DOCSLIB.ORG

2273 Citometry Maquetación 1

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
2273 Citometry Maquetación 1 Anales del Jardín Botánico de Madrid Vol. 68(2): 139-154 julio-diciembre 2011 ISSN: 0211-1322 doi: 10.3989/ajbm.2273 Isolation of plant nuclei suitable for flow cytometry from species with extremely mucilaginous compounds: an example in the genus Viola L. (Violaceae) by Eduardo Cires 1*, Candela Cuesta 2, 3*, María Ángeles Fernández Casado 1, Herminio S. Nava 1, Víctor M. Vázquez 4 & José Antonio Fernández Prieto 1 1 Área de Botánica, Universidad de Oviedo, Departamento de Biología de Organismos y Sistemas, Catedrático Rodrigo Uría s/n, E-33071 Oviedo, Spain. Corresponding author: Phone: +34 985 104835; fax: +34 985 104777; e-mail: [email protected] 2 Department of Plant Systems Biology, VIB, Technologiepark 927, 9052 Gent, Belgium. [email protected] 3 Área de Fisiología Vegetal, Universidad de Oviedo, Departamento de Biología de Organismos y Sistemas, Catedrático Rodrigo Uría s/n, E-33071 Oviedo, Spain 4 Real Instituto de Estudios Asturianos, Plaza de Porlier 9, E-33003 Oviedo, Spain * Both authors contributed equally to this work Abstract Resumen Cires, E., Cuesta, C., Fernández, M.A., Nava, H.S., Vázquez, Cires, E., Cuesta, C., Fernández, M.A., Nava, H.S., Vázquez, V.M. & Fernández, J.A. 2011. Isolation of plant nuclei suitable V.M. & Fernández, J.A. 2011. Aislamiento de núcleos para cito- for flow cytometry from species with extremely mucilaginous metría de flujo en plantas con alto contenido de compuestos compounds: an example in the genus Viola L. (Violaceae). Ana - mucilaginosos: un ejemplo en el género Viola L. (Violaceae). les Jard. Bot. Madrid 68(2): 139-154. Ana les Jard. Bot. Madrid 68(2): 139-154 (en inglés). Flow cytometry analysis has been widely applied in the determi- El análisis mediante citometría de flujo ha sido aplicado de modo nation of nuclear DNA content and ploidy level in many organ- general para determinar el contenido de ADN nuclear y el nivel isms. Despite being the most appropriate method for DNA con- de ploidía en muchos organismos. A pesar de ser el método más tent measurement, flow cytometry also presents some limita- adecuado para medir la cantidad de ADN, esta técnica también tions. A fairly common, but little-studied problem is the effect presenta algunas limitaciones. Un problema bastante común, on measurements of the presence of secondary metabolites. A aunque poco estudiado, es el efecto de los metabolitos secun- good example is the genus Viola, which is composed of 525-600 darios en los resultados obtenidos. Un ejemplo respecto a la pre- species distributed worldwide. These species have proved to be sencia de estos compuestos se encuentra en el género Viola, problematic for flow cytometric analyses due to the release of compuesto por 525-600 especies distribuidas por todo el mun- extremely mucilaginous compounds into the nuclear suspen- do. Las especies de este género ya han sido previamente descri- sion. In this work, the genome size of 13 species of Viola using tas como problemáticas en los análisis de citometría de flujo de- flow cytometry are presented for the first time. Despite obtain- bido a la presencia de compuestos extremadamente mucilagi- ing histograms with high coefficients of variation, we here pre- nosos en las suspensiones de núcleos. En el presente trabajo se sent an optimized protocol to remove cytoplasmic compounds, analiza por primera vez el tamaño genómico de 13 especies del particularly mucilaginous ones, from plant nuclei that pave the genero Viola mediante el empleo de citometría de flujo. A pesar way for its application to estimate the genome size of other de los altos valores mostrados en los coeficientes de variación de species exhibiting similar problems. Statistical analyses revealed los histogramas, se presenta un protocolo optimizado para eli- significant differences between sections Viola and Melanium, minar compuestos citoplasmáticos, y más concretamente muci- and within each section (P < 0.001). Furthermore, statistically laginosos de las suspensiones nucleares, siendo de aplicación en significant differences were not detected among samples of the la estimación del tamaño genómico de plantas con problemas same species. similares. Los análisis estadísticos mostraron diferencias signifi- cativas entre las secciones Viola y Melanium, así como dentro de cada sección (P < 0,001). Además, no se encontraron diferencias significativas entre aquellas muestras pertenecientes a la misma especie. 140 E. Cires & al. Keywords: Chromosome number, flow cytometry, genome Palabras clave: Número cromosomático, citometría de flujo, size, mucilaginous compounds, nuclear DNA content, ploidy tamaño genómico, compuestos mucilaginosos, contenido de levels, section Melanium, section Viola. Abbreviations: CV: ADN nuclear, niveles de ploidía, sección Melanium, sección coefficient of variation; DI: DNA index; FCM: flow cytometry; Viola. PI: propidium iodide. Introduction compounds. On the other hand, Loureiro & al. (2007b), in the study of the genera Ulmus L. and Flow cytometry (FCM) is a fast, sensitive technique Celtis L., presented a protocol which ensured high for measuring specific components in large numbers quality analyses with a low coefficient of variation and of cells and organelles. Plant scientists have been at- minimal background debris as well as nuclear fluores- tracted by the numerous advantages of this technique cence stability. However, even Loureiro & al. (2007b) (e.g. ease of sample preparation, speed of analysis, found similar problems in leaves of Ulmus species, and no requirement for dividing cells) and the num- where they got poor results and had to use an alterna- ber of articles related to this technique has been con- tive tissue (samaras) characterized by lower concen- tinuously increasing over the years (Doležel & al., tration of mucilaginous compounds. 2007a). Estimation of DNA content in cell nuclei is The genus Viola L. includes examples of several one of the most important applications of FCM in species with extremely mucilaginous compounds. This plant sciences. In most plants, analyses of relative genus, the largest of the Violaceae family, comprises DNA content of nuclei isolated from young tissues 525-600 species distributed throughout most frost- yield histograms with good resolution. In principle, free regions of the world (Ballard & al., 1999; Yock- every tissue containing vital nuclei should be suitable teng & al., 2003; Karlsson & al., 2008). It is divided for measurement of nuclear DNA content with FCM. into ca. 14 sections and many infrasectional groups However, the presence or absence of endogenous flu- (Becker 1910a, 1910b, 1925) (for a review of infra- orescence inhibitor substances and coatings of debris generic classification see Ballard & al., 1999). Five of influences the quality of the results (Greilhuber & al., those sections are present in Europe: Chamae me - 2007). Plant cells produce a vast array of secondary lanium Ging., Melanium Ging., Xylinosium W. Bec - metabolites which may interfere with staining of par- ker, Delphiniopsis W. Becker and Viola (Valentine & ticular cell constituents and/or exhibit autofluores- al., 1968); all of which have also been described in the cence, thus hampering quantification of signals from Iberian Peninsula (Fernández Casado, 1982; Muñoz fluorescent probes (Doležel & al., 2007b). Although Garmendia & al., 1993). the interference of secondary metabolites with stain- In the present work, the samples selected belong to ing procedures had been recognized for some time in 2 of these sections: Melanium, which includes the so- cytophotometry (e.g. Greilhuber, 1986), it was not un- called pansies and is a morphologically well-defined til Noirot & al. (2000, 2002, 2003, 2005) and Price & group of about 80-100 species; and Viola, which is a al. (2000) published their findings that this effect was Eurasian group of about 25 species with several sub- taken seriously in plant FCM. Moreover, some species sections described (Valentine & al., 1968; Karlsson contained mucilaginous compounds which interfered & al., 2008), of which the following were studied: with analysis, affecting overall histogram quality and Viola, Rostratae Kupffer and Stolonosae Kupffer nuclear fluorescence stability (e.g. Betulaceae, Vio- [=Plagiostigma Godr.]. Taxa of these sections are well laceae, Lythraceae, Malvaceae, Orchidaceae and sub- known for their taxonomic complexity, apparently merged plants) (Suda, 2004; Loureiro, 2007; Greilhu- caused by several factors: (a) scarcity of reliable diag- ber, 2008). Besides hampering the release of nuclei, nostic morphological characters; (b) high phenotypic these compounds confer a high viscosity to the nu- plasticity; (c) frequent interspecific hybridizations; clear suspension, which makes the filtration step diffi- and (d) assumed past reticulate evolution within the cult to accomplish and consequently results in the re- section and subsection (Ballard & al., 1999; Yockteng covery of a low volume of nuclear homogenate. Some & al., 2003; Hodálová & al., 2008; Mered’a & al., authors have found great difficulty in using FCM due 2008; Van den Hof & al., 2008). to these compounds (e.g. Morgan & Westoby, 2005; A high diversity of chromosome numbers has been Talent & Dickinson, 2005; Loureiro & al., 2007a). To proposed within genus Viola (Moore, 1982; Castro- give an example, Morgan and Westoby (2005) found viejo & Valdés-Bermejo, 1991) by showing different for many species of Myrtaceae and Rutaceae that the ploidy levels (Miyaji, 1930; Karlsson & al., 2008). The ineffective
Load more

Recommended publications
  • VÍAS ROMANAS DE MONTAÑA ENTRE ASTURIAS Y LEÓN. LA INTEGRACIÓN DE LA ASTURIA TRANSMONTANA EN LA RED VIARIA DE HISPANIA Roman Mountain Roads Between Asturias and León
    ISSN: 0514-7336 VÍAS ROMANAS DE MONTAÑA ENTRE ASTURIAS Y LEÓN. LA INTEGRACIÓN DE LA ASTURIA TRANSMONTANA EN LA RED VIARIA DE HISPANIA Roman mountain roads between Asturias and León. The integration of Asturia transmontana in the road network of Hispania David GONZÁLEZ ÁLVAREZ Departamento de Prehistoria. UCM. Correo-e: [email protected] Recepción: 2010-08-27; Revisión: 2010-10-15; Aceptación: 2011-05-31 BIBLID [0514-7336 (2011) LXVII, enero-junio; 171-192] RESUMEN: Se revisa la información disponible sobre los itinerarios de época romana a través de la Cordillera Cantábrica entre las actuales provincias de Asturias y León. El objetivo es cuestionar estos datos a la luz de recientes aportaciones sobre las características técnicas de las vías romanas, apostando así por introducir en las narrativas arqueológicas la mayor concreción que aporta el uso adecuado de los términos técnicos e ingenieriles relacionados con el viario romano en el caso concreto que nos ocupa. De este modo, podremos evaluar en mejor medida el grado de integración de la Asturia transmontana en la realidad administrativa y sociopolítica del Imperio romano. Palabras clave: Vías romanas. Calzada romana. Romanización. Cordillera Cantábrica. ABSTRACT: The available information of the Roman ways across the Cantabrian Mountains is checked, between the present-day provinces of Asturias and León. The aim is to question this information in the light of recent contributions of the technical characteristics of the Roman ways. We bet for introducing in the archaeological narratives a bigger concretion that provides the suitable use of the technical terms related to the Roman system of communications in our case of study.
    [Show full text]
  • Come Home To
    Come home to Guide to discover Asturias asturiastourism.co.uk 2 Vuelve al Paraíso Come home to paradise EDITING: SOCIEDAD PÚBLICA DE GESTIÓN Y PROMOCIÓN TURÍSTICA Y CULTURAL DEL PRINCIPADO DE ASTURIAS, SAU Design: Arrontes y Barrera Estudio de Publicidad Layout: Paco Currás Diseñadores Maps: Da Vinci Estudio Gráfi co Texts: Susana D. Machargo Translation: Morote Traducciones Photography: Front cover: Amar Hernández. Inside pages: Amar Hernández, Armando Álvarez, Benedicto Santos, Casona del Viajante, Comarca de la Sidra, Hotel 3 Cabos, José Ángel Diego García, José Mª Díaz-Formentí, José Ramón Navarro Tudela, Juan de Tury, Juanjo Arrojo, Manuel S. Calvo, Marcos Vega García, María Martín Berciano, Miki López, Museo Bellas Artes, Noé Baranda, Paco Currás SL, Pelayo Lacazette, Pozo Sotón, Incuna, Xurde Margaride and own Files. Printing: Dayton SA - Dec. 19 D.L.: AS 04181-2017 © CONSEJERÍA DE CULTURA, POLÍTICA LLINGÜÍSTICA Y TURISMO DEL PRINCIPADO DE ASTURIAS asturiastourism.co.uk Vuelve al Paraíso 1 The first settlers captured their dreams, fears and emotions on the walls of caves where they took refuge from danger. From these cave paintings, which today enjoy international recognition and protection, to the avant-garde architecture of the Centro Niemeyer in Avilés, centuries of history have passed which have helped to form the landscape of the Principality and the idiosyncrasies of the Asturian people. The Castro culture, the warrior Pelayo and the Kingdom of Asturias, the figures of the Enlightenment of the 18th century, the industrial revolution in a land that provided water and carbon, these are all elements you can still perceive and enjoy. By land, sea or air.
    [Show full text]
  • Las Caldas De Oviedo: De Hospital De Caridad a Balneario De Lujo (1773-1860)
    DEPARTAMENTO DE CIENCIAS BIOMÉDICAS Y DEL DIAGNÓSTICO TESIS DOCTORAL LAS CALDAS DE OVIEDO: DE HOSPITAL DE CARIDAD A BALNEARIO DE LUJO (1773-1860) RAQUEL PÉREZ-BUSTAMANTE ILANDER DIRECTOR: D. Juan Antonio Rodríguez Sánchez SALAMANCA, 2015 2 D. Juan Antonio Rodríguez Sánchez, Profesor Titular del Área de Historia de la Ciencia, Departamento de Ciencias Biomédicas y del Diagnóstico de la Facultad de Medicina de la Universidad de Salamanca CERTIFICA: Que el trabajo titulado “Las Caldas de Oviedo: de hospital de caridad a balneario de lujo (1773-1860)” realizado por Dª Raquel Pérez-Bustamante Ilander, cumple todos los requisitos necesarios para su presentación y defensa para optar al Grado de Doctor por la Universidad de Salamanca. Y para que así conste en cumplimiento de la normativa vigente, firmo el presente certificado en Salamanca a tres de diciembre de dos mil quince. Fdo.: Juan Antonio Rodríguez Sánchez 3 4 Sepan que olvidar lo malo también es tener memoria M. Hernández 5 6 AGRADECIMIENTOS Al profesor D. Juan Antonio Rodríguez Sánchez por la oportunidad brindada y su generosa ayuda. A la profesora doña Josefina San Martín Bacaicoa por su estímulo y aliento para iniciar este trabajo. A la multitud de personas que con su trabajo y ayuda han hecho posible que este proyecto sea una realidad, especialmente al personal de las siguientes instituciones: Archivo del Principado de Asturias, Archivo Histórico de Asturias, Archivo Municipal de Oviedo, Biblioteca del Instituto de Estudios Asturianos, Biblioteca de la Real Academia de Medicina de Madrid, Biblioteca de la Facultad de Medicina de la Universidad Complutense de Madrid, Biblioteca Nacional, Cátedra de Hidrología Médica de la Universidad Complutense de Madrid, Departamento de Ciencias Biomédicas y del Diagnóstico de la Facultad de Medicina de la Universidad de Salamanca.
    [Show full text]
  • Annotated and Updated Checklist of Land and Freshwater Molluscs from Asturias (Northern Spain) with Emphasis on Parasite Transmitters and Exotic Species
    diversity Review Annotated and Updated Checklist of Land and Freshwater Molluscs from Asturias (Northern Spain) with Emphasis on Parasite Transmitters and Exotic Species Omar Sánchez 1 , Jairo Robla 2 and Andrés Arias 1,* 1 Department of Organisms and Systems Biology (Zoology), University of Oviedo, 33071 Oviedo, Spain; [email protected] 2 Department of Conservation Biology, Doñana Biological Station—CSIC. C/Américo Vespucio, 26, 41092 Sevilla, Spain; [email protected] * Correspondence: [email protected] Abstract: Land and freshwater molluscs are the most abundant non-arthropod invertebrates from inland habitats worldwide, playing important ecological roles and some being important pests in agriculture. However, despite their ecological, and even economic and sanitary importance, their local diversity in many European regions is not perfectly understood, with a particularly notableknowledge gap in the northern Iberian malacofauna. This work aims at providing a revised checklist of continental gastropods and bivalves from the Asturias (northern Spain), based on the examination of newly collected and deposited material and on the critical analysis of published and gray literature. A total of 165 molluscan species are recognized. Ten species constitute new records from Asturias and seven from northern Iberian Peninsula. Seventeen species are introduced or Citation: Sánchez, O.; Robla, J.; invasive, evidencing the current increase of the bioinvasion rate in continental molluscs. Furthermore, Arias, A. Annotated and Updated all these exotic species are parasite transmitters or trematode intermediate hosts, and thus represent Checklist of Land and Freshwater a potential bio-sanitary risk for human and other animal health. The provided data strongly suggest Molluscs from Asturias (Northern that the increase of invasive freshwater snail species can lead to an increase in parasitic infections, Spain) with Emphasis on Parasite and this is a crucial point that transcends the merely scientific to the political-social sphere.
    [Show full text]
  • Science for Women in the Spanish Enlightenment 1753-1808
    SCIENCE FOR WOMEN IN THE SPANISH ENLIGHTENMENT 1753-1808 ELENA SERRANO SUPERVISOR AGUSTÍ NIETO-GALAN THIS DISSERTATION IS SUBMITTED FOR THE DEGREE OF DOCTOR Programa de Doctorat en Història de la Ciència Centre d'Història de la Ciència (CEHIC) Universitat Autònoma de Barcelona BARCELONA, May, 2012 A mis padres y a mis hijas que, inexplicablemente, siempre confiaron en mí. 1 SCIENCE FOR WOMEN IN THE SPANISH ENLIGHTENMENT 1753-1808 Elena Serrano CEHIC-UAB This dissertation explores women’s involvement in shaping the Spanish eighteenth-century scientific culture. In particular, it analyses the role of women in knowledge circulation both as actors and as audiences and it puts special emphasis on highlighting the complex relations between science, gender, economy and politics. It is structured around four social sites: the city, the country-house, the nursery-room and the library. In the city, I focus on the activities of the Junta de Damas, the women’s branch of the Madrid Economic Society (Real Sociedad Económica Matritense de Amigos del País) during the 1790s. Backed by the crown, these powerful ladies were involved in chemistry research activities, such as colouring and bleaching textiles, trials on baby-feeding food and air-purification of jail cells. Due to their dual social condition of being women and aristocrats, they were able to create an active network that connected scientific and charitable societies, politicians, doctors, academics, men of letters, and craftsmen. In the country- house, I analyse how women shaped the practices, values and public image of economic agriculture, a broadly defined science which encompassed agricultural knowledge, botany, chemistry, healing practices, domestic economy, artisan skills, and rural architecture.
    [Show full text]
  • Come Home To
    Come home to The Way of St. James routes through Asturias NIO M O UN IM D R T IA A L • P • W L O A I R D L D N H O E M R I E TA IN G O E • PATRIM United Nations Routes of Santiago de Compostela: Educational, Scientific and Camino Francés and Routes of Northern Spain Cultural Organization inscribed on the World Heritage List in 1993 asturiastourism.co.uk 2 #AsturiasUrbana Asturias is on your Way #StJamesWay #Pilgrims EDITING: SOCIEDAD PÚBLICA DE GESTIÓN Y PROMOCIÓN TURÍSTICA Y CULTURAL DEL PRINCIPADO DE ASTURIAS, SAU Design: Arrontes y Barrera Estudio de Publicidad Layout: Paco Currás Diseñadores Maps: Da Vinci Estudio Gráfico Texts: Miguel Barrero Translation: Morote Traduciciones, SL Photography: Front cover: Amar Hernández. Inside pages: Amar Hernández, Own file, Armando Álvarez, Arnaud Späni, Juanjo Arrojo, Juan de Tury, Noé Baranda, Paco Currás Diseñadores and Pelayo Lacazette. Printing: Gráficas Eujoa, SA D.L.: AS 03364-2018 © CONSEJERÍA DE EMPLEO, INDUSTRIA Y TURISMO DEL PRINCIPADO DE ASTURIAS asturiastourism.co.uk #AsturiasUrbana In July 2015, the socalled Northern St. James In this diversity of methods and traditions, Ways received World Heritage classification Asturias plays a key role. Its natural and by Unesco. In receipt of this status are the artistic attractions alone would be reason historic routes that, over the centuries, enough to justify the pilgrims choosing have led millions of pilgrims from their it when planning their journeys to the native lands to Compostela through the apostolic tomb. However, one does not northernmost part of the Iberian Peninsula.
    [Show full text]
  • Bryophytes Collected in Spain During the Tenth I.P.E. in 1953
    Bryophytes collected in Spain during the tenth I.P.E. in 1953 Autor(en): Allorge, V. / Richards, P.W. Objekttyp: Article Zeitschrift: Veröffentlichungen des Geobotanischen Institutes Rübel in Zürich Band (Jahr): 31 (1956) PDF erstellt am: 10.10.2021 Persistenter Link: http://doi.org/10.5169/seals-307971 Nutzungsbedingungen Die ETH-Bibliothek ist Anbieterin der digitalisierten Zeitschriften. Sie besitzt keine Urheberrechte an den Inhalten der Zeitschriften. Die Rechte liegen in der Regel bei den Herausgebern. Die auf der Plattform e-periodica veröffentlichten Dokumente stehen für nicht-kommerzielle Zwecke in Lehre und Forschung sowie für die private Nutzung frei zur Verfügung. Einzelne Dateien oder Ausdrucke aus diesem Angebot können zusammen mit diesen Nutzungsbedingungen und den korrekten Herkunftsbezeichnungen weitergegeben werden. Das Veröffentlichen von Bildern in Print- und Online-Publikationen ist nur mit vorheriger Genehmigung der Rechteinhaber erlaubt. Die systematische Speicherung von Teilen des elektronischen Angebots auf anderen Servern bedarf ebenfalls des schriftlichen Einverständnisses der Rechteinhaber. Haftungsausschluss Alle Angaben erfolgen ohne Gewähr für Vollständigkeit oder Richtigkeit. Es wird keine Haftung übernommen für Schäden durch die Verwendung von Informationen aus diesem Online-Angebot oder durch das Fehlen von Informationen. Dies gilt auch für Inhalte Dritter, die über dieses Angebot zugänglich sind. Ein Dienst der ETH-Bibliothek ETH Zürich, Rämistrasse 101, 8092 Zürich, Schweiz, www.library.ethz.ch http://www.e-periodica.ch — 250 — Bryophytes collected in Spain during the Tenth I. P. E. in 1953 By V. Allorge, Paris, and P. W. Richards, Bangor (North Wales) To those fortunate enough to take part, the excellently planned tour which the Spanish Committee arranged for the Tenth I.
    [Show full text]
  • Download Download
    2273 citometry_Maquetación 1 28/11/2011 11:32 Página 139 Anales del Jardín Botánico de Madrid Vol. 68(2): 139-154 julio-diciembre 2011 ISSN: 0211-1322 doi: 10.3989/ajbm.2273 Isolation of plant nuclei suitable for flow cytometry from species with extremely mucilaginous compounds: an example in the genus Viola L. (Violaceae) by Eduardo Cires 1*, Candela Cuesta 2, 3*, María Ángeles Fernández Casado 1, Herminio S. Nava 1, Víctor M. Vázquez 4 & José Antonio Fernández Prieto 1 1 Área de Botánica, Universidad de Oviedo, Departamento de Biología de Organismos y Sistemas, Catedrático Rodrigo Uría s/n, E-33071 Oviedo, Spain. Corresponding author: Phone: +34 985 104835; fax: +34 985 104777; e-mail: [email protected] 2 Department of Plant Systems Biology, VIB, Technologiepark 927, 9052 Gent, Belgium. [email protected] 3 Área de Fisiología Vegetal, Universidad de Oviedo, Departamento de Biología de Organismos y Sistemas, Catedrático Rodrigo Uría s/n, E-33071 Oviedo, Spain 4 Real Instituto de Estudios Asturianos, Plaza de Porlier 9, E-33003 Oviedo, Spain * Both authors contributed equally to this work Abstract Resumen Cires, E., Cuesta, C., Fernández, M.A., Nava, H.S., Vázquez, Cires, E., Cuesta, C., Fernández, M.A., Nava, H.S., Vázquez, V.M. & Fernández, J.A. 2011. Isolation of plant nuclei suitable V.M. & Fernández, J.A. 2011. Aislamiento de núcleos para cito- for flow cytometry from species with extremely mucilaginous metría de flujo en plantas con alto contenido de compuestos compounds: an example in the genus Viola L. (Violaceae). Ana- mucilaginosos: un ejemplo en el género Viola L.
    [Show full text]
  • Reservas De La Biosfera De Castilla Y León Biosphere Reserves of Castilla Y León Castilla Y León
    Castilla y León Reservas de la Biosfera de Castilla y león Biosphere Reserves of Castilla y León Castilla y León 66 44 33 11 55 22 77 LEÓN BURGOS PALENCIA 10 ZAMORA SORIA VALLADOLID 10 SEGOVIA SALAMANCA Alto Bernesga 9 1 2 Los Ancares Leoneses ÁVILA 8 3 Babia 4 Valle de Laciana Montaña Cantábrica. 5 Los Argüellos Cantabrian Mountains. 6 Picos de Europa 7 Valles de Omaña y Luna Ambiente Mediterráneo. Mediterranean enviroment. 8 Sierras de Béjar y Francia 9 Real Sitio de San Ildefonso Transfronterizas. - El Espinar Cross-border. 10 Meseta Ibérica La información contenida en esta publicación Edita/Published by: Fundación Siglo para el Turismo no puede abarcar la totalidad de la riqueza del y las Artes de Castilla y León. patrimonio cultural de Castilla y León, por este Diseño/Design: Proximia Adcity. motivo se recomienda al lector que quiera ampliar Depósito Legal/Legal Deposit: DL VA 609-2018 la información contenida que acceda a la página Revisión/Revision: Septiembre de 2018 / September 2018 web www.turismocastillayleon.com The information contained in this guide cannot Fotografias y textos/Photograhs and texts: encompass all the cultural wealth and heritage to Reservas de la Biosfera de Castilla y León be discovered in Castilla y León. Readers wishing to / Castilla y León Biosphere Reserves. obtain further information are therefore advised to visit the website www.turismocastillayleon.com. www.turismocastillayleon.com índice | index RESERVAS DE LA BIOSFERA | BIOSPHERE RESERVES LEÓN Alto Bernesga 4 Los Ancares Leoneses 8 Babia 12 Valle
    [Show full text]