INVESTIGADORES
RODRIGUEZ TALOU Julian
congresos y reuniones científicas
Título:
Methyl jasmonate elicitation and reactive oxygen species production in Rubia tinctorum cell suspension cultures
Autor/es:
PERASSOLO, M; QUEVEDO, CV; BUSTO, VD; CARDILLO AB; MARTÍNEZ, CA; GIULIETTI, AM; RODRIGUEZ TALOU , J
Lugar:
Barcelona
Reunión:
Congreso; 14th European Congress on Biotechnology,; 2009
Institución organizadora:
European Federation of Biotechnology
Resumen:
Elicitation is a useful strategy to induce accumulation of secondary metabolites in plants. Besides the production of secondary metabolites, the defense response against elicitation also triggers the generation of reactive oxygen species (ROS), and can also lead to cell death. In particular, methyl jasmonate (MeJ) is a well-described agent that has been widely used to enhance anthraquinone (AQs) production in Rubia tinctorum. AQs biosynthesis involves different metabolic routes: o-succinylbenzoate, precursor of A and B rings, is derived from -ketoglutarate and isochorismic acid. This is produced by the isochorismate synthase from chorismic acid, which is the end-product of shikimate pathway. C ring is derived from the terpenoid pathway. It has been proposed that proline cycle could be coupled to the pentose phosphate pathway (PPP) as the NADP+ generated by proline reduction from glutamate could act as cofactor of the first enzymes of the PPP. This pathway generates erithrose-4-phosphate, the substrate of the shikimate pathway. The aim of the present work was to study the relationship between MeJ elicitation, ROS, proline cycle and anthraquinone production in R. tinctorum cell suspension cultures. Cell cultures were treated with MeJ, diphenyl iodonium (DPI, an inhibitor of H2O2 generation) and a glucose/glucose oxidase system (Glu/GOD, a H2O2 generating-system). After 48 hours of culture, MeJ increased AQs content (53%, P < 0.05) compared to control cells, but not after 24 hours. Treatment with DPI of both control and MeJ-treated cells showed less AQs accumulation at 24 (26% and 33% less) and at 48 hours of culture (18.5% and 51% less, compared to both treatments without DPI). The Glu/GOD system induced AQs production (14 and 48% more than the control treatment, after 24 and 48 hours of culture). MeJ treatment increased proline content at 48 hours of culture (22% more), while it was decreased when DPI was added to MeJ-treated cells (28% less). Relationship between ROS and elicitation is currently under study.Rubia tinctorum. AQs biosynthesis involves different metabolic routes: o-succinylbenzoate, precursor of A and B rings, is derived from -ketoglutarate and isochorismic acid. This is produced by the isochorismate synthase from chorismic acid, which is the end-product of shikimate pathway. C ring is derived from the terpenoid pathway. It has been proposed that proline cycle could be coupled to the pentose phosphate pathway (PPP) as the NADP+ generated by proline reduction from glutamate could act as cofactor of the first enzymes of the PPP. This pathway generates erithrose-4-phosphate, the substrate of the shikimate pathway. The aim of the present work was to study the relationship between MeJ elicitation, ROS, proline cycle and anthraquinone production in R. tinctorum cell suspension cultures. Cell cultures were treated with MeJ, diphenyl iodonium (DPI, an inhibitor of H2O2 generation) and a glucose/glucose oxidase system (Glu/GOD, a H2O2 generating-system). After 48 hours of culture, MeJ increased AQs content (53%, P < 0.05) compared to control cells, but not after 24 hours. Treatment with DPI of both control and MeJ-treated cells showed less AQs accumulation at 24 (26% and 33% less) and at 48 hours of culture (18.5% and 51% less, compared to both treatments without DPI). The Glu/GOD system induced AQs production (14 and 48% more than the control treatment, after 24 and 48 hours of culture). MeJ treatment increased proline content at 48 hours of culture (22% more), while it was decreased when DPI was added to MeJ-treated cells (28% less). Relationship between ROS and elicitation is currently under study.o-succinylbenzoate, precursor of A and B rings, is derived from -ketoglutarate and isochorismic acid. This is produced by the isochorismate synthase from chorismic acid, which is the end-product of shikimate pathway. C ring is derived from the terpenoid pathway. It has been proposed that proline cycle could be coupled to the pentose phosphate pathway (PPP) as the NADP+ generated by proline reduction from glutamate could act as cofactor of the first enzymes of the PPP. This pathway generates erithrose-4-phosphate, the substrate of the shikimate pathway. The aim of the present work was to study the relationship between MeJ elicitation, ROS, proline cycle and anthraquinone production in R. tinctorum cell suspension cultures. Cell cultures were treated with MeJ, diphenyl iodonium (DPI, an inhibitor of H2O2 generation) and a glucose/glucose oxidase system (Glu/GOD, a H2O2 generating-system). After 48 hours of culture, MeJ increased AQs content (53%, P < 0.05) compared to control cells, but not after 24 hours. Treatment with DPI of both control and MeJ-treated cells showed less AQs accumulation at 24 (26% and 33% less) and at 48 hours of culture (18.5% and 51% less, compared to both treatments without DPI). The Glu/GOD system induced AQs production (14 and 48% more than the control treatment, after 24 and 48 hours of culture). MeJ treatment increased proline content at 48 hours of culture (22% more), while it was decreased when DPI was added to MeJ-treated cells (28% less). Relationship between ROS and elicitation is currently under study.+ generated by proline reduction from glutamate could act as cofactor of the first enzymes of the PPP. This pathway generates erithrose-4-phosphate, the substrate of the shikimate pathway. The aim of the present work was to study the relationship between MeJ elicitation, ROS, proline cycle and anthraquinone production in R. tinctorum cell suspension cultures. Cell cultures were treated with MeJ, diphenyl iodonium (DPI, an inhibitor of H2O2 generation) and a glucose/glucose oxidase system (Glu/GOD, a H2O2 generating-system). After 48 hours of culture, MeJ increased AQs content (53%, P < 0.05) compared to control cells, but not after 24 hours. Treatment with DPI of both control and MeJ-treated cells showed less AQs accumulation at 24 (26% and 33% less) and at 48 hours of culture (18.5% and 51% less, compared to both treatments without DPI). The Glu/GOD system induced AQs production (14 and 48% more than the control treatment, after 24 and 48 hours of culture). MeJ treatment increased proline content at 48 hours of culture (22% more), while it was decreased when DPI was added to MeJ-treated cells (28% less). Relationship between ROS and elicitation is currently under study.R. tinctorum cell suspension cultures. Cell cultures were treated with MeJ, diphenyl iodonium (DPI, an inhibitor of H2O2 generation) and a glucose/glucose oxidase system (Glu/GOD, a H2O2 generating-system). After 48 hours of culture, MeJ increased AQs content (53%, P < 0.05) compared to control cells, but not after 24 hours. Treatment with DPI of both control and MeJ-treated cells showed less AQs accumulation at 24 (26% and 33% less) and at 48 hours of culture (18.5% and 51% less, compared to both treatments without DPI). The Glu/GOD system induced AQs production (14 and 48% more than the control treatment, after 24 and 48 hours of culture). MeJ treatment increased proline content at 48 hours of culture (22% more), while it was decreased when DPI was added to MeJ-treated cells (28% less). Relationship between ROS and elicitation is currently under study.2O2 generation) and a glucose/glucose oxidase system (Glu/GOD, a H2O2 generating-system). After 48 hours of culture, MeJ increased AQs content (53%, P < 0.05) compared to control cells, but not after 24 hours. Treatment with DPI of both control and MeJ-treated cells showed less AQs accumulation at 24 (26% and 33% less) and at 48 hours of culture (18.5% and 51% less, compared to both treatments without DPI). The Glu/GOD system induced AQs production (14 and 48% more than the control treatment, after 24 and 48 hours of culture). MeJ treatment increased proline content at 48 hours of culture (22% more), while it was decreased when DPI was added to MeJ-treated cells (28% less). Relationship between ROS and elicitation is currently under study.2O2 generating-system). After 48 hours of culture, MeJ increased AQs content (53%, P < 0.05) compared to control cells, but not after 24 hours. Treatment with DPI of both control and MeJ-treated cells showed less AQs accumulation at 24 (26% and 33% less) and at 48 hours of culture (18.5% and 51% less, compared to both treatments without DPI). The Glu/GOD system induced AQs production (14 and 48% more than the control treatment, after 24 and 48 hours of culture). MeJ treatment increased proline content at 48 hours of culture (22% more), while it was decreased when DPI was added to MeJ-treated cells (28% less). Relationship between ROS and elicitation is currently under study.P < 0.05) compared to control cells, but not after 24 hours. Treatment with DPI of both control and MeJ-treated cells showed less AQs accumulation at 24 (26% and 33% less) and at 48 hours of culture (18.5% and 51% less, compared to both treatments without DPI). The Glu/GOD system induced AQs production (14 and 48% more than the control treatment, after 24 and 48 hours of culture). MeJ treatment increased proline content at 48 hours of culture (22% more), while it was decreased when DPI was added to MeJ-treated cells (28% less). Relationship between ROS and elicitation is currently under study.