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M. A. Bueno - One of the best experts on this subject based on the ideXlab platform.
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Proteomic perspective of Quercus Suber somatic embryogenesis.
Journal of proteomics, 2013Co-Authors: Aranzazu Gomez-garay, M. A. Bueno, Juan Antonio López, Emilio Camafeita, Beatriz PintosAbstract:Abstract Quercus Suber L. is a forest tree with remarkable ecological, social and economic value in the southern Europe ecosystems. To circumvent the difficulties of breeding such long-lived species like Q. Suber in a conventional fashion, clonal propagation of Q. Suber elite trees can be carried out, although this process is sometimes unsuccessful. To help decipher the complex program underlying the development of Q. Suber somatic embryos from the first early stage until maturity, a proteomic approach based on DIGE and MALDI-MS has been envisaged. Results highlighted several key processes involved in the three developmental stages (proliferative, cotyledonary and mature) of Q. Suber somatic embryogenesis studied. Results show that the proliferation stage is characterized by fermentation as an alternative energy source at the first steps of somatic embryo development, as well as by up-regulation of proteins involved in cell division. In this stage reactive oxygen species play a role in proliferation, while other proteins like CAD and PR5 seem to be implied in embryonic competence. In the transition to the cotyledonary stage diverse ROS detoxification enzymes are activated and reserve products (mainly carbohydrates and proteins) are accumulated, whereas energy production is increased probably to participate in the synthesis of primary metabolites such as amino acids and fatty acids. Finally, in the mature stage ethylene accumulation regulates embryo development. Biological significance Quercus Suber L. is a forest tree with remarkable ecological, social and economic value in the southern Europe ecosystems. To circumvent the difficulties of breeding such long-lived species like Q. Suber in a conventional fashion, clonal propagation of Q. Suber elite trees can be carried out, although this process is sometimes unsuccessful. To help decipher the complex program underlying the development of Q. Suber somatic embryos from the first early stage until maturity, in deep studies become necessary. This article is part of a Special Issue entitled: Translational Plant Proteomics.
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Agrobacterium-mediated transformation of cork oak (Quercus Suber L.) somatic embryos
New Forests, 2005Co-Authors: N. Sánchez, Beatriz Pintos, José Antonio Manzanera, M. A. BuenoAbstract:A simple method is presented for the unreported genetic transformation of cork oak (Quercus Suber L.). Pro-embryo masses (PEMs) were induced on immature zygotic embryos applied to medium supplemented with 2.3 μM 2,4-dichlorophenoxyacetic acid. The established PEMs were inoculated with Agrobacterium tumefaciens LBA4404/p35S GUS INT/pCAMBIA 1301 strain. Transformants were selected on hygromycin (HYG) 94 μM-supplemented medium. Viable embryos constituted 13% of those selected on HYG during 4 months. Expression of β-glucuronidase at 4 months following co-cultivation confirmed transformation in 5.8% embryos selected on HYG. This method forms a basis for genetic transformation of cork oak somatic embryos.
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SSR Markers for Quercus Suber Tree Identification and Embryo Analysis
The Journal of heredity, 2001Co-Authors: A. Gómez, José Antonio Manzanera, Beatriz Pintos, E. Aguiriano, M. A. BuenoAbstract:Three Quercus simple sequence repeat (SSR) markers were amplified by polymerase chain reaction (PCR) from nuclear DNA extracts of trees and in vitro-induced haploid embryos from anther cultures of Quercus Suber L. These markers were sufficiently polymorphic to identify 10 of 12 trees located in two Spanish natural areas. The same loci have been analyzed in anther-derived haploid embryos showing the parental tree allele segregation. All the alleles were present in the haploid progeny. The presence of diverse alleles in embryos derived from the same anther demonstrated that they were induced on multiple microspores or pollen grains and they were not clonally propagated. Also, diploid cultures and mixtures of haploid-diploid tissues were obtained. The origin of such cultures, either somatic or gametic, was elucidated by SSR markers. All the embryos showed only one allele, corroborating a haploid origin. Allelic composition of the haploid progeny permitted parental identification among all analyzed trees.
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Somatic and Gamatic Embryogenesis in Quercus Suber L.
Somatic Embryogenesis in Woody Plants, 2000Co-Authors: M. A. Bueno, A. Gómez, José Antonio ManzaneraAbstract:Cork-oak (Quercus Suber L., order Fagales, family Fagaceae) belongs to a group of forest species of outstanding importance in the Boreal hemisphere. Oaks are the dominant species in many ecosystems of the Holartic region.
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stress induced formation of haploid plants through anther culture in cork oak Quercus Suber
Physiologia Plantarum, 1997Co-Authors: M. A. Bueno, Arancha Gomez, Monica Boscaiu, J A Manzanera, Oscar VicenteAbstract:Induction of haploid embryos and regeneration of plantlets have been obtained, for the first time, in cork oak (Quercus Suber L.) by combining a starvation treatment in anther culture with a mild heat shock at 33°C for 5 days, followed by culture at 25°C in a simple agar medium without growth regulators. The same conditions had been shown previously to be optimal for embryogenic induction in isolated microspore cultures of several model species such as tobacco and wheat. These results support the notion that stress, particularly sucrose starvation, a heat shock or a combination of both treatments could be the major and general signal responsible for the inhibition of normal gametophytic development of the microspores and for the induction of the alternative embryogenic pathway. A similar approach may be used for the production of haploid and doubled haploids for plant breeding in other species that, like most forest trees, are still recalcitrant in anther culture.
José Antonio Manzanera - One of the best experts on this subject based on the ideXlab platform.
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Agrobacterium-mediated transformation of cork oak (Quercus Suber L.) somatic embryos
New Forests, 2005Co-Authors: N. Sánchez, Beatriz Pintos, José Antonio Manzanera, M. A. BuenoAbstract:A simple method is presented for the unreported genetic transformation of cork oak (Quercus Suber L.). Pro-embryo masses (PEMs) were induced on immature zygotic embryos applied to medium supplemented with 2.3 μM 2,4-dichlorophenoxyacetic acid. The established PEMs were inoculated with Agrobacterium tumefaciens LBA4404/p35S GUS INT/pCAMBIA 1301 strain. Transformants were selected on hygromycin (HYG) 94 μM-supplemented medium. Viable embryos constituted 13% of those selected on HYG during 4 months. Expression of β-glucuronidase at 4 months following co-cultivation confirmed transformation in 5.8% embryos selected on HYG. This method forms a basis for genetic transformation of cork oak somatic embryos.
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Shoot development in Quercus Suber L. somatic embryos
In Vitro Cellular & Developmental Biology - Plant, 2002Co-Authors: M. Elena González-benito, Gemma García-martín, José Antonio ManzaneraAbstract:N6-Benzyladenine (BA; 0.04–4μM) application to germinated Quercus Suber somatic embryos considerably increased caulinar apex elongation frequency and maintained active growth in the plantlets, although it did not have a significant effect on the percentage of shoots with normal morphology. The addition of 0.5 μM indoleacetic acid together with the cytokinin did not have any effect. The use of a low concentration (0.04 μM) of BA allowed the appropriate radicle elongation in all germinating somatic embryos, but higher concentrations arrested this elongation.
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SSR Markers for Quercus Suber Tree Identification and Embryo Analysis
The Journal of heredity, 2001Co-Authors: A. Gómez, José Antonio Manzanera, Beatriz Pintos, E. Aguiriano, M. A. BuenoAbstract:Three Quercus simple sequence repeat (SSR) markers were amplified by polymerase chain reaction (PCR) from nuclear DNA extracts of trees and in vitro-induced haploid embryos from anther cultures of Quercus Suber L. These markers were sufficiently polymorphic to identify 10 of 12 trees located in two Spanish natural areas. The same loci have been analyzed in anther-derived haploid embryos showing the parental tree allele segregation. All the alleles were present in the haploid progeny. The presence of diverse alleles in embryos derived from the same anther demonstrated that they were induced on multiple microspores or pollen grains and they were not clonally propagated. Also, diploid cultures and mixtures of haploid-diploid tissues were obtained. The origin of such cultures, either somatic or gametic, was elucidated by SSR markers. All the embryos showed only one allele, corroborating a haploid origin. Allelic composition of the haploid progeny permitted parental identification among all analyzed trees.
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Somatic and Gamatic Embryogenesis in Quercus Suber L.
Somatic Embryogenesis in Woody Plants, 2000Co-Authors: M. A. Bueno, A. Gómez, José Antonio ManzaneraAbstract:Cork-oak (Quercus Suber L., order Fagales, family Fagaceae) belongs to a group of forest species of outstanding importance in the Boreal hemisphere. Oaks are the dominant species in many ecosystems of the Holartic region.
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Plant regeneration through somatic embryogenesis in Quercus Suber
Physiologia Plantarum, 1992Co-Authors: M. A. Bueno, R. Astorga, José Antonio ManzaneraAbstract:Cork oak (Quercus Suber L.) zygotic embryos, endosperm and ovules were treated with different concentrations of 2,4-D for induction of somatic embryos. Plant material was collected during the embryo development season, from June to September. Immature embryos proved to be the most reactive initial explant. Callus and somatic embryos developed a few weeks after the beginning of the 2,4-D treatment. For embryo development experiments, different growth regulators and cold and desiccation treatments were tested. Cold storage of somatic embryos matured in vitro at 5°C was the best treatment for breaking dormancy.
Beatriz Pintos - One of the best experts on this subject based on the ideXlab platform.
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Proteomic perspective of Quercus Suber somatic embryogenesis.
Journal of proteomics, 2013Co-Authors: Aranzazu Gomez-garay, M. A. Bueno, Juan Antonio López, Emilio Camafeita, Beatriz PintosAbstract:Abstract Quercus Suber L. is a forest tree with remarkable ecological, social and economic value in the southern Europe ecosystems. To circumvent the difficulties of breeding such long-lived species like Q. Suber in a conventional fashion, clonal propagation of Q. Suber elite trees can be carried out, although this process is sometimes unsuccessful. To help decipher the complex program underlying the development of Q. Suber somatic embryos from the first early stage until maturity, a proteomic approach based on DIGE and MALDI-MS has been envisaged. Results highlighted several key processes involved in the three developmental stages (proliferative, cotyledonary and mature) of Q. Suber somatic embryogenesis studied. Results show that the proliferation stage is characterized by fermentation as an alternative energy source at the first steps of somatic embryo development, as well as by up-regulation of proteins involved in cell division. In this stage reactive oxygen species play a role in proliferation, while other proteins like CAD and PR5 seem to be implied in embryonic competence. In the transition to the cotyledonary stage diverse ROS detoxification enzymes are activated and reserve products (mainly carbohydrates and proteins) are accumulated, whereas energy production is increased probably to participate in the synthesis of primary metabolites such as amino acids and fatty acids. Finally, in the mature stage ethylene accumulation regulates embryo development. Biological significance Quercus Suber L. is a forest tree with remarkable ecological, social and economic value in the southern Europe ecosystems. To circumvent the difficulties of breeding such long-lived species like Q. Suber in a conventional fashion, clonal propagation of Q. Suber elite trees can be carried out, although this process is sometimes unsuccessful. To help decipher the complex program underlying the development of Q. Suber somatic embryos from the first early stage until maturity, in deep studies become necessary. This article is part of a Special Issue entitled: Translational Plant Proteomics.
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Identification of DNA-microsatellite markers for the characterization of somatic embryos in Quercus Suber.
Methods in molecular biology (Clifton N.J.), 2013Co-Authors: Arancha Gómez-garay, Ángeles Bueno, Beatriz PintosAbstract:Nuclear DNA-microsatellite markers led the possibility to characterize individually both Quercus Suber trees and somatic embryos. The genotype inferred by SSR markers opens the possibility to obtain a fingerprint for clonal lines identification. Furthermore, allow to infer the origin of somatic embryos from haploid cells (microspores) or from diploid tissues. Using few SSR markers from other Quercus species and an automatic system based in fluorescence, it is possible to obtain a high discrimination power between genotypes. This method is sufficient to assign tissues to an individual tree with high statistical certainty. Nevertheless, it is necessary to take care to select the adequate DNA extraction method to avoid PCR inhibitors present in diverse Q. Suber tissues.
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Agrobacterium-mediated transformation of cork oak (Quercus Suber L.) somatic embryos
New Forests, 2005Co-Authors: N. Sánchez, Beatriz Pintos, José Antonio Manzanera, M. A. BuenoAbstract:A simple method is presented for the unreported genetic transformation of cork oak (Quercus Suber L.). Pro-embryo masses (PEMs) were induced on immature zygotic embryos applied to medium supplemented with 2.3 μM 2,4-dichlorophenoxyacetic acid. The established PEMs were inoculated with Agrobacterium tumefaciens LBA4404/p35S GUS INT/pCAMBIA 1301 strain. Transformants were selected on hygromycin (HYG) 94 μM-supplemented medium. Viable embryos constituted 13% of those selected on HYG during 4 months. Expression of β-glucuronidase at 4 months following co-cultivation confirmed transformation in 5.8% embryos selected on HYG. This method forms a basis for genetic transformation of cork oak somatic embryos.
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SSR Markers for Quercus Suber Tree Identification and Embryo Analysis
The Journal of heredity, 2001Co-Authors: A. Gómez, José Antonio Manzanera, Beatriz Pintos, E. Aguiriano, M. A. BuenoAbstract:Three Quercus simple sequence repeat (SSR) markers were amplified by polymerase chain reaction (PCR) from nuclear DNA extracts of trees and in vitro-induced haploid embryos from anther cultures of Quercus Suber L. These markers were sufficiently polymorphic to identify 10 of 12 trees located in two Spanish natural areas. The same loci have been analyzed in anther-derived haploid embryos showing the parental tree allele segregation. All the alleles were present in the haploid progeny. The presence of diverse alleles in embryos derived from the same anther demonstrated that they were induced on multiple microspores or pollen grains and they were not clonally propagated. Also, diploid cultures and mixtures of haploid-diploid tissues were obtained. The origin of such cultures, either somatic or gametic, was elucidated by SSR markers. All the embryos showed only one allele, corroborating a haploid origin. Allelic composition of the haploid progeny permitted parental identification among all analyzed trees.
Javier Madrigal - One of the best experts on this subject based on the ideXlab platform.
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Flammability of some companion species in cork oak (Quercus Suber L.) forests.
Annals of Forest Science, 2017Co-Authors: Belkheir Dehane, Carmen Hernando, Mercedes Guijarro, Javier MadrigalAbstract:Key message The high flammability of some companion species in Quercus Suber forests, estimated in laboratory tests, could potentially generate an increase in fire vulnerability and in fire risk.
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Flammability of some companion species in cork oak (Quercus Suber L.) forests
Annals of Forest Science, 2017Co-Authors: Belkheir Dehane, Carmen Hernando, Mercedes Guijarro, Javier MadrigalAbstract:AbstractKey messageThe high flammability of some companion species inQuercus Suberforests, estimated in laboratory tests, could potentially generate an increase in fire vulnerability and in fire risk.ContextRecurrent wildfire is one of the main causes of forest degradation, especially in the Mediterranean region. Increased fire frequency and severity due to global change could reduce the natural resilience of cork oak to wildfire in the future. Hence, it is important to evaluate the flammability of companion species in cork oak forests in the particularly dry bioclimatic conditions of North Africa.AimsThis study aimed to assess and compare flammability parameters at laboratory scale among ten companion frequent species in cork oak forests.MethodsFuel samples were collected in a cork oak (Quercus Suber L) forest in the southern part of the mountains of Tlemcen (Western Algeria). A series of flammability tests were carried out using a Mass Loss Calorimeter device (FTT ®). A cluster analysis to classify flammability of the selected species was conducted using the K-means algorithm.ResultsThe results revealed differences in the four flammability parameters (ignitability, sustainability, combustibility and consumability), in both fresh and dried fine fuel samples from Quercus Suber, Pinus halepensis, Quercus ilex, Quercus faginea, Erica arborea, Arbutus unedo, Pistacia lentiscus, Calicotome spinosa, Juniperus oxycedrus and Tetraclinis articulata. Application of the K-means clustering algorithm showed that C. spinosa, T. articulata, J. oxycedrus and P. halepensis are highly flammable because of their high combustibility and sustainability.ConclusionThe findings identify species that could potentially increase the vulnerability of cork oak forests to forest fires.
Mariano Toribio - One of the best experts on this subject based on the ideXlab platform.
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Cork Oak Quercus Suber L. Embryogenic Liquid Cultures
Step Wise Protocols for Somatic Embryogenesis of Important Woody Plants, 2018Co-Authors: Mar Ruiz-galea, Cristina Celestino, Dolores López-vela, Jesús Jiménez, Nieves Alonso-blázquez, Jesús Alegre, Mariano ToribioAbstract:The cork oak is an evergreen tree species typical of Mediterranean forests. It grows in dense stands but also in low density, in the savanna-like system called “dehesa” in Spain and “montado” in Portugal. The most characteristic production of Quercus Suber is cork, although acorn crops are also important for feeding pigs of the Iberian race. The products obtained from the species justify the accomplishment of genetic improvement programs. Current strategies of forest tree breeding emphasize on the use of vegetative propagation to rapidly capture all the potential of selected individuals and establish them in plantations, performing Multi-Varietal Forestry. Somatic embryogenesis is one of the enabling technologies to carry out that strategy. Our protocol for cloning adult cork oak trees by somatic embryogenesis is robust enough to capture any genotype, and currently it is adequate to produce the limited number of plants needed to establish clonal test. However for large-scale profitable clonal propagation, the scaling-up of plant production using culture in liquid medium has to be developed. Herein we describe step-wise protocols to perform the production of somatic embryos obtained from selected Quercus Suber trees using culture systems based on liquid medium.
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Cork Oak Trees (Quercus Suber L.).
Methods in molecular biology (Clifton N.J.), 2006Co-Authors: Rubén Álvarez, Mariano Toribio, Millán Cortizo, Ricardo-javier Ordás FernándezAbstract:A transformation system for selected mature Quercus Suber L. trees using Agrobacterium tumefaciens has been established. Embryos obtained from recurrent proliferating embryogenic masses are inoculated with AGL1 strain harbouring the plasmid pBINUbiGUSint, which carries the nptII and uidA genes. Evidence of stable transgene integration is obtained by polymerase chain reaction for nptII and uidA genes, Southern blotting and expression of the uidA gene. The transgenic embryos are germinated and successfully transferred to soil.
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Mycorrhization of vitroplants raised from somatic embryos of cork oak (Quercus Suber L.)
Applied Soil Ecology, 2000Co-Authors: Jesús Díez, José Luis Manjón, Gábor M. Kovács, Cristina Celestino, Mariano ToribioAbstract:Abstract The technique described herein allows in vitro ectomycorrhizal synthesis in Quercus Suber vitroplants raised from somatic embryos with Pisolithus tinctorius and Scleroderma polyrhizum strains. Only strains of this species coming from fruit bodies collected in Quercus Suber stands (strain QS241 and strain QS247) formed ectomycorrhizas, and hence these species seem to exhibit host adaptation. The in vitro mycorrhization facilitated the development of secondary roots and the ex vitro weaning of cork oak vitroplants.
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Origin and Early Development of Secondary Embryos in Quercus Suber L.
International Journal of Plant Sciences, 1996Co-Authors: Pere Puigderrajols, Mariano Toribio, B. Fernández-guijarro, Marisa MolinasAbstract:Somatic embryos of the cork oak (Quercus Suber L.) obtained after many generations of recurrent embryogenesis do not show signs of major structural abnormalities except for the appearance of more than two cotyledons or embryos fused at their embryonic axes. Secondary embryogenesis is mainly of multicellular origin and takes place after a slight proliferation of cells in the external layers of the primary embryo root cap. The first sign of secondary embryo formation is the appearance of meristematic primordia on the periphery of the proliferating tissue. They soon develop into bipolar structures and form cotyledonary primordia, and then the new embryos emerge from the proliferation mass. However, the unicellular pathway for secondary embryogenesis cannot be disregarded. In some cases the proliferation mass becomes brownish and takes on a more friable appearance. In this case, single cells, groups of cells, and globular proembryos appear isolated from the surrounding necrotic tissue. However, no further dev...
