Unconformity-bounded stratigraphic units (UBSUs) in an italian alluvial-plain area: recognizing and dating

We studied the coastal zone of the Tavoliere di Puglia plain, (Puglia region, southern Italy) with the aim to recognize the main unconformities, and therefore, the unconformity-bounded stratigraphic units (UBSUs; Salvador 1987, 1994) forming its Quaternary sedimentary fill. Recognizing unconformities is particularly problematic in an alluvial plain, due to the difficulties in distinguishing the unconformities that bound the UBSUs. So far, the recognition of UBSUs in buried successions has been made mostly by using seismic profiles. Instead, in our case, the unavailability of the latter has prompted us to address the problem by developing a methodological protocol consisting of the following steps: I) geological survey in the field; II) draft of a preliminary geological setting based on the field-survey results; III) dating of 102 samples coming from a large number of boreholes and some outcropping sections by means of the amino acid racemization (AAR) method applied to ostracod shells and 14C dating, filtering of the ages and the selection of valid ages; IV) correction of the preliminary geological setting in the light of the numerical ages; definition of the final geological setting with UBSUs; identification of a ‘‘hypothetical’’ or ‘‘attributed time range’’ (HTR or ATR) for each UBSU, the former very wide and subject to a subsequent modification, the latter definitive; V) cross-checking between the numerical ages and/or other characteristics of the sedimentary bodies and/or the sea-level curves (with their effects on the sedimentary processes) in order to restrict also the hypothetical time ranges in the attributed time ranges. The successful application of AAR geochronology to ostracod shells relies on the fact that the ability of ostracods to colonize almost all environments constitutes a tool for correlation, and also allow the inclusion in the same unit of coeval sediments that differ lithologically and paleoenvironmentally. The treatment of the numerical ages obtained using the AAR method required special attention. The first filtering step was made by the laboratory (rejection criteria a and b). Then, the second filtering step was made by testing in the field the remaining ages. Among these, in fact, we never compared an age with a single preceding and/or following age; instead, we identified homogeneous groups of numerical ages consistent with their reciprocal stratigraphic position. This operation led to the rejection of further numerical ages that deviate erratically from a larger, homogeneous age population which fits well with its stratigraphic position (rejection criterion c). After all of the filtering steps, the valid ages that remained were used for the subdivision of the sedimentary sequences into UBSUs together with the lithological and paleoenvironmental criteria. The numerical ages allowed us, in the first instance, to recognize all of the age gaps between two consecutive samples. Next, we identified the level, in the sedimentary thickness that is between these two samples, that may represent the most suitable UBSU boundary based on its lithology and/or the paleoenvironment. The recognized units are: I) Coppa Nevigata sands (NEA), HTR: MIS 20–14, ATR: MIS 17–16; II) Argille subappennine (ASP), HTR: MIS 15–11, ATR: MIS 15–13; III) Coppa Nevigata synthem (NVI), HTR: MIS 13–8, ATR: MIS 12–11; IV) Sabbie di Torre Quarto (STQ), HTR: MIS 13–9.1, ATR: MIS 11; V) Amendola subsynthem (MLM1), HTR: MIS 12–10, ATR: MIS 11; VI) Undifferentiated continental unit (UCI), HTR: MIS 11–6.2, ATR: MIS 9.3–7.1; VII) Foggia synthem (TGF), ATR: MIS 6; VIII) Masseria Finamondo synthem (TPF), ATR: Upper Pleistocene; IX) Carapelle and Cervaro streams synthem (RPL), subdivided into: IXa) Incoronata subsynthem (RPL1), HTR: MIS 6–3; ATR: MIS 5–3; IXb) Marane La Pidocchiosa–Castello subsynthem (RPL3), ATR: Holocene; X) Masseria Inacquata synthem (NAQ), ATR: Holocene. The possibility of recognizing and dating Quaternary units in an alluvial plain to the scale of a marine isotope stage constitutes a clear step forward compared with similar studies regarding other alluvial-plain areas, where Quaternary units were dated almost exclusively using their stratigraphic position. As a result, they were generically associated with a geological sub-epoch. Instead, our method allowed a higher detail in the timing of the sedimentary processes: for example, MIS 11 and MIS 5.5 deposits have been recognized and characterized for the first time in the study area, highlighting their importance as phases of sedimentation.

Interconexión monolítica de módulos fotovoltaicos de silicio amorfo en lámina delgada sobre vidrio mediante ablación láser en régimen de nanosegundos

Esta tesis se centra en el estudio de una secuencia de procesos basados en la tecnología láser y ejecutados en dispositivos fotovoltaicos, que son imprescindibles para el desarrollo en general de las tecnologías fotovoltaicas basadas en lámina delgada y, en particular, de aquellas que utilizan silicio amorfo como absorbente, así como en aplicaciones posteriores de estas tecnologías de alto valor añadido como es la integración arquitectónica de este tipo de dispositivos. En gran parte de las tecnologías FV de lámina delgada, y muy particularmente en la de silicio amorfo, el material se deposita sobre un substrato en un área lo suficientemente grande para que se requiera de un proceso de subdivisión del dispositivo en células de tamaño adecuado, y su posterior conexión en serie para garantizar las figuras eléctricas nominales del dispositivo. Este proceso se ha desarrollado industrialmente hace años, pero no ha habido un esfuerzo científico asociado que permitiera conocer en profundidad los efectos que los procesos en si mismos tiene de forma individualizada sobre los materiales que componen el dispositivo y sus características finales. Este trabajo, desarrollado durante años en el Centro Láser de la UPM, en estrecha colaboración con Centro de Investigaciones Energéticas y Medioambientales (CIEMAT), la Universidad de Barcelona (UB), y la Universidad Politécnica de Cataluña (UPC), se centra justamente en un estudio detallado de dichos procesos, denominados habitualmente P1, P2, P3 y P4 atendiendo al orden en el que se realizan en el dispositivo. Este estudio incluye tanto la parametrización de los procesos, el análisis del efecto que los mismos producen sobre los materiales que componen el dispositivo y su comportamiento fotoeléctrico final, así como la evaluación del potencial uso de fuentes láser de última generación (ultrarrápidas) frente al estándar industrial en la actualidad que es el empleo de fuentes láser convencionales de ancho temporal en el rango de los nanosegundos. En concreto se ha estudiado en detalle las ventajas y limitaciones del uso de sistemas con diferentes rangos espectrales (IR, VIS y UV) y temporales (nanosegundos y picosegundos) para diferentes tipos de configuraciones y disposiciones tecnológicas (entendiendo por estas las habituales configuraciones en substrato y superestrato de este tipo de dispositivos). La caracterización individual de los procesos fue realizada primeramente en células de laboratorio específicamente diseñadas, abriendo nuevos planteamientos y conceptos originales para la mejora de los procesos láser de interconexión y posibilitando el empleo y desarrollo de técnicas y métodos avanzados de caracterización para el estudio de los procesos de ablación en las distintas láminas que conforman la estructura de los dispositivos fotovoltaicos, por lo que se considera que este trabajo ha propuesto una metodología completamente original, y que se ha demostrado efectiva, en este ámbito. Por último el trabajo aborda un tema de particular interés, como es el posible uso de los procesos desarrollados, no para construir los módulos fotovoltaicos en sí, sino para personalizarlos en forma y efectos visuales para potenciar su uso mediante elementos integrables arquitectónicamente, lo que es un ámbito de gran potencial de desarrollo futuro de las tecnologías fotovoltaicas de lámina delgada. En concreto se presentan estudios de fabricación de dispositivos integrables arquitectónicamente y plenamente funcionales no solo en dispositivos de silicio amorfo con efectos de transparencias y generación de formas libres, si no que también se incluye la posibilidad de hacer tales dispositivos con células de silicio cristalino estándar que es la tecnología fotovoltaica de mayor presencia en mercado. Es importante, además, resaltar que la realización de este trabajo ha sido posible gracias a la financiación obtenida con dos proyectos de investigación aplicada, MICROSIL (PSE-120000-2008-1) e INNDISOL (IPT-420000-2019-6), y los correspondientes al Plan Nacional de I+D+I financiados por el ministerio de Ciencia e Innovación y el Ministerio de Economía y Competitividad: CLÁSICO (ENE 2007- 67742-C04-04) y AMIC ENE2010-21384-C04-02. De hecho, y en el marco de estos proyectos, los resultados de este trabajo han ayudado a conseguir algunos de los hitos más importantes de la tecnología fotovoltaica en nuestro país en los últimos años, como fue en el marco de MICROSIL la fabricación del primer módulo de silicio amorfo con tecnología íntegramente española (hecho en colaboración con el CIEMAT), o la fabricación de los dispositivos para integración arquitectónica con geometrías libres que se describen en esta Tesis y que fueron parte de los desarrollos del proyecto INNDISOL. ABSTRACT This thesis focuses on the study of a sequence of laser-based technology and processes executed in photovoltaic devices, which are essential for the overall development of photovoltaic technologies based on thin film and, in particular, those using amorphous silicon as absorbent and subsequent applications of these technologies with high added value such as the architectural integration of such devices. In much of the PV thin film technologies, and particularly in the amorphous silicon material is deposited on a substrate in an area large enough so that it requires a process of subdivision of the device in cells of appropriate size, and subsequent serial connection to ensure nominal device power figures. This process has been industrially developed years ago, but there has been an associate scientific effort that would learn more about the effects that the processes themselves have either individually on the materials that make up the device and its final characteristics. This work, developed over years in the Laser Center of the UPM, in close collaboration with Centre for Energy and Environmental Research (CIEMAT), the University of Barcelona (UB) and the Polytechnic University of Catalonia (UPC)., Focuses precisely in a detailed study of these processes, usually they called P1, P2, P3 and P4 according to the order in which they perform on the device. This study includes both the parameters of the processes, the analysis of the effect they produce on the materials making up the device and its final photoelectric behavior as well as the potential use of EVALUATION of next-generation laser sources (ultrafast) versus standard industry today is the use of conventional laser sources temporal width in the range of nanoseconds. In particular we have studied in detail the advantages and limitations of using systems with different spectral ranges (IR, UV and VIS) and time (nanosecond and picosecond) for different configurations and technological provisions (meaning these typical configurations in substrate and superstrate such devices). Individual characterization of the processes was conducted primarily in laboratory cells specifically designed, opening new approaches and original concepts for improving laser interconnection processes and enabling the use and development of advanced techniques and characterization methods for studying the processes ablation in the different sheets making up the structure of the photovoltaic devices, so it is considered that this work has proposed a completely original methodology, which has proven effective in this area. Finally, the paper addresses a topic of particular interest, as is the possible use of lso developed processes, not to build the photovoltaic modules themselves but to customize fit and visual effects to enhance their use by integrated architectural elements, which is an area of great potential for future development of thin film photovoltaic technologies. Specifically studies manufacture of integrated architecturally and fully functional not only in amorphous silicon devices with transparency effects and generating freeform devices occur, if not also include the ability to make such devices with cells of standard crystalline silicon photovoltaic technology is more visible in the market. It is also important to note that the completion of this work has been possible thanks to the financing obtained with two applied research projects, Microsil (PSE-120000- 2008-1) and INNDISOL (IPT-420000-2019-6), and those for the National R & D funded by the Ministry of Science and Innovation and the Ministry of Economy and Competitiveness: CLASSIC (ENE 2007-67742-C04-04) and AMIC ENE2010-21384-C04- 02. In fact, within the framework of these projects, the results of this work have helped get some of the most important milestones of photovoltaic technology in our country in recent years, as it was under Microsil making the first module Amorphous silicon technology with entirely Spanish (made in collaboration with CIEMAT), or the manufacture of devices for architectural integration with free geometries that are described in this thesis and that were part of the project Inndisol developments.

Nagele: un nuevo asentamiento en un paisaje artificial

Nagele es un asentamiento urbano situado en el Noordoostpolder, territorio neerlandés ganado al mar. Fue diseñado por arquitectos de los grupos De 8 en Opbouw entre los que destacaron Rietveld, Van Eesteren, Van Eyck, Bakema, Stam y Ruys. El proyecto se desarrolló entre 1947 y 1956, un periodo de tiempo con formas de proyectar muy ricas en interpretaciones. Los arquitectos pusieron en crisis los planteamientos historicistas de las nuevas poblaciones de los pólderes. Propusieron un nuevo prototipo, una morfología compacta y concéntrica que transmitiría igualdad a una comunidad agrícola, entendida como una sociedad urbana del siglo XX. La administración apoyó la propuesta que convertiría el proyecto en un arriesgado reto por su falta de antecedentes. La vigencia de las formulaciones permanece hoy en día en la ciudad construida, aunque con alteraciones. En los dibujos del proceso se encuentran los principales enunciados teóricos que este trabajo pretende descubrir. El trabajo aborda aspectos no suficientemente explorados, como su relación con el pólder, la evolución de las estrategias proyectivas, la ordenación paisajista y los elementos urbanos. El Noordoostpolder es la culminación de una serie de experiencias multidisciplinares en el reclamo de tierras a gran escala. Se estudia su estructura urbana policéntrica, la parcelación agrícola que origina el proyecto urbano y la vinculación de la vegetación con la infraestructura, proporcionando orientación, protección climática y escala humana, conceptos que impregnan las estrategias del proyecto urbano. La primera fase de la ordenación configuró áreas monofuncionales que respondían a cada una de las cuatro necesidades básicas del método científico de la ciudad higienista. El acontecimiento que marcó el final de la primera fase fue su presentación en el séptimo CIAM de 1949, cuyo título fue Aplicación de la Carta de Atenas. El programa residencial se dividió en clusters organizados en torno a una pradera vecinal central, vinculándose el orden vecinal, urbano y territorial. La segunda fase fue un catalizador de nuevos planteamientos. El proyecto se transformó en un In-between Realm, un escenario teórico donde coexisten fenómenos tradicionalmente antagónicos que Van Eyck denominó Twin Phenomena, convirtiéndose la ciudad en una réplica formal de la ambivalencia de la mente humana. La indefinición espacial no programada en la propuesta anterior se transformó en un conjunto de espacios urbanos, con límites y dimensiones adaptados a la escala humana. El proyecto es anterior a la obra escrita de Van Eyck por lo que estimuló sus enunciados teóricos. Unas ideas también reconocidas en los tres CIAM posteriores en los que también se expuso el proyecto. El diseño paisajista se integra en el proyecto urbano desde sus orígenes. El límite se compone de una barrera boscosa que protege climáticamente, proporciona escala humana y control visual frente a las llanuras infinitas del pólder. Van Eyck sintetizó el proyecto como una habitación verde sin techo, afirmación que dilucida su equivalencia con el de un interior doméstico. Exteriormente la ciudad se convierte en una unidad autónoma del territorio. Interiormente, un sistema jerarquizado de vegetación vinculado a la arquitectura y la infraestructura constituye espacios urbanos de diferentes escalas. La propuesta fue transformada por Boer y Ruys en un nuevo espacio urbano único, no asimilando los conceptos anteriores. El proyecto y construcción de los primeros elementos urbanos consistió en un reparto de tareas a De 8 en Opbouw, hecho que estimula estudiar su relación con el proyecto urbano. La estructura policéntrica organiza las aulas de las escuelas de Van Eyck, las diferentes áreas confesionales del cementerio de Ruys y las unidades residenciales, diseñadas por Stam, Rietveld y Stam-Besse. Los Twin Phenomena alcanzan un acuerdo en el corredor comercial, diseñado por Bakema y Van der Broek. La generación de espacios dentro de otros aparece también en el cementerio, a través de una nueva barrera boscosa y en el sistema de pliegues del muro que configura la iglesia de Bakema y Van der Broek. El proyecto se vincula a un planteamiento holístico, mediante el que el diseño de cada uno de sus elementos tiene en cuenta las estrategias proyectivas del todo del cual forma parte, convirtiéndose, al igual que las obras de De Stijl, en parte de una composición infinita que acerca arte y diseño en la vida cotidiana de la sociedad. La diversidad generacional e ideológica de estos arquitectos convirtió el proyecto en un tablero de juego sobre el que se aplicaron diferentes formas de proyectar la ciudad, ubicando a Nagele en un punto de inflexión del Movimiento Moderno. ABSTRACT The research focuses on the Nagele project, a Dutch urban settlement located in the Noordoostpolder, a territory which was entirely reclaimed from the IJsselmeer lake. It was designed by a group of architects from the De 8 and Opbouw teams, the leading protagonists being Rietveld, Van Eesteren, Van Eyck, Bakema, Stam y Ruys. It was designed from 1947 to 1956, a fruitful period in urban planning. These architects questioned the traditionalist urban design applied to the new populations in the IJsselmeer polders. Facing their principles, the work group proposed a new prototype; a compact and concentric urban pattern to foster equality in a new community of farm labourers, which was recognized by the architects as a twentieth century urban society. The government supported their new proposals. The lack of implementation of the innovatory conceptual statements subjected the project into a high-risk challenge. However, in spite of these difficulties, the basic concepts remain though partially transformed, in the actual city. The project drawings reflect the principle concepts that this work aims to discover. Some approaches that have not been sufficiently studied are tackled in this thesis. Firstly, the project´s relationship with the polder. Secondly, the evolution of projective strategies during the period of urban planning, the landscape design and the design of urban elements. The Noordoostpolder is the culmination of a series of multi-disciplinary experiences in large scale land reclamation, whose polycentric urban structure and agricultural subdivision provide the framework of Nagele. Linking the vegetation to infrastructure fostered orientation, climate protection and human scale; strategies which were repeated, though on a smaller scale, in the actual city. The first phase of the project was composed of mono-functional urban areas which responded to each of the four basic human needs indicated by the scientific method of the functional city. The presentation of the project at the seventh CIAM in 1949 was the event which marked the end of the first phase of the planning. This congress was entitled Implementation of the Athens Charter. The residential program was divided into housing clusters surrounding a central prairie, a pattern which was related to its urban and territorial whole. The second phase of the plan was subjected to a new theoretical approach. The urban planning became an In-between Realm, a theoretical scenario where traditionally antagonistic concepts coexist. Van Eyck named these concepts Twin Phenomena. The city thus conceived of as a counterform of the ambivalence of the human mind where spatial indefinition in the previous proposals was transformed into a Bunch of Places with defined boundaries and dimensions, all of which reflecting human scale. The landscape design was integrated into the urban project from its inception. The limits consist of a green wind-barrier which not only provides climate protection but also provides human scale and visual control towards the unlimited plains of the polder. Van Eyck summarised the project as a green room without a roof. This statement elucidates its equivalence to a domestic interior. Outwardly, the city becomes an autonomous unit on the territory. Inwardly a hierarchical vegetation system is linked to architecture and infrastructure. Together, they configure different scales of urban spaces. The proposal was transformed by Boer and Ruys into a unique urban space without assimilating Van Eyck´s concepts. The study of the Nagele landscape project of Nagele and the writings of Van Eyck verify the fact that many of his theoretical foundations (In-between Realm, Twin Phenomena, Bunch of Places, Right Scale) can be applied not only to architecture and city but also to landscape design. The application of these principles led the Nagele project to become a counterform of Van Eyck´s thinking. The design and construction of the first urban elements involved a distribution of tasks to De 8 en Opbouw, which stimulated their relationship with the urban project. The polycentric structure organised the school classrooms outlined by Van Eyck, the different areas of the cemetery planned by Ruys and the housing clusters designed by Stam, Rietveld and Stam-Besse. The Twin Phenomena concept can be applied in Van der Broek´s shopping corridor. The concept space within another space is also implemented in the cemetery surrounded by a new green barrier, and in the church built by Van der Broek and Bakema, whose spaces are configured by a folding wall. The project takes a holistic approach, which considers the design of each element within the strategies of the whole, where they become parts of an infinite composition, as in the art works of De Stijl fostering art and design to ordinary people´s daily lives. The generational and ideological diversity of these architects turned the project into a game board on which different ways of planning the city were played, obtaining Nagele the distinction of being a turning point of Modernism.

A dimeric crystal structure for the N-terminal two domains of intercellular adhesion molecule-1

The 3.0-Å structure of a 190-residue fragment of intercellular adhesion molecule-1 (ICAM-1, CD54) reveals two tandem Ig-superfamily (IgSF) domains. Each of two independent molecules dimerizes identically with a symmetry-related molecule over a hydrophobic interface on the BED sheet of domain 1, in agreement with dimerization of ICAM-1 on the cell surface. The residues that bind to the integrin LFA-1 are well oriented for bivalent binding in the dimer, with the critical Glu-34 residues pointing away from each other on the periphery. Residues that bind to rhinovirus are in the flexible BC and FG loops at the tip of domain 1, and these and the upper half of domain 1 are well exposed in the dimer for docking to virus. By contrast, a residue important for binding to Plasmodium falciparum-infected erythrocytes is in the dimer interface. The presence of A′ strands in both domains 1 and 2, conserved hydrogen bonds at domain junctions, and elaborate hydrogen bond networks around the key integrin binding residues in domain 1 make these domains suited to resist tensile forces during adhesive interactions. A subdivision of the intermediate (I) set of IgSF domains is proposed in which domain 1 of ICAM-1 and previously described I set domains belong to the I1 set and domain 2 of ICAM-1, ICAM-2, and vascular cell adhesion molecule-1 belong to the I2 set.

The Vibrio cholerae genome contains two unique circular chromosomes

Vibrio cholerae, the etiologic agent of the diarrheal disease cholera, is a Gram-negative bacterium that belongs to the γ subdivision of the family Proteobacteriaceae. The physical map of the genome has been reported, and the genome has been described as a single 3.2-Mb chromosome [Majumder, R., et al. (1996) J. Bacteriol. 178, 1105–1112]. By using pulsed-field gel electrophoresis of genomic DNA immobilized in agarose plugs and digested with the restriction enzymes I-CeuI, SfiI, and NotI, we have also constructed the physical map of V. cholerae. Our analysis estimates the size of the genome at 4.0 Mb, 25% larger than the physical map reported by others. Our most notable finding is, however, that the V. cholerae chromosome appears to be not the single chromosome reported but two unique and separate circular megareplicons.

A conserved mode of head segmentation in arthropods revealed by the expression pattern of Hox genes in a spider

Chelicerates constitute a basic arthropod group with fossil representatives from as early as the Cambrian period. Embryonic development and the subdivision of the segmented body region into a prosoma and an opisthosoma are very similar in all extant chelicerates. The mode of head segmentation, however, has long been controversial. Although all other arthropod groups show a subdivision of the head region into six segments, the chelicerates are thought to have the first antennal segment missing. To examine this problem on a molecular level, we have compared the expression pattern of Hox genes in the spider Cupiennius salei with the pattern known from insects. Surprisingly, we find that the anterior expression borders of the Hox genes are in the same register and the same relative segmental position as in Drosophila. This contradicts the view that the homologue of the first antennal segment is absent in the spider. Instead, our data suggest that the cheliceral segment is homologous to the first antennal segment and the pedipalpal segment is homologous to the second antennal (or intercalary) segment in arthropods. Our finding implies that chelicerates, myriapods, crustaceans, and insects share a single mode of head segmentation, reinforcing the argument for a monophyletic origin of the arthropods.

Complete genomic sequence of Pasteurella multocida,Pm70

We present here the complete genome sequence of a common avian clone of Pasteurella multocida, Pm70. The genome of Pm70 is a single circular chromosome 2,257,487 base pairs in length and contains 2,014 predicted coding regions, 6 ribosomal RNA operons, and 57 tRNAs. Genome-scale evolutionary analyses based on pairwise comparisons of 1,197 orthologous sequences between P. multocida, Haemophilus influenzae, and Escherichia coli suggest that P. multocida and H. influenzae diverged ≈270 million years ago and the γ subdivision of the proteobacteria radiated about 680 million years ago. Two previously undescribed open reading frames, accounting for ≈1% of the genome, encode large proteins with homology to the virulence-associated filamentous hemagglutinin of Bordetella pertussis. Consistent with the critical role of iron in the survival of many microbial pathogens, in silico and whole-genome microarray analyses identified more than 50 Pm70 genes with a potential role in iron acquisition and metabolism. Overall, the complete genomic sequence and preliminary functional analyses provide a foundation for future research into the mechanisms of pathogenesis and host specificity of this important multispecies pathogen.

A cell cycle-regulated bacterial DNA methyltransferase is essential for viability.

The CcrM adenine DNA methyltransferase, which specifically modifies GANTC sequences, is necessary for viability in Caulobacter crescentus. To our knowledge, this is the first example of an essential prokaryotic DNA methyltransferase that is not part of a DNA restriction/modification system. Homologs of CcrM are widespread in the alpha subdivision of the Proteobacteria, suggesting that methylation at GANTC sites may have important functions in other members of this diverse group as well. Temporal control of DNA methylation state has an important role in Caulobacter development, and we show that this organism utilizes an unusual mechanism for control of remethylation of newly replicated DNA. CcrM is synthesized de novo late in the cell cycle, coincident with full methylation of the chromosome, and is then subjected to proteolysis prior to cell division.

Polymorphic simple sequence repeat regions in chloroplast genomes: applications to the population genetics of pines.

Simple sequence repeats (SSRs), consisting of tandemly repeated multiple copies of mono-, di-, tri-, or tetranucleotide motifs, are ubiquitous in eukaryotic genomes and are frequently used as genetic markers, taking advantage of their length polymorphism. We have examined the polymorphism of such sequences in the chloroplast genomes of plants, by using a PCR-based assay. GenBank searches identified the presence of several (dA)n.(dT)n mononucleotide stretches in chloroplast genomes. A chloroplast (cp) SSR was identified in three pine species (Pinus contorta, Pinus sylvestris, and Pinus thunbergii) 312 bp upstream of the psbA gene. DNA amplification of this repeated region from 11 pine species identified nine length variants. The polymorphic amplified fragments were isolated and the DNA sequence was determined, confirming that the length polymorphism was caused by variation in the length of the repeated region. In the pines, the chloroplast genome is transmitted through pollen and this PCR assay may be used to monitor gene flow in this genus. Analysis of 305 individuals from seven populations of Pinus leucodermis Ant. revealed the presence of four variants with intrapopulational diversities ranging from 0.000 to 0.629 and an average of 0.320. Restriction fragment length polymorphism analysis of cpDNA on the same populations previously failed to detect any variation. Population subdivision based on cpSSR was higher (Gst = 0.22, where Gst is coefficient of gene differentiation) than that revealed in a previous isozyme study (Gst = 0.05). We anticipate that SSR loci within the chloroplast genome should provide a highly informative assay for the analysis of the genetic structure of plant populations.