PROYECTOS Y METODOLOGÍA

 

 


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Institution:                 University of Alcala

                                   Department of Plant Biology

                                   28871 Alcalá de Henares, Madrid

 

Contact Person:         Carmen Díaz-Sala

e-mail:carmen.diazsala@uah.es

 

Contribution: Expertise in molecular biology and physiology of forest trees, especially in studies of developmetal and morphogenic processes involved in forest productivity: maturation, adventitious rooting  and morphogenesis. Library construction, large-scale gene expression profiling technologies, gene cloning, temporal and spatial expression of genes and proteins, genetic transformation in Arabidopsis etc. Large-scale micropropagation and in vitro tissue culture of forest species. Fully equipped for molecular biology and molecular genetics analysis at laboratory level including growth chambers and a greenhouse. In addition, University Central Research Facilities and Equipment include: a sequencing (two sequencers) facility, a molecular biology unit, a plant production unit, a mass spectrometry facility, a TEM, SEM and confocal microscopy (and additional equipment),  an isotope unit, a cell culture unit, a X-Ray difraction facility and a NMR spectroscopy facility.

 

Participation in several projects and networks in the field of molecular biology of forest trees and forest genomics. C. Díaz-Sala is the Spanish representative to the Management Committee and member of the Steering Committee in Genosilva: European Forest Genomics Network (COST Action E-28) and chairperson of the COST Action E-28 Working Group2 ”Functional Genomics of Tree Maturation and Reproduction”

 

Scientists involved: Carmen Díaz-Sala, Dolores Abarca, Alicia Solé

 

Publications (three):

 

Day M, Greenwood MS, Díaz-Sala C. 2002. Age-and size-related trends in woody plant shoot development: regulatory pathways and evidence for genetic control. Tree Physiology 22:507-513.

 

Díaz-Sala C, Garrido G, Sabater B. 2002. Age-related loss of rooting capability in Arabidopsis thaliana and its reversal by peptides containing the Arg-Gly-Asp (RGD) motif. Physiologia Plantarum 114:601-607.

 

Hutchison KW, Singer PB, McInnis S, Díaz-Sala C, Greenwood MS. 1999. Expansins are conserved in conifers and expressed in hypocotyls in response to exogenous auxin. Plant Physiology 120:827-831.

 

 


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Institution:                 University of Córdoba

                                   Department of Forestry and Department of Molecular Biology

                                   34071, Córdoba

 

Contact Person:         Rafael Navarro and Jesús Jorrín

                                   e-mail: ir1nacer@uco.es, bf1jonoj@uco.es

 

 

Contribution: Expertise in large-scale protein analysis of forest species by 2D gel-electrophoresis and mass spectrometry, especially in studies of abiotic stress. Forestation and field performance of Pinus. Facilities include: greenhouse, cultivate chamber and ecophysiological equipment (CIRAS 1; Fluorimeter, pressure chamber), and other basic vegetal biology equipment. Participation in several European (INCO EU project), and National projects (CICYT), as well as international Consortiums. 

 

 

Scientists involved: Rafael Navarro, Jesús Jorrín

 

 


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Institution:                 University of Girona

                                   Department of Biology

                                   17071 Girona

 

Contact  Person:        Marisa Molinas

                                   e-mail: marissa.molinas@udg.es

 

Contribution: Expertise in molecular biology, ecophysiology and anatomy of forest trees specially cork-oak: ecophysiology, anatomy, development, cork and phellem anatomy and abiotic stress. Library contruction, gene cloning, gene and protein expression patterns, heterologous expression in yeast and E.coli models. Micro propagation and repetitive embryogenic lines and tissue culture. Expertise in in vivo functional assays in E.coli system. Search of candidate genes involved in phellem development. Fully equiped for histology and molecular biology : optical and electron microscopy, two automatic sequencers and  a Real-time PCR machine. Participation in serveral projects and networks in forestry and forest genomics.

 

Scientists involved: Mercè Figueras, Gemma Huguet , Dolors Verdaguer and Marissa Molinas

 

Publications (three):

 

Pla M, Jofré A, Martell M, Molinas M, Gómez J. 2000. Large accumulation of mRNA

and DNA point mutations in a plant senescent tissue. FEBS Letters, 472: 14-16.

 

Puigderrajols P, Jofré A,  Mir G, Pla M, Molinas M. 2002. Temporal and spatial distribution patterns of developmental and stress induced heat shock proteins in cork oak somatic embryos. J. Expt. Bot. 53 (373): 1-8 (june 2002)

 

Verdaguer D, Aranda X, Jofré A, El Omari B, Molinas M, Fleck I. 2003. Expression of low molecular weight heat-shock proteins (sHsps) and total antioxidant activity in the Mediterranean woody Quercus ilex L. in relation to seasonal and diurnal changes in physiological parameters. Plant, Cell @ Environmet  (in press)

 

 

 


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Institution:                 University of Malaga

                                   Department of Molecular Biology and Biochemistry

                                   29071 Málaga

 

Contact Person:        
(1) Francisco J. Cantón, e-mail: frcanton@uma.es
(2) M. Gonzalo Claros Díaz, e-mail: claros@uma.es
(3) Fernando Gallardo, e-mail: fgallardo@uma.es
(4) Francisco Cánovas, e-mail: canovas@uma.es
(5) Concepción Ávila Sáez, e-mail: cavila@uma.es

 

 

 

(1) Contribution: Expertise in molecular biology of forest trees, especially in studies of functional genomics of wood formation and marker development. Library construction, development of EST collections, large-scale gene expression profiling, gene cloning, temporal and spatial expression of genes and proteins, heterologous expression, genetic transformation and reverse genetics in Arabidopsis etc. Fully equipped for molecular biology and molecular genetics analysis including sequencing facilities.

Scientists involved: Francisco R. Cantón, Concepción Avila.

Publications (three):

1.-  Cantón FR, Quail PH. 1999. Both phyA and phyB mediate light-imposed repression of PHYA gene expression in Arabidopsis . Plant Physiol., 121: 1207-1215.

2.- Avila C, Cantón FR, Barnestein P, Suárez MF, Marraccini P, Rey M, Humara J, Ordás R, Cánovas FM. 2001. The promoter of a cytosolic glutamine synthetase gene from the conifer Pinus sylvestris is active in cotyledons of germinating seeds and light regulated in transgenic Arabidopsis thaliana . Physiologia Plantarum , 112: 388-396

3.- De la Torre F, García-Gutiérrez A, Crespillo R, Cantón FR, Avila C, Cánovas FM. 2002. Functional expression of two pine glutamine synthetase genes in bacteria reveals that they encode cytosolic holoenzymes with different molecular and catalytic properties. Plant Cell Physiol., 43(7): 802-809


(2)
Contribution: Expertise in molecular biology of trees (pine and olive), as well as in bioinformatics. Special attention paid on development of molecular markers based on unspecifc and specific sequences. Genomic library constructuion of pine to map nitrogen metabolism genes. Development of bioinformatics tools to analyse sequences and to store sequences in databases to extract unigene, clusters, gene families etc. Large-scale gene expression profiling, gene cloning, temporal and spatial expression of genes and proteins, heterologous expression, and genetic transformation. Fully equipped for molecular biology and molecular genetics analysis including sequencing facilities.

 

Scientists involved: M.G. Claros, F.M. Cánovas, Rocío Bautista, G. Pérez-Trabado

 

Publications (three):

1.- S. Funes, E. Davidson, M.G. Claros, R. Van Lis, X. Pérez-Martínez, M. Vázquez-Acevedo, M.P. King, D. González-Halpen (2002)  The tipically mitochondrial DNA-encoded ATP6 subunit of the mitochondrial F1F0-ATPase is encoded by a nuclear gene  in Chlamydomonas reinhardtii. J. Biol. Chem 277: 6051-6058

2.- R. Bautista, R. Crespillo, F. M. Canovas, M.G. Claros (2003) Identification of olive-tree cultivars with SCAR markers. Euphytica 129: 33-41

 

3.- R. Bautista, F. M. Canovas, M.G. Claros (2003) Genomic evidence for a repetitive nature of the RAPD polymorphisms in Olea europaea (olive-tree). Euphytica in press

 

(3) Contribution: Expertise in genetic transformation of forest trees. Biolistic technologies. Analysis and maintanance of transgenic plants. Fully equipped for molecular biology and transgenic plant analysis including extensive transgene-secure chamber facilities and a unit for production of transgenic plants. Capabilities for maintaining 50-60 independently transgenic lines.               

Scientists involved: Fernando Gallardo

Publications (three):

1.- Gallardo F, Fu J, Cantón FR, García-Gutiérrez A, Cánovas FM, Kirby EG (1999) Expression of a conifer glutamine synthetase gene in transgenic poplar. Planta 210:19-26.

2.- Fu J, Sampalo R, Gallardo F, Cánovas FM , Kirby EG (2003) Assembly of a cytosolic pine glutamine synthetase holoenzyme in leaves of transgenic poplar leads to enhaced vegetative growth in young plants. Plant Cell and Environment.  In press.

3.-Gallardo F, Fu J, Jing ZP, Kirby EG, Cánovas FM (2003) Genetic modification of amino acid metabolism in woody plants. Plant Physiology and Biochemistry Special Issue, In press.

   

(5) Concepción Ávila Sáez, e-mail: cavila@uma.es
(5) Contribution: Expertise in molecular biology of trees, especially in studies of functional genomics of nitrogen assimilation in conifers. Mainly involved in the study of transcriptional regulation of genes. Isolation of promoters and transient expression studies on pine protoplasts, yeast and heterologous systems, characterization of cis elements and isolation of transfactors.
Scientist involved: Concepción Ávila Sáez
Publications (three):
Gómez-Maldonado J, Cánovas FM and Ávila C. Molecular analysis of the 5' upstream region of a gibberellin-inducible cytosolic glutamine synthetase gene (GS1b) expressed in the pine vascular tissue.Planta(2004) 218: 1036-1045
Gómez-Maldonado J, Ávila C, Barnestein P, Crespillo R and  Cánovas.  Interaction of cis-acting elements in the expression of a gene encoding cytosolic glutamine synthetase in pine seedlings. Physiologia Plantarum (2004) 121:537-545.
Gómez-Maldonado J, Avila C, de la Torre F, Cañas R, Cánovas FM, and Campbell M. Functional interactions between a glutamine synthetase promoter and MYB proteins. Plant Journal (2004) 39:513-526.

 

 


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 Institution:                Universidad Politecnica de Madrid (1)

                                   Departamento de Biotecnologia . E.T.S.Ingenieros de Montes

                                   28040 Madrid

 

Contact Person:         (1) Isabel Allona, e-mail: iallona@montes.upm.es

(2) Carmen Collada, e-mail: collada@montes.upm.es

 

(1) Contribution: Expertise in molecular biology of forest trees, especially in studies of wood formation, cell wall development, biotic and abiotic stress, and tree dormancy. Library construction, development of EST collections, substracted cDNA libraries, PCR substration, large-scale gene expression profiling, gene cloning, temporal and spatial expression of genes and proteins, heterologous expression. Fully equipped for molecular biology and molecular genetics analysis including sequencing facilities in the Department.

Scientists involved: Isabel Allona, Cipriano Aragoncillo y Rosa Casado

Publications (three):

1.- I. Allona, M. Quinn, E. Shoop, K. Swope, S. St. Cyr, J. Carlis, J. Riedl, E. Retzel, M. Campbell, R. Sederoff y R. W. Whetten. “Analysis of xylem formation in pine by cDNA sequencing” Proceedings of the National Academy of Science USA, 95: 9693-9698 (1998)

2.- Y. Zhang, R.R. Sederoff y I. Allona. “Differential expression of genes encoding cell wall proteins in vascular tissues from vertical and bent loblolly pine trees” Tree Physiology, 20: 457-466 (2000)

3.- L. Gomez, I. Allona, R. Casado y C. Aragoncillo. “Seed chitinases” Seed Science Research, 12:217-230 (2002)

 

(2) Contribution: Expertise in genetic analysis of forest trees. Marker development including nuclear micro-satellites (SSR) as well as PCR derived markers resulting from EST and gene sequence conversion, to be used in candidate gene approaches. Marker transfer among species. Construction of saturated linkage maps using dominant (AFLPs and SAMPLs) and co-dominant markers (SSR and P-ESTs/SNPs). Candidate-based and neutral marker-based analysis. Comparative and associative mapping. Population genetics, gene flow and structural analysis of natural populations. Fully equipped for molecular biology and molecular genetics analysis, including growth chamber and a greenhouse. Facilities include: automatic sequencers for fragment analysis, PCR machines, and other basic molecular biology equipment. In addition, University Central Facilities.

Participation in several European (TREESNPS I+D EU project) as bilateral (with INRA- France and University of Davis –USA), and National projects as well as international Consortiums. 

Scientists involved: Carmen Collada

Publications (three):

1.- Gómez A, González Martínez SC, Collada C, Climent J and Gil L. 2003. Complex population genetic structure in the endemic Canary Island pine revealed usig chloroplast microsatellite markers. Theorical and Applied Genetics (in press)

 

2.- García –Casado G, Collada C, Allona I, Soto A, Casado R, Rodríguez Cerezo E, Gómez L and Aragoncillo C. 2000. Characterization of an apoplastic basis thaumatin-like protein from recalcitrant chestnut seeds. Physiologia Plantarum 110: 172-180.

3.- Sanchez-Monge R, Blanco C, Díaz-Perales A, Collada C, Carrillo T, Aragoncillo C and Salcedo G. 2000. Class I Chitinases, the panallergens responsible for the latex-fruit syndrome, are induced by ethylene treatment and inactivated by heating. J. of Allergy Clin Immunol. 106: 190-195


 

 


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Institution:                 University of Oviedo

                                   Department B.O.S.

                                   33071, Oviedo

 

Contact Person:         (1) Roberto Rodríguez

                                   e-mail: rrodri@correo.uniovi.es

 

(2) Ricardo Ordás

                                    e-mail: rordas@correo.uniovi.es.

 

(1) Contribution: Expertise in epigenetic regulation of tree development. Genomic DNA methylation by High Performance Capillary Electrophoresis, exon and promoter methylation (MSP), immunodetection of methylated bases, MSAPs (Methylation-Sensitive Amplification Polimorphisms). Large-scale in vitro tissue culture of forest trees. Fully equipped for DNA methylation analysis and tissue culture facilities.

 

Scientists involved: Roberto Rodríguez, Maria Jesús Cañal, Mario F. Fraga, Borja Diego, María Berdasco, Rodrigo Hansbun, Ricardo Hernández, Carlos Noceda.

 

Publications (three):

Fraga F. Mario, Uriol Esther, Diego L. Borja, Berdasco María, Esteller Manel, Cañal María Jesús, Rodríguez Roberto. 2002. High-performance capillary electrophoresis method for the quantification of 5-methyl 2-deoxycytidine in genomic DNA: Application to plant,animal and human cancer tissues. Electrophoresis 23, 1677-1681.

 

Fraga F Mario, Cañal María Jesús, Rodríguez Roberto. 2002. Phase-change related epigenetic and physiological changes in Pinus radiata D. Don. Planta.42, 795-784.

 

Fraga F Mario, Cañal María Jesús, Rodríguez Roberto. 2002. Genomic DNA methylation-demethylation during aging and reinvigoration of Pinus radiata. Tree Physiology 22,813-816.

    

           

(2) Contribution: Expertise in plant development regulation and genetic transformation of forest trees. Large-scale tissue culture and fully equipped for biochemical and molecular analysis. Biolistic technologies. Analysis and maintenance of transgenic plants. Fully equipped for transgenic plant analysis including chamber facilities and capabilities for maintaining independently transgenic lines. 

 

Scientists involved: Ricardo Ordás, Manuel Rey, Victoria González, Mariam López, Belén Fernández, Ana Rodríguez, Mariluz Centeno.

 

 

Publications (three):

 

HUMARA, J.M. LOPEZ,M; ORDAS,R.J. 1999. Agrobacterium tumefacions-mediated transformation of Pinus pinea L. Cotyledons: an assessment of factors influencing the effficiency of uidA gene transfer. Plant Cell Reports, 19:  51-58.

HUMARA, J.M.; MARIN, M.S., PARRA, F.; ORDAS,R.J. 1999. Improved efficiency of uida gene transfer in Pinus pinea l. cotyledons using a modified binary vector. Canadian Journal of Forest Research, 73(5):1627-1632.

LÓPEZ, M.; HUMARA,J.M., RODRÍGUEZ, R.; ORDÁS, R.J. 2000. Factors involved in Agrobacterium tumefaciens-mediated gene transfer into Pinus nigra Arn. ssp. salzmannii (Dunal) Franco. Euphytica114:195-203.

 

 

 


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Institution:                 University of Salamanca

                                   Department of Plant Physiology

                                   37007 Salamanca

 

Contact Person:         Dolores Rodriguez and Carlos Nicolas

                                   e-mail: mdr@usal.es,    cnicolas@usal.es

 

 

Contribution: Expertise in molecular biology and physiology of forest trees, especially in studies of seed dormancy and germination. Library construction, gene cloning, temporal and spatial expression of genes and proteins, heterologous expression, genetic transformation etc. Fully equipped for molecular biology and molecular genetics analysis including growth chambers, sequencing and confocal microscopy facilities.

Participation in several projects in the field of molecular biology of forest trees.

 

Scientists involved: Dolores Rodríguez, Carlos Nicolás

 

Publications (three):

 

Lorenzo O, Nicolás C, Nicolás G, Rodríguez D (2002). GA3-induced expression of a new functional AAA-ATPase (FsA1) is correlated with the onset of germination in Fagus sylvatica L. seeds (beechnuts). Plant Cell Physiol 43: 27-34

 

Lorenzo, O., Nicolás, C., Nicolás, G. and  Rodríguez, D. (2002) Molecular cloning of a functional protein phosphatase 2C (FsPP2C2) with unusual features and synergistically up-regulated by ABA and calcium in dormant seeds of Fagus sylvatica. Physiol Plant 114: 482-490.

 

Lorenzo, O., Rodríguez, D., Nicolás, G., Rodríguez, P.L. and Nicolás, C. (2001) A new protein phosphatase 2C (FsPP2C1) induced by abscisic acid is specifically expressed in dormant beechnut seeds. Plant Physiol 125: 1949-1956


 

GRUPO BIOLOGÍA DE SEMILLAS. DEPARTAMENTO DE FISIOLOGÍA VEGETAL. UNIVERSIDAD DE SALAMANCA:

 

Plantilla:

Gregorio Nicolás Rodrigo

Dolores Rodríguez Martín

Carlos Nicolás Rodríguez

Becarios:

Mary Paz González García

David Reyes Martín

 

PUBLICACIONES

 

AUTORES : C. NICOLAS, G. NICOLAS, D. RODRIGUEZ.

TÍTULO: “Transcripts of a gene, encoding a small GTP-binding protein from Fagus sylvatica, are induced by ABA and accumulated in embryo axis of dormant seeds.”

REF. REVISTA/LIBRO: Plant Mol. Biol. 36:487-491. 1998.                                 CLAVE: A

                                                                                                                                                                                                                                            

AUTORES : C. NICOLAS, J.M. dePRADA, O. LORENZO, G. NICOLAS, D. RODRIGUEZ.

TÍTULO: “Abscisic acid and stress regulate the expression of calmodulin in germinating chick-pea seeds.”

REF. REVISTA/LIBRO: Physiol. Plant. 104: 379-384. 1998.                                CLAVE: A

 

AUTORES : O. LORENZO, D. RODRIGUEZ, C. NICOLAS, G. NICOLAS.

TÍTULO: “Characterization and expression of two protein kinase and an EIN3-like genes, which are regulated by ABA and GA3 in dormant Fagus sylvatica seeds”.

REF. REVISTA/LIBRO: “Seed Biology: Advances and Applications”. Eds. M. Black, K.J. Bradford and J. Vázquez-Ramos. Pg. 329-340. 2000.

EDITORIAL: CAB International                                                                             CLAVE: CL

 

AUTORES: D. RODRIGUEZ, O. LORENZO, C. NICOLAS.

TÍTULO:“El ácido abscísico y la dormición de semillas.”.

REF. REVISTA/LIBRO: “Fisiología Hormonal”. Ed. A. Matilla. Pg, 41-51. 2000

EDITORIAL:            Servicio de Publicaciones de la Universidad de Santiago de Compostela                                                                                                                                           CLAVE: R

 

AUTORES : O. LORENZO, D. RODRIGUEZ, G. NICOLAS, PL RODRIGUEZ, C. NICOLAS.

TÍTULO: “A new protein phosphatase 2C (FsPP2C1) induced by ABA is specifically expressed in

dormant beechnut seeds.”.

REF. REVISTA/LIBRO: Plant Physiol 125: 1949-1956.  2001.                             CLAVE: A

 

AUTORES : O. LORENZO,  C. NICOLAS, G. NICOLAS, D RODRIGUEZ.

TÍTULO: “Molecular cloning of a functional protein phosphatase 2C (FsPP2C2) with unusual features and synergistically up-regulated by ABA and calcium in dormant seeds of Fagus sylvatica”.

REF. REVISTA/LIBRO: Physiol Plant 114: 482-490                      2002 .             CLAVE: A

 

AUTORES : O. LORENZO,  C. NICOLAS, G. NICOLAS, D RODRIGUEZ.

TÍTULO: “GA3-induced expression of a new functional AAA-ATPase (FsA1) is correlated with the onset of germination in Fagus sylvatica L. seeds (beechnuts).

REF. REVISTA/LIBRO: Plant and Cell Physiology 43:27-34.     2002.                  CLAVE: A

AUTORES : MP GONZÁLEZ, D RODRÍGUEZ, C NICOLÁS, PL RODRÍGUEZ, G NICOLÁS, O LORENZO

TÍTULO: Negative regulation of ABA-signaling by the Fagus sylvatica FsPP2C1 plays a role in seed dormancy regulation and promotion of seed germination

REF. REVISTA/LIBRO: Plant Physiol (en prensa)                                      CLAVE: A

 

AUTORES : AP CALVO, JA JIMÉNEZ, C NICOLÁS, G NICOLÁS, D RODRÍGUEZ

TÍTULO: Isolation and characterization of genes related with the breaking of beechnuts dormancy and putatively involved in ethylene signal perception and transduction

REF. REVISTA/LIBRO: The Biology of Seeds: Recent Research Advances. Eds: G Nicolás, KJ Bradford, D Come, HW Pritchard. (en prensa)

EDITORIAL: CAB International                                                                             CLAVE: CL

 

AUTORES : O. LORENZO,  C. NICOLAS, G. NICOLAS, D RODRIGUEZ.

TÍTULO: Characterization of a dual plant protein kinase (FsPK1) up-regulated by ABA and calcium and specifically expressed in dormant seeds of Fagus sylvatica L.

REF. REVISTA/LIBRO: Enviado a Seed Sci Res    .                                              CLAVE: A

 

AUTORES : LC MORTENSEN, D. RODRÍGUEZ, G. NICOLÁS, EN ERIKSEN, C NICOLAS

TÍTULO: Changes in a seed-specific abscisic acid-responsive glycine rich protein (GRPF1) mRNA may reflects the release of seed dormancy in Fagus sylvatica during moist prechilling

REF. REVISTA/LIBRO: Enviado a Seed Sci Res    .                                              CLAVE: A

 


 

PROYECTOS DE INVESTIGACION FINANCIADOS

 

TITULO DEL PROYECTO : Estudio de la dormición en semillas forestales: Papel regulador del ácido abscísico, ácido giberelico y etileno en la dormición de semillas de Fagus sylvatica y Pseudotsuga menziesii.

ENTIDAD FINANCIADORA DGES

IDENTIFICACION: PB96-1313

DURACION; 1997    HASTA; 2000

INVESTIGADOR PRINCIPAL: Gregorio Nicolás Rodrigo

NUMERO DE INVESTIGADORES PARTICIPANTES: 4

IMPORTE TOTAL DEL PROYECTO: 15.005.000 pts

 

TITULO DEL PROYECTO : Regulación de la dormición de semillas de haya (Fagus sylvatica) por ácido abscísico, giberelinas y etileno.

ENTIDAD FINANCIADORA: Junta de Castilla y León.

IDENTIFICACION : SA60/99

DURACION; 1999    HASTA; 2001

INVESTIGADOR PRINCIPAL: Mª Dolores Rodríguez

NUMERO DE INVESTIGADORES PARTICIPANTES: 5

IMPORTE TOTAL DEL PROYECTO: 3.095.000 pts

 

TÍTULO: Caracterización de la dormición de semillas de Fagus sylvatica. Regulación del proceso por etileno, ácido abscísico y ácido giberélico

ENTIDAD: Ministerio de Ciencia y Tecnología. (BFI2000-1361)

DURACION, DESDE: 2000            HASTA: 2003

INVESTIGADOR RESPONSABLE: Dr. Gregorio Nicolás Rodrígo

NUMERO DE INVESTIGADORES PARTICIPANTES: 3

IMPORTE TOTAL DEL PROYECTO: 21.000.000 pts

 

TÍTULO: Caracterización y expresión de proteína fosfatasas (PP1, PP2A y PP2C) relacionadas con la dormición de semillas de haya (Fagus sylvatica L.)

ENTIDAD: Junta de Castilla y León. (SA010/02)

DURACION, DESDE: 2002            HASTA: 2004

INVESTIGADOR RESPONSABLE: Dr. Carlos Nicolás Rodríguez.

NUMERO DE INVESTIGADORES PARTICIPANTES: 4

IMPORTE TOTAL DEL PROYECTO: 19.423 EUROS

 

 


TESIS DOCTORALES DIRIGIDAS

 

TITULO: “Regulación hormonal de la dormición de semillas de haya (Fagus sylvatica): Expresión de proteínas específicas relacionadas con la transducción de señales.”

DOCTORANDO: OSCAR LORENZO SANCHEZ

UNIVERSIDAD: Salamanca                        FACULTAD/ESCUELA: F. de Biología

DIRECTOR/ES: Dra. D.Rodríguez y Dr. C. Nicolás

AÑO: 2000                                                    CALIFICACION: Apto cum laude

                                                                                                                                

TITULO: “Regulación de la dormición de semillas de haya (Fagus sylvatica): aislamiento y caracterización de clones implicados en la biosíntesis de etileno y giberelinas y en las respuestas a estas hormonas.”

DOCTORANDO: ANGEL PABLO CALVO BULLON

DIRECTOR/ES: Dr. G. Nicolás y Dra. D.Rodríguez 

UNIVERSIDAD: Salamanca                        FACULTAD/ESCUELA: F. de Biología

AÑO: 2002                                                   CALIFICACION: Sobresaliente cum laude

 

TÍTULO: "Caracterización bioquímica y molecular de la 1-aminociclopropano-1-carboxílico y la endopoligalacturonasa en semillas de nabiza (Brassica rapa L. cv. Rapa).

DOCTORANDO: MARÍA DEL CARMEN RODRÍGUEZ GACIO

DIRECTOR/ES: Dra. A. Matilla y Dr. C. Nicolás

UNIVERSIDAD: Santiago de Compostela   FACULTAD/ESCUELA: F. de Farmacia

AÑO: 2002                CALIFICACION: Sobresaliente cum laude

                                                                                                                                                   

TITULO: “Regulación por ácido abscísico y giberelinas de la expresión de proteínas relacionadas con la dormición de semillas de haya (Fagus sylvatica L.)”.

DOCTORANDO: JESUS ANGEL JIMENEZ NIETO

UNIVERSIDAD: Salamanca                        FACULTAD/ESCUELA: F. de Biología

DIRECTOR/ES: Dra. D.Rodríguez y Dr. C. Nicolás

AÑO: Fecha lectura aproximada, 2003          CALIFICACION: En realización.

                                                                                                                                             

TITULO: “Regulación de la expresión de proteín-fosfatasas relacionadas con la dormición de semillas de haya (Fagus sylvatica L.)”.

DOCTORANDO: Mª PAZ GONZALEZ GARCIA

UNIVERSIDAD: Salamanca                        FACULTAD/ESCUELA: F. de Biología

AÑO: Fecha lectura aproximada, 2004          CALIFICACION: En realización.

 

 


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Institution:                 University of Valencia

                                   Department of Plant Biology

                                   46100 Valencia

 

Contact Person:         Isabel Arrillaga

                                   e-mail: isabel.arrillaga@uv.es

 

Contribution: Expertise in large-scale tissue culture. Protocols to in vitro propagation of tree species (Sorbus domestica, Fraxinus ornus, Robinia pseudoacacia, Juniperus oxycedrus, Cedrus libani and Cedrus atlantica). Expertise in genetic transformation of forest trees. Biolistic technologies. Analysis and maintanance of transgenic plants. Two fully equipped laboratories for molecular biology and transgenic plant analysis including extensive transgene-secure chamber facilities. Capabilities for maintaining independently transgenic lines (in vitro or in the greenhouse). In vitro tests of stress tolerance. In addition, sequencing, TEM, SEM microscopy, and mass spectrometry facilities for supporting other groups involved in the project.

 

TRANSGENIC LINES AVAILABLE

- Herbicide tolerant Robinia pseudoacacia (black locust) transgenic lines

- Transgenic lines of the aromatic shrub Lavandula latifolia

- Transgenic tobacco lines

Role in the project: Objective 3. Development of transformation technologies. In vitro tests of stress tolerance.

 

Scientists involved: Isabel Arrillaga, Juan Segura and Jesus Muñoz

 

Publications (three):

- Gisbert C, Rus AM, Bolarin MC, Coronado JM, Arrillaga I, Montesinos C, Caro M, Serrano R, Moreno V. 2000. The yeast HAL1 gene improves salt tolerance of transgenic tomato. Plant Physiology, 123: 393- 402.

- Nebauer SG, Arrillaga I, del Castillo-Agudo L, Segura J. 2000. .Agrobacterium tumefaciens-mediated transformation of the aromatic shrub Lavandula latifolia: Long-term stable transgene expression and segregation in the progeny: Molecular Breeding, 6: 539-552.

- Zaragozá C; Muñoz, J Arrillaga I. 2003. Regeneration of herbicide tolerant black locust transgenic plants by the SAAT method. (under revision)

- Sales E, Segura J, Arrillaga I. 2003. Agrobacterium tumefaciens-mediated genetic transformation of the cardenolide-producing plant Digitalis minor L. Planta Medica. 69: 143-147

 

 

 

 


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Institution:                 University of Valladolid

                                   Escuela Técnica Superior de Ingenierías Agrarias
                                   Department of Plant Production and Silvopasciculture
                                   Avda de Madrid,44

                                   34071, Palencia

 

Contact Person:         Rosario Sierra

                        e-mail: rsierra@pvs.uva.es

 

Contribution: Expertise in genetics and improvement of forest trees.
Large-scale tissue culture, development of biochemical and molecular markers. Fully equipped for tissue culture, biochemical and molecular analysis.

 

Scientists involved: Rosario Sierra, Elena Hidalgo, Dolores Cristóbal, Teresa Martín.

 

Publications (three):

 

 Hidalgo, E.; Palacios, JM.; Murillo, J. y Ruiz-Argüeso, T . "Nucleotide sequence and characterization of four additional genes of the hydrogenase structural operon from Rhizobium leguminosarum".

M.T. Martin, M.T. Cervera & J.A. Garcia Properties of the active plum pox potyvirus RNA polymerase complex indefined glycerol gradient fractions.
Virus Res. 37, 127-137. (1995)

Sierra-de-Grado, R.; Moulia, B.; Fournier, M.; Alía, R.; Díez-Barra, R. Genetic control of stem form in Pinus pinaster Ait. seedlings exposed tolateral light. Trees, Structure and Function 11:455-461.(1997)

 

 

 


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Institution:                 University of Vigo

                                   Department of Plant Biology and Soil Science

                                   33071 Vigo

 

University of Santiago de Compostela

Department of Plant Physiology

27002 Lugo

 

Contact person:         Manuel Rey

                                   e-mail: mrey@uvigo.es

 

Contribution: Expertise in genetic transformation and molecular biology in woody species, especially in PCR-based marker development (RAPD and AFLP), their conversion into SCAR-type markers, and in gene expression profiling technology (cDNA-AFLP). Maintenance of transgenic lines and molecular characterization and gene expression analysis of transgenic plants. Laboratory facilities include basic molecular biology equipment, growth chambers and greenhouse, and real-time PCR machine Universities central research facilities include microscopy services (TEM, SEM, and confocal microscopy), mass spectrometry facilities, proteomic services and extensive growth chambers.

 

Scientists involved: Manuel Rey, Mª Victoria González, Mª Ángeles López

 

Publications (three):

 

Rey, M.; González, M.V.; Ordás, R.J.; Ancora, G.; Tavazza, R. 1996. Factors affecting transient gene expression in cultured radiata pine cotyledons following particle bombardment. Physiologia Plantarum 96: 630-636.

López, M.; Humara, J.M.; Rodríguez, R.; Ordás, R.J. 2000. Transient uidA gene expression in electroporated cotyledonary protoplasts of Pinus nigra Arn. subsp. salzmannii (Dunal) Franco and in bombarded cotyledons. Canadian Journal of Forest Research 30: 448-455.

Ávila, C.; Cantón, F.R.; Barnestein, P.; Suárez, M.-F.; Marraccini, P.; Rey, M.; Humara, J.M.; Ordás, R.; Cánovas, F.M. 2001. The promoter of a cytosolic glutamine synthetase gene from the conifer Pinus sylvestris is active in cotyledons of germinating seeds and light-regulated in transgenic Arabidopsis thaliana. Physiologia Plantarum 112: 388-396.

 

 

 


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 Institution:                IIAG-CSIC

                                   Department of Plant Physiology

                                   28040 Santiago de Compostela

 

Contact Person:         M. Concepcion Sánchez

                                   e-mail: conchi@cesga.iiag.es

 

Contribution: Expertise in molecular biology of forest trees, especially in studies of phase change, adventitious rooting and morphogenic capacity of adult tissues. Library construction, gene cloning, temporal and spatial expression of genes and proteins. Genetic transformation of forest species and analysis of transgenic lines. Large scale micropropagation of forest trees and in vitro tissue culture facilities. Plant regeneration by adventitious buds and somatic embryogenesis in different forest tree species. Fully equipped for molecular biology and molecular genetics analysis, including microscopy and histology facilities.

Participation in different National projects in the field of molecular biology, genetic transformation, and plant regeneration by somatic embryogenesis.

 

Scientists involved: Concepción Sánchez, Antonio Ballester, Ana Mª Vieitez.

 

Publications (three):

 

Gil B, Pastoriza E, Ballester A, Sánchez C. 2003. Isolation and characterization of a cDNA differentially expressed in juvenile-like and mature shoots of Quercus robur. Tree Physiology. In press.

 

Corredoira E, Vieitez AM, Ballester A. 2002. Somatic embryogenesis in elm. Annals of Botany, 89: 637-644.

 

Corredoira E, Ballester A, Vieitez AM. 2003. Proliferation and maturation of Castanea sativa somatic embryos originated from leaf explants. Annals of Botany. In press.


 

 

 


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 Institution:                INIA-CIFOR

Department of Forest Systems and Resources
Ctra. La Coruña. 28040, Madrid

 

Contact Person:       

(1) Unit of Forest Genetics and Ecophysiology:
Genomics: Maria Teresa Cervera e-mail: cervera@inia.es
Population Genetics: Santiago González-Martínez. e-mail: santiago@inia.es
Quantitative Genetics: Ricardo Alía. e-mail: alia@inia.es
Ecophysiology: Ismael Aranda. e-mail: aranda@inia.es

(2) Unit of Biotechnology
Mª Angeles Bueno e-mail: bueno@inia.es

(

(1) Contribution:

Genomics:Analysis of pine adaptation to drought stress: Identification of genes involved in response to drought stress (construction of EST, SSH libraries), construction of functional linkage maps. Comparative and association analysis. QTL analysis. Marker development including nuclear micro-satellites and gene-based markers.
Participation in several European (TREESNPS R&D EU project; EVOLTREE NoE), bilateral (with INRA-France and University of Göttingen–Germany), and National projects as well as international Consortiums.

Scientists involved: María Teresa Cervera, M. Ángeles Guevara, Nuria de María, Carmen Collada (Joint Unit INIA-UPM) and Álvaro Soto (Joint Unit INIA-UPM).

Publications (three):
Cervera M-T, Storme V, Liu B, Steenackers M, Ivens B, Michiels B, Gusmao J, van Montagu M and Boerjan W. 2001. Dense genetic linkage maps of three Populus species (P. deltoides, P. nigra and p. trichocarpa) based on AFLP, and microsatellite markers. Genetics 158: 787-809.

L Gil; P Fuentes-Utrilla; A. Soto; M.T. Cervera; C. Collada. 2004. English elm is a 2.000-year-old Roman clone. Nature 431: 1053-1053

S.C. González- Martínez, J. J. Robledo-Arnuncio, C. Collada, A. Díaz, C. G. Williams, R. Alía, M.T. Cervera. 2004. Cross-amplification and sequence variation of microsatellite loci in Eurasian hard pines. Theoretical and Applied Genetics 109: 103-111

 

(2) Contribution: Expertise in Biotechnology, Genetics and improvement of forest trees, specially in organogenesis and embryogenesis. Large-scale micropropagation, rooting, response to stress, maturation, dormancy, germination, genetic transformation and development of neutral-based markers for analysing plant quality. Genetic analysis of forest trees by isozyme, RAPD and microsatellite markers, characterisation, population genetics and quantitative genetics. Induction of haploidy, double haploids and homozygous plant lines by anther, microspore and pollen culture. Fully equipped laboratory for tissue culture and genetic characterisation, including two laminar flow cabinets (1 vertical flow, 1 horizontal), fluorescence microscope, flow cytometer, PCR termocycler, electrophoresis equipment, growth chamber facilities, greenhouse and capabilities for maintaining independently transgenic/cultured lines. Participation in several projects and networks in the field of Biotechnology of forest trees and forest genetics at European (FOREST I+D EU projects: Upgrading of Southern Pines; Adaptation and Selection of Mediterranean Pinus and Cedrus for sustainable Afforestation of marginal lands; European Research on cork oak and cork), bilateral (with BOKU-Vienna, Austria, and the Institut für Obstzüchtung of Dresden–Germany), and National projects as well as international Consortiums: Biotechnological approach for utilisation of gametic cells (European COST Action 824), and co-ordinator of several research projects.

 

Scientists involved: M.A. Bueno, José Antonio Manzanera

 

Publications (three):

 

BUENO M.A, GOMEZ A, MANZANERA J.A.  Propagation and DNA markers characterization of Populus tremula L and Populus alba L. 2003 IN: Jain SM, Ishii K, (eds.). Micropropagation of Woody Trees and Fruits. Forestry Sciences. Vol. 75, 37-74.Kluwer Academic Publishers. Dordrecht. The Netherlands ISBN 1-4020-1135-0

 

BUENO M.A, MANZANERA J.A.  Oak anther culture. 2003 IN: Maluszynski M, Kasha K.J , Foster B.P. and Szarejko I (Eds). Doubled Haploid Production in Crops Plants. A Manual. Kluwer Academic Publishers, Dordrecht. Vienna.

 

GÓMEZ, A., PINTOS, B., AGUIRIANO, E., MANZANERA, J.A., BUENO, M.A. 2001 SSR markers for Quercus suber tree identification and embryo analysis. Vol 92 (3). 292-295. J. of Heredity.

 

 

 


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 Institution:                NEIKER

                                   Apartado 46

                                   01080 Alava

 

Contact Person:         Enrique Ritter

                                   e-mail: eritter@neiker.net

 

Contribution:

NEIKER is a public research institute depending on the Basque Government in Northern Spain. The institute consists of three departments carrying  out basic and applied research in agriculture (plants and animals). Between 100 to 120 scientists, technical  assistants and other staff members, including undergraduate and graduate students are active in the three departments and administration.

NEIKER is involved in general agricultural research including forest species. Pinus radiata repre­sents the predominant species in the productive forests of northern Spain. The scientific staff is  experienced in forest breeding and is responsible for the ongoing local forest breeding pro­gram.

Molecular techniques are well established at the institute. Within other national and EU projects different biotechnological techniques were developed and are optimised continuously for forest species but also for other crop species. These techniques include all kinds of PCR and hybridisation based techniques, electrophoretic techniques, sequencing, cloning and others.

Particularly, NEIKER has cordinated the related EU project FAIR3 CT96-1445 "Development, validation and application of molecular, morphological and physiological markers for juvenile and mature state characterization in woody plant species" (see http://www.neiker.net/PhaseDB for details) in which different marker types and indicators for phase changes (juvenil-adult; vegetative-generative) were developed which can be used for predicting rooting capacities or flowering behaviour. Currently, NEIKER coordinates the EU project QLK5-1999-01159: "Construction and application of a multifunctional and saturated genetic map for coniferous species".

From this project an ultra-high density linkage map (UHD map) of  Pinus pinaster based on nearly 2000 AFLP markers and numerous EST and SSR markers is available. This map is exploited to full extent  for comparative genome and QTL analysis in different genetic backgrounds. Other published linkage maps in forest species have been partially aligned with the reference map. Satellite maps based on a common subset of  AFLP primer combinations and EST and SSR markers have been obtained in this project for P. radiata, P. sylvestris, Picea abies and Pseudotsuga menziesii, allowing to perform comparative genome analysis in different pine species and related gymnosperms. QTL analysis for important characters such as growth traits, wood quality and physiological characters have been performed in reference and satellite mapping populations. Allele-specific markers for important QTLs have been developed and their variability in local germplasm and breeding material of different forest species is currently analysed (see http://www.neiker.net/UHDfor for details).

Within the frame of the EU-Project QLK5-CT2002-01849 "Developing a physical and functional map of potato: creating new sources for molecular markers to breed cultivars with multiple resistances and quality traits (APOPHYS)" and a National Project RTA03-001-C2 "Obtención de un mapa genético, físico y funcional y detección de genes para estrés biótico y abiótico en la vid (GENOVID)", transcriptome maps are being constructed in potato and grapewine, respectively. On the other hand differential cDNA display and candidate gene expression related to drought resistance is currently being performed in these species within the mentioned projects. 

 

Scientists involved: Enrique Ritter, Santiago Espinel, Pablo Goicoetchea, Ana Aragonés.

 

Publications (three):

Ritter E; Aragones A; Markussen T; Acheré V; Espinel S; Fladung M, Wrobel S; Faivre-Rampant P; Jeandroz S; Favre JM (2002). Towards construction of an ultra high density linkage map for Pinus pinaster. Annals of Forest Science 59: 637-643.

 

Markussen, T., Fladung, M., Achere, V., Favre, J.M., Faivre-Rampant, P., Aragones, A.,  Da Silva Perez, D., Harvengt, L., Espinel, S., Ritter, E. (2003) Identification of QTLs controlling growth, chemical and physical wood property traits in Pinus pinaster (Ait.) Silvae Genetica (in press)

 

Ritter E, , Espinel S, Fladung M, Favre JM (2003). High density linkage maps in conifer species and their potential application. In: Molecular Genetics and Breeding of Forest  Trees. Eds: Kumar S & Fladung M., The Haworth Press Inc. (in press)

           

 


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