35 results on '"Marum, Liliana"'
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2. Periderm differentiation: a cellular and molecular approach to cork oak
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Faustino, Ana, Pires, Rita Costa, and Marum, Liliana
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- 2023
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3. Laser Microdissection of Woody and Suberized Plant Tissues for RNA-Seq Analysis
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Pires, Rita Costa, Ferro, Ana, Capote, Tiago, Usié, Ana, Correia, Bárbara, Pinto, Glória, Menéndez, Esther, and Marum, Liliana
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- 2023
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4. Cryopreservation of Hybrid Pinus elliottii × P. caribaea
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Marum, Liliana, Nunes, Sandra, Almeida, Tânia, Pereira, Vanessa Tolentino, Farinha, Nelson, Dias, Maria Celeste, Santos, Conceição, Jain, S.Mohan, Series Editor, Häggman, Hely, Series Editor, Jain, Shri Mohan, editor, and Gupta, Pramod, editor
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- 2018
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5. Efficient protocol for in vitro mass micropropagation of slash pine
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Nunes, Sandra, Sousa, Diana, Pereira, Vanessa T., Correia, Sandra, Marum, Liliana, Santos, Conceição, and Dias, Maria C.
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- 2018
6. Cryopreservation of Hybrid Pinus elliottii × P. caribaea
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Marum, Liliana, primary, Nunes, Sandra, additional, Almeida, Tânia, additional, Pereira, Vanessa Tolentino, additional, Farinha, Nelson, additional, Dias, Maria Celeste, additional, and Santos, Conceição, additional
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- 2018
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7. Somatic embryogenesis of hybrid Pinus elliottii var. elliottii × P. caribaea var. hondurensis and ploidy assessment of somatic plants
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Nunes, Sandra, Marum, Liliana, Farinha, Nelson, Pereira, Vanessa T., Almeida, Tânia, Sousa, Diana, Mano, Nélia, Figueiredo, Joana, Dias, Maria C., and Santos, Conceição
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- 2017
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8. Laser Microdissection of Woody and Suberized Plant Tissues for RNA-Seq Analysis
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Pires, Rita Costa, primary, Ferro, Ana, additional, Capote, Tiago, additional, Usié, Ana, additional, Correia, Bárbara, additional, Pinto, Glória, additional, Menéndez, Esther, additional, and Marum, Liliana, additional
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- 2022
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9. Ferramentas moleculares para a seleção de cultivares de amendoeira com teor de vitamina E diferenciador.
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João, Sérgio, Faustino, Ana, Pires, Rita C., Martins, Rita, and Marum, Liliana
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- 2023
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10. Quercus suber Transcriptome Analyses: Identification of Genes and SNPs Related to Cork Quality
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Mendes, Bruna, primary, Usié, Ana, additional, Capote, Tiago, additional, Meireles, Brígida, additional, Almeida, Tânia, additional, Marum, Liliana, additional, Gonçaves, Sónia, additional, and Ramos, António, additional
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- 2022
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11. Pinus elliottii and P. elliottii x P. caribaea hybrid differently cope with combined drought and heat episodes
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Dias, Maria Celeste, primary, Ferreira de Oliveira, José Miguel P., additional, Marum, Liliana, additional, Pereira, Vanessa, additional, Almeida, Tânia, additional, Nunes, Sandra, additional, Araújo, Márcia, additional, Moutinho-Pereira, José, additional, Correia, Carlos M., additional, and Santos, Conceição, additional
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- 2022
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12. An epigenetic view of plant cells cultured in vitro: somaclonal variation and beyond
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Miguel, Célia and Marum, Liliana
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- 2011
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13. Chemical and Genetic Relationships of Cynara cardunculus L. (Cardoon) in Southern Portugal
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Paulino, Ana, primary, Brás, Teresa, additional, Rosa, Daniela, additional, Pires, Rita Costa, additional, Santos, Jacqueline, additional, Pereira, Margarida, additional, Paulo, Octávio S., additional, Marum, Liliana, additional, and Duarte, Mª Fátima, additional
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- 2021
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14. Transcriptome Characterization of Different Tissues of Stone Pine (Pinus pinea L.): De Novo Assembly
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Usié, Ana, primary, Mendes, Bruna, additional, Antunes, Marta, additional, Leão, Célia, additional, Marum, Liliana, additional, and Ramos, António Marcos, additional
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- 2021
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15. Quantification and Immunolocalization of Auxin in Prunus dulcis (Mill.) D. A. Webb Micrografts
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Caeiro, Sandra, primary, Lopes, Tércia, additional, Pedrosa, Ana, additional, Caeiro, André, additional, Pires, Rita Costa, additional, Faustino, Ana, additional, Rosa, Armindo, additional, Marreiros, António, additional, Canhoto, Jorge, additional, Marum, Liliana, additional, and Correia, Sandra, additional
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- 2021
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16. Transcriptome Analysis in Cork Oak Using Laser Microdissection and RNA-Seq
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Pires, Rita Costa, primary, Capote, Tiago, additional, Ferro, Ana, additional, and Marum, Liliana, additional
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- 2021
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17. Establishment and Optimization of Micrografting Assays with Almond (Prunus dulcis) Portuguese Varieties
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Faustino, Ana, primary, Pires, Rita Costa, additional, Caeiro, Sandra, additional, Rosa, Armindo, additional, Marreiros, António, additional, Canhoto, Jorge, additional, Correia, Sandra, additional, and Marum, Liliana, additional
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- 2021
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18. Flow cytometric and morphological analyses of Pinus pinaster somatic embryogenesis
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Marum, Liliana, Loureiro, João, Rodriguez, Eleazar, Santos, Conceição, Oliveira, M. Margarida, and Miguel, Célia
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- 2009
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19. Genetic diversity and population structure of Cynara cardunculus L. in southern Portugal
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Castro, Maria Miguel, primary, Rosa, Daniela, additional, Ferro, Ana M., additional, Faustino, Ana, additional, Paulino, Ana, additional, Brás, Teresa, additional, Machado, Eliana, additional, Cruz, Carla Pinto, additional, Belo, Anabela D. F., additional, Nozes, Paula, additional, Portugal, João, additional, Ramôa, Sofia, additional, Mendonça, Diogo, additional, Simões, Fernanda, additional, Duarte, Maria F., additional, and Marum, Liliana, additional
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- 2021
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20. Determining DNA strand breakage from embryogenic cell cultures of a conifer species using the single-cell gel electrophoresis assay
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Costa, Pedro M., Milhinhos, Ana, Simões, Marta, Marum, Liliana, Oliveira, Ana Maria, Costa, Maria H., and Miguel, Célia
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- 2012
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21. Analysis of genetic stability at SSR loci during somatic embryogenesis in maritime pine (Pinus pinaster)
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Marum, Liliana, Rocheta, Margarida, Maroco, João, Oliveira, M. Margarida, and Miguel, Célia
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- 2009
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22. Identification of GPAT acyltransferases in cork oak
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Marum Liliana, Miguel Andreia, Ricardo Pinto C, and Miguel Célia
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Medicine ,Science - Published
- 2011
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23. Somatic Embryogenesis from 20 Open-Pollinated Families of Portuguese Plus Trees of Maritime Pine
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Miguel, Célia, Gonçalves, Sónia, Tereso, Susana, Marum, Liliana, Maroco, João, and Margarida Oliveira, M.
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- 2004
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24. Plant regeneration from ploidy-stable cryopreserved embryogenic lines of the hybrid Pinus elliottii x P. caribaea
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Nunes, Sandra, primary, Marum, Liliana, additional, Farinha, Nelson, additional, Pereira, Vanessa Tolentino, additional, Almeida, Tânia, additional, Dias, Maria Celeste, additional, and Santos, Conceição, additional
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- 2017
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25. Physiological characterization and true-to-typeness evaluation of in vitro and ex vitro seedlings of Pinus elliottii : A contribution to breeding programs
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Nunes, Sandra, primary, Santos, Conceição, additional, Moutinho-Pereira, José, additional, Correia, Carlos, additional, Oliveira, Helena, additional, Ferreira de Oliveira, José Miguel, additional, Pereira, Vanessa Tolentino, additional, Almeida, Tânia, additional, Marum, Liliana, additional, and Dias, Maria Celeste, additional
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- 2016
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26. Somatic embryogenesis of hybrid Pinus elliottii var. elliottii × P. caribaea var. hondurensis and ploidy assessment of somatic plants.
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Nunes, Sandra, Marum, Liliana, Farinha, Nelson, Pereira, Vanessa T., Almeida, Tânia, Sousa, Diana, Mano, Nélia, Figueiredo, Joana, Dias, Maria C., and Santos, Conceição
- Abstract
A protocol is described for somatic embryogenesis (SE) Pinus elliottii × P. caribaea, an interspecific pine hybrid of major economic importance. Different embryogenesis conditions were tested, from time of initiation to plant regeneration. Immature megagametophytes containing zygotic embryos of five open pollinated (OP) mother trees were used for SE initiation. An established rate of 10.2% was achieved for explants inoculated on modified Litvay's medium (mLV), supplemented with BAP (4.5 µM) and 2,4-D (9 µM). Initiation of SE was influenced by the mother-tree genotype however, modification of culture conditions enabled establishment of embryogenic cell lines (ECLs) from all OP seed families. Fifty-two percent of somatic embryos developed to the cotyledonary stage on mLV, supplemented with 40 µM ABA and 6% w/v sucrose. Cotyledonary stage somatic embryos converted to plants at 45-86%. Four months after acclimatization in a greenhouse, plant survival rates ranged between 78 and 94%. Flow cytometry analysis of ploidy, did not show any variation in ECLs and somatic seedlings. [ABSTRACT FROM AUTHOR]
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- 2018
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27. Characterization of the cork oak transcriptome dynamics during acorn development
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Miguel, Andreia, primary, de Vega-Bartol, José, additional, Marum, Liliana, additional, Chaves, Inês, additional, Santo, Tatiana, additional, Leitão, José, additional, Varela, Maria Carolina, additional, and Miguel, Célia M., additional
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- 2015
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28. A comprehensive assessment of the transcriptome of cork oak (Quercus suber) through EST sequencing
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Pereira-Leal, José B, primary, Abreu, Isabel A, additional, Alabaça, Cláudia S, additional, Almeida, Maria, additional, Almeida, Paulo, additional, Almeida, Tânia, additional, Amorim, Maria, additional, Araújo, Susana, additional, Azevedo, Herlânder, additional, Badia, Aleix, additional, Batista, Dora, additional, Bohn, Andreas, additional, Capote, Tiago, additional, Carrasquinho, Isabel, additional, Chaves, Inês, additional, Coelho, Ana, additional, Costa, Maria, additional, Costa, Rita, additional, Cravador, Alfredo, additional, Egas, Conceição, additional, Faro, Carlos, additional, Fortes, Ana M, additional, Fortunato, Ana S, additional, Gaspar, Maria, additional, Gonçalves, Sónia, additional, Graça, José, additional, Horta, Marília, additional, Inácio, Vera, additional, Leitão, José M, additional, Lino-Neto, Teresa, additional, Marum, Liliana, additional, Matos, José, additional, Mendonça, Diogo, additional, Miguel, Andreia, additional, Miguel, Célia M, additional, Morais-Cecílio, Leonor, additional, Neves, Isabel, additional, Nóbrega, Filomena, additional, Oliveira, Maria, additional, Oliveira, Rute, additional, Pais, Maria, additional, Paiva, Jorge A, additional, Paulo, Octávio S, additional, Pinheiro, Miguel, additional, Raimundo, João AP, additional, Ramalho, José C, additional, Ribeiro, Ana I, additional, Ribeiro, Teresa, additional, Rocheta, Margarida, additional, Rodrigues, Ana, additional, Rodrigues, José C, additional, Saibo, Nelson JM, additional, Santo, Tatiana E, additional, Santos, Ana, additional, Sá-Pereira, Paula, additional, Sebastiana, Mónica, additional, Simões, Fernanda, additional, Sobral, Rómulo S, additional, Tavares, Rui, additional, Teixeira, Rita, additional, Varela, Carolina, additional, Veloso, Maria, additional, and Ricardo, Cândido PP, additional
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- 2014
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29. Correction: Reference Gene Selection for Quantitative Real-time PCR Normalization in Quercus suber
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Marum, Liliana, primary, Miguel, Andreia, additional, Ricardo, Cândido P., additional, and Miguel, Célia, additional
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- 2012
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30. Reference Gene Selection for Quantitative Real-time PCR Normalization in Quercus suber
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Marum, Liliana, primary, Miguel, Andreia, additional, Ricardo, Cândido P., additional, and Miguel, Célia, additional
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- 2012
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31. Determining DNA strand breakage from embryogenic cell cultures of a conifer species using the single-cell gel electrophoresis assay
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Costa, Pedro M., primary, Milhinhos, Ana, additional, Simões, Marta, additional, Marum, Liliana, additional, Oliveira, Ana Maria, additional, Costa, Maria H., additional, and Miguel, Célia, additional
- Published
- 2011
- Full Text
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32. Characterization, micropropagation and preservation of Pinus genotypes
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Nunes, Sandra Sofia Cachulo, Santos, Maria da Conceição, Dias, Maria Celeste Pereira, and Marum, Liliana Maria Bota
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hybrid, micropropagation ,Biologia ,plant performance ,photosynthesis ,Micropropagação vegetal ,breeding programs ,cryoprotectant ,flow cytometry ,somatic embryogenesis ,cryopreservation ,Coníferas ,Pinus elliottii ,genetic stability ,Criopreservação ,Pinus elliottii x Pinus caribaea ,slash pine ,slow cooling storage ,coniferous - Abstract
Doutoramento em Biologia Slash Pine (Pinus elliottii var. elliottii) and the hybrid (Pinus elliottii var. elliottii x Pinus caribaea var. hondurensis) have a great economic value due to their high growth ratio and resin production. Therefore, it is important to achieve a strategy to propagate this species and the hybrid more rapidly maintaining their characteristics. This project aims to preserve/enlarge the Pinus germplasm collection, provided by the company KLÓN, Innovative Technologies from Cloning, by micropropagation and cryopreservation techniques and analyze the putative changes (genetic stability) in micropropagated plants by flow cytometry, to study plant survival rates, growth and photosynthetic performance after acclimatization. In Chapter I, general aspects of the species and hybrid under study are presented, as well as a brief description of the breeding programs in Pinus spp. and the contribution that innovative techniques of in vitro propagation can give, leading to decades of anticipation on the breeding programs results. It was also described some major aspects to the different micropropagation and cryopreservation techniques, always presenting a review of current knowledge on the use of these techniques in the genus Pinus. Finally, the research objectives of this thesis are presented. Chapter II is dedicated to the application of a micropropagation protocol by proliferation of axillary shoots in the specie Pinus elliottii var. elliottii. This Chapter was divided in two sections. In section II.1 a protocol was optimized for large-scale P. elliottii micropropagation, which describes all the steps from disinfection and seed germination to the production of seedlings in vitro, which were used as explants for shoot induction. Various conditions for induction, shoot elongation and rooting were tested, and a protocol enabling the production of micropropagated plantlets 20 to 22 weeks after germination in vitro has been established. In section II.2 was performed the genetic and physiological characterization of P.elliottii plants micropropagated by the methodology developed in the previous section, in comparison with seedlings. The physiological performance of the plants was evaluated by determination of various parameters associated with photosynthesis and carbon metabolism, such as: chlorophyll a fluorescence; relative water content; gas exchange; pigment and carbohydrate contents. In turn, the genetic characterization was performed by analysis by flow cytometry of putative alterations in DNA content, ploidy level and in cell cycle dynamics. The results indicate that the developed micropropagation protocol for P. elliottii did not induce significative changes, both at physiological and genetic level, in the plants. Chapter III focuses on the optimization of a somatic embryogenesis process for the hybrid Pinus elliottii var. elliottii x Pinus caribaea var. hondurensis, from the initiation to the plants regeneration produced from somatic embryos. For initiation of embryogenic cultures of this hybrid, immature megagametophytes obtained from five open-pollinated plus trees were used as explants. To optimize the process, the effect of genotype in both the initiation and maturation, as the influence of different formulations of basal media and growth regulators in the various stages of the process were evaluated. Throughout the process was assessed the genetic stability of embryogenic masses with different time in culture, and at the end of the produced emblings, in comparison with the mother-trees needles. This protocol allows the production of emblings from somatic embryos not having been detected variability in the DNA content and ploidy level. Chapter IV is dedicated to the preservation of germplasm bank produced for the hybrid under study. Cryopreservation of embryogenic masses is beneficial not only for the preservation of germplasm during the breeding programs development, as well as to avoid the loss of the potential of the embryogenic masses. For the optimization of an embryogenic masses cryopreservation protocol using the slow freezing method, different variations were tested in pretreatments and in the duration of slow freezing step. Pretreatments to which the embryogenic tissue was subjected, did not compromise the maturation capacity of cryopreserved masses. On the contrary, cryopreservation had in some genotypes a beneficial effect. The optimized protocol allowed the regeneration of plants from cryopreserved masses and the process did not induce major genetic changes (embryogenic masses cryo and non cryopreserved were analyzed by flow cytometric). Finally, Chapter V presents the final conclusions of this PhD thesis, gathering the results of this thesis on the propagation and preservation methodologies for the species and hybrid in study and discussion this contribution to the state of art in this field. Future challenges for further research in these areas are presented in this Chapter. Pinus elliottii var. elliottii e o híbrido, Pinus elliottii var. elliottii x Pinus caribaea var. hondurensis, têm um grande valor económico, devido à sua elevada taxa de crescimento e produção de resina. Surge assim a necessidade de desenvolver estratégias de propagação mais eficientes desta espécie e do híbrido, mantendo as suas características. Este estudo tem como objetivos preservar / aumentar o banco de germoplasma de Pinus, fornecido pela empresa KLÓN, Innovative Technologies from Cloning, utilizando técnicas de micropropagação e de criopreservação, analisar possíveis alterações na estabilidade genética das plantas micropropagadas através de citometria de fluxo, estudar a taxa de sobrevivência de plantas, crescimento e desempenho fotossintético após aclimatização. No Capítulo I foram abordados aspetos gerais da espécie e híbrido em estudo, assim como uma breve descrição dos programas de melhoramento em Pinus spp. e da contribuição que técnicas inovadoras de propagação in vitro podem dar levando a décadas de antecipação dos resultados dos respectivos programas de melhoramento. Foram também descritos alguns aspetos mais importantes relativos às diferentes técnicas de micropropagação e criopreservação existentes, apresentando sempre uma revisão do conhecimento atual sobre o uso destas técnicas no género Pinus. Para finalizar este capítulo, os objetivos de investigação desta Tese são apresentados. O Capítulo II é dedicado à aplicação de uma metodologia de micropropagação (por proliferação de rebentos axilares) da espécie Pinus elliottii var. elliottii. Este capítulo foi dividido em duas secções. Na secção II.1 foi otimizado um protocolo para micropropagação em larga escala de P. elliottii desde a desinfecção e germinação de sementes para a produção de plântulas in vitro. Estas por sua vez são utilizadas como explantes para a indução de rebentos. Foram testadas várias condições para a indução, alongamento e enraizamento de rebentos, tendo sido estabelecido um protocolo que permite a produção de plântulas micropropagadas 20 a 22 semanas após germinação in vitro. Na secção II.2 foi realizada a caracterização genética e fisiológica de plantas de P. elliottii micropropagadas pela metodologia desenvolvida na secção anterior, em comparação com plantas provenientes de sementeira. O desempenho fisiológico das plantas foi avaliado pela determinação de diversos parâmetros relacionados com a fotossíntese e o metabolismo de carbono, tais como a fluorescência de clorofila a, o teor relativo em água, as trocas gasosas, o teor de pigmentos e de carbohidratos. Por sua vez a caracterização genética foi realizada pela análise do conteúdo em DNA e nível de ploidia, e ainda as dinâmicas do ciclo célular, com recurso à citometria de fluxo. Os resultados obtidos indicam que o protocolo de micropropagação desenvolvido para P. elliottii não provoca alterações significativas tanto a nível fisiológico como genético nas plantas. O Capítulo III centra-se na otimização de um processo de embriogénese somática para o híbrido Pinus elliottii var. elliottii x Pinus caribaea var. hondurensis, desde a iniciação até à regeneração de plantas produzidas a partir de embriões somáticos. Para a iniciação de culturas embriogénicas deste híbrido foram utilizados como explantes megagametófitos imaturos obtidos a partir de polinização aberta de cinco árvores plus. Para a otimização do processo foi avaliado o efeito do genótipo tanto na iniciação como na maturação, tal como o influência de diferentes formulações de meios basais e reguladores de crescimento nas diversas fases do processo. Ao longo do processo foi avaliada a estabilidade genética das massas embriogénicas com diferentes tempos de cultura, e no final das plântulas produzidas, em comparação com as agulhas das árvores-mãe, concluindo-se que foi desenvolvido um protocolo que permite a produção de plantas provenientes de embriões somáticos não tendo sido detectada variabilidade ao nível de conteúdo em DNA e nível de ploidia. O Capítulo IV é dedicado à preservação do banco de germoplasma produzido para o híbrido em estudo. A criopreservação de massas embriogénicas é benéfica não só para a preservação de germoplasma durante o desenvolvimento de programas de melhoramento, como para evitar a perda do potencial embriogénico das massas. Para a otimização de um protocolo de criopreservação de massas embriogénicas pelo método de congelamento lento, foram testadas diferentes variações nos pré-tratamentos e na duração do passo de congelação lenta. Os pré-tratamentos a que o tecido embriogénico foi submetido não influenciaram negativamente a capacidade de maturação das massas criopreservadas, apresentando-se a criopreservação até em alguns genótipos com um efeito benéfico. O protocolo otimizado permitiu a regeneração de plantas a partir de massas criopreservadas, para as quais se comprovou que o processo não provocou alterações genéticas, através da análise por citometria de fluxo de massas embriogénicas crio e não criopreservadas. Finalmente, no Capítulo V são apresentadas as conclusões finais da Tese de Doutoramento, onde são realçados os avanços realizados como resultado desta tese nas metodologias de propagação e preservação para a espécie e híbrido em estudo. Neste capítulo são também apresentados os desafios futuros para a continuação da investigação nas áreas de propagação e preservação de Pinus.
- Published
- 2016
33. Caracterização, micropropagação e preservação de genótipos de Pinus
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Nunes, Sandra Sofia Cachulo, Santos, Maria da Conceição, Dias, Maria Celeste Pereira, and Marum, Liliana Maria Bota
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hybrid, micropropagation ,Biologia ,plant performance ,photosynthesis ,Micropropagação vegetal ,breeding programs ,cryoprotectant ,flow cytometry ,somatic embryogenesis ,cryopreservation ,Coníferas ,Pinus elliottii ,genetic stability ,Criopreservação ,Pinus elliottii x Pinus caribaea ,slash pine ,slow cooling storage ,coniferous - Abstract
Doutoramento em Biologia Submitted by Alexandra Bastos (alexandrabastos@ua.pt) on 2016-12-13T18:53:22Z No. of bitstreams: 1 tese.pdf: 60532906 bytes, checksum: fc5742c6a5fee74a8015abb3472a607a (MD5) Made available in DSpace on 2016-12-13T18:53:22Z (GMT). No. of bitstreams: 1 tese.pdf: 60532906 bytes, checksum: fc5742c6a5fee74a8015abb3472a607a (MD5) Previous issue date: 2016-03-14
- Published
- 2016
34. Untitled.
- Author
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Nunes, Sandra, Sousa, Diana, Pereira, Vanessa T., Correia, Sandra, Marum, Liliana, Santos, Conceição, and Dias, Maria C.
- Abstract
An efficient protocol was developed for large-scale micropropagation of slash pine ( Pinus elliottii Engelm. var.elliottii ). For that, explants consisting of shoot apices 1.5–2.0 cm in length from 4 wk-old seedlings were cultured on two different basal media (Murashige & Skoog (MS) and Westvaco WV5 (WV5)). All media were supplemented with 6-benzylaminopurine (BAP), to stimulate the formation of axillary buds. Best bud induction was achieved, after 4 wk, on Westvaco WV5 medium (with 10-μM BAP) with rates close to 100%, and an average number of ~ 7 new buds formed per explant. Elongation took place for 6 wk on Westvaco WV5 medium containing activated charcoal (0.2% (w /v )) and without growth regulators. Rooting took place on half-strength Westvaco WV5 medium containing 9.8-μM indole-3-butyric acid (IBA). After 6 wk, root primordia were visible in ~ 43% of shoots. The acclimatization protocol was also optimized by controlling relative humidity, light/photoperiod, temperature, and nutrition, which led to an acclimatization success of ~ 89%. Flow cytometry analysis of DNA-ploidy did not show any variation between micropropagated plants and seedlings. With the protocol here described, it is possible to obtain a high number of genetically uniform plants per explant, 1 yr afterin vitro germination of slash pine seeds.An efficient protocol was developed for large-scale micropropagation of slash pine ( Pinus elliottii Engelm. var.elliottii ). For that, explants consisting of shoot apices 1.5–2.0 cm in length from 4 wk-old seedlings were cultured on two different basal media (Murashige & Skoog (MS) and Westvaco WV5 (WV5)). All media were supplemented with 6-benzylaminopurine (BAP), to stimulate the formation of axillary buds. Best bud induction was achieved, after 4 wk, on Westvaco WV5 medium (with 10-μM BAP) with rates close to 100%, and an average number of ~ 7 new buds formed per explant. Elongation took place for 6 wk on Westvaco WV5 medium containing activated charcoal (0.2% (w /v )) and without growth regulators. Rooting took place on half-strength Westvaco WV5 medium containing 9.8-μM indole-3-butyric acid (IBA). After 6 wk, root primordia were visible in ~ 43% of shoots. The acclimatization protocol was also optimized by controlling relative humidity, light/photoperiod, temperature, and nutrition, which led to an acclimatization success of ~ 89%. Flow cytometry analysis of DNA-ploidy did not show any variation between micropropagated plants and seedlings. With the protocol here described, it is possible to obtain a high number of genetically uniform plants per explant, 1 yr afterin vitro germination of slash pine seeds.An efficient protocol was developed for large-scale micropropagation of slash pine ( Pinus elliottii Engelm. var.elliottii ). For that, explants consisting of shoot apices 1.5–2.0 cm in length from 4 wk-old seedlings were cultured on two different basal media (Murashige & Skoog (MS) and Westvaco WV5 (WV5)). All media were supplemented with 6-benzylaminopurine (BAP), to stimulate the formation of axillary buds. Best bud induction was achieved, after 4 wk, on Westvaco WV5 medium (with 10-μM BAP) with rates close to 100%, and an average number of ~ 7 new buds formed per explant. Elongation took place for 6 wk on Westvaco WV5 medium containing activated charcoal (0.2% (w /v )) and without growth regulators. Rooting took place on half-strength Westvaco WV5 medium containing 9.8-μM indole-3-butyric acid (IBA). After 6 wk, root primordia were visible in ~ 43% of shoots. The acclimatization protocol was also optimized by controlling relative humidity, light/photoperiod, temperature, and nutrition, which led to an acclimatization success of ~ 89%. Flow cytometry analysis of DNA-ploidy did not show any variation between micropropagated plants and seedlings. With the protocol here described, it is possible to obtain a high number of genetically uniform plants per explant, 1 yr afterin vitro germination of slash pine seeds.An efficient protocol was developed for large-scale micropropagation of slash pine ( Pinus elliottii Engelm. var.elliottii ). For that, explants consisting of shoot apices 1.5–2.0 cm in length from 4 wk-old seedlings were cultured on two different basal media (Murashige & Skoog (MS) and Westvaco WV5 (WV5)). All media were supplemented with 6-benzylaminopurine (BAP), to stimulate the formation of axillary buds. Best bud induction was achieved, after 4 wk, on Westvaco WV5 medium (with 10-μM BAP) with rates close to 100%, and an average number of ~ 7 new buds formed per explant. Elongation took place for 6 wk on Westvaco WV5 medium containing activated charcoal (0.2% (w /v )) and without growth regulators. Rooting took place on half-strength Westvaco WV5 medium containing 9.8-μM indole-3-butyric acid (IBA). After 6 wk, root primordia were visible in ~ 43% of shoots. The acclimatization protocol was also optimized by controlling relative humidity, light/photoperiod, temperature, and nutrition, which led to an acclimatization success of ~ 89%. Flow cytometry analysis of DNA-ploidy did not show any variation between micropropagated plants and seedlings. With the protocol here described, it is possible to obtain a high number of genetically uniform plants per explant, 1 yr afterin vitro germination of slash pine seeds.An efficient protocol was developed for large-scale micropropagation of slash pine ( Pinus elliottii Engelm. var.elliottii ). For that, explants consisting of shoot apices 1.5–2.0 cm in length from 4 wk-old seedlings were cultured on two different basal media (Murashige & Skoog (MS) and Westvaco WV5 (WV5)). All media were supplemented with 6-benzylaminopurine (BAP), to stimulate the formation of axillary buds. Best bud induction was achieved, after 4 wk, on Westvaco WV5 medium (with 10-μM BAP) with rates close to 100%, and an average number of ~ 7 new buds formed per explant. Elongation took place for 6 wk on Westvaco WV5 medium containing activated charcoal (0.2% (w /v )) and without growth regulators. Rooting took place on half-strength Westvaco WV5 medium containing 9.8-μM indole-3-butyric acid (IBA). After 6 wk, root primordia were visible in ~ 43% of shoots. The acclimatization protocol was also optimized by controlling relative humidity, light/photoperiod, temperature, and nutrition, which led to an acclimatization success of ~ 89%. Flow cytometry analysis of DNA-ploidy did not show any variation between micropropagated plants and seedlings. With the protocol here described, it is possible to obtain a high number of genetically uniform plants per explant, 1 yr afterin vitro germination of slash pine seeds.An efficient protocol was developed for large-scale micropropagation of slash pine ( Pinus elliottii Engelm. var.elliottii ). For that, explants consisting of shoot apices 1.5–2.0 cm in length from 4 wk-old seedlings were cultured on two different basal media (Murashige & Skoog (MS) and Westvaco WV5 (WV5)). All media were supplemented with 6-benzylaminopurine (BAP), to stimulate the formation of axillary buds. Best bud induction was achieved, after 4 wk, on Westvaco WV5 medium (with 10-μM BAP) with rates close to 100%, and an average number of ~ 7 new buds formed per explant. Elongation took place for 6 wk on Westvaco WV5 medium containing activated charcoal (0.2% (w /v )) and without growth regulators. Rooting took place on half-strength Westvaco WV5 medium containing 9.8-μM indole-3-butyric acid (IBA). After 6 wk, root primordia were visible in ~ 43% of shoots. The acclimatization protocol was also optimized by controlling relative humidity, light/photoperiod, temperature, and nutrition, which led to an acclimatization success of ~ 89%. Flow cytometry analysis of DNA-ploidy did not show any variation between micropropagated plants and seedlings. With the protocol here described, it is possible to obtain a high number of genetically uniform plants per explant, 1 yr afterin vitro germination of slash pine seeds.An efficient protocol was developed for large-scale micropropagation of slash pine ( Pinus elliottii Engelm. var.elliottii ). For that, explants consisting of shoot apices 1.5–2.0 cm in length from 4 wk-old seedlings were cultured on two different basal media (Murashige & Skoog (MS) and Westvaco WV5 (WV5)). All media were supplemented with 6-benzylaminopurine (BAP), to stimulate the formation of axillary buds. Best bud induction was achieved, after 4 wk, on Westvaco WV5 medium (with 10-μM BAP) with rates close to 100%, and an average number of ~ 7 new buds formed per explant. Elongation took place for 6 wk on Westvaco WV5 medium containing activated charcoal (0.2% (w /v )) and without growth regulators. Rooting took place on half-strength Westvaco WV5 medium containing 9.8-μM indole-3-butyric acid (IBA). After 6 wk, root primordia were visible in ~ 43% of shoots. The acclimatization protocol was also optimized by controlling relative humidity, light/photoperiod, temperature, and nutrition, which led to an acclimatization success of ~ 89%. Flow cytometry analysis of DNA-ploidy did not show any variation between micropropagated plants and seedlings. With the protocol here described, it is possible to obtain a high number of genetically uniform plants per explant, 1 yr afterin vitro germination of slash pine seeds.An efficient protocol was developed for large-scale micropropagation of slash pine ( Pinus elliottii Engelm. var.elliottii ). For that, explants consisting of shoot apices 1.5–2.0 cm in length from 4 wk-old seedlings were cultured on two different basal media (Murashige & Skoog (MS) and Westvaco WV5 (WV5)). All media were supplemented with 6-benzylaminopurine (BAP), to stimulate the formation of axillary buds. Best bud induction was achieved, after 4 wk, on Westvaco WV5 medium (with 10-μM BAP) with rates close to 100%, and an average number of ~ 7 new buds formed per explant. Elongation took place for 6 wk on Westvaco WV5 medium containing activated charcoal (0.2% (w /v )) and without growth regulators. Rooting took place on half-strength Westvaco WV5 medium containing 9.8-μM indole-3-butyric acid (IBA). After 6 wk, root primordia were visible in ~ 43% of shoots. The acclimatization protocol was also optimized by controlling relative humidity, light/photoperiod, temperature, and nutrition, which led to an acclimatization success of ~ 89%. Flow cytometry analysis of DNA-ploidy did not show any variation between micropropagated plants and seedlings. With the protocol here described, it is possible to obtain a high number of genetically uniform plants per explant, 1 yr afterin vitro germination of slash pine seeds.An efficient protocol was developed for large-scale micropropagation of slash pine ( Pinus elliottii Engelm. var.elliottii ). For that, explants consisting of shoot apices 1.5–2.0 cm in length from 4 wk-old seedlings were cultured on two different basal media (Murashige & Skoog (MS) and Westvaco WV5 (WV5)). All media were supplemented with 6-benzylaminopurine (BAP), to stimulate the formation of axillary buds. Best bud induction was achieved, after 4 wk, on Westvaco WV5 medium (with 10-μM BAP) with rates close to 100%, and an average number of ~ 7 new buds formed per explant. Elongation took place for 6 wk on Westvaco WV5 medium containing activated charcoal (0.2% (w /v )) and without growth regulators. Rooting took place on half-strength Westvaco WV5 medium containing 9.8-μM indole-3-butyric acid (IBA). After 6 wk, root primordia were visible in ~ 43% of shoots. The acclimatization protocol was also optimized by controlling relative humidity, light/photoperiod, temperature, and nutrition, which led to an acclimatization success of ~ 89%. Flow cytometry analysis of DNA-ploidy did not show any variation between micropropagated plants and seedlings. With the protocol here described, it is possible to obtain a high number of genetically uniform plants per explant, 1 yr afterin vitro germination of slash pine seeds.An efficient protocol was developed for large-scale micropropagation of slash pine ( Pinus elliottii Engelm. var.elliottii ). For that, explants consisting of shoot apices 1.5–2.0 cm in length from 4 wk-old seedlings were cultured on two different basal media (Murashige & Skoog (MS) and Westvaco WV5 (WV5)). All media were supplemented with 6-benzylaminopurine (BAP), to stimulate the formation of axillary buds. Best bud induction was achieved, after 4 wk, on Westvaco WV5 medium (with 10-μM BAP) with rates close to 100%, and an average number of ~ 7 new buds formed per explant. Elongation took place for 6 wk on Westvaco WV5 medium containing activated charcoal (0.2% (w /v )) and without growth regulators. Rooting took place on half-strength Westvaco WV5 medium containing 9.8-μM indole-3-butyric acid (IBA). After 6 wk, root primordia were visible in ~ 43% of shoots. The acclimatization protocol was also optimized by controlling relative humidity, light/photoperiod, temperature, and nutrition, which led to an acclimatization success of ~ 89%. Flow cytometry analysis of DNA-ploidy did not show any variation between micropropagated plants and seedlings. With the protocol here described, it is possible to obtain a high number of genetically uniform plants per explant, 1 yr afterin vitro germination of slash pine seeds.An efficient protocol was developed for large-scale micropropagation of slash pine ( Pinus elliottii Engelm. var.elliottii ). For that, explants consisting of shoot apices 1.5–2.0 cm in length from 4 wk-old seedlings were cultured on two different basal media (Murashige & Skoog (MS) and Westvaco WV5 (WV5)). All media were supplemented with 6-benzylaminopurine (BAP), to stimulate the formation of axillary buds. Best bud induction was achieved, after 4 wk, on Westvaco WV5 medium (with 10-μM BAP) with rates close to 100%, and an average number of ~ 7 new buds formed per explant. Elongation took place for 6 wk on Westvaco WV5 medium containing activated charcoal (0.2% (w /v )) and without growth regulators. Rooting took place on half-strength Westvaco WV5 medium containing 9.8-μM indole-3-butyric acid (IBA). After 6 wk, root primordia were visible in ~ 43% of shoots. The acclimatization protocol was also optimized by controlling relative humidity, light/photoperiod, temperature, and nutrition, which led to an acclimatization success of ~ 89%. Flow cytometry analysis of DNA-ploidy did not show any variation between micropropagated plants and seedlings. With the protocol here described, it is possible to obtain a high number of genetically uniform plants per explant, 1 yr afterin vitro germination of slash pine seeds.[ABSTRACT FROM AUTHOR] - Published
- 2018
- Full Text
- View/download PDF
35. Recovery of cryopreserved embryogenic cultures of maritime pine--effect of cryoprotectant and suspension density.
- Author
-
Marum L, Estêvão C, Oliveira MM, Amâncio S, Rodrigues L, and Miguel C
- Subjects
- Carbohydrates pharmacology, Cell Line, Dimethyl Sulfoxide pharmacology, Nitrogen, Pinus cytology, Polyethylene Glycols pharmacology, Seeds cytology, Cryopreservation methods, Cryoprotective Agents pharmacology, Pinus growth & development, Seeds growth & development
- Abstract
Embryogenic cultures were obtained from seeds of open-pollinated maritime pine trees representing part of a breeding population. The aim of the present study was to develop and optimize a protocol for cryopreservation of Pinus pinaster somatic embryogenic tissue. The density of the embryogenic suspension and the type of pre-treatment significantly affected the recovery of cryopreserved embryogenic cultures, as evaluated by their survival and regrowth rate. An initial density of the suspension culture of 250 mg/ml improved the regrowth rate of the embryogenic lines. Pre-treatment with maltose 0.4 M significantly increased the regrowth rate when compared to the other tested carbohydrates. Also, the addition of DMSO in a mixture of PEG 4000 and sucrose (PSD solution), instead of DMSO alone, at the same final concentration, was clearly beneficial for recovery of cryopreserved tissues. This improved method for the cryopreservation of P pinaster embryogenic of cultures allowed the successful recovery of 97 percent of the lines stored in liquid of nitrogen.
- Published
- 2004
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