ABSTRACTS
ten days of culture (P < 0.01). These findings could be correlated
with PPP stimulation.
doi:10.1016/j.nbt.2009.06.807
4.1.26
Methyl jasmonate elicitation and reactive oxygen species
production in Rubia tinctorum cell suspension cultures
M. Perassolo ∗ , C.V. Quevedo, V.D. Busto, A.B. Cardillo,
C.A. Martínez, A.M. Giulietti, J. Rodríguez Talou
Facultad de Farmacia y Bioquimica, UBA, Buenos Aires, Argentina
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 H2 O2 generation) and a glucose/glucose oxidase system (Glu/GOD, a H2 O2 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.
doi:10.1016/j.nbt.2009.06.808
S334
www.elsevier.com/locate/nbt
New Biotechnology · Volume 25S · September 2009
4.1.27
Overexpression of the Arabidopsis thaliana squalene synthase gene in Withania coagulans hairy root cultures
increases the biosynthesis of phytosterols and withanolides
M. Hossein Mirjalili 1,∗ , M. Bonfill 3 , E. Moyano 2 , R.M. Cusido 3 ,
J. Palazón 3
1
Department of Agriculture, Medicinal Plants and Drugs Research Institute,
Shahid Beheshti University, G. C., Evin, Tehran, Iran
2
Departament de Ciencies Experimentals i de la Salut, Universitat Pompeu
Fabra, Barcelona, Spain
3
Laboratori de Fisiologia Vegetal, Facultat de Farmacia, Universitat de
Barcelona, Barcelona, Spain
Squalene synthase (SS) is able to dimerize two molecules of farnesyl
diphosphate to synthesize squalene, a shared precursor in steroid
and triterpenoid biosynthesis in plants. The SS1 gene encoding
SS from Arabidopsis thaliana was introduced in Withania coagulans
under the control of the CaMV35S promoter together with the TDNA of Agrobacterium rhizogenes A4. The engineered hairy roots
were studied for withanolide production and phytosterol accumulation and the results were compared with those obtained from
control roots harbouring only the T-DNA from pRiA4. Both kinds
of roots were able to biosynthesize the most important bioactive
withanolides, withanolide A and withaferin A, the latter only in
very low concentrations. The engineered root line SS135 exhibited the highest withanolide content (4.93 g/g DW), more than
doubling the most productive control (C26), and thereby showing the effectiveness of the transgene in increasing withanolide
production. This root type also exhibited a greater capacity than
the control (nearly threefold) for accumulating squalene-derived
primary metabolites such as phytosterols.
doi:10.1016/j.nbt.2009.06.809
4.1.28
Pathway activity profiling (papi): an integration system
for metabolomics data
R. Aggio ∗ , K. Ruggiero, S. Villas-Boas
The Auckland University, Auckland, New Zealand
Metabolomics has gained increased popularity in the last ten years.
This popularity comes from its use as a functional genomics tool
and its diverse range of potential applications. The ultimate goal
for metabolomics is its integration with other omics-related data.
However, today there is no straightforward method able to combine the results from different omics. These type of data are usually
complex, difficult to interpret and even more difficult to correlate. As a consequence, the data analysis requires sophisticated
bioinformatic tools coupled to strong background knowledge on
chemistry, biochemistry and molecular biology. Metabolomics
data, in particular, are the most difficult to interpret because of
the non-direct linkage between metabolites and genes. In this work
we present a new method to correlate metabolite levels to potential metabolic pathways likely to be active inside the cells. This