ABSTRACT
Cell suspension cultures of Thevetia peruviana were established under dark for 19 days to investigate kinetic behavior related to biomass, substrate, cardiac glycoside, polyphenols, reactive oxygen species and anti-oxidant activity. The results showed high biomass production (18.80gDW/L) as well as sucrose consumption in 7 days. Preferential glucose over fructose consumption was observed. Intracellular production of cardiac glycosides reached 2.58mg DE/gDW at day 19. Highest extracellular production was reached between day 2 and 7 (6.19mg DE/L). Highest extracellular phenolic content was 80.61 ± 5.16mg GAE/L at day 7. Intracellular phenolic content increased to 2.76 ± 0.14mg GAE/gFW at day 7 and remained constant until day 19. ROS production at day 7 could be related to sucrose and glucose total consumption. Pearson Product-Moment Correlation Coefficient (ρ) showed that the phenolic compounds in cell suspension cultures of T. peruviana were responsible for the observed anti-oxidant activity. All together, these results give the first steps in metabolic behavior in cell suspension cultures of T. peruviana.
Se establecieron cultivos en suspensión de la especie vegetal Thevetia peruviana en oscuridad, durante 19 días, para estudiar el comportamiento cinético de producción de biomasa, consumo de sustrato, producción de glicósidos cardiotónicos, polifenoles, especies reactivas de oxígeno y actividad antioxidante. Los resultados mostraron una alta producción de biomasa (18,80g PS/L), al igual que consumo total de sacarosa, a los 7 días de cultivo. Se observó un consumo preferencial de glucosa sobre fructosa durante todo el cultivo. La producción de glicósidos cardiotónicos intracelulares alcanzó valores de 2,58mg ED/g PS, al día 19. La mayor producción extracelular (6,19mg ED/L), se alcanzó entre los días 2 y 7. El mayor contenido de compuestos fenólicos extracelular fue de 80,61 ± 5,16mg GAE/L, en el día 7. El contenido de compuestos fenólicos intracelulares incrementó a 2,76 ± 0,14mg AGE/gPF, al día 7 y se mantuvo constante, hasta el día 19. La producción de EROs, al día 7, puede estar relacionada con el consumo total de sacarosa y glucosa. El coeficiente de correlación producto-momento de Pearson (ρ) indicó que los compuestos fenólicos en cultivos celulares en suspensión de T. peruviana eran los responsables de la actividad antioxidante observada. En conjunto, estos resultados brindan las primeras bases relacionadas al comportamiento metabólico de cultivos celulares en suspensión, de T. peruviana.
ABSTRACT
OBJECTIVE@#To evaluate the impact of plant growth regulators including kinetin (KN), benzyl adenine and naphthalene acetic acid, yeast extract and casein hydrolyzate on biomass accumulation of Vietnamese ginseng Panax vietnamensis (P. vietnamensis) in cell suspension culture.@*METHODS@#Cell suspension cultures were established from friable calluses derived from leaves and petioles of 3-year-old in-vitro P. vietnamensis plants. The cell suspension cultures were grown in Murashige and Skoog basal media supplemented with various concentrations of KN, benzyl adenine, naphthalene acetic acid, and yeast extract and casein hydrolyzate.@*RESULTS@#All tested factors generated an increase in the cell biomass of P. vietnamensis in suspension culture, but the impact of each varies depended on the factor type, concentration, and incubation period. Addition of 2.0 mg/L KN resulted in the largest biomass increase after 24 d, (57.0 ± 0.9) and (3.1 ± 0.1) mg/mL fresh and dry weight, respectively, whereas addition of benzyl adenine or naphthalene acetic acid produced optimum levels of Panax cell biomass at 1.0 and 1.5 mg/L, respectively. Addition of the elicitor yeast extract led to a 1.4-2.4 fold increase in biomass of P. vietnamensis, while addition of casein hydrolyzate enhanced biomass accumulation 1.8-2.6 fold.@*CONCLUSIONS@#The addition of each factor causes significant changes in biomass accumulation of P. vietnamensis. The largest biomass accumulation is from cultures grown in MS media containing 2.0 mg/L KN for 24 d. The outcome of the present study provides new insights into the optimal suspension culture conditions for studies on the in vitro cell biomass production of P. vietnamensis.
ABSTRACT
Objective To evaluate the impact of plant growth regulators including kinetin (KN), benzyl adenine and naphthalene acetic acid, yeast extract and casein hydrolyzate on biomass accumulation of Vietnamese ginseng Panax vietnamensis (P. vietnamensis) in cell suspension culture. Methods Cell suspension cultures were established from friable calluses derived from leaves and petioles of 3-year-old in-vitro P. vietnamensis plants. The cell suspension cultures were grown in Murashige and Skoog basal media supplemented with various concentrations of KN, benzyl adenine, naphthalene acetic acid, and yeast extract and casein hydrolyzate. Results All tested factors generated an increase in the cell biomass of P. vietnamensis in suspension culture, but the impact of each varies depended on the factor type, concentration, and incubation period. Addition of 2.0 mg/L KN resulted in the largest biomass increase after 24 d, (57.0 ± 0.9) and (3.1 ± 0.1) mg/mL fresh and dry weight, respectively, whereas addition of benzyl adenine or naphthalene acetic acid produced optimum levels of Panax cell biomass at 1.0 and 1.5 mg/L, respectively. Addition of the elicitor yeast extract led to a 1.4–2.4 fold increase in biomass of P. vietnamensis, while addition of casein hydrolyzate enhanced biomass accumulation 1.8–2.6 fold. Conclusions The addition of each factor causes significant changes in biomass accumulation of P. vietnamensis. The largest biomass accumulation is from cultures grown in MS media containing 2.0 mg/L KN for 24 d. The outcome of the present study provides new insights into the optimal suspension culture conditions for studies on the in vitro cell biomass production of P. vietnamensis.
ABSTRACT
Hormones are the main materials which regulate the plant growth and the secondary metabolites formation of plant. The effects of hormones on cell biomass and metabolites content and the application progress of honrmones in plant cell suspension culture is reviewed. It covers the influence of exogenous hormones’ categories, concentration and proportion on cell biomass and metabolites content in plant cell suspension culture, the developmental course of the endogenous hormones determination, the changes of endogenous hormones content and its effects on the cell biomass and metabolites content, the relation of interaction between exogenous hormones and endogenous hormones, the study on the effect of new hormone varieties on the culture.