Análisis de la expresión del gen fitoeno sintasa (psy) en la biosíntesis de carotenoides en Dunaliella sp. expuesta a condiciones de estrés mediante RT-PCR semicuantitativa
DOI:
https://doi.org/10.34117/bjdv12n2-005Keywords:
Dunaliella, fitoeno sintasa, carotenoides, RT-PCR semicuantitativaAbstract
El mecanismo regulador de los carotenoides en Dunaliella sp. no se conoce bien, pero se sabe que el gen psy es responsable de activar la vía metabólica de los carotenoides. En este estudio, se comparó la expresión del gen psy en la biosíntesis de carotenoides en cultivos vegetativos y estresados de Dunaliella sp. (aislado PM018) utilizando RT-PCR semicuantitativa. Para ello, el aislado se cultivó en medio Guillard F/2 y se sometió a 200 µmol de fotones m⁻² s⁻¹ con privación de nutrientes durante 24 días, y se analizó la expresión relativa del gen inducido mediante RT-PCR semicuantitativa utilizando densitometría normalizada con el gen 18S rRNA. Los resultados revelaron que el cultivo vegetativo alcanzó una densidad de 19 x 105 células/mL, observándose cambios morfológicos (de ovoide a redondo) y cambios de color (de verde a amarillo) tras la fase de estrés. Antes de los niveles de expresión, se verificó la calidad del ARN, obteniendo una concentración de 454,65 ± 279,02 ng/µL en crecimiento vegetativo y 989 ± 191,73 ng/µL bajo estrés. Finalmente, en la fase vegetativa se obtuvo una expresión relativa del gen psy de 0,16 ± 0,002, y se obtuvo una expresión más alta de 2,72 ± 0,001 en condiciones de estrés. Se concluye que la transcripción del gen psy está influenciada por la combinación de factores de estrés, y que la técnica de RT-PCR semicuantitativa permitió la detección de alta sensibilidad en la expresión génica.
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