Pathways towards the integration of periurban agrarian ecosystems into the spatial planning system

The potential of periurban agrarian ecosystems is recognized as one of the cornerstones for improving urban sustainability; however, this potential has been disregarded in spatial planning and decision making. The main goal of the PAEc-SP project was to assess the feasibility of integrating periurban agricultural areas into spatial planning by developing a viable ecosystem services-based methodology

Plant β-diversity i in human-altered forest ecosystems: the importance of the structural, spatial, and topographical characteristics of stands in patterning plant species assemblages

An understanding of spatial patterns of plant species diversity and the factors that drive those patterns is critical for the development of appropriate biodiversity management in forest ecosystems. We studied the spatial organization of plants species in human- modified and managed oak forests (primarily, Quercus faginea) in the Central Pre- Pyrenees, Spain. To test whether plant community assemblages varied non-randomly across the spatial scales, we used multiplicative diversity partitioning based on a nested hierarchical design of three increasingly coarser spatial scales (transect, stand, region). To quantify the importance of the structural, spatial, and topographical characteristics of stands in patterning plant species assemblages and identify the determinants of plant diversity patterns, we used canonical ordination. We observed a high contribution of ˟-diversity to total -diversity and found ˟-diversity to be higher and ˞-diversity to be lower than expected by random distributions of individuals at different spatial scales. Results, however, partly depended on the weighting of rare and abundant species. Variables expressing the historical management intensities of the stand such as mean stand age, the abundance of the dominant tree species (Q. faginea), age structure of the stand, and stand size were the main factors that explained the compositional variation in plant communities. The results indicate that (1) the structural, spatial, and topographical characteristics of the forest stands have the greatest effect on diversity patterns, (2) forests in landscapes that have different land use histories are environmentally heterogeneous and, therefore, can experience high levels of compositional differentiation, even at local scales (e.g., within the same stand). Maintaining habitat heterogeneity at multiple spatial scales should be considered in the development of management plans for enhancing plant diversity and related functions in human-altered forests

Towards workflow ecosystems through standard representations

Workflows are increasingly used to manage and share scientific computations and methods. Workflow tools can be used to design, validate, execute and visualize scientific workflows and their execution results. Other tools manage workflow libraries or mine their contents. There has been a lot of recent work on workflow system integration as well as common workflow interlinguas, but the interoperability among workflow systems remains a challenge. Ideally, these tools would form a workflow ecosystem such that it should be possible to create a workflow with a tool, execute it with another, visualize it with another, and use yet another tool to mine a repository of such workflows or their executions. In this paper, we describe our approach to create a workflow ecosystem through the use of standard models for provenance (OPM and W3C PROV) and extensions (P-PLAN and OPMW) to represent workflows. The ecosystem integrates different workflow tools with diverse functions (workflow generation, execution, browsing, mining, and visualization) created by a variety of research groups. This is, to our knowledge, the first time that such a variety of workflow systems and functions are integrated.

Native bradyrhizobial symbionts of Lupinus mariae-josephae, a unique endemism thriving in alkaline soils in Eastern Spain

Lupinus mariae-josephae (Lmj) es una especie de lupino endémica de una pequeña y específica área de Comunidad Valenciana (Este de España), donde prospera en suelos alcalinoscalcáreos, un hábitat singular para los altramuces, que crecen preferentemente en suelos ácidos o neutros. Esto hace de Lmj una especie de lupino única. Cuando se inició este trabajo, la extensión conocida de este endemismo abarcaba unos 700 kilómetros cuadrados, confinados en la provincia de Valencia. En esta área, Lmj prospera en pequeñas poblaciones aisladas que contienen un número reducido de plantas por lo que se la consideró una especie en peligro de extinción. Todos los esfuerzos, utilizando estrategias clásicas dirigidas a ampliar el área de crecimiento de Lmj y garantizar su conservación, han tenido un éxito limitado. El trabajo que se presenta está dirigido a mejorar el conocimiento de la ecología de Lmj, en particular la interacción simbiótica que establece con bacterias del suelo denominadas rizobios y se centra en la caracterización fenotípica, filogenética y genómica de esos rizobios. También se investiga la posible contribución de la simbiosis en mejorar la conservación de Lmj. Para este fin, se han estudiado diferentes aspectos que se describen a continuación. El primero objetivo se centró en aislar y estudiar de la diversidad genética de las bacterias endosimbióticas de Lmj. . Se realizó un análisis filogenético de genes esenciales que mostró que las cepas de Lmj pertenecen al género Bradyrhizobium y que presentan una gran diversidad con características fenotípicas y simbióticas diferentes de cepas de Bradyrhizobium que nodulan otras especies de lupinos nativos de España (cepas ISLU). Las cepas estudiadas se dividieron en dos grupos (Clado I y Clado II). El Clado I, incluye a las cepas Lmj, definiendo un nuevo linaje, filogenéticamente relacionado con otras especies de Bradyrhizobium, como B. jicamae y B. elkanii. El Clado II contiene cepas ISLU relacionadas con cepas de B. canariense y B. japonicum que establecen simbiosis con lupinos de suelos ácidos. Otro análisis filogenético basado en genes simbióticos, distribuyó las cepas de Lmj en sólo dos grupos diferentes. La singularidad y gran diversidad de estas cepas en una pequeña área geográfica, hacen de este, un atractivo sistema para el estudio de la evolución y adaptación de las bacterias simbióticas a su respectiva planta huésped. Adicionalmente, se estudio la presencia de bacterias capaces de nodular Lmj en suelos básicos de Chiapas, México. Sorprendentemente, estos suelos contienen bacterias capaces establecer interacciones simbióticas eficientes con Lmj en ensayos de invernadero. A continuación se investigó la taxonomía de los endosimbiontes de Lmj analizando la secuencia de cuatro genes esenciales (16S rRNA, recA, glnII y atpD) y el promedio de identidad de nucleótidos de genomas completos de algunas cepas representativas de la diversidad (ANIm). Se identificaron nuevas especies de Bradyrhizobium dentro del Clado I y se definió una de ellas: 'Bradyrhizobium valentinum' sp. nov (cepa tipo LmjM3T = CECT 8364T, LMG 2761T). También se abordó cómo conservar Lmj en su hábitat natural mediante inoculación con alguna de las cepas aisladas. Se demostró la ausencia de bacterias capaces de nodular Lmj en suelos rojos alcalinos o ‘‘terra rossa’’ de la Península Ibérica y Baleares. Dos cepas, altamente eficientes en cuanto a la fijación de nitrógeno, LmjC y LmjM3T, fueron seleccionadas para ser empleadas como inoculantes. Dos experimentos de campo llevados a cabo en años consecutivos en áreas con características edafoclimáticas similares a las que presentan las poblaciones de Lmj, lograron la reproducción exitosa de la planta. Se concluyó que un ciclo reproductivo exitoso de Lmj es absolutamente dependiente de la inoculación con sus simbiontes naturales y que la simbiosis debe ser considerada un factor esencial en estrategias de conservación de leguminosas en peligro. La obtención de varias secuencias genómicas de cepas aisladas de Lmj y de otras cepas de Bradyrhizobium reveló una alta similitud entre los genomas de las cepas del Clado I, y permitió la identificación de cinco posibles nuevas especies. Además, se estudiaron tres agrupaciones de genes relacionados con la simbiosis (nod, nif y fix) definiendo un nuevo linaje para las cepas de Lmj, diferente del symbiovar “genistearum” de B. canariense y B. japonicum. La baja diversidad encontrada en el análisis filogenético de los genes simbióticos contrasta con la gran diversidad asociada a genes esenciales. La presencia de plásmidos en cepas del género Bradyrhizobium ha sido descrita en muy pocas ocasiones, sin embargo el análisis de la secuencia genómica de la cepa ISLU101, aislada de Lupinus angustifolius, reveló la presencia de un origen de replicación extracromosómico homólogo al operón repABC, presente en el plásmido de Bradyrhizobium sp BTAi1. Gracias a esta secuencia se identificaron genes homólogos en 19 de 72 cepas ISLU. Filogenéticamente, las secuencias de repABC se agruparon en un grupo monofilético con las de pBTAi1 y separadas de los rizobios de crecimiento rápido. Finalmente, se identificaron sistemas de secreción de proteínas de tipo III (T3SS) en nueve genomas de cepas de Lmj. Los T3SS pueden inyectar proteínas efectoras al interior de células vegetales. Su presencia en rizobios se ha relacionado con la gama de hospedador que pueden nodular y puede tener un efecto beneficioso, neutro o perjudicial en la simbiosis. Los T3SS de las cepas de Lmj codifican para una proteína efectora similar a NopE, un efector dependiente de T3SS descrito en B. diazoefficiens USDA 110T. La proteína NopE de la cepa LmjC se ha caracterizado bioquímicamente. ABSTRACT Lupinus mariae-josephae (Lmj) is a lupine species endemic of a unique small area in Valencia region (Eastern Spain) where the lupine plants thrive in alkaline-limed soils, which preferentially grow in acid or neutral soils. This is the type of soils native lupines of Spain. When this work was initiated, the extension of the endemic area of Lmj was of about 700 squared kilometers confined to the Valencia province. In this area, Lmj thrives in small, isolated patches containing a reduced number of plants, and points to an endemism that can easily became endangered or extinct. Consequently, the Valencia Community authorities gave a ‘‘microreserve” status for conservation of the species. All efforts, using classical strategies directed to extend the area of Lmj growth and ensure its conservation have been so far unsuccessful. The work presented here is directed to improve our knowledge of Lmj ecology and it is centered in the characterization of the rhizobial symbiosis by phenotypic, phylogenetic and genomic analysis as well as in investigate the potential contribution of the symbiosis to improve its conservation. To this end, five different topics have been studied, and results are briefly described here. Extensive details can be followed en the attached, published articles. The first topic deals with the indigenous rhizobial symbionts of the Lmj endemism, and its genetic diversity was investigated. The Lmj root symbionts belong to the Bradyrhizobium genus, and phylogenetic analysis based on core genes identified a large diversity of Bradyrhizobium strains with phenotypic and symbiotic characteristics different from rhizobia nodulating other Lupinus spp. native of Spain. The strains were split in two clades. Clade II contained strains close to classical B. canariense and B. japonicum lineages that establish symbioses with lupines in acid soils of the Mediterranean area. Clade I included Lmj strains that define a new lineage, close to other Bradyrhizobium species as B. jicamae and B. elkanii. The phylogenetic analysis based on symbiotic genes identified only two distinct clusters. The singularity and large diversity of these strains in such a small geographical area makes this an attractive system for studying the evolution and adaptation of the rhizobial symbiont to the plant host. Additionally, the presence of bacteria able to nodulate Lmj in basic soils from Chiapas, Mexico was investigated. Surprisingly, these soils contain bacteria able to effectively nodulate and fix nitrogen with Lmj plants in greenhouse assays. In the second topic, the taxonomic status of the endosymbiotic bacteria of Lmj from Valencia endemism and Chiapas was investigated. Results from phylogenetic analysis of core genes and Average Nucleotide Identity (ANIm) using draft genomic sequences identified new Bradyrhizobium species within strains of Clade I of Lmj endosymbiotic bacteria. Only one of these potentially new species has been defined, meanwhile the others are under process of characterization. The name ‘Bradyrhizobium valentinum’ sp. nov. was proposed for the defined species (type strain LmjM3T= CECT 8364T, LMG 2761T). The third topic was directed to conservation of endangered Lmj in its natural habitat. The relevant conclusion of this experimentation is that the symbiosis should be considered as a relevant factor in the conservation strategies for endangered legumes. First, we showed absence of bacteria able to nodulate Lmj in all the inspected ‘‘terra rossa’’ or alkaline red soils of the Iberian Peninsula and Balearic Islands. Then, two efficient nitrogen fixing strains with Lmj plants, LmjC and LmjM3T, were selected as inoculum for seed coating. Two planting experiments were carried out in consecutive years under natural conditions in areas with edapho-climatic characteristics identical to those sustaining natural Lmj populations, and successful reproduction of the plant was achieved. The relevant conclusion from these assays was that the successful reproductive cycle was absolutely dependent on seedling inoculation with effective bradyrhizobia The forth topic deep into the analysis of the genomic of Lmj representative strains. To this end, draft genomic sequences of selected Lmj strains and type strains of Bradyrhizobium spp. were assembled. The comparison analysis of the draft genomic sequences of Lmj strains and related Bradyrhizobium species grouped in Clade I, revealed a high genomic homology among them, and allowed the definition of five potentially new species of Lmj nodulating bacteria. Also, based on the available draft genomic sequences, only three clusters of nod, fix and nif genes from Lmj strains were identified and showed to define a new symbiotic lineage, distant from that of B. canariense and B. japonicum bv. genistearum. The low diversity exhibited by the phylogenetic analysis of symbiotic genes contrast with the large diversity of strains as regards the housekeeping genes analyzed. Besides, the genomic analysis of a Lupinus angustifolius strain ISLU101, revealed the presence of an extrachromosomal replication origin homologous to repABC cluster from plasmid present in Bradyrhizobium spp BTAi1. This repABC cluster gene sequence allowed the identification of extrachromosomic replication origin in 19 out of 72 Bradyrhizobium strains from Lupinus spp., a highly significant result since the absence of plasmids in the Bradyrhizobium genus was traditionally assumed. The repABC gene sequences of these strains grouped them in a unique monophyletic group, related to B. sp. BTAi1 plasmid, but differentiated from the repABC gene cluster of plasmids in fast growing rhizobium strains. The last topic was focused on characterization of type III secreted effectors present in Lmj endosymbiotic bacteria. Type III secretion systems (T3SS) are specialized protein export machineries which can deliver effector proteins into plant cells. The presence of T3SS in rhizobia has frequently been related to the symbiotic nodulation host-range and may have a beneficial or detrimental effect on the symbiosis with legumes. In this context, the presence of T3SS in genomes of nine Lmj strains was investigated, and it was shown the presence of clusters encoding NopE type III-secreted protein similar to the NopE1 and NopE2 of B. diazoefficiens USDA 110T. The putative NopE protein of LmjC strain is at present being characterized regarding its structure and function.

Laser remote sensing applications (LiDAR) in forest hydrology and the management of river ecosystems.

La utilización de sensores láser desde plataformas aéreas (LiDAR) ofrece nuevas posibilidades en el cartografiado de sistemas fluviales, tanto en áreas densamente cubiertas por vegetación, como en aquellas que presentan una escasa cubierta. La información topográfica de alta resolución que se obtiene a partir de las medidas láser puede ser utilizada en el análisis y estimación de diversas variables hidrológicas, y en el estudio de diferentes componentes del medio fluvial. Entre éstas, cabe citar la vegetación riparia, la morfología fluvial, el régimen hidrológico o el grado de alteración de los ecosistemas debido a las presiones de origen antrópico. La gestión del medio fluvial puede ser mejorada en gran medida gracias a la precisión y fiabilidad de esta información. En muchas ocasiones, el escaso relieve de los valles fluviales y la densa cubierta vegetal que existe en ellos han dificultado la aplicación de otras técnicas de teledetección. Sin embargo, los datos obtenidos mediante altimetría láser son especialmente aconsejables para estos trabajos, mediante análisis numéricos o a través de la simple interpretación de las imágenes obtenidas. Este artículo muestra las posibilidades de uso de los datos LiDAR en hidrología forestal y en la gestión de zonas húmedas, a lo largo de tramos con condiciones climáticas bien diferenciadas. En todas ellas, se comparan los resultados obtenidos mediante la aplicación de distinto software, con el fin de mostrar la mejor metodología de tratamiento de la información láser. Asimismo, se muestra la diferencia con otras técnicas de teledetección, y se muestra la fácil integración de los datos LiDAR con otras herramientas y metodologías de estudio de las variables citadas.

Impacts of global change on iberian forest ecosystems inferred from the spanish national forest inventory = Impactos del cambio global en ecosistemas forestales ibéricos a partir del inventario forestal nacional

Los efectos del cambio global sobre los bosques son una de las grandes preocupaciones de la sociedad del siglo XXI. Algunas de sus posibles consecuencias como son los efectos en la producción, la sostenibilidad, la pérdida de biodiversidad o cambios en la distribución y ensamblaje de especies forestales pueden tener grandes repercusiones sociales, ecológicas y económicas. La detección y seguimiento de estos efectos constituyen uno de los retos a los que se enfrentan en la actualidad científicos y gestores forestales. En base a la comparación de series históricas del Inventario Forestal Nacional Español (IFN), esta tesis trata de arrojar luz sobre algunos de los impactos que los cambios socioeconómicos y ambientales de las últimas décadas han generado sobre nuestros bosques. En primer lugar, esta tesis presenta una innovadora metodología con base geoestadística que permite la comparación de diferentes ciclos de inventario sin importar los diferentes métodos de muestreo empleados en cada uno de ellos (Capítulo 3). Esta metodología permite analizar cambios en la dinámica y distribución espacial de especies forestales en diferentes gradientes geográficos. Mediante su aplicación, se constatarán y cuantificarán algunas de las primeras evidencias de cambio en la distribución altitudinal y latitudinal de diferentes especies forestales ibéricas, que junto al estudio de su dinámica poblacional y tasas demográficas, ayudarán a testar algunas hipótesis biogeográficas en un escenario de cambio global en zonas de especial vulnerabilidad (Capítulos 3, 4 y 5). Por último, mediante la comparación de ciclos de parcelas permanentes del IFN se ahondará en el conocimiento de la evolución en las últimas décadas de especies invasoras en los ecosistemas forestales del cuadrante noroccidental ibérico, uno de los más afectados por la invasión de esta flora (Capítulo 6). ABSTRACT The effects of global change on forests are one of the major concerns of the XXI century. Some of the potential impacts of global change on forest growth, productivity, biodiversity or changes in species assembly and spatial distribution may have great ecological and economic consequences. The detection and monitoring of these effects are some of the major challenges that scientists and forest managers face nowadays. Based on the comparison of historical series of the Spanish National Forest Inventory (NFI), this thesis tries to shed some light on some of the impacts driven by recent socio-economic and environmental changes on our forest ecosystems. Firstly, this thesis presents an innovative methodology based on geostatistical techniques that allows the comparison of different NFI cycles regardless of the different sampling methods used in each of them (Chapter 3). This methodology, in conjunction with other statistical techniques, allows to analyze changes in the spatial distribution and population dynamics of forest species along different geographic gradients. By its application, this thesis presents some of the first evidences of changes in species distribution along different geographical gradients in the Iberian Peninsula. The analysis of these findings, of species population dynamics and demographic rates will help to test some biogeographical hypothesis on forests under climate change scenarios in areas of particular vulnerability (Chapters 3, 4 and 5). Finally, by comparing NFI cycles with permanent plots, this thesis increases our knowledge about the patterns and processes associated with the recent evolution of invasive species in the forest ecosystems of North-western Iberia, one of the areas most affected by the invasion of allien species at national scale (Chapter 6).

Deforestación y Pérdida de hábitat en Bosques de montaña en la Cuenca alta del Río Zamora (Loja, Ecuador)

Los bosques húmedos de montaña se encuentran reconocidos como uno de los ecosistemas más amenazados en el mundo, llegando inclusive a ser considerado como un “hotspot” por su alta diversidad y endemismo. La acelerada pérdida de cobertura vegetal de estos bosques ha ocasionado que, en la actualidad, se encuentren restringidos a una pequeña fracción de su área de distribución histórica. Pese a esto, los estudios realizados sobre cual es efecto de la deforestación, fragmentación, cambios de uso de suelo y su efecto en las comunidades de plantas presentes en este tipo de vegetación aún son muy escuetos, en comparación a los realizados con sus similares amazónicos. En este trabajo, el cual se encuentra dividido en seis capítulos, abordaremos los siguientes objetivos: a) Comprender cuál es la dinámica que han seguido los diferentes tipos de bosques montanos andinos de la cuenca del Rio Zamora, Sur de Ecuador durante entre 1976 y 2002. b) Proveer de evidencia de las tasas de deforestación y fragmentación de todos los tipos diferentes de bosques montanos andinos presentes en la cuenca del Rio Zamora, Sur de Ecuador entre 1976 y 2002. c) Determinar qué factores inducen a la fragmentación de bosques de montaña en la cuenca alta del río Zamora entre 1976 y 2002. d) Determinar cuáles son y cómo afectan los factores ambientales y socioeconómicos a la dinámica de la deforestación y regeneración (pérdida y recuperación del hábitat) sufrida por los bosques de montaña dentro de la zona de estudio y e) Determinar si la deforestación y fragmentación actúan sobre la diversidad y estructura de las comunidades de tres tipos de organismos (comunidades de árboles, comunidades de líquenes epífitos y comunidades de hepáticas epífitas). Este estudio se centró en el cuenca alta del río Zamora, localizada al sur de Ecuador entre las coordenadas 3º 00´ 53” a 4º 20´ 24.65” de latitud sur y 79º 49´58” a 78º 35´ 38” de longitud oeste, que cubre alrededor de 4300 km2 de territorio situado entre las capitales de las provincias de Loja y Zamora-Chinchipe. Con objeto de predecir la dinámica futura de la deforestación en la región de Loja y cómo se verán afectados los diferentes tipos de hábitat, así como para detectar los factores que más influyen en dicha dinámica, se han construido modelos basados en la historia de la deforestación derivados de fotografías aéreas e imágenes satelitales de tres fechas (1976, 1989 y 2002). La cuantificación de la deforestación se realizó mediante la tasa de interés compuesto y para la caracterización de la configuración espacial de los fragmentos de bosque nativo se calcularon índices de paisaje los cuales fueron calculados utilizando el programa Fragstats 3.3. Se ha clasificado el recubrimiento del terreno en forestal y no forestal y se ha modelado su evolución temporal con Modelos Lineales Generalizados Mixtos (GLMM), empleando como variables explicativas tanto variables ambientales espacialmente explícitas (altitud, orientación, pendiente, etc) como antrópicas (distancia a zonas urbanizadas, deforestadas, caminos, entre otras). Para medir el efecto de la deforestación sobre las comunidades modelo (de árboles, líquenes y hepáticas) se monitorearon 11 fragmentos de vegetación de distinto tamaño: dos fragmentos de más de cien hectáreas, tres fragmentos de entre diez y noventa ha y seis fragmentos de menos de diez hectáreas. En ellos se instalaron un total de 38 transectos y 113 cuadrantes de 20 x 20 m a distancias que se alejaban progresivamente del borde en 10, 40 y 80 m. Nuestros resultados muestran una tasa media anual de deforestación del 1,16% para todo el período de estudio, que el tipo de vegetación que más alta tasa de destrucción ha sufrido, es el páramo herbáceo, con un 2,45% anual. El análisis de los patrones de fragmentación determinó un aumento en 2002 de más del doble de fragmentos presentes en 1976, lo cual se repite en el análisis del índice de densidad promedio. El índice de proximidad media entre fragmentos muestra una reducción progresiva de la continuidad de las áreas forestadas. Si bien las formas de los fragmentos se han mantenido bastante similares a lo largo del período de estudio, la conectividad entre estos ha disminuido en un 84%. Por otro lado, de nuestros análisis se desprende que las zonas con mayor probabilidad de deforestarse son aquellas que están cercanas a zonas previamente deforestadas; la cercanía a las vías también influye significativamente en la deforestación, causando un efecto directo en la composición y estructura de las comunidades estudiadas, que en el caso de los árboles viene mediado por el tamaño del fragmento y en el caso del componente epífito (hepáticas y líquenes), viene mediado tanto por el tamaño del fragmento como por la distancia al borde del mismo. Se concluye la posibilidad de que, de mantenerse esta tendencia, este tipo de bosques desaparecerá en corto tiempo y los servicios ecosistémicos que prestan, se verán seriamente comprometidos. ABSTRACT Mountain rainforests are recognized as one of the most threatened ecosystems in the world, and have even come to be considered as a “hotspot” due to their high degree of diversity and endemism. The accelerated loss of plant cover of these forests has caused them to be restricted today to a small fraction of their area of historic distribution. In spite of this, studies done on the effect of deforestation, fragmentation, changes in soil use and their effect on the plant communities present in this type of vegetation are very brief compared to those done on their analogues in the Amazon region. In this study, which is divided into six chapters, we will address the following objectives: a) To understand what the dynamic followed by the different types of Andean mountain forests in the Zamora River watershed of southern Ecuador has been between 1976 and 2002. b) To provide evidence of the rates of deforestation and fragmentation of all the different types of Andean mountain forests existing in the upper watershed of the Zamora River between 1976 and 2002. c) To determine the factors that induces fragmentation of all different types of Andean mountain forests existing in the upper watershed of the Zamora River between 1976 and 2002. d) To determine what the environmental and anthropogenic factors are driving the dynamic of deforestation and regeneration (loss and recuperation of the habitat) suffered by the mountain forests in the area of the study and e) To determine if the deforestation and fragmentation act upon the diversity and structure of three model communities: trees, epiphytic lichens and epiphytic liverworts. This study is centered on the upper Zamora River watershed, located in southern Ecuador between 3º 00´ 53” and 4º 20´ 24.65 south latitude and 79º 49´ 58” to 78º 35´ 38” west longitude, and covers around 4,300 km2 of territory located between Loja and Zamora-Chinchipe provinces. For the purpose of predicting the future dynamic of deforestation in the Loja region and how different types of habitats will be affected, as well as detecting the environmental and socioeconomic factors that influence landscape dynamics, models were constructed based on deforestation history, derived from aerial photographs and satellite images for three dates (1976, 1989 and 2002). Quantifying the deforestation was done using the compound interest rate; to characterize the spatial configuration of fragments of native forest, landscape indices were calculated with Fragstats 3.3 program. Land cover was classified as forested and not forested and its evolution over time was modeled with Generalized Linear Mixed Models (GLMM), using spatially explicit environmental variables (altitude, orientation, slope, etc.) as well as anthropic variables (distance to urbanized, deforested areas and roads, among others) as explanatory variables. To measure the effects of fragmentation on three types of model communities (forest trees and epiphytic lichen and liverworts), 11 vegetation fragments of different sizes were monitored: two fragments of more than one hundred hectares, three fragments of between ten and ninety ha and six fragments of fewer than ten hectares . In these fragments, a total of 38 transects and 113 20 x 20 m quadrats were installed at distances that progressively moved away from the edge of the fragment by 10, 40 and 80 m. Our results show an average annual rate of deforestation of 1.16% for the entire period of the study, and that the type of vegetation that suffered the highest rate of destruction was grassy paramo, with an annual rate of 2.45%. The analysis of fragmentation patterns determined the number of fragments in 2002 more than doubled the number of fragments present in 1976, and the same occurred for the average density index. The variation of the average proximity index among fragments showed a progressive reduction of the continuity of forested areas. Although fragment shapes have remained quite similar over the period of the study, connectivity among them has diminished by 84%. On the other hand, it emerged from our analysis that the areas of greatest probability of deforestation were those that are close to previously deforested areas; proximity to roads also significantly favored the deforestation causing a direct effect on the composition of our model communities, that in the case of forest trees is determined by the size of the fragment, and in the case of the epiphyte communities (liverworts and lichens), is determined, by the size of the fragment as well as the distance to edge. A subject under discussion is the possibility that if this tendency continues, this type of forest will disappear in a short time, and the ecological services it provides, will be seriously endangered.

Environmental niche variation and evolutionary diversification of the Brachypodium distachyon grass complex species in their native circum-Mediterranean range

PREMISE OF THE STUDY: We conducted environmental niche modeling (ENM) of the Brachypodium distachyon s.l. complex, a model group of two diploid annual grasses ( B. distachyon , B. stacei ) and their derived allotetraploid ( B. hybridum) , native to the circum-Mediterranean region. We (1) investigated the ENMs of the three species in their native range based on present and past climate data; (2) identifi ed potential overlapping niches of the diploids and their hybrid across four Quaternary windows; (3) tested whether speciation was associated with niche divergence/conservatism in the complex species; and (4) tested for the potential of the polyploid outperforming the diploids in the native range. M ETHODS: Geo-referenced data, altitude, and 19 climatic variables were used to construct the ENMs. We used paleoclimate niche models to trace the potential existence of ancestral gene fl ow among the hybridizing species of the complex. KEY RESULTS: Brachypodium distachyon grows in higher, cooler, and wetter places, B. stacei in lower, warmer, and drier places, and B. hybridum in places with intermediate climatic features. Brachypodium hybridum had the largest niche overlap with its parent niches, but a similar distribution range and niche breadth. C ONCLUSIONS: Each species had a unique environmental niche though there were multiple niche overlapping areas for the diploids across time, suggesting the potential existence of several hybrid zones during the Pleistocene and the Holocene. No evidence of niche divergence was found, suggesting that species diversifi cation was not driven by ecological speciation but by evolutionary history, though it could be associated to distinct environmental adaptations.

Proceedings of the third USA-USSR symposium on the effects of pollutants upon aquatic ecosystems.

3rd.

Proceedings of the third USA-USSR symposium on the effects of pollutants upon aquatic ecosystems.

1st & 2nd

Structure and function of the Bacillus hybrid enzyme GluXyn-1: Native-like jellyroll fold preserved after insertion of autonomous globular domain

The 1,3–1,4-β-glucanase from Bacillus macerans (wtGLU) and the 1,4-β-xylanase from Bacillus subtilis (wtXYN) are both single-domain jellyroll proteins catalyzing similar enzymatic reactions. In the fusion protein GluXyn-1, the two proteins are joined by insertion of the entire XYN domain into a surface loop of cpMAC-57, a circularly permuted variant of wtGLU. GluXyn-1 was generated by protein engineering methods, produced in Escherichia coli and shown to fold spontaneously and have both enzymatic activities at wild-type level. The crystal structure of GluXyn-1 was determined at 2.1 Å resolution and refined to R = 17.7% and R(free) = 22.4%. It shows nearly ideal, native-like folding of both protein domains and a small, but significant hinge bending between the domains. The active sites are independent and accessible explaining the observed enzymatic activity. Because in GluXyn-1 the complete XYN domain is inserted into the compact folding unit of GLU, the wild-type-like activity and tertiary structure of the latter proves that the folding process of GLU does not depend on intramolecular interactions that are short-ranged in the sequence. Insertion fusions of the GluXyn-1 type may prove to be an easy route toward more stable bifunctional proteins in which the two parts are more closely associated than in linear end-to-end protein fusions.

Antibodies to several conformation-dependent epitopes of gp120/gp41 inhibit CCR-5-dependent cell-to-cell fusion mediated by the native envelope glycoprotein of a primary macrophage-tropic HIV-1 isolate

The β-chemokine receptor CCR-5 is essential for the efficient entry of primary macrophage-tropic HIV-1 isolates into CD4+ target cells. To study CCR-5-dependent cell-to-cell fusion, we have developed an assay system based on the infection of CD4+ CCR-5+ HeLa cells with a Semliki Forest virus recombinant expressing the gp120/gp41 envelope (Env) from a primary clade B HIV-1 isolate (BX08), or from a laboratory T cell line-adapted strain (LAI). In this system, gp120/gp41 of the “nonsyncytium-inducing,” primary, macrophage-tropic HIV-1BX08 isolate, was at least as fusogenic as that of the “syncytium-inducing” HIV-1LAI strain. BX08 Env-mediated fusion was inhibited by the β-chemokines RANTES (regulated upon activation, normal T cell expressed and secreted) and macrophage inflammatory proteins 1β (MIP-1β) and by antibodies to CD4, whereas LAI Env-mediated fusion was insensitive to these β-chemokines. In contrast soluble CD4 significantly reduced LAI, but not BX08 Env-mediated fusion, suggesting that the primary isolate Env glycoprotein has a reduced affinity for CD4. The domains in gp120/gp41 involved in the interaction with the CD4 and CCR-5 molecules were probed using monoclonal antibodies. For the antibodies tested here, the greatest inhibition of fusion was observed with those directed to conformation-dependent, rather than linear epitopes. Efficient inhibition of fusion was not restricted to epitopes in any one domain of gp120/gp41. The assay was sufficiently sensitive to distinguish between antibody- and β-chemokine-mediated fusion inhibition using serum samples from patient BX08, suggesting that the system may be useful for screening human sera for the presence of biologically significant antibodies.

Structural mimicry of a native protein by a minimized binding domain

The affinity between molecules depends both on the nature and presentation of the contacts. Here, we observe coupling of functional and structural elements when a protein binding domain is evolved to a smaller functional mimic. Previously, a 38-residue form of the 59-residue B-domain of protein A, termed Z38, was selected by phage display. Z38 contains 13 mutations and binds IgG only 10-fold weaker than the native B-domain. We present the solution structure of Z38 and show that it adopts a tertiary structure remarkably similar to that observed for the first two helices of B-domain in the B-domain/Fc complex [Deisenhofer, J. (1981) Biochemistry 20, 2361–2370], although it is significantly less stable. Based on this structure, we have improved on Z38 by designing a 34-residue disulfide-bonded variant (Z34C) that has dramatically enhanced stability and binds IgG with 9-fold higher affinity. The improved stability of Z34C led to NMR spectra with much greater chemical shift dispersion, resulting in a more precisely determined structure. Z34C, like Z38, has a structure virtually identical to the equivalent region from native protein A domains. The well-defined hydrophobic core of Z34C reveals key structural features that have evolved in this small, functional domain. Thus, the stabilized two-helix peptide, about half the size and having one-third of the remaining residues altered, accurately mimics both the structure and function of the native domain.