Goicoa Mangado, Tomás
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Goicoa Mangado
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Tomás
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Estadística, Informática y Matemáticas
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InaMat2. Instituto de Investigación en Materiales Avanzados y Matemáticas
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Publication Open Access Induction of radiata pine somatic embryogenesis at high temperatures provokes a long-term decrease in DNA methylation/hydroxymethylation and differential expression of stress-related genes(MDPI, 2020) Castander Olarieta, Ander; Pereira, Catia; Sales, Ester; Meijón, Mónica; Arrillaga, Isabel; Goicoa Mangado, Tomás; Ugarte Martínez, María Dolores; Cañal, María Jesús; Moncaleán, Paloma; Montalbán, Itziar A.; Estadística, Informática y Matemáticas; Estatistika, Informatika eta Matematika; Institute for Advanced Materials and Mathematics - INAMAT2Based on the hypothesis that embryo development is a crucial stage for the formation of stable epigenetic marks that could modulate the behaviour of the resulting plants, in this study, radiata pine somatic embryogenesis was induced at high temperatures (23◦ C, eight weeks, control; 40◦ C, 4 h; 60◦ C, 5 min) and the global methylation and hydroxymethylation levels of emerging embryonal masses and somatic plants were analysed using LC-ESI-MS/ MS-MRM. In this context, the expression pattern of six genes previously described as stress-mediators was studied throughout the embryogenic process until plant level to assess whether the observed epigenetic changes could have provoked a sustained alteration of the transcriptome. Results indicated that the highest temperatures led to hypomethylation of both embryonal masses and somatic plants. Moreover, we detected for the first time in a pine species the presence of 5-hydroxymethylcytosine, and revealed its tissue specificity and potential involvement in heat-stress responses. Additionally, a heat shock protein-coding gene showed a down-regulation tendency along the process, with a special emphasis given to embryonal masses at first subculture and ex vitro somatic plants. Likewise, the transcripts of several proteins related with translation, oxidative stress response, and drought resilience were differentially expressed.Publication Open Access High temperature and water deficit cause epigenetic changes in somatic plants of Pinus radiata D. Don(Springer, 2022) Marqués do Nascimento, Antonia Maiara; Montalbán, Itziar A.; Llamazares de Miguel, Diego; Goicoa Mangado, Tomás; Ugarte Martínez, María Dolores; Moncaleán, Paloma; Estadística, Informática y Matemáticas; Estatistika, Informatika eta MatematikaCurrent climate changes imply an imminent risk for forest species. In this context, somatic embryogenesis is a valuable tool to study the response of plants to different abiotic stresses. Based on this, we applied a high-temperature regime (50 °C, 5 min) during the maturation of Pinus radiata D. Don embryogenic masses in order to evaluate the development of an epigenetic memory months later. Therefore, somatic plants (SP) resulting from somatic embryos (ses) maturated at control temperature and cultivated in a greenhouse were submitted to heat stress (40 °C, 2 h, 10 days; 23 °C, 10 days) or at a control temperature (23 °C, 20 days); while another 20 SP resulting from ses maturated in the two temperature regimes and cultivated in the greenhouse were submitted to drought stress or weekly irrigated. All plants were evaluated for relative water content, water potential, electrolyte leakage, stomatal conductance, transpiration, methylation (5-mC) and hydroxymethylation (5-hmC) levels. The results showed that the SP obtained from ses maturated at 50 °C showed an adaptation to drought stress based on water potential and transpiration. Furthermore, SP kept under heat stress in a greenhouse showed lower 5-hmC levels than SP kept at 23 °C. Furthermore, the 5-hmC and 5-hmC/5-mC ratio showed a significantly negative correlation with changes in water potential; and a significantly negative correlation was observed between the levels of stomatal conductance and 5-mC. We conclude that the manipulation of conditions during the maturation process in somatic embryogenesis modulates the physiological characteristics of the SP obtained.