RESUMEN
BACKGROUND: Theobroma cacao is a major source of flavonoids such as catechins and their monomers proanthocyanidins (PAs), widely studied for their potential benefits in cardiovascular diseases. Light has been shown to promote plant secondary metabolite production in vitro. In this study, cacao cells cultured in 7.5 L stirred tank photobioreactors (STPs) were exposed to a change of white to blue LED lights for 28 days (d). RESULTS: Transcriptomic analyses were performed in three time points comparing changing expression patterns, after cell exposure to white light (d0-VS-d14), after a shift from white to blue light (d14-VS-d15), and after an extended period of blue light for the following 15 days (d15-VS-d28). Under white light, there was enrichment in metabolic pathways associated with cell growth (carbon, glycolysis, and amino acid biosynthesis) accompanied by a significant increase in the PAs content. In the shift to blue light, further increase in PAs content was observed concomitantly with the significant expression of TWO-COMPONENT RESPONSE REGULATOR genes involved in the early stress responses via circadian clock and hormone pathways. Under blue light exposure, we observed a depletion of PAs content associated with ROS-mediated stress pathways. CONCLUSIONS: Light effects on large-scale cell cultures in photobioreactors are complex and pleiotropic; however, we have been able to identify key regulatory players upstream cacao flavonoid biosynthesis in STPs, including TWO-COMPONENT SYSTEM and ROS-signaling genes. The crosstalk between flavonoid biosynthesis and regulatory networks led to understand the dynamics of flavonoid production and degradation in response to light-driven ROS signals. This can be used to optimize the time, and the yield of in vitro targeted metabolites in large-scale culture systems.
Asunto(s)
Cacao , Cacao/genética , Flavonoides , Regulación de la Expresión Génica de las Plantas , Fotobiorreactores , TranscriptomaRESUMEN
Las Zamiaceas son plantas relictuales consideradas fósiles vivientes. En Colombia, el 65% de esta familia se encuentra en alguna categoría de amenaza, por la destrucción del hábitat e intensa recolección. Teniendo en cuenta que entre las ventajas de la propagación in vitro está la conservación ex situ de germoplasma, el presente trabajo tuvo como objetivo evaluar el potencial de regeneración de plantas de Z. incognita a partir de explantes foliares y embriones cigoticos. Se evaluó el efecto de diferentes combinaciones de Auxinas (2,4-D y ANA) y citoquininas (KIN, BAP y TDZ) sobre la formación de callo y la regeneración de brotes (directa o indirecta), utilizando como medio basal MS (MB1) y medio basal B5 modificado (MB2). La formación de callo se presentó sobre un amplio rango de concentraciones de 2,4-D con KIN y 2,4-D con BAP, independientemente del medio basal, pero no en los explantes tratados con ANA más KIN o TDZ. Para los explantes foliares no hubo respuesta a la formación de embriones somáticos y/o brotes con las combinaciones y concentraciones hormonales evaluadas, no obstante los callos inducidos en MB2 con 2,4-D (0,22 mg/l) y BAP (0, 1, 2, 3 mg/l) fueron diferentes, su aspecto nodular, color crema y apariencia proembriogénica coincidió con una gran cantidad de células meristemáticas potenciales para el proceso de regeneración. A partir de embriones cigoticos inmaduros se logró la formación de embriones somáticos en el medio MB2 exento de reguladores o conteniendo 2,4-D solo (0,22 mg/l) y en combinación con BAP (1 mg/l), sin lograr el proceso de conversión a plántulas.
Zamiaceas are relict plants considered living fossils. In Colombia, 65% of this family is under some threat category due to their habitat destruction and their intense collection. Given that the advantages of in vitro propagation is ex situ conservation of germoplasm, this study aimed to evaluate the regeneration potential of Z incognita plants from leaf explants and zygotic embryos. The effect of different combinations of auxin (2.4-D and NAA) and cytokinins (KIN, BAP and TDZ) was evaluated on the formation of callus and shoot regeneration (direct or indirect), using MS (MB1) basal medium and B5 (MB2) basal modified medium. The callus formation was presented over a wide concentration range of 2.4-D with KIN and 2.4-D with BAP, regardless of the basal medium, but not in explants treated with ANA more TDZ or KIN. For leaf explants there was no response to the formation of somatic embryos or shoots with hormonal combinations and concentrations evaluated; however, MB2 calluses induced with 2.4-D (0.22 mg / l) and BAP (0. 1 , 2. 3 mg / l) were different, their nodular aspect, cream color and pro-embryogenic appearance coincided with a lot of potential meristematic cells for the regeneration process. From immature zygotic embryos, somatic embryo formation in the MB2 medium was achieved without growth regulators or containing 2.4-D alone (0.22 mg /l) or 2.4-D in combination with BAP (1 mg/l) without achieving the conversion process to seedlings.