propagación vegetativa es una potente herramienta de los programas de mejora, por lo que es preciso desarrollar protocolos de regeneración somática en encina. La embriogénesis somática está considerada como la modalidad más adecuada de regeneración basada en técnicas de cultivo de tejidos vegetales utilizada en biotecnología forestal. Este trabajo se centra en el estudio de determinados aspectos de la embriogénesis somática para la regeneración clonal de encinas adultas. La memoria de esta tesis se ha dividido en capítulos que se corresponden con diferentes aspectos del sistema embriogénico. La embriogénesis somática se indujo en tegumentos maternos de óvulos en desarrollo procedentes de bellotas inmaduras de encinas adultas. A pesar de las bajas frecuencias de inducción, las líneas embriogénicas generadas se amplificaron mediante embriogénesis secundaria observándose cierta pérdida de la capacidad de diferenciación con el tiempo. Tanto el genotipo como la formulación del medio de cultivo influyeron en la respuesta embriogénica, concluyendo que la formulación de macronutrientes de Schenk y Hildebrant del medio sin reguladores de crecimiento fue la combinación más efectiva en la inducción. Los resultados sugirieron la existencia de una ventana en el desarrollo del óvulo más sensible a la inducción. El genotipo in[luyó en la capacidad proliferativa de los cultivos y en la conversión de los embriones somáticos, que se incrementó suplementando el medio con ácido indol-3-butírico y 6-benciladenina. El cultivo en medio líquido de líneas embriogénicas en condiciones de inmersión transitoria incrementó el crecimiento, dependiendo del genotipo, con respecto al cultivo en medio semisólido. Sin embargo, no mejoró la capacidad de diferenciar embriones cotiledonares aislados. Se estableció un protocolo de inicio y mantenimiento de cultivos en suspensión para varias líneas embriogénicas mediante inoculación en alta densidad de agregados embrionarios procedentes del medio semisólido. Para evitar la pérdida de vigor y la capacidad morfogénica debida al cultivo prolongado se desarrolló un protocolo de crioconservación de líneas embriogénicas mediante vitrificación. Al determinar la influencia de los agentes crioprotectores antes y después de su inmersión en nitrógeno líquido se concluyó que las respuestas de capacidad de crecimiento y de diferenciación del material embriogénico son independientes, además de estar bajo influencia del genotipo y el tipo de material crioconservado. La combinación de sacarosa y PVS2 previa a la inmersión en nitrógeno líquido proporcionó la mayor tasa de recuperación. Cuando las líneas fueron crioconservadas 30 días la capacidad de diferenciación se perdió en todas ellas. El análisis de SSR detectó variación somaclonal en el material crioconservado a corto plazo. SSR y RAPD mostraron importantes diferencias genéticas entre los árboles donantes y el material embriogénico que dependieron del genotipo. El grado de detección dependió del marcador empleado. Ambos marcadores revelaron baja inestabilidad intraclonal. Los RAPD revelaron variación genética intra-individuo en las encinas donantes. Se discuten la variación genética pre-existente en encina, su aparición durante las primeras fases de la inducción de embriogénesis, y la presencia de tejidos provenientes de la fertilización en el explanto materno. Esto hace preciso definir la identidad genética del material donante y acometer ensayos de detección precoz de variación somaclonal. ABSTRACT Holm oak (Quercus ilex L.) is one of the most important Mediterranean forest species. It conforms the tree layer of dehesas or montados, it produces acorns to feed the livestock and it establishes symbiosis with profitable mycorrhizal fungi. Holm oak is considered as recalcitrant species in terms of seed conservation and morphogenic capacities, which complicates the development of genetic conservation and improvement programs. Vegetative propagation is one of the mightiest tools for breeding programs therefore; developing protocols for clonal regeneration of holm oak is essential. Somatic embryogenesis is considered the best tissue culture-based way of plant regeneration in forest biotechnology. The present study is focused on the study of certain aspects of somatic embryogenesis for clonal regeneration of mature holm oak. This thesis manuscript is divided into several chapters that match with different aspects of the embryogenic system. Somatic embryogenesis induction was achieved on maternal teguments of developing ovules from immature acorns of adult holm oak trees. Despite the low induction frequencies, the generated embryogenic lines were amplified by secondary embryogenesis. A decline in the differentiation capacity over time was also observed. It was concluded that both genotype and culture media formulation influenced the embryogenic response, being the Schenk and Hildebrandt´s macronutrients formulation from culture medium and the lack of plant growth regulators the most effective combination for the induction of the embryogenic response. It has been suggested the existence of a developmental window in which ovules are prone to induction. Genotype influenced the proliferation capacity and the plant conversion of somatic embryos, which was also favoured by the presence of indol-3-butyric acid and 6-bencyladenine. The use of temporary immersion systems as proliferation in liquid culture of the embryogenic lines increased the growth depending on genotype, when compared to semisolid cultures. However, it did not improve the differentiation of single cotyledonary embryos. A protocol for the initiation and maintenance of embryogenic suspension cultures was established for several embryogenic lines with highly dense inoculi of embryogenic clusters from proliferating semisolid cultures. In order to avoid the loss of vigour and morphogenic ability of embryogenic lines due to prolonged cultures, a cryopreservation protocol for embryogenic lines of holm oak has been developed. During the determination of the influence of cryoprotective agents on the growth and differentiation capacities before and after liquid nitrogen immersion, it was concluded that both responses were independent from each other and also under the influence of genotype and the type of cryopreserved material. The combination of sucrose and PVS2 prior liquid nitrogen immersion provided higher recovery rates. When the same embryogenic lines were cryopreserved for 30 days, none was able to differentiate. The SSRs analysis of the short-term cryopreserved material detected somaclonal variation. Both SSR and RAPD markers showed high sensitivity to detect genetic differences between the donor trees and the generated embryogenic material. Nevertheless, the degree of instability detection depended on the marker. The SSR analysis indicated a relationship between genotype, the studied loci and the located polymorphisms. Also, both markers revealed low intraclonal genetic variation. The RAPD detected genetic variation within the donor trees. The presence of pre-existent genetic variation within mature trees, in addition to its occurrence during the early stages of the embryogenic induction, and the presence of tissues of fertilisation origin within the maternal explants are all discussed. Nonetheless, the determination of the genetic identity of donor material is required, in addition to early detection methods of somaclonal variation.

Development of somatic embryogenesis for cloning and conservation of mature holm oak trees (Quercus ilex L.)

La encina (Quercus ilex L.) es una de las especies forestales mediterráneas más importantes. Constituye gran parte del estrato arbóreo de dehesas o montados, produce bellota como alimento del ganado y establece simbiosis con hongos micorrizógenos de gran valor económico. La encina está considerada como una especie recalcitrante en términos de conservación de semillas y capacidad morfogénica, lo que dificulta los programas de conservación de recursos genéticos y la mejora de la especie. La propagación vegetativa es una potente herramienta de los programas de mejora, por lo que es preciso desarrollar protocolos de regeneración somática en encina. La embriogénesis somática está considerada como la modalidad más adecuada de regeneración basada en técnicas de cultivo de tejidos vegetales utilizada en biotecnología forestal. Este trabajo se centra en el estudio de determinados aspectos de la embriogénesis somática para la regeneración clonal de encinas adultas. La memoria de esta tesis se ha dividido en capítulos que se corresponden con diferentes aspectos del sistema embriogénico. La embriogénesis somática se indujo en tegumentos maternos de óvulos en desarrollo procedentes de bellotas inmaduras de encinas adultas. A pesar de las bajas frecuencias de inducción, las líneas embriogénicas generadas se amplificaron mediante embriogénesis secundaria observándose cierta pérdida de la capacidad de diferenciación con el tiempo. Tanto el genotipo como la formulación del medio de cultivo influyeron en la respuesta embriogénica, concluyendo que la formulación de macronutrientes de Schenk y Hildebrant del medio sin reguladores de crecimiento fue la combinación más efectiva en la inducción. Los resultados sugirieron la existencia de una ventana en el desarrollo del óvulo más sensible a la inducción. El genotipo in[luyó en la capacidad proliferativa de los cultivos y en la conversión de los embriones somáticos, que se incrementó suplementando el medio con ácido indol-3-butírico y 6-benciladenina. El cultivo en medio líquido de líneas embriogénicas en condiciones de inmersión transitoria incrementó el crecimiento, dependiendo del genotipo, con respecto al cultivo en medio semisólido. Sin embargo, no mejoró la capacidad de diferenciar embriones cotiledonares aislados. Se estableció un protocolo de inicio y mantenimiento de cultivos en suspensión para varias líneas embriogénicas mediante inoculación en alta densidad de agregados embrionarios procedentes del medio semisólido. Para evitar la pérdida de vigor y la capacidad morfogénica debida al cultivo prolongado se desarrolló un protocolo de crioconservación de líneas embriogénicas mediante vitrificación. Al determinar la influencia de los agentes crioprotectores antes y después de su inmersión en nitrógeno líquido se concluyó que las respuestas de capacidad de crecimiento y de diferenciación del material embriogénico son independientes, además de estar bajo influencia del genotipo y el tipo de material crioconservado. La combinación de sacarosa y PVS2 previa a la inmersión en nitrógeno líquido proporcionó la mayor tasa de recuperación. Cuando las líneas fueron crioconservadas 30 días la capacidad de diferenciación se perdió en todas ellas. El análisis de SSR detectó variación somaclonal en el material crioconservado a corto plazo. SSR y RAPD mostraron importantes diferencias genéticas entre los árboles donantes y el material embriogénico que dependieron del genotipo. El grado de detección dependió del marcador empleado. Ambos marcadores revelaron baja inestabilidad intraclonal. Los RAPD revelaron variación genética intra-individuo en las encinas donantes. Se discuten la variación genética pre-existente en encina, su aparición durante las primeras fases de la inducción de embriogénesis, y la presencia de tejidos provenientes de la fertilización en el explanto materno. Esto hace preciso definir la identidad genética del material donante y acometer ensayos de detección precoz de variación somaclonal. ABSTRACT Holm oak (Quercus ilex L.) is one of the most important Mediterranean forest species. It conforms the tree layer of dehesas or montados, it produces acorns to feed the livestock and it establishes symbiosis with profitable mycorrhizal fungi. Holm oak is considered as recalcitrant species in terms of seed conservation and morphogenic capacities, which complicates the development of genetic conservation and improvement programs. Vegetative propagation is one of the mightiest tools for breeding programs therefore; developing protocols for clonal regeneration of holm oak is essential. Somatic embryogenesis is considered the best tissue culture-based way of plant regeneration in forest biotechnology. The present study is focused on the study of certain aspects of somatic embryogenesis for clonal regeneration of mature holm oak. This thesis manuscript is divided into several chapters that match with different aspects of the embryogenic system. Somatic embryogenesis induction was achieved on maternal teguments of developing ovules from immature acorns of adult holm oak trees. Despite the low induction frequencies, the generated embryogenic lines were amplified by secondary embryogenesis. A decline in the differentiation capacity over time was also observed. It was concluded that both genotype and culture media formulation influenced the embryogenic response, being the Schenk and Hildebrandt´s macronutrients formulation from culture medium and the lack of plant growth regulators the most effective combination for the induction of the embryogenic response. It has been suggested the existence of a developmental window in which ovules are prone to induction. Genotype influenced the proliferation capacity and the plant conversion of somatic embryos, which was also favoured by the presence of indol-3-butyric acid and 6-bencyladenine. The use of temporary immersion systems as proliferation in liquid culture of the embryogenic lines increased the growth depending on genotype, when compared to semisolid cultures. However, it did not improve the differentiation of single cotyledonary embryos. A protocol for the initiation and maintenance of embryogenic suspension cultures was established for several embryogenic lines with highly dense inoculi of embryogenic clusters from proliferating semisolid cultures. In order to avoid the loss of vigour and morphogenic ability of embryogenic lines due to prolonged cultures, a cryopreservation protocol for embryogenic lines of holm oak has been developed. During the determination of the influence of cryoprotective agents on the growth and differentiation capacities before and after liquid nitrogen immersion, it was concluded that both responses were independent from each other and also under the influence of genotype and the type of cryopreserved material. The combination of sucrose and PVS2 prior liquid nitrogen immersion provided higher recovery rates. When the same embryogenic lines were cryopreserved for 30 days, none was able to differentiate. The SSRs analysis of the short-term cryopreserved material detected somaclonal variation. Both SSR and RAPD markers showed high sensitivity to detect genetic differences between the donor trees and the generated embryogenic material. Nevertheless, the degree of instability detection depended on the marker. The SSR analysis indicated a relationship between genotype, the studied loci and the located polymorphisms. Also, both markers revealed low intraclonal genetic variation. The RAPD detected genetic variation within the donor trees. The presence of pre-existent genetic variation within mature trees, in addition to its occurrence during the early stages of the embryogenic induction, and the presence of tissues of fertilisation origin within the maternal explants are all discussed. Nonetheless, the determination of the genetic identity of donor material is required, in addition to early detection methods of somaclonal variation.

Effect of the season on the free phytoprostanes content in Cornicabra extra virgin olive oil from deficit irrigated olive trees

BACKGROUND: The effect of regulated deficit irrigation (RDI) on the phytoprostane (PhytoP) content in extra virgin olive (Olea europaea L., cv. Cornicabra) oil (EVOO) was studied. During the 2012 and 2013 seasons, T0 plants were irrigated at 100% ETc, while T1 and T2 plants were irrigated avoiding water deficit during phases I and III of fruit growth and saving water during the non-critical phenological period of pit hardening (phase II), developing amore severewater deficit in T2 plants. In 2013, a fourth treatment (T3) was also performed, which was similar to T2 except that water saving was from the beginning of phase II to 15 days after the end of phase II. RESULTS: 9-F1t-PhytoP, 9-epi-9-F1t-PhytoP, 9-epi-9-D1t-PhytoP, 9-D1t-PhytoP, 16-B1-PhytoP and 9-L1-PhytoP were present in Cornicabra EVOO, and their contents increased in the EVOO fromRDI plants. CONCLUSION: Deficit irrigation during pit hardening or for a further period of 2 weeks thereafter to increase irrigation water saving is clearly critical for EVOO composition because of the enhancement of free PhytoPs, which have potential beneficial effects on human health. The response of individual free PhytoPs to changes in plant water status was not as perceptible as expected, preventing their use as biomarkers of water stress.

Bootstrap confidence levels for phylogenetic trees

Evolutionary trees are often estimated from DNA or RNA sequence data. How much confidence should we have in the estimated trees? In 1985, Felsenstein [Felsenstein, J. (1985) Evolution 39, 783–791] suggested the use of the bootstrap to answer this question. Felsenstein’s method, which in concept is a straightforward application of the bootstrap, is widely used, but has been criticized as biased in the genetics literature. This paper concerns the use of the bootstrap in the tree problem. We show that Felsenstein’s method is not biased, but that it can be corrected to better agree with standard ideas of confidence levels and hypothesis testing. These corrections can be made by using the more elaborate bootstrap method presented here, at the expense of considerably more computation.

Matchings and phylogenetic trees

This paper presents a natural coordinate system for phylogenetic trees using a correspondence with the set of perfect matchings in the complete graph. This correspondence produces a distance between phylogenetic trees, and a way of enumerating all trees in a minimal step order. It is useful in randomized algorithms because it enables moves on the space of trees that make random optimization strategies “mix” quickly. It also promises a generalization to intermediary trees when data are not decisive as to their choice of tree, and a new way of constructing Bayesian priors on tree space.

Visualization of Cavitated Vessels in Winter and Refilled Vessels in Spring in Diffuse-Porous Trees by Cryo-Scanning Electron Microscopy1

Xylem cavitation in winter and recovery from cavitation in the spring were visualized in two species of diffuse-porous trees, Betula platyphylla var. japonica Hara and Salix sachalinensis Fr. Schm., by cryo-scanning electron microscopy after freeze-fixation of living twigs. Water in the vessel lumina of the outer three annual rings of twigs of B. platyphylla var. japonica and of S. sachalinensis gradually disappeared during the period from January to March, an indication that cavitation occurs gradually in these species during the winter. In April, when no leaves had yet expanded, the lumina of most of the vessels of both species were filled with water. Many vessel lumina in twigs of both species were filled with water during the period from the subsequent growth season to the beginning of the next winter. These observations indicate that recovery in spring occurs before the onset of transpiration and that water transport through twigs occurs during the subsequent growing season. We found, moreover, that vessels repeat an annual cycle of winter cavitation and spring recovery from cavitation for several years until irreversible cavitation occurs.

Bootstrap confidence levels for phylogenetic trees.

Evolutionary trees are often estimated from DNA or RNA sequence data. How much confidence should we have in the estimated trees? In 1985, Felsenstein [Felsenstein, J. (1985) Evolution 39, 783-791] suggested the use of the bootstrap to answer this question. Felsenstein's method, which in concept is a straightforward application of the bootstrap, is widely used, but has been criticized as biased in the genetics literature. This paper concerns the use of the bootstrap in the tree problem. We show that Felsenstein's method is not biased, but that it can be corrected to better agree with standard ideas of confidence levels and hypothesis testing. These corrections can be made by using the more elaborate bootstrap method presented here, at the expense of considerably more computation.