Abstracts

P03 Biochemistry and Molecular Biology of Plants

P03-24
Natural rubber and antioxidants in Euphorbia
characias latex
D. Spano`1, F. Pintus1, C. Mascia1, A. Macone2, G. Floris1 and
R. Medda1
1
Department of Sciences of life and environment University of
Cagliari (Italy), Monserrato (CA), Italy, 2Department of
Biochemical Sciences A. Rossi Fanelli, University of Rome La
Sapienza (Italy), Rome, Italy
We have selected the mediterranean shrub Euphorbia characias as
an experimental model to study the complexity of plant latex
chemistry. Latex is a mixture with diversied composition, that
includes alkaloids, terpenoid compounds, polymeric substances
such as resins and gums, starch, oil and a large number of proteins and enzymatic activities. The aim of the present study is to
contribute to the knowledge of this plant product evaluating the
antioxidant properties of extracts of E. characias and searching
for polymeric substances as natural rubber.
We analyzed different extracts from the latex of E. characias
and performed a new extraction method (involving the use of trichloacetic acid, TCA) that turned out to be easier, faster and
higher reproducible if compared to common extraction methods
involving organic solvents like methanol, ethanol, and petroleum
ether/methanol.
TCA extract of E. characias latex exhibits antioxidant activities
determined as total content of free-radical scavenging, polyphenols and total avonoids. GC-MS analysis conrms the presence
of several compound identied as antioxidant molecules.
E. characias latex contains a natural rubber. The optimum rubber extraction is achieved with acetic acid followed by cyclohexane/ethanol treatment. The rubber content was shown to be 14%
(w/v) of the E. characias latex and the gel content is 2.5% of the
rubber weight.
E. characias natural rubber showed a molecular weight of
93000 and Mw/Mn of 2.9. On the basis of 1H NMR, 13C NMR
and FT-IR spectroscopy, the structure of this rubber can be identied as cis-1,4-polyisoprene.
This study was partially supported by a grant from Regione
Autonoma della Sardegna, Progetti di ricerca di base CRP2-22.

P03-25
Identification and characterization of SlCIPK6
phosphorylation targets involved in plant
innate immunity
Y. Pareja-Jaime, E. Gutierrez-Beltran and O. del Pozo
Instituto de Bioqumica Vegetal y Fotosntesis (CSIC), Sevilla,
Spain
Plants respond to pathogen attack inducing an immune response,
which has two layers. The rst layer, called PTI (Pathogen Associated Molecular Patterns [PAMPs]-Triggered Immunity), was
traditionally known as basal immunity and is effective against
whole species of microbes. The ETI response (Effector-Triggered
Immunity) constitutes a second layer of immunity, more intense
and powerful, and frequently accompanied by a form of
programmed cell death (PCD) in infected cells to limit pathogen
proliferation.
Both responses share signal transduction pathways resulting in
a sequence of common molecular events, such as the increase in
cytosolic Ca2+, the production of reactive oxygen species (ROS)
and the activation of mitogen-activated protein kinase (MAPK)
cascades.
Previously, two genes involved in PCD associated with ETI
were identied in a random Virus Induced Gene Silencing

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(VIGS) screening in Nicotiana benthamiana. Later, we characterized that the proteins encoded by these genes, the calcium sensor
NbCBL10 (Calcineurine B-Like protein 10) and the intracellular
protein kinase NbCIPK6 (Calcineurine B-Like Interacting Protein
Kinase 6), constitute a Ca2+ signaling module. To identify phosphorylation targets of CIPK6 in biotic stress, a yeast two hybrid
screening was carried out. Here we describe the biochemical and
physiological analysis of two of the CIPK6-Interactor Proteins
(CIPs) identied, named CIP70 and CIP90. These proteins seem
to play an important role in the cellular processes regulated by
CBL10/CIPK6 signaling module.

P03m-26
Implication of granule-bound starch synthase
(GBSS) in photoperiodic signaling
M. I. Ortiz1, T. Albi1, E. Lucas-Reina1, F. J. Romero-Campero2, F. E. Said1, B. Cano1, M. T. Ruiz1, J. M. Romero1 and
F. Valverde1
1
Instituto de Bioqumica Vegetal y Fotosntesis, CSIC, Sevilla,
Spain, 2Departamento de Ciencia Computacional e Inteligencia
Articial, Grupo de Investigacion en Computacion Natural,
Universidad de Sevilla, Sevilla, Spain
Starch is the main carbon reserve in higher plants and constitutes
the mayor energy reserve component of some of the most important crops for human consumption. Starch is synthesized by
ADP glucose pyrophosphorylase and starch synthase enzymes as
GBSS. GBSS enzyme catalyzes amylose synthesis, the linear component of the glucose polymer in starch. The circadian-regulated
gene CONSTANS (CO) plays a central role in the photoperiodic
control of the oral transition. Recently, a CO homologue gene
present in the genome of the unicellular green alga Chlamydomonas reinhardtii (CrCO) was identied. CrCO has a conserved
role in the coordination of processes regulated by photoperiod
and the circadian clock (1). To study the rate of conservation of
the photoperiodic signaling, starch and hexoses levels were analyzed in different Arabidopsis thaliana mutants implied in photoperiod regulation, both before and after blooming. In addition,
amylose amount were quantied. AsGBSS expression is controlled byCrCO in Chlamydomonas reinhardtii (2), in order to
study the same effect in Arabidopsis thaliana, we followed the
expression of the GBSS gene in several mutants in short and long
day conditions (3). Moreover, GBSS was fused to GFP to study
its tisular localization during a 24 hour period. CO-overexpressing and CO mutant plants were crossed to GBSS mutant and
their oral phenotype observed. The effect of CO on GBSS could
be part of a new regulatory mechanism connecting photoperiodic
signaling with carbon metabolism in plants (4).
References
[1] Serrano, G. et al., (2009). Curr Biol 19: 359368.
[2] Tenorio, G. et al., (2003) Plant Mol Biol 51: 949958.
[3] Valverde, F. et al., (2004) Science 303: 10031006.
[4] Romero, J.M. et al., (2009) Plant Sig Behav 4:7, 642644.

P03r-27
A new redox component in the mechanisms
for adaptation of plants to light stress
B. Naranjo1, D. Hornero2, G. Lourdes2, F. J. Cejudo1 and
M. Lindahl1
1
Instituto de Bioqumica Vegetal y Fotosntesis (Universidad de
Sevilla y CSIC), Sevilla, Spain, 2Departamento de Biotecnologa
de Alimentos, Instituto de la Grasa (CSIC), Sevilla, Spain
When the light energy absorbed by photosystem II (PSII) exceeds
its capacity for utilization, the concomitant production of reac-

FEBS Journal 279 (Suppl. 1) (2012) 52576 2012 The Authors FEBS Journal 2012 FEBS