Open Access
The effect of changing temperature and agar concentration at proliferation stage in the final success of Aleppo pine somatic embryogenesis
(Instituto Nacional de Investigacion y Tecnologia Agraria y Alimentaria (INIA), 2017) Pereira, Catia; Montalbán, Itziar A.; Goicoa Mangado, Tomás; Ugarte Martínez, María Dolores; Correia, Sandra; Canhoto, Jorge M.; Moncaleán, Paloma; Estadística e Investigación Operativa; Estatistika eta Ikerketa Operatiboa
Aim of the study: The effect of physical and chemical conditions at proliferation stage was evaluated in order to elucidate if this stage is the determinant phase to induce a marked effect in Pinus halepensis somatic embryogenesis. Area of study: The study was conducted in research laboratories of Neiker (Arkaute, Spain). Material and methods: Pinus halepensis embryonal masses from ten embryogenic cell lines subjected to nine treatments (tissues cultured at three temperatures on media supplemented with three agar concentrations) at proliferation stage. Main results: Significant differences were observed among different proliferation conditions months later at the end of maturation, germination and acclimatization stages. Research highlights: Aleppo pine embryonal masses are cultured under standard conditions on a culture medium supplemented with 4.5 g/L Gelrite® at 23ºC. However, better results in terms of plantlet production can be obtained proliferating the embryonal masses at 18ºC in a culture media with significantly lower water availability.
Open Access
Hybrid pine (Pinus attenuata × Pinus radiata) somatic embryogenesis: what do you prefer, mother or nurse?
(MDPI, 2021) Montalbán, Itziar A.; Castander Olarieta, Ander; Hargreaves, Cathy L.; Gough, Keiko; Reeves, Cathie B.; Ballekom, Shaf van; Goicoa Mangado, Tomás; Ugarte Martínez, María Dolores; Moncaleán, Paloma; Estatistika, Informatika eta Matematika; Institute for Advanced Materials and Mathematics - INAMAT2; Estadística, Informática y Matemáticas; Universidad Pública de Navarra / Nafarroako Unibertsitate Publikoa
Development of hybrid pines of Pinus radiata D. Don for commercial forestry presents an opportunity to diversify the current resource of plant material. Climate change and different land uses pose challenges, making alternative species necessary to guarantee wood and non-wood products in the future. Pinus radiata var. cedrosensis × Pinus attenuata hybrid possesses different attributes, such as tolerance to drought conditions, better growth and resistance to snow damage at higher altitudes, and more importantly, different wood quality characteristics. Embryogenic cell lines were successfully initiated reciprocal hybrids using as initial explants megagametophytes, excised zygotic embryos and excised zygotic embryos plus nurse culture. However, the questions raised were: does the initiation environment affect the conversion to somatic plantlets months later? Does the mother tree or the cross have an effect on the conversion to somatic plantlets? In the present work we analysed the maturation rate, number of somatic embryos, germination rate, and the ex-vitro growth in cell lines derived from different initiation treatments, mother tree species, and crosses. Differences were not observed for in vitro parameters such as maturation and germination. However, significant differences were observed due to the mother tree species in relation with the ex-vitro growth rates observed, being higher those in which P. radiata acted as a mother. Moreover, embryogenic cell lines from these hybrids were stored at −80◦C and regenerated after one and five years.
Open Access
Temperature and water availability during maturation affect the cytokinins and auxins profile of radiata pine somatic embryos
(Frontiers Media, 2018) Moncaleán, Paloma; García Mendiguren, Olatz; Novák, Ondrej; Strnad,Miroslav; Goicoa Mangado, Tomás; Ugarte Martínez, María Dolores; Montalbán, Itziar A.; Estadística, Informática y Matemáticas; Estatistika, Informatika eta Matematika
Somatic embryogenesis (SE) provides us a potent biotechnological tool to manipulate the physical and chemical conditions (water availability) along the process and to study their effect in the final success in terms of quantity of somatic embryos produced. In the last years, our research team has been focused on the study of different aspects of the SE in Pinus spp. One of the main aspects affecting SE is the composition of culture media; in this sense, phytohormones play one of the most crucial roles in this propagation system. Many studies in conifers have shown that different stages of SE and somatic embryo development are correlated with distinct endogenous phytohormone profiles under the stress conditions needed for the process (i.e., cytokinins play a regulatory role in stress signaling, which it is essential for radiata pine SE). Based on this knowledge, the aim of this study was to test the effect of different temperatures (18, 23, and 28°C) and gelling agent concentrations (8, 9, and 10 gL-1) during the maturation stage of Pinus radiata SE in maturation and germination rates. Parallel, phytohormone profile of somatic embryos developed was evaluated. In this sense, the highest gellan gum concentration led to significantly lower water availability. At this gellan gum concentration and 23°C a significantly higher number of somatic embryos was obtained and the overall success of the process increased with respect to other treatments assayed. The somatic embryos produced in these conditions showed the highest concentration of iP-type cytokinins and total ribosides. Although, the different conditions applied during maturation of somatic embryos led to different hormonal profiles, they did not affect the ex vitro survival of the resulting somatic plants, where no significant differences were observed.

Goicoa Mangado, Tomás

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