Caracterización e inducción de embriogénesis somática en Araucaria araucana (molina) k. koch.: especie en peligro (EN).
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Date
2024
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Publisher
Universidad de Concepción
Abstract
Araucaria araucana, especie endémica de gran importancia ecológica y cultural en Chile, enfrenta amenazas atribuibles al cambio climático y la acción antrópica, principalmente la recolección indiscriminada de su semilla, provocando pérdida de su biodiversidad. A través de la embriogénesis somática buscamos masificar genotipos de interés y propagarlos con fines de conservación. Identificamos por primera vez las condiciones de cultivo que permiten desencadenar la inducción de proembriones somáticos a partir de semillas inmaduras de tres orígenes, costero (Villa Las Araucarias), Cordillera de Nahuelbuta (Trongol Alto) y Cordillera de los Andes (Malalcahuello). Los resultados muestran que existen líneas celulares embriogénicas con potencial embriogénico de las tres procedencias. La inducción de estructuras embriogénicas se obtiene a los 45 días cuando se utilizó el medio BM suplementado con NAA (11 µM), 6-BA (2,8 µM) y Kin (2,8 µM). Se evaluaron por 105 días siete líneas celulares donde, las mejores tasas de proliferación se obtuvieron en el medio MP1 suplementado con (5 µM 2,4-D + 2,8 µM 6-BA y Kin). Además de cuantificar PEM en estas líneas, se relacionó con los niveles de poliaminas libres y razón Put:Spd cuantificadas a partir de muestras de los tratamientos de proliferación, donde apreciamos una marcada diferencia en la bioquímica de las ECLs entre las tres líneas celulares evaluadas y el crecimiento gradual de los cultivos, lo que sugiere que la determinación de PAs es un eficiente marcador bioquímico para testear ECLs en A. araucana.
La transición de PEM a estadios embrionarios posteriores sigue bajo investigación; sin embargo, se observaron estadios tempranos de embriones somáticos en el tratamiento con maltosa (3% p/v), PEG 3350 (7% p/v) y ABA (34 y 68 µM). La maduración completa de los embriones somáticos aún está en proceso, lo que demuestra que la técnica puede ser una alternativa viable en la conservación de esta especie.
Araucaria araucana, an endemic species of great ecological and cultural importance in Chile, faces threats attributable to climate change and anthropic action, mainly the indiscriminate collection of its seed, causing loss of its biodiversity. Through somatic embryogenesis we seek to massify genotypes of interest and propagate them for conservation purposes. We identified for the first time the culture conditions that allow triggering the induction of somatic proembryos from immature seeds from three origins, coastal (Villa Las Araucarias), Cordillera de Nahuelbuta (Trongol Alto) and Cordillera de Los Andes (Malalcahuello). The results show that there are embryogenic cell lines with embryogenic potential from the three origins. The induction of embryogenic structures was obtained at 45 days when the BM medium supplemented with NAA (11 µM), 6-BA (2.8 µM) and Kin (2.8 µM) was used. Seven cell lines were evaluated for 105 days where the best proliferation rates were obtained in MP1 medium supplemented with (5 µM 2,4-D + 2.8 µM 6-BA and Kin). In addition to quantifying PEM in these lines, it was related to the levels of free polyamines and Put:Spd ratio quantified from samples of the proliferation treatments, where we appreciated a marked difference in the biochemistry of the ECLs between the three cell lines evaluated and the gradual growth of the cultures, which suggests that the determination of PAs is an efficient biochemical marker to test ECLs in A. araucana. The transition from PEM to later embryonic stages remain under investigation; However, early stages of somatic embryos were observed in treatment with maltose (3% w/v), PEG 3350 (7% w/v) and ABA (34 and 68 µM). The complete maturation of the somatic embryos is still in process, which demonstrates that the technique can be a viable alternative in the conservation of this species.
Araucaria araucana, an endemic species of great ecological and cultural importance in Chile, faces threats attributable to climate change and anthropic action, mainly the indiscriminate collection of its seed, causing loss of its biodiversity. Through somatic embryogenesis we seek to massify genotypes of interest and propagate them for conservation purposes. We identified for the first time the culture conditions that allow triggering the induction of somatic proembryos from immature seeds from three origins, coastal (Villa Las Araucarias), Cordillera de Nahuelbuta (Trongol Alto) and Cordillera de Los Andes (Malalcahuello). The results show that there are embryogenic cell lines with embryogenic potential from the three origins. The induction of embryogenic structures was obtained at 45 days when the BM medium supplemented with NAA (11 µM), 6-BA (2.8 µM) and Kin (2.8 µM) was used. Seven cell lines were evaluated for 105 days where the best proliferation rates were obtained in MP1 medium supplemented with (5 µM 2,4-D + 2.8 µM 6-BA and Kin). In addition to quantifying PEM in these lines, it was related to the levels of free polyamines and Put:Spd ratio quantified from samples of the proliferation treatments, where we appreciated a marked difference in the biochemistry of the ECLs between the three cell lines evaluated and the gradual growth of the cultures, which suggests that the determination of PAs is an efficient biochemical marker to test ECLs in A. araucana. The transition from PEM to later embryonic stages remain under investigation; However, early stages of somatic embryos were observed in treatment with maltose (3% w/v), PEG 3350 (7% w/v) and ABA (34 and 68 µM). The complete maturation of the somatic embryos is still in process, which demonstrates that the technique can be a viable alternative in the conservation of this species.
Description
Tesis presentada para optar al grado académico de Magister en Ciencias Forestales
Keywords
Araucaria, Biodiversidad, Biotecnología vegetal