p An - MAP — MAP

 Prog
grama In
nter-Univ
versitário
o de Dou
utorame nto MAP
P em
Biologia de
e Planta s – Bioplant
MAP Jo
oint Doc
ctoral Pro
ogramme in Plan
nt Biolog
gy - Biop
plant
4
4ª Works
shop
p An
nual Biop
plan
nt
18 e 1
19 Julho
o 2013
PROGRA
AMA / P
PROGRAM
MME
Liivro de resumos
r
/ Book of abstra
acts
4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 Bem
m vindos
s à 4ª Wo
orkshop A
Anual BioP
Plant organizada pela
la 4ª ediçã
ão do
Programa Interr-Universitá
ário de Dou
utoramento
o em Biolog
gia de Plan
ntas – Biop
plant,
das U
Universidad
des do Minh
ho, Aveiro e Porto (prrograma MA
AP).
Wellcome to the 1st Ann
nual BioPlan
nt Worksho
op organize
ed by the 1sst edition of the
Jointt Doctoral Programm
me in Plantt Biology – BioPlant, from the Universities of
Minh
ho, Aveiro and
a
Porto (M
MAP progra
amme).
Con
nteúdos / Conten
nts
___
______
_______
______
______
_______
______
_______
__
Programa / Pro
ogramme
2
Confferências Co
onvidadas / Invited Le
ectures
5
Comunicações Orais
O
/ Oral Communiications
9
Poste
ers
18
8
Lista
a de Particip
pantes / Lis
st of Particip
ipants
38
8
Apo
oios / Su
upport
O B
BioPlant tem
m o privillégio de u
usufruir de
e um financiamento da Fundação
ouste Gulb
benkian ao
a abrigo do seu Prrograma de
e Reforço da Capacidade
Calo
Cienttífica para Projectos Inter-Univerrsitários de
e Doutoram
mento.
The BioPlant has
h
the priv
vilege of b
benefiting from
f
the financing
f
o
of the Calo
ouste
benkian Fo
oundation
n under its Programm
me of Reinfforcement o
of the Scie
entific
Gulb
Capa
acity for Intter-Univers
sitary Docto
oral Program
mmes.
Org
ganização
o da Workshop / Organiz
zation off the worrkshop
Maria
ana Sottom
mayor
Paula
a Melo
Juliana Oliveira
Departam
mento de Biologia,
B
FC
CUP
Departam
mento de Biologia,
B
FC
CUP
Secretarriado Biopla
ant
Com
missão Científica
C
a Bioplan
nt / Scien
ntific Com
mmittee Bioplant
Rui T
Tavares, UM
M
Hern
nâni Gerós, UM
Conc
ceição Santtos, UA
Antó
ónio Calado, UA
Maria
ana Sottom
mayor UP, Diretora
D
Bio
oplant 2013
3
Paula
a Melo, UP
www.mapp.edu.pt/bioplant 1 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 Pro
ograma / Prog
gramme
e
___
______
_______
______
______
_______
______
_______
__
18 Julho / July 18th
1
09:0
00
Abertu
ura / Open
ning
José Lu
uís Santos, Vice-Direto
or da FCUP
P
09:3
30
Conferrência de abertura / Opening
g lecture
Excitation about sex in pllants: mer
rging biop
physics witth genetic
cs on
the po
ollen tube system CC
C1
José Fe
eijó, FCUL / IGC
10:3
30
Coffee
e Break - Posters
P
11:0
00
AGP6 and AGP11 biologic
cal mode of
o action in
n Arabidop
psis pollen
n
and po
ollen tube growth C
CC2
Sílvia Coimbra,
C
FC
CUP / BioFIIG
11:3
30
RNASe
eq analysis of the Q
Quercus su
uber root response
r
tto droughtt O1
Herlând
der Azeved
do, DB-UM / BioFIG
11:4
45
SUMO proteases
s control d
developme
ent and st
tress respo
onses in
Arabid
dopsis thaliana O2
Pedro Humberto
H
Castro,
C
DB--UM / BioFIIG
12:0
00
Posterrs
13:0
00
Almoç
ço / Lunch
h
14:0
00
Legacy
y and Eme
erging Con
ntaminantts in Plants
s: selectio
on and utillity
of Biom
markers CC3
C
Conceição Santos
s, DB-UA / C
CESAM
14:3
30
Anti-b
bacterial potential off silver na
anoparticle
es green s
synthesise
ed
using Withania somnifera
a aqueous leaf extra
act O3
Marslin
n Gregory, DB-UM / CIITAB - Alun
no Bioplant
14:4
45
Antiox
xidant cap
pacity of M
Melia azeda
arach under water d
deficit
condittions O4
Carla Azevedo,
A
DB-UA / CES
SAM
15:0
00
argoseira: I.
egradadas
Reflorrestação de zonas de
s pela seca com ama
crescim
mento e fo
otossíntes
se O5
Maria da
d Costa, DB-UA
D
/ CES
SAM - Alun
na Bioplant
www.mapp.edu.pt/bioplant 2 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 15:1
15
Coffee
e Break - Posters
P
15:4
45
Funga
al diversity
y in chestn
nut orchar
rds: a com
mparison b
between
above- and belo
ow-ground
d diversity
y CC4
Teresa Lino Neto, DB-UM / B
BioFIG
16:1
15
Dot-bllot validattion of nov
vel DNA markers
m
for
r detection
n and
genoty
yping of Ralstonia
R
s
solanacear
rum specie
es comple
ex O6
Pedro Albuquerqu
A
ue, FCUP / C
CIBIO
16:3
30
Contribution of the plant rhizosphe
ere system
m to the
phytorremediatio
on of cadm
mium in es
stuarine areas O7
Cristina
a Almeida, FCUP / CIIIMAR
16:4
45
Deciph
hering intr
racellular protein tr
rafficking pathways
s CC5
José Pissarra, FCU
UP / BioFIG
G
17:1
15
The Pllant Speciffic Insert and Its Molecular Role
R
in Pro
otein
Sorting O8
Bruno Peixoto, DB
B-UP / BioF
FIG
17:3
30
Posterrs
___
______
_______
______
______
_______
______
_______
__
19 Julho / July 19th
1
09:3
30
Purific
cation an
nd functio
onal
aquap
porin VvSIP1 CC6
Hernân
ni Gerós, DB-UM / CIT
TAB
00
10:0
Photos
synthesis in grape b
berry tissu
ues: an in vivo apprroach by
imagin
ng PAM flu
uorometry
y O10
Ana Cu
unha, DB-UM / CITAB
10:1
15
Unravelling the
e molecu lar netwo
ork regullating zin
nc conten
nt in
plants
s: from Ara
abidopsis to rice CC7
Ana As
ssunção, CIIBIO
10:4
45
Coffee
e Break - Posters
P
11:1
15
Coppe
er transpor
rters in grrapevine O11
O
Viviana
a Martins, DB-UM
D
/ CIITAB - Aluna Bioplant
11:3
30
Identification off heavy m
metal trans
sporters in
n Solanace
eae plants O13
Alberto
o Pessoa, FCUP / BioFIIG
char
racterization
www.mapp.edu.pt/bioplant of
tthe
grape
evine
3 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 11:4
45
A high
hly efficien
nt leaf pro
otoplast ex
xpression system fo
or the stud
dy of
anticancer alkalloid metab
bolism in Catharant
C
thus roseu
us O9
Patrícia
a Duarte, IB
BMC
12:0
00
Posterrs
13:0
00
Almoç
ço / Lunch
h
14:0
00
A mole
ecular anttagonism tthat shape
es the flow
wer CC8 Manuela Costa, DB-UM
D
/ Bio
oFIG 14:3
30
Major Research Topics att CITAB CC
C9
Francis
sco Manuel Pereira Peiixoto, UTAD
D / CITAB
15:0
00
Desen
nvolvimentto Susten
ntável e Ecoinovaç
ção - ferrramentas
s de
comba
ate à crise
e O12
Joaquim
m Manuel Soares
S
Gue
edes, Ecoins
side
15:1
15
Coffee
e Break - Posters
P
15.45
Unravelling the metabolis
sm and tra
ansmembr
rane trans
sport of th
he
highly
y valuable medicinall alkaloids
s from Catharanthus
s roseus (L) G.
Don. CC10
C
Mariana Sottomay
yor, FCUP / IBMC
16:4
45
s Posters
Atribu
uição de pr
rémios aos
www.mapp.edu.pt/bioplant 4 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 Con
nferência
as Convid
dadas
Inv
vited Lectures
___
______
_______
______
______
_______
______
_______
__
CC1
1 – José Feijó
Ex
xcitation about sex in
i plants: merging biophysic
b
s with gen
netics on the
polle
en tube sy
ystem
Institu
uto Gulbenkian
n de Ciência (IGC),
(
Oeiras
Deparrtment of Plan
nt Biology, Fac
culty of Sciencces, University
y of Lisbon
CC2
2 – Sílvia
a Coimbr
ra
AGP6 and AGP11
A
bio
ological m
mode of ac
ction in Arrabidopsiss pollen an
nd
polle
en tube grrowth
BioFIG
G, Departmen
nt of Biology, Faculty
F
of Scie
ences, Univers
sity of Porto
Arabiinogalactan proteins (AG
GPs) are cel l wall proteo
oglycans that were show
wn to be imp
portant
for po
ollen develop
pment. An Arabidopsis
A
d
double null mutant
m
for tw
wo pollen-speecific AGPs (agp6
agp11) showed re
educed polle
en tube grow
wth and compromised res
sponse to geermination cues in
nd the mode
e of action of these AGPs
s, an Affymettrix ATH1 ge
enome
vivo. So, in order to understan
6 agp11 doub
ble null muta
ant pollen tub
be was performed. The laack of two sp
pecific
arrayy in the agp6
AGPss induced a meaningful shift
s
of the g
gene express
sion profile. CalciumC
andd signaling-re
elated
genes were found
d to be altere
ed, which givves support to
t the known
n roles of succh genes in pollen
e presumed involvemen t of AGPs in
n signaling cascades
c
waas also reinfo
orced.
tube growth. The
eine-rich proteins have been
b
propose
ed to play a role in reco
ognition and fertilization, and it
Cyste
was tthus quite re
elevant that such
s
genes w
were found to
t be differen
ntially expresssed. The pu
utative
involvvement of AGPs
A
in sign
naling casca
ades through
h calmodulin
n and proteinn degradatio
on via
ubiqu
uitin was found. Also, strress related genes were
e found to be
e affected, w
which supporrts the
recog
gnized simila
arities betwe
een signaling
g pathways in both defense and poollen tube growth
g
(Costta et al., 2013).
Yeasst two-hybrid experiments
s gave furth
her support to
t these sign
naling pathw
ways and rev
vealed
nd AGP11 in
nteractors im
mplicated in the
t process of recycling by endocyto
osis of
putative AGP6 an
membrane co
omponents, through
t
clath
hrin-mediated endosome
es and multivvesicular bod
dies. A
cell m
mode
el for AGP6 and
a AGP11 biological
b
mo
ode of action
n in pollen tub
be growth is presented.
Coimbra, S. 2013.
a, M., Nobre, S., Becker, J., Masiero,, S., Amorim, M.I., Pereirra, L.G. and C
Costa
On ha
and, putative
e ligands for arabinogala ctan proteins
s in Arabidop
psis pollen ddevelopment.. BMC
Plantt Biology, 13::7.
3 – Conce
eição Sa
antos
CC3
Legacy and
d Emergin
ng Contam
minants in Plants: se
election aand utility of
B
Biomarkerrs
Deparrtment of Biolo
ogy & CESAM
M, University A
Aveiro 3810 19
93 Aveiro
The selection off adequate plant endpo
oints, having
g the sensittivity and sppecificity tha
at are
oxicology, iss being supported by complementa
c
ary fields off e.g.,
required in modern plant to
moleccular biologyy, -omics, systems
s
biol ogy, etc. Ho
owever, other challengees emerge during
d
selecction, includin
ng the dema
and for a high
h predictive ability of any
y putative enndpoint. More
eover,
bioma
arkers prope
erties are often affected by experime
ental variable
es, which maay jeopardize
e their
www.mapp.edu.pt/bioplant 5 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 valida
ation as relia
able endpoin
nts in plant (eco)toxicolo
ogical studies. Some casse studies will
w be
explo
ored concern
ning selected
d biomarkerss (and related techniques
s) that are ggaining imporrtance
in asssessing plan
nt responses
s to legacy a
and emerging
g contaminants. In particcular, examp
ples of
metals and metal-based nano
oparticles efffects on the plant genom
me structuree and functio
on and
on th
he cell funcctioning will be discusssed. Some techniques used in thee multiparametric
appro
oaches expllored here include gen
ne expressio
on (qRT-PCR), metaboonomics, cyttomics
(FCM
M), COMETS, catastrophe
e mitosis, ele
ectron param
magnetic reso
onance (EPR
R) and others
s. The
comb
bination of th
his multiple information aims at pro
oviding, as most
m
compleete as possible, a
functiional networrk of the pla
ant cell’s ressponse. Bes
sides using more innovaative approa
aches,
there
e is also a ne
eed to refocus strategies and methodologies, including, for exxample risk-b
benefit
balan
nce analysess. This comp
prehensive o
overview is an example of the channges of para
adigm
within
n modern pla
ant toxicolog
gy. Interestin
ngly, and de
espite plant toxicologists
t
have today more
tools and knowle
edge to study
y multiple m echanisms of
o functional effects of cchemicals on
n plant
cells, many cha
allenges rem
main in mod
dern toxicology, i.e, “h
how to use this pletho
ora of
inform
mation?” or “how
“
to defin
ne best endp
points?”.
4 – Teres
sa Lino Neto
N
CC4
Fun
ngal divers
sity in che
estnut orc
chards: a compariso
c
on betweeen above -and
below--ground diversity
Baptissta P1, Reis F2, Pereira E1, Tavares R2, L
Lino-Neto T2
1
CIMO
O / School off Agriculture, Polytechnic
P
In
nstitute of Bra
agança, Camp
pus de Santa Apolónia, Ap
partado
1172, 5301-854 Bra
agança, Portu
ugal.
2
Centtre for Biodive
ersity Function
nal and Integrrative Genom
mics (BioFIG), Plant Functioonal Biology Centre,
C
Unive
ersity of Minho
o, Campus de Gualtar, 4710
0-057 Braga, Portugal.
P
Chesstnut (Castan
nea sativa) groves have a significant impact in the
e Portuguesee economy, due
d to
the p
production off fruits and wood,
w
as welll as to relate
ed activities, such as huunt and mush
hroom
collecction. The fu
ungal comm
munity presen
nt in chestn
nut orchards was studie d by a traditional
surve
ey of abovve-ground mushrooms
m
and by a metabarco
oding approoach using 454pyrossequencing of
o soil DNA samples. Th
he fungal div
versity present in chestn ut orchards found
by bo
oth analyses will be compared and re
eliability of both approaches discusseed. Results will
w be
also discussed ta
aking into ac
ccount the Im
mplications of
o specific fu
ungal interacctions for che
estnut
orcha
ard sustainab
bility.
CC5
5 – José Pissarra
a
Deciphering intracellu
ular protein trafficking pathwaays
,
BioFIG
G, Departmen
nt of Biology, Faculty
F
of Scie
ences, Univers
sity of Porto
CC6
6 – Hernâ
âni Geró
ós
Pu
urification and functional cha
aracterizattion of the
e grapevin
ne aquapo
orin
VvSIP1
1
Henriq
que Noronha1,2
, Ana Paula Martins3, Gra ça Soveral3, François
F
Chau
umont4 and Heernâni Gerós1,2
1
Centtro de Investig
gação e de Te
ecnologias Agrro-ambientais e Biológicas CITAB-UM,
C
Po
Portugal
Gru
upo de Invesstigação em Biologia Veg
getal Aplicada
a e Inovação
o Agroalimenntar (AgroBio
oPlant),
Deparrtamento de Biologia,
B
Escolla de Ciênciass Universidade
e do Minho, Braga, Portugaal
3
Ressearch Institu
ute for Mediicines and P
Pharmaceutica
al Sciences (iMed.UL) aand Departme
ent of
Biochemistry and Human
H
Biology
y, Faculty of P
Pharmacy, Uniiversity of Lisb
bon, Lisbon, P
Portugal
4
Unité
é de Biochimie
e Physiologiqu
ue, Institut de s Science de la Vie, Univerrsité Catholiquue de Louvain
n, Croix
du Su
ud 2-20, B-134
48 Louvain-la-Neuve, Belgiu
um
2
Wate
er is transporrted through biological m embranes by aquaporins
s, members of the wides
spread
Majorr Intrinsic Proteins (MIP
Ps). In plantss, aquaporin
ns are group
ped in five ssub-families, PIPs
ma membra
(plasm
ane intrinsic proteins), TIIPs (tonoplast intrinsic proteins),
p
NIP
Ps (nodulin2
26-like
intrinssic proteins), XIPs (X intrinsic Protein
ns) and SIPs
s (small and basic intrins ic proteins). In the
prese
ent study the
e localization
n, expression
n and functio
onal charactterization of a VvSIP from the
www.mapp.edu.pt/bioplant 6 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 grape
e berry were performed. VvSIP1 is e
expressed in leaves and berries, withh higher number of
transcripts in the
e mature sta
age, and co
olocalizes at the ER in transformedd yeast cells
s with
VvSIP
P1-GFP. ER
R membran
ne vesicles purified fro
om yeast overexpressin
o
ng VvSIP1 were
chara
acterized by stopped flow
w technique for their cap
pacity to tran
nsport waterr. The protein was
purifie
ed to homog
geneity after VvSIP1-his tag heterolo
ogous expres
ssion in yeaast followed by
b ER
purificcation, mem
mbrane solubilization and Ni-NTA affinity chromattography. Waater transport was
confirrmed after reconstitution
r
n of the purrified protein
n in phospha
atidylethanol amine liposo
omes.
VvSIP
P1 expressiion is rema
arkably up regulated by
b heat, as shown by Real-time PCR,
sugge
esting a role in stress res
sponse in gra
apevine.
Acknowledgmentts: This work
k was suppo
orted by Eurropean Union Funds (FE
EDER/COMP
PETEOperrational Competitiveness Programme
e) and by na
ational funds (FCT-Portugguese Found
dation
for S
Science and
d Technolog
gy) under the projects FCOMP-0
01-0124-FED
DER-022692
2 and
Europ
pean COST action FA 1106
1
and the
e research projects
p
PTD
DC/AGR-ALI//100636/2008 and
PTDC
C/AGR-AAM/099154/200
08. HN was ssupported by
y the PhD grrant no. SFR
RH/BD/75257
7/2010
and A
APM by the PhD
P
grant no
o. SFRH/BD//65046/2009
CC7
7 – Ana Assunçã
ão
Unrave
elling the molecular
m
network regulating
r
g zinc con tent in pla
ants:
from Arrabidopsis
s to rice
Joana
a G. Guedes1, Diego Almeid
da2, Mark G.M
M. Aarts3, Nels
son J. M. Saibo
o2, Ana G.L. A
Assunção1
1
Plantt Evolution Grroup, CIBIO/In
nBIO, Universid
idade do Porto
o, Portugal
Geno
omics of Plantt Stress Laborratory, ITQB, U
Universidade Nova
N
de Lisbo
oa, Portugal
3
Labo
oratory of Genetics, Wageniingen Universiity, The Nethe
erlands
2
As a plant micro
onutrient, zin
nc is of ma
ajor interest in agronom
my and for hhuman and cattle
nutrition. Zinc deficient soils are widespre
ead all over the world and the risk oof inadequatte diet
and zzinc malnutrition is estimated to affecct one third of
o the global human popuulation, i.e. around
a
2 billlion people
e. Developin
ng countriess, where people depend on cereeal-rich dietts for
suste
enance, are
e the most affected. Improvemen
nt of crop efficiency in zinc de
eficient
enviro
onments, an
nd bio-fortific
cation to all eviate huma
an nutrition are plant-baased solutions as
sourcce of micron
nutrients. An
n incomplete
e understand
ding of zinc homeostasiis mechanisms in
plantss is howeverr a limitation.. Recently th e first transc
cription factorrs regulatingg zinc homeo
ostasis
in Ara
abidopsis we
ere identified
d (Assunção et al. 2010).. The AtbZIP
P19 and AtbZ
ZIP23 were shown
s
to be essential for the adaptattion to zinc d
deficiency. Trranscript profiling revealeed only a small set
of ge
enes, to be induced in wild-type p lants in response to zinc deficienccy, but not in the
bzip1
19bzip23 Ara
abidopsis double mutant line. A sugg
gested conse
ervation of thhe zinc defic
ciency
respo
onse regulatory network within the p
plant kingdom makes it very attractiive to explore the
moleccular basis of
o such netwo
ork in rice (O
Oriza sativa L.).
L Rice is on
ne of the moost important crops
world
dwide and it is the staple food for morre than half of
o the world’s
s population. Additionally
y it is a
very relevant crop in Portuga
al. Transferrin
ng the know
wledge from Arabidopsis
A
to rice is of major
imporrtance. We present ou
ur preliminarry results in
n the identification andd analysis of
o the
bZIP1
19/bZIP23 ricce orthologs.
CC8
8 – Manu
uela Costta
A molecu
ular antago
onism tha
at shapes the
t flowerr
BioFIG
G, Departmen
nt of Biology, Faculty
F
of Scie
ences, Univers
sity of Porto
The a
action of anttagonistic pro
oteins on the
e transcriptio
onal activity of developm
mental key genetic
progrrammes is an important regulatory m
mechanism in
n the genera
ation of differrent morpho
ologies
that ccharacterise multicellular organisms.
In An
ntirrhinum majus,
m
the do
orsoventral asymmetry of the flowe
ers is estabblished throu
ugh a
moleccular antago
onism involv
ving two MY
YB-like transcription fac
ctors, RADIA
ALIS (RAD)), and
DIVA
ARICATA (DIV). RAD is expressed
e
do
orsally in the floral primorrdia and prom
motes dorsal petal
identiity, whilst DIV is expressed in the wh
hole floral primordia, desp
pite having a phenotypic effect
only in more ven
ntral regions of the flowe
er. Genetic and
a
molecular studies hhave revealed that
RAD antagonisess the activitty of DIV th
hrough an in
ndirect intera
action with another MY
YB-like
prote
ein identified on a yeastt two-hybrid screen using RAD as bait (DRIFss, DIV and RADintera
acting protein
ns).
www.mapp.edu.pt/bioplant 7 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 To be
etter understtand the role of the DRIF
F proteins in the
t molecula
ar antagonism
m that RAD exerts
over DIV function, the DNA--binding actiivity of DIV was analysed by gel-shhift assays in the
prese
ence of the DRIF
D
and RA
AD proteins. The subcellular co-localisation of theese three prroteins
was determined using fluore
escent tagge
ed proteins in
i transient assays in toobacco epid
dermal
cells. The expresssion pattern of the DRIF genes was analysed by
y RT-PCR annd the evolutionary
historry of these new MYB-like
e proteins stu
udied by phylogenetic analysis.
Our rresults sugge
est that the molecular an
ntagonism th
hat RAD has
s over DIV iss mediated by
b the
DRIF
F proteins. In the ventral domain of th
he flower DIV
V and DRIFs
s interact insside the nucle
eus in
orderr to determin
ne the ventra
al identity. H owever, in the dorsal do
omain, wheree RAD is pre
esent,
RAD interacts witth the DRIF in
i the cytopla
asm and pre
events their nuclear
n
localiisation, preventing
DIV ffrom interactting with the RIPs, which
h results in flower
f
dorsal identity. Thhese results along
with tthe fact that the DRIF, RAD
R
and DIV
V proteins are
a present in such a vaariety of land
d plant
speciies, suggest that this mo
odule of interractions may
y have been implicated inn a broader set of
functiions throughout plant evo
olution.
9 – Franc
cisco Peiixoto
CC9
Major Rese
earch Topics at CITA
AB
CITAB
B/UTAD
CITA
AB research is focused on
o the agro-ffood and forrestry system
ms using thee production--chain
appro
oach as a whole.
w
This Research Unit is com
mposed by a multidiscipplinary team
m with
experrtise ranging
g from funda
amental scie
ences such as
a biology and
a
chemistrry to agrono
omists,
foresttry engineerss and ecolog
gists.
CITA
AB was orga
anized with the objectivve of movin
ng a step forward
f
in tthe research
h and
devellopment of the
t
agricultu
ural sciencess bringing in
nnovation to
o the convenntional production
chain
ns in the field
ds of agricultu
ure and foresstry and resp
pective relatio
ons to enviroonment.
CITA
AB is divided
d in three areas; Ecointe
egrity, Susta
ainable Agro--food Chainss and Biosystems
Engin
neering. Eacch of these groups is sttructured in different lines of resea rch and the main
sustainable ecosystems and
leadin
ng areas arre: Increasin
ng resource efficiency; biodiversity,
b
ecosyystem servicces. Added value
v
of agrri-food and forestry
f
co-p
products. Opptimization off agrifood chains. Innovative tech
hnologies; b
biobased ma
aterials, agri--food and fo
forestry. It will
w be
prese
ented some of the main results ach ieved by the
e projects cu
urrently undeer developm
ment in
CITA
AB.
CC1
10 – Marriana Sotttomayorr
Un
nravelling the metab
bolism and transme
embrane transport
t
o
of the highly
m
s from Cattharanthus
s roseus ( L) G. Don.
valuable medicinal
alkaloids
IBMC
C, Department of Biology, Fa
aculty of Scien
nces, Universiity of Porto
www.mapp.edu.pt/bioplant 8 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 Com
municaçõ
ões Orais
s
Ora
al Commu
unication
ns
___
______
_______
______
______
_______
______
_______
__
O1 RNASeq analysis
a
of
o the Que
ercus sube
er root res
sponse to drought
Azeve
edo H, Magalh
hães A, Martin
ns I, Lino-Neto
o T, Tavares RM
R
Cente
er for Biodiversity, Function
nal & Integra
ative Genomic
cs (BioFIG), Plant
P
Functionnal Biology Center,
C
Unive
ersity of Minho
o, Campus de Gualtar, 4710
0-057 Braga, Portugal
P
Cork oak (Querccus suber L.) is a typica
al species fro
om the Portuguese Medditerranean forest,
f
usuallly dissemina
ated in dry environments
e
s. Mainly due
e to its high economic vaalue, cork oa
ak has
been considered
d a protecte
ed plant sp
pecies and a national species of interest. Climate
altera
ations, in parrticular, incre
eased temperrature, high light intensity
y, drought annd air pollutio
on are
factorrs that directtly affect the developmen
nt of plants and reduce th
heir productivvity. In the prresent
work,, the effect of soil water content
c
was studied in Q. suber seed
dlings, as twoo-month-old plants
were subject to different
d
wate
er stress reg imes to impo
ose drought stress. Phottosynthetic activity
a
meassured by PAM
M fluorometrry and photo
osynthetic pigment conte
ent were useed as physiological
indica
ators of stresss. Subsequently, the tra
anscriptome of Q. suber roots in respponse to mod
derate
and ssevere droug
ght stress was
w analysed
d by Next Generation Se
equencing uusing 454 (R
Roche)
techn
nology. Data
a was then mapped
m
aga
ainst the rece
ently established Q. subber transcrip
ptome,
and R
RNASeq ana
alysis was performed
p
to
o establish differential
d
ex
xpression. T
This allowed us to
identiify over 300 effector and
d regulatory proteins tha
at are likely to
t play a keyy role in Q. suber
droug
ght tolerance
e at root level, establishin
ng a fundame
ental basis fo
or new functi onal studies.
Proje
ect funding byy FCT (SOBREIRO/0033
3/2009 and PTDC/AGR-G
P
GPL/118505//2010).
O2 SUMO pro
oteases co
ontrol dev
velopmentt and stres
ss respon
nses in
A
Arabidops
sis thalian
na
Castrro PH, Couto D,
D Freitas S, Ruiz-Albert
R
J, Tavares RM, Bejarano ER, Azevedo H
Cente
er for Biodiversity, Function
nal & Integra
ative Genomic
cs (BioFIG), Plant
P
Functionnal Biology Center,
C
Unive
ersity of Minho
o, Campus de Gualtar, 4710
0-057 Braga, Portugal
P
(P.H.C., D.C., S.F.,, R.M.T., H.A.))
Institu
uto de Hortofr
fruticultura Su
ubtropical y M
Mediterránea “La
“
Mayora”, Universidad de Málaga-C
Consejo
Superrior de Investiigaciones Cien
ntíficas (IHSM
M-UMA-CSIC),, Dept. Biolog
gía Celular, Geenética y Fisiiología,
Unive
ersidad de Mállaga, Campus
s Teatinos, 290
071 Málaga, Spain
S
(P.H.C., J.R.-A., E.R.B
B.)
Living
g organisms are subjecte
ed to consta
antly changin
ng environme
ents that com
mpromise survival.
As se
essile organisms, plants have optimiized a myria
ad of strategies to adaptt, avoid or to
olerate
stresss conditions.. A general type of mech
hanism assoc
ciated to env
vironmental rresponses is
s posttranslational mod
dification (PT
TM), which a
acts as a fas
st and revers
sible regulatoor of key pro
oteins.
Relevvant PTMs includes sma
all peptides such as ubiquitin and ubiquitin-like modifiers, namely
the S
Small Ubiquittin-like Modiffier (SUMO) . SUMO ma
ay exert different effects depending on
o the
targe
et protein, either
e
controlling its con
nformation, or
o even cre
eating or bloocking interracting
interffaces that ultimately modulate its activvity. SUMO attachment
a
or
o sumoylatioon requires SUMO
S
peptid
des to first be
b processed
d by SUMO p
proteases (U
ULP/SENP fa
amily), and tthen conjuga
ated to
a targ
get’s lysine via
v SUMO E1 activase a
and SUMO E2
E conjugase
e, aided by S
SUMO E3 lig
gases.
Deco
onjugation of the SUMO peptide can subsequently be carried
d out by the SUMO prote
eases.
A gre
eat number of SUMO ta
argets are in
nvolved in nuclear-relate
n
ed functionss, and are part
p
of
essen
ntial cellular processes. This is particcularly evide
ent in plants since loss-of
of-function mutants
of m
most SUMO componentts display embryonic lethality or pleiotropic defects. SUMOconju
ugation levelss vary in res
spect to grow
wth condition
ns, tissues an
nd developm
mental stages
s. It is
there
efore expecte
ed that highly dynamic S
SUMO conjugation and deconjugatio
d
on machinery
y is at
play. Unlike ubiqu
uitin, SUMO conjugation machinery components
c
are less abuundant in the
e plant
genome. SUMO proteases (ULPs) consttitute a fairly
y large gene
e family andd are the potential
sourcces of specifficity within the
t pathwayy by displayin
ng different enzymatic aactivity, subcellular
localization and expression levels. Base
ed on a reverse genettics strategyy, several se
ets of
Arabiidopsis ULP T-DNA inse
ertion mutan
nts have bee
en characterrized. Resultts place ULPs as
funda
amental regu
ulators of both plant deve
elopment and
d the respons
se to environnmental stres
ss.
Work w
was supported by
b FCT/FEDER
R (refs. PTDC/BIIA-PLA/3850/20
012 and FCOMP
P-01-0124-FED
DER-028459).
www.mapp.edu.pt/bioplant 9 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 O3 Anti-bacte
erial poten
ntial of sil ver nanop
particles green
g
syntthesised
u
using With
hania som
mnifera aq
queous lea
af extract
2,3
Grego
ory Marslin 1,, Selvakesavan
n R.K1, Franklilin G1, Bruno Sarmento
S
, Alberto
A
C.P. D
Dias 1
ntre for the Research
R
and Technology o
of Agro-Envirronment and Biological
B
Sciiences (CITAB
B-UM),
1 Cen
AgroB
BioPlant Group
p, Department of Biology, U
University of Minho,
M
Portuga
al
2 INE
EB – Instituto de
d Engenharia
a Biomédica, U
University of Porto,
P
Portugall
3 CIC
CS, Departmen
nt of Pharmac
ceutical Sciencces, Instituto Superior
S
de Ciências
C
da Saaúde-Norte, Gandra,
G
Portug
gal
In the present study,
s
we re
eport on gre
een synthesis of silver nanoparticlees (AgNPs) using
Witha
ania somnife
era aqueous
s leaf extractts as reduciing agent an
nd their chaaracterization
n. The
forma
ation, size and shape off green syntthesized AgN
NPs were co
onfirmed by physical-che
emical
techn
niques such as UV-Visib
ble spectros copy, laser Doppler ane
emometry, S
Scanning Ele
ectron
Micro
oscopy (SEM
M), Atomic Fo
orce Microsccopy (AFM), X-ray diffrac
ction (XRD) and X-ray energy
e
dispe
ersive spectroscopy (ED
DX). The S
SEM and AFM images confirmed that the siize of
synth
hesised AgN
NPs ranged between 70
0-110 nm, mainly
m
spherrical and soome in hexa
agonal
shape
e. AgNPs exxhibited significantly high
her antibacterial activity (up to 200x) against hum
man as
well as plant pa
athogens (E
Escherichia coli, Pseud
domonas aeruginosa annd Agrobactterium
tumefaciens) com
mpared to AgNO3 sol ution and W. somniferra leaf extrract. The cellular
intera
action study coupled witth SEM ana
alysis reveale
ed the effec
ctive uptake of AgNPs by
b the
bacte
eria. The co
ompounds responsible
r
for the na
anoparticle formation
f
w
were identifie
ed as
phenolics and wh
hitanolids pre
esent in the e
extract, as re
evealed by HPLC analysiss.
ant capacitty of Melia
a azedarac
ch under water
w
defiicit condittions
O4 Antioxida
Azeve
edo C, Dias MC*,
M
Costa M, Pinto G and S
Santos C
Deparrtment of Biollogy and CES
SAM – Centre for Environme
ental and Marrine Studies, U
University of Aveiro,
A
Camp
pus Universitário de Santiag
go, 3810-193 A
Aveiro, Portug
gal.
*Corre
espondig auth
hor: celeste.dia
[email protected]
This w
work aims to
o evaluate th
he effects of water stress
s (WS) on th
he antioxidannt capacity and
a on
the photosynthetic apparatus of the mediccinal plant Melia
M
azedara
ach L. Two m
month old pla
ants of
M. azzedarach we
ere exposed to WS (plan
nts at 20% off field capacity) during 220 days. Afte
er this
period, plant pe
erformance was evalua
ated through the measurrement of thhe water potential,
plant growth, chllorophyll a fluorescence
, gas excha
f
ange, pigment content, H 2O2, proline, the
activities of antioxxidant enzym
mes and anttioxidant mettabolites, cell membranee permeabilitty and
lipid p
peroxidation. IRGA analy
ysis demonsstrated that WS
W induced stomatal cloosure, reduce
ed the
net C
CO2 assimilation rate (A)) and decrea
ased the CO
O2 availability
y in the interrcellular spac
ces of
meso
ophyll cells (Ci). Howeve
er, WS did n ot affect the
e photosynthetic efficienccy of PSII orr plant
growtth (dry weight accumulation and pla
ant height). WS
W increased cell membbrane perme
eability
and induced an up-regulation
u
n of the antio
oxidant enzy
ymes, CAT, APX
A
and Grr and also an
n over
produ
uction of anttioxidant metabolites. De
espite these effects on photosyntheetic apparatus and
increa
ased antioxid
dant capacitty, plant grow
wth was not affected, supporting thatt this species
s may
be u
used in re/a
afforestation programs ffor drought prone habiitats. Moreoover, it wou
uld be
d increase of
intere
esting to ana
alyse if the sttress-induced
o the antioxid
dant capacity
ty may be us
sed as
a possitive strategyy to increase
e antioxidant properties of
o this species.
www.mapp.edu.pt/bioplant 10 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 O5 Refloresta
ação de zo
onas degrradadas pela
p
seca com
c
amarrgoseira:
II. crescimento e fottossíntese
e
1
Maria da Costa 1,2, Glória Pinto1, Conceição Sa
antos
1
2
Depa
artamento de Biologia,
B
Univ
versidade de A
Aveiro, 3810-1
193, Aveiro Po
ortugal
Minisstério da Educcação de Timo
or-Leste, Dili, T
Timor-Leste
Melia
a azedarach,, ou amargoseira, é uma
a espécie len
nhosa que produz
p
madeeira com inte
eresse
come
ercial, e apre
esenta proprriedades med
dicinais utiliz
zadas na tra
adição etnobbotânica de TimorT
Leste
e. Esta espé
écie existe em quase ttodas as regiões do pa
aís –litorais e montanho
osas-.
Contu
udo, Timor-L
Leste apres
senta proble mas cresce
entes de deg
gradação dee solos dev
vido a
perío
odos de seca
a, agricultura
a itinerante, desmatamen
nto descontrrolado, etc. A necessidade de
valoriizar recurso
os florestais,, proteger p
práticas etno
obotânicas e combaterr a desertificação
justifica a introduçção da amarrgoseira na a
arborização de solos deg
gradados pella seca em TimorT
Leste
e. Esta medida permitiria
a melhorar a qualidade de vida e auxiliaria
a
a im
mplementação do
Progrrama Estraté
égico para o Desenvolvim
mento (PED) do Governo
o Leste-Timoorense.
Para esta estraté
égia é neces
ssário avalia r o grau de suscetibilida
ade da amarrgoseira a sttresse
hídricco. Para isso, usaram-s
se sementess e plantas com 2-3 me
eses de idadde, a cresce
er em
estufa
a. Lotes de sementes e de plantass foram exp
postos a condições norm
mais de irrig
gação
(controlos) e outrros lotes de sementes e plantas foram exposto
os a stresse hídrico (20%). O
stressse hídrico attrasou a gerrminação dass sementes e afetou de forma difereente as raíze
es e a
porçã
ão aérea (eg
g., maior pe
eso das raízzes). A sobre
evivência da
as plantas nnão foi afetada. O
stressse diminuiu a eficiência fotossintética
f
a, e reduziu o índice de abertura
a
estoomática, limitando
as tro
ocas gasosa
as. Diminuiu
u também o conteúdo de
d pigmentos, o que poode justificarr uma
meno
or eficácia na
a captação de
d luz peloss PSI/PSII. Houve
H
uma te
endência paara aumentarr o F0
enquanto Fv e Fv/Fm
F
aprese
entaram uma
a tendência de decréscimo com o sstresse. Sen
ndo os
clorop
plastos um organelo muito
m
sensívvel a stresse, analisou
u-se também
m a degrad
dação
membranar (EL). Os dados preliminares
p
ssugerem que
e a fotossíntese na amarrgoseira é affetada
pelo sstresse hídrico, embora não
n seja sufiiciente para compromete
er a viabilidadde das plantas.
www.mapp.edu.pt/bioplant 11 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 O6 Dot-blot validation
v
of novel D
DNA mark
kers for de
etection an
nd
g
genotypin
ng of Ralstonia sola
anacearum
m species complex
2
Albuq
querque P1, Ca
aridade CMR2,3
, Marçal ARS
S2,4, Tavares F1,5
BIO- Centro de Investigaç
ção em Biodiiversidade e Recursos Ge
enéticos, Univversidade do Porto,
1- CIB
Camp
pus Agrário de
e Vairão, Vairã
ão, Portugal
2- CIC
CGE-Centro de
d Investigaçã
ão em Ciência
as Geo-Espac
ciais, Faculdade de Ciênciaas, Universida
ade do
Porto,, Porto, Portug
gal
3- ISE
EC-Instituto Su
uperior de Eng
genharia de C
Coimbra, Coim
mbra, Portugal
4- CM
MUP-Centro de
d Matemátic
ca da Universsidade do Porto, Faculdad
de de Ciênciaas, Universida
ade do
Porto,, Porto, Portug
gal
5- FC
CUP- Faculdad
de de Ciência
as, Departame
ento de Biolo
ogia, Universid
dade do Portoo, Edifício FC
C4, Via
Panorrâmica nº 36, Porto, Portuga
al
Ralsttonia solanaccearum (Rs)) is a Gram--negative rod
d that cause
es systemic wilting disea
ase in
severral crops, namely potato, tomato, ban
nana and tob
bacco, causin
ng importantt economic lo
osses.
In ord
der to enforcce large-scale monitoring
g and quaran
ntine phytosa
anitary proceedures there is the
need to develop specific
s
dete
ection and ty ping method
ds for this pathogen, partticularly due to the
high genotypic diversity obse
erved within the Rs spe
ecies comple
ex. In this reegard, DNA-based
methods presen
nt several advantages
a
over culturre-based methods of detection, mainly
m
conce
erning their specificity,
s
de
etection reso
olution and assay time.
In this work, nove
el Rs specific
c markers, a
able to detec
ct and discrim
minate Rs sttrains belong
ging to
different races and biovars, were selectted and valiidated. Rs-specific proteein domains were
identiified using the
t
Pfam da
atabase and
d a follow-u
up BLAST analysis
a
of tthe correspo
onding
nucle
eotide seque
ences, as prreviously de
escribed [1]. The 17 novel putative Rs-specific
c DNA
markers were asssessed for th
heir specificitty and consistency acros
ss diverse Rss strains and
d nontarge
et species ussing PCR an
nd hybridizattion-based approaches as
a detailed bby Albuquerq
que et
al. [2]. The data obtained sho
owed that w
while some markers
m
were
e broad specctrum, i.e. prresent
throughout the different teste
ed Rs strainss, the majorrity displayed
d strain-speccific patterns
s. The
simulltaneous use
e of broad spectrum
s
an
nd strain-spe
ecific markerrs in high-thhroughput as
ssays,
such as inverted dot blots, prrovides discr iminatory hy
ybridization patterns
p
usefful for presum
mptive
infrassubspecific discrimination
d
n of Rs isolattes. Ultimate
ely, this work
k might contrribute to dete
ermine
epide
emiologic pa
atterns which
h are importa
ant to implement suitablle control meeasures and
d infer
disse
emination mo
odels.
Referrences:
1- Viieira J, Men
ndes MV, Albuquerque P, Moradas
s-Ferreira P,, Tavares F
F. (2007) A novel
appro
oach for the identification of bacteria
al taxa-specific molecula
ar markers. LLett. App. Microb.
M
44: 506-512.
P Caridade CMR, Rodrig
gues AS, Ma
arcal ARS, Cruz J, et al. ((2012)
2- Allbuquerque P,
Evolu
utionary and experimenta
al assessmen
nt of novel markers
m
for de
etection of X
Xanthomonas
s
euvessicatoria in plant
p
samples
s. PLoS ONE
E 7(5): e37836. doi:10.13
371/journal.ppone.003783
36
www.mapp.edu.pt/bioplant 12 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 e plant rhizzosphere system to
o the phyto
oremediattion
O7 Contributtion of the
o
of cadmiu
um in estuarine area
as Marta
a Nunes da Silva
S 1, Ana P. Mucha2, A. Cristina Roch
ha1, Catarina Teixeira2, Caarlos R. Gome
es1, C.
2
Marisa
a R. Almeida
MAR/CIIMAR, Faculdade de
e Ciências, Un
niversidade do
o Porto, Rua do Campo Allegre, s/n, 416
69-007
1 CIM
Porto,, Portugal.
2 CIM
MAR/CIIMAR – Centro Interrdisciplinar de
e Investigação
o Marinha e Ambiental,
A
Uniiversidade do Porto,
Rua d
dos Bragas, 28
89, 4050-123 Porto, Portuga
al.
Some
e salt marsh
h plants have already sh
hown to hav
ve potential for metal phhytoremediattion in
estua
arine areas [1], being imp
portant to st udy and testt strategies to
t enhance tthat potentia
al. The
aim o
of this work was to eva
aluate how t he rhizosphere of Juncu
us maritimuss and Phrag
gmites
austrralis plants in
nfluenced Cd
d removal fro
om sediments
s favoring en
nvironmentall decontamin
nation.
For th
his study, pla
ants of both species, col lected togeth
her with the sediment invvolving their roots,
were placed in ve
essels and maintained
m
in
n greenhous
ses, exposed
d to natural eenvironmenta
al and
light conditions. A nutritive saline
s
solutio
on was add
ded to all ve
essels throu gh an autom
mated
irrigattion system (2 daily cyc
cles of flood / draught) to
t mimic the
e tides and m
maintain pla
ants at
optim
mum nutrition
nal condition
ns. After 2 w
weeks of acclimation, all vessels weere spiked with
w
a
saline
e Cd solution
n, which was
s in contact w
with the plantt rhizosphere
e for about 6hh. Afterwards
sa
solutiion containin
ng an autoch
hthonous en
nriched cons
sortium of microorganism
ms resistant to Cd
(prep
pared in the laboratory) was
w added tto half of the
e vessels. Vessels
V
weree maintained
d for 2
months in the abovementio
a
ned conditio
ons, being afterwards
a
disassemble.
d
. For that, plants
above
eground tisssues were se
eparated from
m belowgrou
und structures, which in tturn were carefully
separrated from the
t
sediment. Cadmium was determ
mined as beffore [1]. Ressults sugges
st that
both plants werre able to accumulate considerab
ble amounts
s of Cd, esspecially in their
below
wground tissu
ues, without showing sig
gns of toxicity
y. The additio
on of the miccrobial conso
ortium
seem
med to prom
mote Cd tran
nslocation to
o P. australlis stems, in
ncreasing thhe phytoextrraction
poten
ntial of this plant. In ad
ddition, the phytostabiliz
zation potenttial of J. maaritimus was
s also
enhanced. There
efore, the add
dition of auto
ochthonous microorganisms resistannt to Cd see
ems to
be a valuable strrategy to pottentiate the a
ability of the
ese plants to
o phytoremeddiate this me
etal in
saltm
marshes, bein
ng useful for the recoveryy of moderate
ely impacted
d estuaries.
Acknowledgmentts: To FCT, Portugal,
P
for PTDC/MAR//099140/2008.
[1] Allmeida C. M.. R., Mucha A.
A P., Vasco
oncelos M. T.
T S. D. (2011
1). Role of d ifferent salt marsh
m
plantss on metal retention
r
in an
a urban esttuary (Lima estuary, NW
W Portugal). Estuarine Coastal
and S
Shelf Science
e, 91, 243-24
49.
www.mapp.edu.pt/bioplant 13 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 O8 The Plantt Specific Insert and
d Its Molec
cular Role
e in Protein
n Sorting
Peixoto B.1,2, Pereirra C.1, Veríssiimo P.2, Pissa
arra J.1
1
BioFIIG – Centre for Biodivers
sity, Function
nal and Integrrative Genom
mics, Departam
mento de Biiologia,
dade de Ciênccias da Univerrsidade do Po
Faculd
orto, Rua do Campo
C
Alegre, s/nº, 4169-0007, Porto, Porrtugal.
2
Mole
ecular Biotechnology Laborratory, Centerr for Neuroscie
ence and Celll Biology, Unniversity of Co
oimbra,
3004--517 Coimbra, Portugal.
A parrticular chara
acteristic of plant
p
asparticc proteinases is the pres
sence of an aapproximately 100
amino
o-acids long insertion, highly homolo
ogous to botth saposins and saposinn-like protein
ns and
whosse physiological function is currentlyy unknown – the Plant Specific
S
Inseert (PSI). Thiis PSI
doma
ain is characcterized by a closely paccked globula
ar structure comprised
c
byy five amphipathic
α-helices linked to
t each othe
er by three d
disulfide bridg
ges.This dom
mains’ imporrtance in vac
cuolar
trafficcking has alrready been demonstrate
ed in transie
ent expressio
on experimennts using tobacco
proto
oplasts expre
essing a PSI--lacking phyttepsin. Howe
ever, additionally to the PSI’s involve
ement
in pro
otein sorting to the plant vacuole, th
his domain’s
s properties in inducing vvesicle leaka
age in
vitro have been demonstrated, a resultt that sugge
ests plant aspartic proteeinases mig
ght be
bifuncctional moleccules, acting both as mem
mbrane-desttabilizing age
ents and protteinases.
Rece
ently, a nove
el AP has be
een discoverred in Chlam
mydomonas reinhardtii,
r
itts characterization
has rrevealed a se
eries of intrig
guing feature s, such as an 80 amino-a
acid long alaanine-rich ins
sertion
in the
e PSI domain
n, as well as a chloroplasstidial subce
ellular localiza
ation, both oof which had never
been reported forr typical aspa
artic proteasses, turning this novel pro
oteinase, chllapsin, into a most
promising model for studying the moleccular mecha
anisms asso
ociated with the PSI’s role
r
in
prote
ein sorting. Recent
R
work
k by our gro up, with the
e objective of
o further chaaracterizing these
doma
ains, from a bioinformatics, biochem
mical and cellular perspec
ctives have recently identified
functiional diversiity associate
ed with PSI--mediated protein
p
sortin
ng routes, w
which could prove
usefu
ul from a biottechnological point of view
w.
O9 A highly efficient
e
le
eaf protop
plast expre
ession sys
stem for th
he study of
o
a
anticance
er alkaloid metabolis
sm in Cath
haranthus
s roseus
Patríccia Duarte1*, Diana
D
Ribeiro1,2, Inês Carque
eijeiro1,3 and Mariana
M
Sottomayor1,3,*
1
IBM
MC – Instituto de Biologia Molecular
M
eC
Celular, Univerrsidade do Po
orto, Rua do C
Campo Alegre
e, 823,
4150--180 Porto, Po
ortugal
2
Dep
partamento de
e Biologia, Uniiversidade do Minho, Camp
pus de Gualtarr, 4710-057 Brraga, Portugal
3
Dep
partamento de
e Biologia, Fac
culdade de Ciê
iências da Uniiversidade do Porto, Rua doo Campo Aleg
gre s/n,
4169--007 Porto, Po
ortugal
Catha
aranthus rosseus leaves accumulate in low levels
s the antican
ncer terpeno id indole alk
kaloids
(TIAss) vinblastine
e and vincrristine, and intense research has uncovered a number of
o TIA
biosyynthetic step
ps and regu
ulatory gene
es. Howeverr, there are still enorm
mous gaps in
i the
know
wledge on the
e TIA biosyntthetic pathwa
ay, its regula
ation and TIA
A transmembbrane transport. At
prese
ent, several C. roseus transcriptomiic projects are
a unveiling a high num
mber of cand
didate
genes, urging the
t
develop
pment of e
easy and efficient
e
mo
olecular toools for func
ctional
chara
acterization. Leaves are
e the single
e C. roseus
s organ wh
here the bioosynthesis of
o the
antica
ancer TIAs is
i completed
d, and thereffore mesoph
hyll cells are the ideal taarget for TIA
A gene
functiional analysiis. Here, a highly
h
efficien
nt method fo
or C. roseus mesophyll pprotoplast iso
olation
and ttransformatio
on is reported
d, enabling tto reproducib
bly test TIA candidate
c
geenes in the precise
cells where the biosynthesis
s and accum
mulation of the
t
anticance
er TIAs occcur. As a proof of
princiiple of the application of this mole
ecular tool, the subcellular sorting determinatiion of
CroP
Prx1, a classs III peroxid
dase involve
ed in TIA biosynthesis,
b
was investtigated. Transient
expre
ession of diffe
erent CroPrx
x1-GFP fusio
ons revealed that CroPrx1 is sorted too the vacuole
e by a
C-terminal peptiide signal. After custtomized op
ptimization of
o protoplasst isolation, the
ems to be ea
asily adaptable to differen
nt species, suuggesting that this
transfformation procedure see
is a plant-transfe
erable tool that may b
be applied to
t non-mode
el species ffor the func
ctional
chara
acterization of
o the wealth of candidate
e genes eme
erging from next
n
generati on sequencing.
www.mapp.edu.pt/bioplant 14 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 O10
0 Photosy
ynthesis in
n grape be
erry tissue
es: an in vivo
v
appro
oach by
imaging PAM fluorometry
ard Breia1,2, Só
ónia Vieira3,4, Jorge Marque
es da Silva3, Hernâni
H
Gerós1,2 and Ana Cuunha1,2
Richa
1
Centtro de Investig
gação e de Te
ecnologias Agrro-ambientais e Biológicas CITAB-UM,
C
Po
Portugal
Gru
upo de Invesstigação em Biologia Veg
getal Aplicada
a e Inovação
o Agroalimenntar (AgroBio
oPlant),
Deparrtamento de Biologia,
B
Escolla de Ciênciass Universidade
e do Minho, Braga, Portugaal
3
Dep
partamento de
e Biologia Ve
egetal e Cen
ntro de Biodiv
versidade, Ge
enómica Integgrativa e Fun
ncional
(BioFIIG), Faculdad
de de Ciências
s da Universida
ade de Lisboa
a, Lisboa, Porttugal
4
Centtro de Oceano
ografia, Faculd
dade de Ciênccias da Univerrsidade de Lis
sboa, Lisboa, P
Portugal
2
Photo
osynthesis iss not exclusiv
ve of green lleaves. In fact, some sink tissues, likke fruits, whic
ch are
chloro
ophyllous du
uring early developmen
ntal phases, could exhib
bit photosynnthetic activitty. To
chara
acterize the photosynthetic propertiies of the green
g
grape
e berry, thee pulse amp
plitude
chloro
ophyll fluore
escence imaging (imagin
ng-PAM) wa
as used. Also the effectt of the satu
uration
pulse
e (SP) inten
nsity on pho
otochemical parameters was tested
d. Results reevealed a tissuespeciific distribution pattern of photosyynthesis and
d a strong effect of S
SP on max
ximum
fluore
escence (Fm) and in ra
apid light cu
urves (RLC) parameters
s, such as F’m and efffective
quanttum yield (Φ
ΦII). After RLC
R
modelin
ng, the exoc
carp showed the higheest photosyn
nthetic
capaccity and the lowest pho
otoinhibition ssusceptibility
y and the mesocarp,
m
revvealed the lowest
fluore
escence sign
nals and ph
hotochemica l competenc
ces. Notably
y, the seed outer integu
ument
prese
ented a high
h photochemical capacityy, similar to the exocarp, although itt is more pro
one to
photo
oinhibition. Globally,
G
all tissues satura
ated the PSII reaction ce
enters at relaatively low SP
P light
intenssities. Above
e 5000 μmol m-2 s-1, Fm, F
F’m and ΦII were
w
significa
antly affectedd, but inner tissues
under light-adaptted condition
ns were sussceptible at lower
l
satura
ating light (32200 μmol m-2 s-1)
indica
ating a phottoinhibitory in
nteraction be
etween SP and actinic light intensitties and rep
petitive
expossure to saturrating pulses
s (1). These results open
n the way to further studiies concernin
ng the
involvvement of tisssue-specific
c photosynth
hesis in the highly comp
partmentalizeed production and
accum
mulation of organic
o
comp
pounds durin
ng grape berrry development.
1 - B
Breia et al. 2013. Map
pping Grape Berry Phottosynthesis by Chlorophhyll Fluorescence
Imaging: The Eff
ffect of Satu
urating Pulse
e Intensity in Different Tissues. Phhotochemistry
y and
Photo
obiology, 89:: 579–585.
Acknowledgmentts: This work
k was suppo
orted by Eurropean Union Funds (FE
EDER/COMP
PETEe) and by na
ational funds (FCT-Portugguese Found
dation
Operrational Competitiveness Programme
for S
Science and
d Technolog
gy) under the projects FCOMP-0
01-0124-FED
DER-022692
2 and
Europ
pean COST action FA 1106
1
and th
he research project PTD
DC/AGR-ALI//100636/2008. RB
was ssupported th
hrough a rese
earch grant fellowship (U
UMINHO/BI/177/2011) asssociated wiith the
project PTDC/AG
GR-ALI/10063
36/2008.
www.mapp.edu.pt/bioplant 15 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 O11 Copper transporte
ers in grap
pevine
na Martins1,2, António
A
Teixeira1,2, Elias Ba
assil3, Eduardo
o Blumwald3, Hernâni Geróós1,2
Vivian
1
Centtro de Investig
gação e de Tecnologias
T
A
Agro-Ambienta
ais e Biológica
as (CITAB), P
Portugal; 2Gru
upo de
Investtigação em Biiologia Vegeta
al Aplicada e In
nvestigação Agroalimentar
A
– Agrobioplannt, Departame
ento de
Biolog
gia, Escola Ciências,
C
Univ
versidade do Minho, Cam
mpus de Gualtar, Braga, 44710-057, Po
ortugal;
3
Depa
artment of Plant Sciences, University
U
of C
California, One
e Shields Ave, Davis, CA 955616, USA
Fungicides based
d on copper salts
s
have be
een extensiv
vely used in viticulture
v
sinnce the late 1800s
upon the develop
pment of the
e ‘Bordeaux mixture’, a broad spectrrum fungicidde that consists of
coppe
er sulfate an
nd lime. Des
spite being a
an essential element for plant healtth, excess copper
c
may b
be a cause of
o toxicity, an
nd several m echanisms such
s
as chela
ation and com
mpartmentattion of
metal ions have evolved to avoid
a
negativve effects att cell level while
w
insuringg their appro
opriate
delive
ery within the cell compa
artments. Prrevious studies have sho
own that thee viability of grape
cells decreases with
w the incre
ease in coppe
er concentra
ation in a dos
se-dependennt manner an
nd that
n of the mettal ion in the
e vacuole may
m
constitutte an effectiive mechanism to
the ssequestration
avoid
d its inherent toxicity. Co
opper transp
porters (COP
PT/Ctr) operate at the pplasma mem
mbrane
level and in interrnal membra
anes, such a
as the tonoplast. These high-affinity transporters
s have
mans and in some
s
plant species.
s
In tthe present study,
been mostly charracterized in yeast, hum
eight putative Vitiis vinifera Co
opper Transp
porters (VvC
CTrs) were id
dentified and the express
sion of
VvCT
Tr1 was inve
estigated in grapevine ttissues throu
ughout deve
elopment andd ripening and
a
in
respo
onse to Bord
deaux mixturre. Furthermo
ore, the loca
alization of VvCTr1
V
was assessed in
n plant
cells after transie
ent transform
mation with fu
usion protein
ns of VvCtr1 with GFP aand RFP. VvCTr1
V
succe
essfully resto
ored the grow
wth-defect o
of yeast muta
ants lacking Ctr transporrters, validating its
functiion as a cop
pper transpo
orter and giviing insights on its contribution for coopper mobilization
within
n the cell.
Acknowledgmentts: This work is support
rted by Euro
opean Union
n Funds (FE
EDER/COMP
PETEOperrational Competitiveness Programme
e) and by po
ortuguese na
ational funds (FCT-Portug
guese
Foundation for Science
S
and Technology)) under the project FCO
OMP-01-01244-FEDER-02
22692,
the
project
research
FCOMP-01-0
F
0124-FEDER
R-008760 (Refª.
FC
CT
PTDC//AGRALI/100636/2008), the projec
ct FCT/5955//27/5/2013/S
S - scientific cooperationn Portugal-Tu
unisia,
and tthe PhD gra
ant no. SFR
RH/BD/6458 7/2009 to VM.
V
This wo
ork is also supported by
b the
Europ
pean projectt INNOVINE
E (ref. 31177
75), by the networking activities
a
witthin the euro
opean
funde
ed COST ACTION
A
(FA
A1106 “Qua lityFruit" and FA1003 "East-West Collaboratio
on for
Grapevine Diverssity Exploratio
on and Mobi lization of Ad
daptive Traits
s for Breedinng”).
O12 - Desenv
volvimento
o Sustentá
ável e Eco
oinovação - ferrameentas de
combatte à crise
Ecoinside
www.mapp.edu.pt/bioplant 16 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 O13 Identifica
ation of he
eavy meta
al transporrters in So
olanaceae plants
A.M. P
Pessoa1,2, S. Pereira1,2 an
nd J.Pissarra1
1,2
o, Faculty of Sciences, Bio
ology Departm
ment, Edifício FC4,
F
Rua do Campo Alegrre, s/n,
1Univversity of Porto
4169--007 Porto, Po
ortugal;
2BioF
FIG - Centerr for Biodiverrsity, Function
nal & Integra
ative Genomics, Plant M
Molecular Biollogy &
Biotecchnology Lab, University of Porto, Portug
gal;
The S
Solanaceae include commonly edible
e plants such
h as Potato (Solanum
(
tubberosum), To
omato
(Sola
anum lycoperrsicum) and Eggplant (S olanum melo
ongena). This plant taxoon is considered to
be on
ne of the mo
ost economic
cally importa
ant and the most valuab
ble regardingg vegetable crops.
The ttwo model plants
p
tomatto and potatto had their genome rec
cently sequeenced and is
s now
publiccly available.
Metal transport and uptake of metals are essential fo
or plant growth, developm
ment, nutritio
on and
signa
al transductio
on. In addition, plant me
etal transpo
orts systems are a very important part
p
of
mech
hanisms for decrease
d
the
e toxicity of ce
ertain metal ions.
nown to tran
There
efore, many metal transp
porters are kn
nsport toxic cations
c
(suchh as heavy metals)
m
along
g with cationic nutrients.
The Z
ZIP family (Z
ZRT, IRT-like
e proteins) pllays promine
ent roles in metal
m
ions upptake, transp
porting
Zn, F
Fe and Mn from outside the cell in
nto the cyto
oplasm. The CDF familyy (cation difffusion
facilittators) transsports a wide
w
range of metal ions and is known to promotte Zn
comp
partmentaliza
ation by pum
mping the me
etal out of the
e cell or into
o the lumen oof organelles
s. The
NRAM
MPs (natural resistance-associated m
macrophage proteins) are
e known to pparticipate in
nto the
transport and reg
gulation of Fe
e. All of thesse three families of transporters are kknown to po
ossess
Cd tra
ansport abilitty.
A larrge number of heavy metal
m
transpo
orters have already bee
en describedd for Thlasp
pi and
Arabiidopsis as well
w as for other
o
plant sspecies spec
cies, but only
y a limited nnumber has been
identiified for Solanaceae. However,
H
the
ere are sev
veral inconsistencies in the heavy metal
transporter seque
ences published and alsso many una
annotated So
olanaceae seequences in NCBI
datab
bases (unkn
nown DNA clones,
c
EST
Ts, GSSs, genome
g
seq
quences) thaat may reprresent
putative metal tra
ansporters.
Using
g these anno
otated seque
ences along
g with the se
equence info
ormation from
m NCBI, and
d data
minin
ng and gene
e prediction techniques,
t
several puta
ative transpo
orters from tthe ZIP, CDF and
NRAM
MP family were
w
identifie
ed and chara
acterized in tomato and potato, as w
well as com
mpared
with o
other plant species, such
h as Thlaspi a
and Arabidop
psis.
Our cconserved primers
p
desig
gn tool, Prim
merIdent (http://primerident.up.pt), w
was then use
ed for
desig
gning suitable
e primers tha
at allowed no
ot only to con
nfirm the pre
edicted trans porter seque
ences,
but to
o identify them in other Solanaceae p
plants as well.
We e
expect that th
hese results will
w be a step
pping stone in the unders
standing of m
metal homeo
ostasis
in So
olanaceae pla
ants.
www.mapp.edu.pt/bioplant 17 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 Pos
sters
___
______
_______
______
______
_______
______
_______
__
P1 P
Polyol transport and metabo
olism adjus
stments in
n Vitis vin
nifera in
rresponse to water stress
s
Artur Conde1,2, Ana
a Regalado3, Diana
D
Rodrigu
ues2, M. Manu
uela Chaves3,4 and Hernâni Gerós1,2
1
Centtro de Investig
gação e de Te
ecnologias Agrro-ambientais e Biológicas CITAB-UM,
C
Po
Portugal
Gru
upo de Invesstigação em Biologia Veg
getal Aplicada
a e Inovação
o Agroalimenntar (AgroBio
oPlant),
Deparrtamento de Biologia,
B
Escolla de Ciênciass Universidade
e do Minho, Braga, Portugaal
3
Instittuto de Tecnollogia Química e Biológica, A
Apartado 127, 2781-901 Oe
eiras, Portugall
4
Instittuto Superior de Agronomia
a, Universidad
de Técnica de
e Lisboa, Tap
pada da Ajudaa, 1349-017 Lisboa,
L
Portug
gal
2
Polyo
ols, or sugarr alcohols, arre widely disstributed com
mpounds am
mong plants aand frequenttly act
as ossmoprotectan
nts. In Olea europaea, m
mannitol is an
n important osmoprotect
o
tant under drrought
and ssalinity, where mannitol transport an
nd intracellula
ar oxidation are well cooordinated (1)). GCTOF--MS analysiss revealed th
hat several p
polyols are present
p
in gra
apevine (Vitiis vinifera) mature
m
leave
es and berryy mesocarp in significan
nt concentrations. Severral polyols aaccumulate during
d
waterr deficit in the
e final maturration stagess of berry dev
velopment. A significant inhibition (50
0%) of
sorbittol oxidation to fructose via
v sorbitol d
dehydrogena
ases (VvSDH
Hs) in mesoccarp is clear in the
full m
mature stage of grape be
erries under drought. Ma
annitol dehyd
drogenase (V
VvMTD) activity is
also severely repressed (4-ffold) in the mature and
d full mature stages unnder water deficit
condiitions. These
e data are we
ell in line witth our previous observations regardinng polyol tran
nsport
by VvvPLT1. The combination of metabo
olomic, mole
ecular biolog
gy and tradittional bioche
emical
appro
oaches proviided new ins
sights on the
e role of poly
yols in grape
evine defensse against drrought
stresss. Additionally, increased
d concentratiions of polyo
ols in grape berries may constitute a novel
bioma
arker for dro
ought-toleran
nt grapevine cultivars and a bioindica
ator of plantt health/adap
ptation
under water deficcit.
(1) Conde et al. (2
2011) Plant Cell
C Physiol 52(10):1766
6-75
Acknowledgmentts: This work
k was suppo
orted by Eurropean Union Funds (FE
EDER/COMP
PETEOperrational Competitiveness Programme
e) and by na
ational funds (FCT-Portugguese Found
dation
for S
Science and
d Technolog
gy) under the projects FCOMP-0
01-0124-FED
DER-022692
2 and
Europ
pean COST action FA 1106
1
and the
e research projects
p
PTD
DC/AGR-ALI//100636/2008 and
PTDC
C/AGR-AAM/099154/200
08. AC was ssupported by
y the PhD gra
ant no. SFRH
H/BD/47699//2008.
www.mapp.edu.pt/bioplant 18 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 Reporting
g On Ende
emic Enda
angered Pllant Species Using Predictive
e
P2 R
M
Models
2,3,4
Rita S
S. Silva 1, Paullo Alves 2, Anttoine Guisan 3, João Honrad
do 1,2, Angela Lomba
L
1
Facu
uldade de Ciê
ências, Universidade do Po
orto, Rua do Campo Alegrre, s/n, 4169--007 Porto, Po
ortugal
Cen
ntro de Investiigação em Bio
odiversidade e Recursos Genéticos
G
(CIB
BIO), Rua do Campo Alegrre, s/n,
4169--007 Porto, Po
ortugal
3
Lab
boratoire de Biologie
B
de la
l Conservatiion (LBC), Département
D
d'Ecologie
d
et d'Evolution (DEE),
Unive
ersité de Lausa
anne, Bâtimen
nt de Biologie,, CH-1015 Lau
usanne, Switz
zerland
4
Alterrra Wageninge
en University and Research
h Centre, NL-6
6708 Wagenin
ngen, The Neth
therlands
2
Curre
ent patterns of species distributions a
are largely determined by
y changes inn climate and
d land
uses,, with its im
mpacts on biodiversity and associated ecosystem servicces being widely
w
ackno
owledged. Rare
R
species are a growiing concern in the context of biodiveersity conserrvation
world
dwide, since these specie
es are assum
med to unde
ergo higher risk of extincttion. Rare sp
pecies
are characterized
d by restricted
d geographicc ranges, habitat specialization and/oor small popu
ulation
sizess. Even thoug
gh datasets on rare speccies distributtion usually have
h
few obsservations, limited
spatia
models that relate
al accuracy and lack of valid absen
nces, the dev
velopment of predictive m
speciies occurrences to enviro
onmental de scriptors hav
ve been prov
ved to be paarticularly useful to
realisstically captu
ure a large
e part of th
hese species ecological niche. Veeronica micrrantha
Hoffm
manns. & Link is an en
ndangered p
plant species
s, endemic of the Iberiaan Peninsula
a and
prote
ected under the EU Habitats Directive
e. Under Artic
cle 17 of this
s Directive, E
EU Member States
S
are o
obliged to un
ndertake mo
onitoring, asssessment an
nd reporting of conservaation status for all
habita
ats and speccies of Comm
munity intere
est. Thereby, this study aimed to asseess the usefu
ulness
of pre
edictive mod
dels as asse
essment toolls to effectiv
vely fulfil the reporting obbligations, and as
tools to identify environmenttal condition
ns within wh
hich populations can bee maintained
d. The
poten
ntial distributtion of Veron
nica micranth
ha was estim
mated based
d on two diffeerent approa
aches:
minim
mum convexx polygon and
a
speciess distribution
n modelling. The formeer resulted in an
overe
estimation of
o the species distributi on, whereas the latter gave a m
more conserv
vative,
presu
umably more
e realistic, as
ssessment o
of the distribution of the species whiile also allow
wing a
better understand
ding of the ecology and
d dynamics of the spec
cies. Therefoore, the proposed
appro
oach constittutes a step
p forward in
n the study of the disttribution pattterns of endemic
endangered plan
nt species, as
a well as iin the use of
o these mo
ore informattive projectio
ons to
impro
ove the reporrting and mo
onitoring of th
hese at-risk species.
s
A. Lo
omba is supp
ported by the
e Portugues e Science and Technolo
ogy Foundatiion (FCT) th
hrough
Post Doc Grant SFRH/BPD/
S
1. This study
y was partiallly funded byy FEDER th
hrough
80747/2011
“Prog
grama Opera
acional Facttores de Co
ompetitividade – COMPE
ETE” and byy the Portug
guese
Gove
ernment through “FCT – Fundação p
para a Ciênc
cia e a Tecn
nologia”, in thhe form of project
p
grantt “EcoSensin
ng: indicato
ors, methodss, and prottocols for re
eporting andd monitoring the
condiition of biodivversity and ecosystems
e
i n rural lands
scapes” (PTD
DC/AGR-AAM
M/104819/20
008).
www.mapp.edu.pt/bioplant 19 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 Plant spec
cies assem
mblages a
along a grradient of forest
f
natturalness in
P3 P
d
dairy-farm
mlands of the
t Northw
western Portugal
P
Ana S
Sofia Vaz1, Joã
ão Honrado1,22 and Angela L
Lomba1,3,4
1
Centtro de Investig
gação em Biodiversidade e Recursos Ge
enéticos, Univ
versidade do Porto, Edifício
o FC4,
Rua C
Campo Alegre
e s/n, 4169-007 Porto, Portu
ugal
2
Facu
uldade de Ciê
ências (Departtamento de B
Biologia), Univ
versidade do Porto, Edifícioo FC4, Rua Campo
C
Alegre
e s/n, 4169-00
07 Porto, Portu
ugal
3
Alterr
rra Wageninge
en University and
a Research Centre, NL-6
6708 Wagening
gen, The Nethherlands
4
Dépa
artment d’Eco
ologie et d’Evolution (DEE)), Université de
d Lausanne, Bâtiment dee Biologie, CH
H-1015
Lausa
anne, Switzerlland
Know
wledge of plant assemblages is esssential to co
omprehend ecological pprocesses, and
a
to
enhance efficient managemen
nt and conse
ervation mea
asures across
s forests worrldwide.
A tota
al of 50 stan
nds were surrveyed along
g a gradient of forest natturalness in dairy-farmlands in
Portu
ugal, and cla
assified cons
sidering theirr dominant tree:
t
Eucalyp
ptus globuluus, Pinus pin
naster,
and m
mixed stands with both species. Va
ascular plants were recorded within 10m2, 100m
m2 and
1000m2 squares centred in the
t stand ce
entroid, and within the whole
w
stand. Species rich
hness,
assem
mblages and
d species-are
ea relationsh
hips were evaluated acro
oss forest typpes. For the whole
comm
munity, plantt assemblage
es were asssessed calcu
ulating co-occ
currence inddices. For the tree
comm
munity, assemblages were explored w
within 100m2 squares, where trees w
with ≥1.30m height
were mapped an
nd analysed through po
oint pattern analyses.Pin
a
nus stands eexhibited sm
mallest
areass and highesst species ric
chness. For all forest typ
pes, species richness inccreased with
h area
within
n each stand
d. Segregatio
on patterns p
prevailed in plant commu
unities regarrdless of the scale
and fforest type. Still, random patterns were found
d in mixed and
a
Eucalypptus stands. Tree
assem
mblages revvealed trends
s for regular patterns in Pinus
P
stands
s, and aggreegation patte
erns in
the o
other forests. Overall, re
esults converrge with pre
evious research evaluatinng the impa
acts of
forestt naturalnesss on plant div
versity. Pinu
us stands sho
owed indepe
endent relatioons between
n area,
comm
munity speccies richnes
ss and asssemblages, possibly due
d
to higgh environm
mental
heterrogeneity an
nd low disturrbance levells related to
o forest man
nagement. T
Tree assemb
blages
appeared to refle
ect shifts in the manag
gement inten
nsity of each forest, witth close-to-n
nature
regularities in Pinus,
P
and fine-scale
f
ag
ggregations in mixed and
a
Eucalypptus stands.. This
sugge
ests that stru
uctural changes within sttands could determine plant diversityy and interac
ctions,
and e
ecosystem processes and
d services.
Resu
ults highlight forest natu
uralness and
d manageme
ent as key drivers
d
of pplant diversity and
assem
mblages, so extensively managed ssemi-natural stands mustt be promoteed for sustainable
provissions of ecossystem services from pla
anted forests in intensive farmlands.
Acknowledgemen
nts: A. Lom
mba is supp
ported by th
he Portuguese Science and Techn
nology
Foundation (FCT
T) through Post Doc Gra
ant SFRH/BP
PD/ 80747/2
2011. This sstudy was pa
artially
funde
ed by FEDER
R through “P
Programa Op
peracional Factores de Competitivida
C
ade – COMP
PETE”
and b
by the Portug
guese Government throu
ugh “FCT – Fundação
F
pa
ara a Ciênciaa e a Tecnologia”,
in the
e form of pro
oject grant “EcoSensing : indicators, methods, an
nd protocolss for reporting and
monittoring the condition
c
of biodiversity and ecosys
stems in rural landscappes” (PTDC//AGRAAM//104819/200
08).
www.mapp.edu.pt/bioplant 20 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 Lipoxygen
nase in Viitis Viniferra culture cells and grape berrry
P4 L
Antón
nio Teixeira1,2, Diogo Araújo
o2, Henrique N
Noronha1,2, Jos
sé Eiras Dias3, Hernâni Gerrós1,2
1
Centtro de Investig
gação e de Tec
cnologias Agro
ro-Ambientais e Biológicas CITAB-UM,
C
Poortugal
Grup
po de Investtigação em Biologia
B
Veg
getal Aplicada
a e Inovação
o Agroalimenntar (AgroBio
oPlant),
Deparrtamento de Biologia,
B
Escolla de Ciênciass, Universidad
de do Minho, Braga,
B
Portugaal
3
Instittuto Nacional de
d Investigaçã
ão Agrária INIA
IA, Quinta da Almoinha,
A
256
65-191 Dois P
Portos, Portuga
al
2
Vitis vinifera is a major crop worldwide o
of extreme socioeconom
s
mic importancce. Environm
mental
factorrs like heat, drought, soil salinity or pathogen in
nfections can
n affect berryy compositio
on and
thus compromisin
ng quality. Lipoxygenase
L
es (LOXs) are a group of
o dioxygenaases that ca
atalyse
oxyge
enation of polyunsaturat
p
ted fatty aciids (PUFAs)) and lipids, and initiatee the formation of
biolog
gically active
e compound
ds known a
as oxylipins.. LOXs can play severral roles in plant
metabolism and the
t products of its reactio
on are signa
aling moleculles in woundding and path
hogen
attackk, inducers of cell death, inhibitors of mycotox
xin synthesis
s, and signaaling molecu
ules in
severral other strresses like water defici t. Thus, its study may provide infformation on
n how
grape
evine respon
nds to enviro
onment. In g rape genome (Vitis vinife
fera L.) LOX family consists of
18 individual mem
mbers. While
e the expresssion of VvLO
OXC and -D is evenly diistributed between
seedss, pulp and skin, and VvLOXO
V
is mostly expre
essed in the
e seed, VvLLOXA is the most
abundant isoform
m in berry skins in all devvelopmental stages. In the present sstudy LOX activity
a
was m
Sauvignon Berry)
measured in
n grape cells
s (Cabernet S
B
subjec
cted to salt, osmotic and
d heat
stresss, and to tre
eatments with
h stress rela
ated hormones (salicylic acid - SA aand abscisic acid ABA)). LOX activitty was also measured
m
in grapes (gre
een pea, vera
aison and maature stages) from
the ccv. Alvarinho
o, sampled in
n two differe
ent Portugue
ese regions: Demarcatedd Region of Vinho
Region (Instituto
Verde
e (Estação Vitivinícola Amândio G
Galhano, EV
VAG) and Estremadura
E
Nacio
onal de Inve
estigação Agrária, INIA). Results sho
owed that ex
xcept for saltt treatment, which
decre
eased enzym
me activity by
b 25%, all tthe treatmen
nts caused a significantt increase off LOX
activity, in particu
ular ABA and
d SA. The h
hormone ABA
A is likely to
o stimulate LLOX activity at the
transcriptional levvel because the increassed enzyme activity in grape
g
cells ccorrelated with
w an
increa
ase of VvLOXA transcripts measu
ured by qPC
CR. Concern
ning measu rements in intact
berrie
es, results sh
howed that LOX
L
activity picked at the
e veraison stage in both cultivars and was
highe
er in INIA tha
an in the north.
Acknowledgmentts: This work is support
rted by Euro
opean Union
n Funds (FE
EDER/COMP
PETEmpetitiveness Programme
e), European
n COST action FA 11066, and by na
ational
Operrational Com
fundss (FCT-Portu
uguese Foundation for Sccience and Technology)
T
under the prroject FCOM
MP-010124-FEDER-022
2692 and the
e research prroject PTDC//AGR-ALI/10
00636/2008.
www.mapp.edu.pt/bioplant 21 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 Photomixotrophic grape
g
berrry cell sus
spensions
s: a suitab
ble cell mo
odel
P5 P
ffor the stu
udy of abio
otic stress
ses in fruiit photosy
ynthesis
1,2
Richa
ard Breia1,2, Fá
ábio Ramalho2, João Serôdi
dio3, Hernâni Gerós
G
and An
na Cunha1,2
1 Cen
ntro de Investig
gação e de Te
ecnologias Ag
gro-ambientais
s e Biológicas CITAB-UM, P
Portugal
2 Gru
upo de Invesstigação em Biologia Veg
getal Aplicad
da e Inovação Agroalimenntar (AgroBio
oPlant),
Deparrtamento de Biologia,
B
Escolla de Ciênciass Universidade
e do Minho, Braga, Portugaal
3 Dep
partamento de
e Biologia e CESAM – Ce
entro de Estu
udos do Ambiiente e do Maar, Universida
ade de
Aveiro
o, Campus de
e Santiago, Av
veiro, Portugall (richardgonca
[email protected]
minho.pt)
Fruit are essentia
ally sink organs, so fruit p
photosynthes
sis has been seldom overrlooked. How
wever,
recen
nt findings prroved that so
ome grape b
berry tissues exhibit high photochemiical activity, which
may contribute to fruit and
d seed devvelopment (1). In the present stuudy, two lines of
photo
ed from CS
omixotrophic suspension
n cells derive
SB (Cabernet Sauvignonn Berry) cells and
exoca
arps of grape
e berries from cv. “Alvarrinho” were used
u
to deterrmine the efffect of coope
er and
waterr stress in ce
ell growth, ch
hlorophyll con
ntent and ma
aximum phottochemical qquantum effic
ciency
(Fv/Fm). While cop
pper-based fungicides
f
ha
ave been widely used ag
gainst severa
ral grapevine
e (Vitis
vinife
era L.) disea
ases since the late 18
800s, althou
ugh its intensive appliccation has raised
phyto
otoxicity conccerns, water deficit is pro
obably the most
m
important environmeental stress factor
in agrriculture, ressponsible for major reducctions in plan
nt productivity
y. Several sttudies have shown
s
the n
negative imp
pact of thes
se stresses in leaf pho
otosynthesis in many sspecies, including
grape
evine, but litttle informatio
on is still ava
ailable conce
erning fruit photosynthes
p
sis. In the prresent
studyy, water deficcit conditions
s (Ψw = -0.4
4 MPa and -0.8
MPa) ca
aused a reduuction in the
e fresh
weigh
ht and an inccrease in dry
y weight seve
en days after sub-culture
e of both celll lines. Chlorrophyll
conte
ent was reduced in CSB-derived phottomixotrophic cell culture
es, but maxim
mal photoche
emical
poten
ntial (Fv/Fm) decreased in both cell lines. When 40 or 80 μM CuSO4 w
were added to the
culturre media, cell growth, chlorophyll ccontent and Fv/Fm values were redduced in a dosedependent mann
ner in the CSB-derived
C
photomixottrophic cultu
ure cells, buut none of these
param
meters were
e affected in exocarp-derrived culture
e cells. Resu
ults are discuussed in terrms of
putative differential effects of copper and drought over the photochemical activvity of grape
e berry
meso
ocarp and exxocarp tissue
es.
1 - Breia et al. 2013. Mapping Grape Berry Photosynthes
sis by Chlorophyll Fluoresccence Imaging
g: The
otochemistry and Photobioloogy, 89: 579–5
585.
Effectt of Saturating Pulse Intensity in Different Tissues. Pho
Ackno
owledgments: This work wa
as supported by European Union Funds (FEDER/COM
MPETE-Operrational
Comp
petitiveness Programme)
P
and
a
by natio
onal funds (F
FCT-Portugues
se Foundatioon for Scienc
ce and
Techn
nology) underr the projects FCOMP-01-0
0124-FEDER-022692 and European
E
CO
OST action FA
A 1106
and tthe research project PTDC
C/AGR-ALI/10
00636/2008. RB
R was supp
ported throughh a research
h grant
fellow
wship (UMINHO
O/BI/177/2011
1) associated w
with the project PTDC/AGR
R-ALI/100636/22008.
www.mapp.edu.pt/bioplant 22 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 P6 U
Unravellin
ng the molecular ne
etwork reg
gulating zinc conten
nt in plants:
ffrom Arab
bidopsis to
o rice
Joana
a G. Guedes1, Diego Almeid
da2, Mark G.M
M. Aarts3, Nels
son J. M. Saibo
o2, Ana G.L. A
Assunção1
1
Plantt Evolution Group,
G
CIBIO//InBIO, Unive
ersidade do Porto,
P
R. Pad
dre Armando Quintas, 448
85-661
Vairão
o, Portugal
2
Geno
omics of Plant Stress Labo
oratory, ITQB, Universidade
e Nova de Lisboa, Av. da R
República, 278
80-157
Oeirass, Portugal
3
Labo
oratory of Genetics, Wageningen Unive
ersity, Droeve
endaalsesteeg
g 1, 6708 PB
B Wageningen
n, The
Nethe
erlands
As a plant micro
onutrient, zin
nc is of ma
ajor interest in agronom
my and for hhuman and cattle
nutrition. Zinc deficient soils are widespre
ead all over the world and the risk oof inadequatte diet
and zzinc malnutrition is estimated to affecct one third of
o the global human popuulation, i.e. around
a
2 billion people. Developing countrie
es, where people
p
depe
end on cerreal-rich diets for
enance, are
e the most affected. Improvemen
nt of crop efficiency in zinc de
eficient
suste
enviro
onments, an
nd bio-fortific
cation to all eviate huma
an nutrition are plant-baased solutions as
sourcce of micron
nutrients. An
n incomplete
e understand
ding of zinc homeostasiis mechanisms in
plantss is howeverr a limitation.. Recently th e first transc
cription factorrs regulatingg zinc homeo
ostasis
in Ara
abidopsis we
ere identified
d (Assunção et al. 2010).. The AtbZIP
P19 and AtbZ
ZIP23 were shown
s
to be essential for the adaptattion to zinc d
deficiency. Trranscript profiling revealeed only a small set
of ge
enes, to be induced in wild-type p lants in response to zinc deficienccy, but not in the
bzip1
19bzip23 Ara
abidopsis double mutant line. A sugg
gested conse
ervation of thhe zinc defic
ciency
respo
onse regulatory network within the p
plant kingdom makes it very attractiive to explore the
moleccular basis of
o such netwo
ork in rice (O
Oriza sativa L.).
L Rice is on
ne of the moost important crops
world
dwide and it is the staple food for morre than half of
o the world’s
s population. Additionally
y it is a
very relevant crop in Portuga
al. Transferrin
ng the know
wledge from Arabidopsis
A
to rice is of major
imporrtance. We present ou
ur preliminarry results in
n the identification andd analysis of
o the
bZIP1
19/bZIP23 ricce orthologs.
P7 N
Níveis tax
xonómicos
s Superiorres (e.g. género
g
e fa
amília) com
mo
IIndicadore
es de Riqu
ueza Espe
ecífica de Briófitas
Cristia
ana Alves1, He
elena Hespanhol2, Cristiana
a Vieira2 & Ru
ubim Almeida da
d Silva1,2
1
2
Facu
uldade de Ciên
ncias da Unive
ersidade do Po
Porto
Centtro de Investig
gação em Biod
diversidade e R
Recursos Gen
néticos, CIBIO
O-UP
Devid
do ao rápido declínio da biodiversida
ade que se te
em vindo a acentuar
a
nass últimas déc
cadas,
é neccessária uma
a monitorizaç
ção dos habiitats para ava
aliar o estado dos ecoss istemas.
Por isso, uma das
d
soluçõe
es que tem vindo a se
er utilizada para descrrever padrõe
es de
veis taxonóm
micos superiores como
o indicadorees do número de
diverssidade é o uso de nív
espéccies. Este método
m
é van
ntajoso, uma
a vez que, a identificaçã
ão até ao géénero ou fam
mília é
mais fácil e men
nos demorad
da do que a identificaçã
ão de espécies. O potenncial de utilização
desta
a metodolog
gia para prrever padrõ
ões de riqu
ueza especíífica para ddiferentes grupos
g
taxon
nómicos tem sido bastante explorado
o porém, aind
da não foi alv
vo de investiigação no ca
aso do
grupo
o das briófita
as. O objec
ctivo deste e
estudo foi an
nalisar se níveis taxonóómicos superriores,
como
o o género ou família, po
odem ser utillizados como
o indicadores
s fiáveis da rriqueza espe
ecífica
de brriófitas. Os dados foram recolhidos
r
e m áreas perttencentes à rede NATUR
RA 2000, no Norte
e Centro de Porttugal, em do
ois tipos de habitats: sa
axícolas e flu
uviais. Nos hhabitats sax
xícolas
foram
m encontrada
as 129 espécies que perrtencem a 73
7 géneros de
d briófitas, eenquanto qu
ue nos
habita
ats fluviais fo
oram enconttradas 175 e
espécies de briófitas distribuídas por 99 géneros. Para
os da
ados dos ha
abitats saxíc
colas foi usa
ada a correlação de Pe
earson e paara os dados dos
habita
ats fluviais foi
f usada co
orrelação de Spearman. Nos dois tipos de habiitats estudad
dos, a
corre
elação entre a riqueza específica e o número de géneros
g
e famílias é posiitiva e significativa
(p <0
0.01) e o coeficiente
c
de
d correlaçã
ão é alto (rr >0.8). Este estudo suugere que níveis
taxon
nómicos supe
eriores, pode
em ser bons indicadores de riqueza específica
e
dee briófitas e, como
tal, p
pode ser co
onsiderado um método rápido, efic
ciente e de
e baixo cust
sto que pod
de ser
imple
ementado em
m futuros estu
udos da dive
ersidade de briófitas.
b
www.mapp.edu.pt/bioplant 23 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 P8 A Trait-Ba
ased Apprroach to A
Assess Dune Vegeta
ation
R
Response
es to Chan
nges in Co
oastal Dyn
namics
1
José A
António Mace
edo1,2, João Tereso
T
, Paulo
o Alves1, Renato Henriques
s3,4, Franciscoo Ignacio Pug
gnaire5,
1,2
João H
Honrado
1
Centro de Investig
gação em Biod
diversidade e Recursos Ge
enéticos, Unive
ersidade do P
Porto, Edif. FC
C4, Rua
po Alegre s/n, 4169-007 Porrto, Portugal
Camp
2
Facu
ências da Uniiversidade do
o Porto, Edif. FC4,
F
Rua Campo Alegre ss/n, 4169-007 Porto,
uldade de Ciê
Portug
gal
3
Centro de Geolog
gia da Universiidade do Porto
o, Rua do Cam
mpo Alegre, 687,
6
4169-0077 Porto, Portug
gal
4
Dep
partamento de
e Ciências da
a Terra, Unive
versidade do Minho,
M
Campus de Gualtaar, 4710-057 Braga,
Portug
gal
5
Esta
ación Experim
mental de Zo
onas Áridas, C
Consejo Supe
erior de Investigaciones C
Científicas, Cttra. de
Sacra
amento s/n, La
a Cañada de San
S Urbano, A
Almería, Spain
n
Rese
earch on pla
ant traits has
s become a successful way to und
derstand speecies coexis
stence
patterns as a ressult of ecolo
ogical filtering
g processes. Stability, an
a important property of biotic
comm
munities, is promoted by speciess’ traits mediating their responsess to changes in
enviro
onmental strress and/or disturbance
d
e
events. Also
o, theory pred
dicts the rest
striction of tra
aits by
assem
mbly rules and
a
thereby a limit to t he similarity
y of coexistin
ng species associated to
t the
partition of space and resourc
ces. Thereforre, non-rando
om patterns of trait (dis)ssimilarity are a
comm
mon feature
e of biotic communitiess. Moreoverr, trait-based
d analyses are essenttial to
understand the relationship between
b
envvironmental gradients an
nd species ssorting, whic
ch can
then support po
owerful ecolo
ogical assesssment fram
meworks.Coa
astal erosionn is a prev
vailing
proce
ess in much
h of the Po
ortuguese co
oastline, particularly wh
hen dunes aare the dom
minant
geom
morphologic elements. Aligned
A
with
h the theorretical frame
ework descrribed above
e, this
resea
arch addressses patterns of plant tra
aits in coasta
al foredunes, which are dynamic systems
subje
ect to severe
e stress and frequent dissturbance. Specifically,
S
we
w compareed prevailing
g plant
traits between dune
d
system
ms under ero
rosive vs. meta-stable
m
conditions.
c
W
We analyze
ed the
variab
bility of distinct functiona
al metrics (e
e.g. trait variance and dissimilarity, fu
functional div
versity
ociative patterns related to the distin ctive conditio
indice
es), which re
evealed non--random asso
ons of
coasttal dynamicss. Understanding patterrns of plantt trait distrib
bution and vvariation in dune
vegettation may help
h
to anticipate function
nal modificattions driven by environm
mental chang
ges on
dune ecosystemss.
Acknowledgemen
nts: José An
ntónio Mace
edo is supp
ported by a PhD scholaarship (Refe
erence
SFRH
H/BD/48610//2008) from FCT,
F
the Porrtuguese Fou
undation for Science andd Technology
y.
www.mapp.edu.pt/bioplant 24 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 P9 P
Padrões de
d Coocorrrência de
e Espécies
s da Vegettação Dun
nar no Estu
udo
d
dos Proce
essos de Dinâmica
D
Costeira e na Monittorização Ecológica
a
António Mace
edo1,2, Ana Soffia Vaz1, Ânge
ela Lomba1, Paulo
P
Alves1, Renato
R
Henriqques 3,4,
José A
5
1, 2
Franccisco Ignacio Pugnaire
P
, Joã
ão Honrado
1
Centro de Investig
gação em Biod
diversidade e Recursos Ge
enéticos, Unive
ersidade do P
Porto, Edif. FC
C4, Rua
po Alegre s/n, 4169-007 Porrto, Portugal
Camp
2
Facu
uldade de Ciê
ências da Uniiversidade do Porto, Edif. FC4,
F
Rua Cam
mpo Alegre ss/n, 4169-007 Porto,
Portug
gal
3
Centtro de Geologia da Universiidade do Porto
o, Rua do Cam
mpo Alegre, 687, 4169-007 Porto, Portug
gal
4
Dep
partamento de
e Ciências da
a Terra, Unive
ersidade do Minho,
M
Campus de Gualtaar, 4710-057 Braga,
Portug
gal
5
Esta
ación Experim
mental de Zo
onas Áridas, C
Consejo Supe
erior de Investigaciones C
Científicas, Cttra. de
Sacra
amento s/n, La
a Cañada de San
S Urbano, A
Almería, Espan
nha
As in
nterações en
ntre as espé
écies, e desttas com as condições do
d respetivo meio em que se
insere
em, constitu
uem aspeto
os centrais nos estudo
os ecológico
os. Nas zoonas costeirras, a
distrib
buição das espécies ve
egetais ao lo
ongo dos prrincipais grad
dientes ambbientais apre
esenta
padrõ
ões característicos, relac
cionados com
m a ação dos
s diversos fa
atores ambie ntais, sob a forma
de grradientes dirrecionais, e das suas intterações. Po
or outro lado
o, os modeloos de organização
das ccomunidadess vegetais prreveem que a coexistênc
cia entre as espécies é ffunção do ba
alanço
de interações bió
óticas como a competiçã
ão e facilitaç
ção, cujo pre
edomínio é ddeterminado pelas
condiições ambie
entais. Os div
versos fatore
res de stress
se e de pertturbação asssumem assim um
papel crucial na
a regulação dos padrõe
es de ocorrência que dependem
d
ddeste sistem
ma de
intera
ações. Os sistemas dunares
d
cossteiros apre
esentam co
ondições dee adversida
ade e
instab
bilidade amb
bientais acrescidas parra as espéc
cies vegetais
s que neless ocorrem, sendo
s
expecctável uma prevalência
p
dos
d fenómen
nos de facilita
ação sobre os
o de compeetição, em sintonia
com a teoria eco
ológica. No entanto,
e
nesstes ambienttes extremos
s, alteraçõess importante
es nas
condiições e nos recursos dis
sponíveis (e..g. relacionad
das com alte
erações na ddinâmica cos
steira)
podem modificar o balanço entre
e
interaçõ
ões positivas
s e negativa
as. Deste moodo, a deteç
ção de
padrõ
ões de coo
ocorrência ou
o de exclu
usão mútua entre espé
écies poderáá apresenta
ar um
assin
nalável caráccter diagnóstiico para dete
ectar precoce
emente alterrações ecolóógicas em curso.
Conssiderando esste quadro teórico gera
al, neste tra
abalho foram
m analisadoss os padrõe
es de
variaçção destes processos
p
in
nterativos na
a vegetação de dunas frrontais (tipicaamente dom
minada
por A
Ammophila arenaria), co
omparando situações de
d dinâmica transgressiiva (erosiva)) com
situaçções meta-e
estáveis. A análise de coocorrênc
cia entre es
spécies, util izando diferrentes
métricas indicado
oras de tendências de agregação versus segrregação, revvelou padrões de
assocciação não-a
aleatórios, re
elacionados ccom as distin
ntas condiçõ
ões de dinâm
mica costeira
a. Esta
avalia
ação dos pa
adrões espac
ciais permite
e uma identifficação mais
s robusta doos efeitos iso
olados
ou co
ombinados dos
d
fatores abióticos e bióticos. O conhecimento detalhaddo das regras de
estrutturação desstas comunidades consstitui assim uma importtante ferram
menta na de
eteção
preco
oce de varia
ações nos processos d
de dinâmica
a costeira, bem
b
como na monitorização
ecoló
ógica dos hab
bitats dunare
es.
Agrad
decimentos: José António Macedo é financiado pela FCT – Fundação ppara a Ciênc
cia e a
Tecnologia, atravvés da Bolsa de Doutoram
mento com a Referência SFRH/BD/448610/2008.
www.mapp.edu.pt/bioplant 25 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 0 Class III peroxidas
ses perox
xidase cata
alytic intermediatess CoI and CoII
P10
are reduced by ara
abinogala
actan prote
eins
Sara B
Bettencourt1,2 and Mariana Sottomayor1,22
1
IBMC
C – Instituto de
d Biologia Molecular
M
e Ce
elular, Univers
sidade do Porto, Rua do C
Campo Alegre
e, 823,
4150--180 Porto, Po
ortugal
2
Depa
artamento de Biologia, Facu
uldade de Ciê
ências da Universidade do Porto, Rua doo Campo Aleg
gre s/n
4169--007 Porto, Po
ortugal
Classs III peroxid
dases (Prxs)) are typica
al plant enzy
ymes that have
h
been implicated in key
proce
esses determ
mining the architecture
a
a
and defence
e properties of the plantt cell wall, mainly
m
through H2O2 dep
pendent oxidation activitie
es leading to
o the cross-linking of cell wall components.
Previously, we ha
ave obtained
d indirect evi dence that arabinogalac
a
ctan proteinss (AGPs) are
e an in
vivo ssubstrates off Prx, and that Prxs are iinvolved on the
t appearance /disappeearance of se
everal
speciific AGP glyccosidic epitop
pes implicate
ed in importa
ant aspects of
o plant deveelopment and
d plant
cell p
physiology. In
n this work, the capacity o
of purified AGPs from the
e leaves of thhe medicinal plant
Catha
aranthus roseus to red
duce the ca
atalytic interrmediates of
o class III peroxidases
s was
invesstigated. C. roseus
r
perox
xidase 1 (Cro
oPrx1), the main
m
Prx present in the leeaves of the plant,
was p
purified and its oxidation by H2O2 and
d consequen
nt interaction with AGPs sshowed that these
prote
eoglycans arre suitable electron do
onors for th
he oxidizing intermediattes of CroP
Prx1 comp
pound I (CoI) and comp
pound II (Co
oII). Similarly, AGPs we
ere also cappable of red
ducing
horse
eradish peroxidase-type II CoI and C
CoII, indicating that the capacity
c
to ooxidize AGP
Ps is a
general feature of
o Prxs. The
e Km for AG
GP oxidation by CoII wa
as lower for CroPrx1 tha
an for
HrPrxxII, indicating
g a higher affinity of Cro
oPrx1 for AG
GPs coming from
f
the sam
me species/o
organ.
More
eover, immunofluorescen
nce co-labe
elling of lea
af sections with a monnoclonal antibody
recog
gnizing a sp
pecific AGP glycosidic e
epitope and an anti-Prx antibody cl early showe
ed colocalization with a punctuated
d distribution in plasma membrane
m
an
nd tonoplast of mesophylll cells
from C. roseus. This data strongly ind
dicates that AGPs are in vitro andd possibly in
n vivo
substtrates of cla
ass III peroxidases, with potential im
mplication for the develoopmental an
nd cell
physiiology functio
ons attributed
d to AGPs.
Acknowledgmentts: Project funding by i) F
FEDER funds
s through the
e Operationaal Competitiveness
Progrramme – CO
OMPETE and
d by Nationa
al Funds thro
ough FCT – Fundação ppara a Ciênc
cia e a
Tecnologia underr the project FCOMP-01 -0124-FEDE
ER-022718 (P
PEst-C/SAU//LA0002/201
11), ii)
po Jerónimo
o Martins, and
a
iii) the FCT schola
arship
a Sccientific Meccenate Grant from Grup
SFRH
H/BD/48283//2008 www.mapp.edu.pt/bioplant 26 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 1 Impact analysis
a
of riversca pes fragm
mentation on the con
nservation
n of
P11
bryophytte commu
unities
2
Ana P
Paula Portela
a1, Cristiana Vieira
V
, Helen
na Hespanholl2, Bruno Marrcos2, João H
Honrado1,2 & Rubim
1,2
Almeiida da Silva
1
2
Facu
uldade de Ciên
ncias da Universidade do P
Porto
Centtro de Investig
gação em Biod
diversidade e Recursos Gen
néticos, CIBIO
O-UP
The objective wa
as to condu
uct a region
nal assessment of impacts caused by fragmen
ntation
eleme
ents (dams, small hyd
dropower scchemes, wiind farms, roads and railway) on the
riversscapes, iden
ntifying simplle or cumula
ative impact zones and potential zoones of ecological
prote
ection, of fluvial bryophy
yte commun
nities.We use
ed a compilation of daatabases including
inform
mation on bryophytic co
ommunities ((2000-2013), from 257 watercoursees in the No
orth of
Portu
ugal. Floristicc composition was classsified (TWINS
SPAN, CAP)) and typifiedd (SIMPER, CAP)
and e
ecological pa
arameters off sites statisstically explored (DCA an
nd CCA, CA
ANOCO; SPS
SS) to
obtain
n the region
nal commun
nities types and establis
sh species-e
environment relations. Spatial
S
inform
mation analyyses were pe
erformed in G
GIS program (ArcMap 10
0.1). We spaatialized bryo
ophytic
comm
munities in th
he watershe
eds, establish
hed buffers of impact pe
er fragmentaation elemen
nt and
analyysed the losss of potentia
al occurrencce of commu
unities.We obtained 8 tyypes of bryo
ophytic
comm
munities cha
aracterized by 1 to 6 core taxa and foun
nd indicatorr pseudo-sp
pecies
(TWIN
NSPAN) co
oncordant with
w
many o
of the taxa
a responsible for the cohesion of
o the
group
ps.Communities were spatialized
s
according to
t their fluv
vial Strahlerr order, su
ummer
pitation, ann
precip
nual averag
ge temperatture, local slope, altitude and hil lshade; areas of
occurrrence were calculated fo
or each comm
munity. The impacts are mainly due tto dams and small
hydro
opower sche
emes and the
e areas of lo
oss and potential preserv
vation of eacch community
y type
were calculated and
a
presented in a deta
ailed map th
hat allows regional and llocal conserrvation
plann
ning. Bryophyytic commun
nities are spa
atially restrictted in the con
ntext of Portuuguese, Iberrian
and E
European biodiversity, where
w
comm unities show
w high species richness and diversitty and
manyy taxa of Atlantic
A
distribution. Esp
pecially in mountain
m
areas, these communitie
es are
vulne
erable becau
use of fragm
mentation cau
used by sma
all hydropow
wer schemess, dams and
d wind
farmss implementation.These
e results al lowed us to
t analyse the impactt of energy
y and
comm
munication elements
e
on
n the assesssment of cu
umulative im
mpacts and identify are
eas of
cumu
ulative impacct and conserrvation.
P12
2 Effect off different factors o n the callu
us induction of Pinu
us elliottiii and
Pinus ellliottii x Pin
nus cariba
aea
2
A. Alm
meida1, G. Pin
nto1*, C. Dias1, A. Costa1, V. Pereira2, L. Marum
M
, S. Co
orreia2, C. Sanntos1
1
Depa
artment of Bio
ology,& CESAM – Centre fo
for Environmental and Marin
ne Studies , U
University of Aveiro,
A
Camp
pus Universitário de Santiag
go, 3810-193 A
Aveiro, Portug
gal. E-mail: *gp
[email protected]
2
KLÓN
N, Innovative Technologies
s from Cloning
g, Biocant Pa
ark, Núcleo 4, Lote 4A, 30660-197 Cantan
nhede,
Portug
gal
Pinuss elliottii and the hybrid Pinus ellliottii x Pinu
us caribaea have signifficant comm
mercial
imporrtance. Therrefore it urge
es the need to develop a battery of in vitro cullture protoco
ols for
micro
opropagation
n and for general researrch (eg, phy
ysiology, molecular bioloogy, etc.) of these
conife
ers. The main objective of
o this studyy was to analyze the effect of variouss endogenou
us and
exoge
enous factorrs on the induction of: a) callus tissue (importa
ant in fundaamental stud
dies of
physiiology, mainttenance of germplasm, m
morphogenes
sis, etc.); b) embryogenicc callus (imp
portant
for p
plant regeneration by so
omatic embrryogenesis, cryopreserv
vation studiees, etc). Forr that,
mature zygotic embryos
e
(MZ
ZE) and cottyledons of Pinus elliottii and Pinuus elliottii x Pinus
cariba
aea were exxposed to different grow
wth regulators
s (2,4-D, NA
AA and TDZ)). First callus was
visible after 15 days.
d
The mo
ost efficient growth regu
ulators condiitions in induucing callus were:
2,4-D
D with 85% induction of calli in MZE ; 2,4-D and
d NAA in cottyledons, botth leading to
o 55%
inducction of calli. MZE were
e subjected to different growth reg
gulators (2,44-D+24-epiBrr; 2,4D+BA
AP), carbon sources (sucrose and m
maltose) and
d stress facto
ors, in particcular heat (80 °C),
cold (-15 °C), sa
alicylic acid, proline, H2O 2 and putre
escine. The most efficieent combinattion of
growtth regulatorss was 2,4-D
D+BAP with approximate
ely 30% rate
e of calli indduction. The more
efficie
ent stress conditions
c
were
w
cold a
and putrescine. These results suppport the po
ossible
inducction of calluss in these tw
wo conifers, a
and supports
s preliminary data towardds the optimization
of efffective protoccols for SE.
www.mapp.edu.pt/bioplant 27 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 P13
3 Relação entre pad
drões amb
bientais lo
ocais e a distribuiçã
d
ão de espé
écies
de líquen
nes domin
nantes no Vale do Côa
C (NE Po
ortugal)
dia Oliveira1, Joana
J
Marques
s1,2, Rubim Allmeida1,2
Cláud
1
Dep
partamento de
e Biologia, Fa
aculdade de Ciências da Universidade do Porto, Eddifício FC4, Rua
R
do
Camp
po Alegre s/n, 4169-007 Porrto, Portugal.
2
CIB
BIO, Centro de
e Investigação em Biodive
ersidade e Recursos Genétticos, Campuss Agrário de Vairão,
V
4485--661 Vairão, Portugal
P
O Va
ale do Côa (Vila Nova de Foz Côa, Nordeste de
e Portugal) tornou-se coonhecido em finais
dos a
anos 90 apóss a descoberrta da maior e mais impo
ortante coleç
ção de arte ruupestre ao ar
a livre
do Pa
aleolítico Sup
perior, atualm
mente classiificada como
o património mundial pelaa UNESCO.D
Desde
então
o têm sido re
ealizados div
versos estud
dos de carac
cterização da
as dinâmicass de meteorização
dos a
afloramentoss rochosos e superfíciess gravadas, integrando dados ambieentais, geo-físicoquímicos e biológ
gicos, que visam o dese nvolvimento de ferramen
ntas de gesttão e conserrvação
da arrte rupestre.O
Os líquenes dominam
d
ass comunidade
es que colon
nizam as supperfícies grav
vadas
e dessempenham um papel importante n
no processo
o de deteriorração dessaas superfície
es. No
entan
nto, pouco se
e sabe sobre
e os padrõe s de variaçã
ão dos fatore
es ambientaiis à escala lo
ocal e
sobre
e a forma como estes atu
uam sobre a composição
o destas com
munidades. E
Estudos ante
eriores
indica
am que Leccanora pseu
udistera é a espécie dominante
d
nas
n
superfíccies orientad
das a
noroe
este, Aspicillia hoffmania
ana a sudesste e Calop
placa irrubes
scens, igualm
mente abund
dante,
parecce ser indife
erente à exp
posição. Com
m este estudo pretende
e-se avaliar de que form
ma os
padrõ
ões de variação de fa
atores ambie
entais à escala local afetam
a
a suua abundân
ncia e
distrib
buição desta
as três espé
écies na áre
ea do Parqu
ue Arqueológ
gico do Valee do Côa. Foram
F
recolh
hidos dados de tempera
atura e humid
dade relativa
a de dois anos consecuttivos recorre
endo a
três e
estações me
eteorológicas
s e onze sen sores distrib
buídos pela área
á
de estuddo a altitude
es que
variam
m entre os 136
1 e os 332
2 metros, orie
entados a no
orte (3), sul (6), este (1) e oeste (1). Será
discu
utida a relaçã
ão entre os padrões
p
de d
distribuição destas
d
espéc
cies e os paddrões de variação
local dos fatores ambientais registados, e respetivas
s implicações
s para a connservação da arte
rupesstre no Vale do Côa.
P14
4 Effect off Cr III in Lactuca
L
sa
ativa L. ph
hysiology
1,2
Raísssa CS Faria1,2, Willian GD Bedo
B
,C Dias 1, G Pinto1, C Santos1
1
2
Depa
artment of Biollogy & CESAM
M, University o
of Aveiro, 3810-193 Aveiro, Portugal
FHO
O|Uniararas - Fundação Hermínio Ometto
o, 055 19) 354
43-1400, Brasil
Cr(III) is an esse
ential nutrien
nt to humanss, being inge
ested through different fo
food sources
s (eg.,
cropss, nuts, mea
at and eggs
s). Despite ttheir essentiality to hum
mans Cr (inddependently of its
valen
nce) is curren
ntly considered to be a to
oxic element to plants. Most studies oon Cr phytoto
oxicity
are b
based on the
e Cr(VI) valen
nce, while C
Cr(III) effects remain less studied. On e of the mos
st well
studie
ed crops for Cr(VI) toxicity is Lactuca
a sativa, a worldwide con
nsumed cropp, and an exc
cellent
mode
el in ecotoxiccology. Lettu
uce plants w
were grown hydroponicall
h
y on Hoaglaands (1/3 strrengh)
for 15
5 days and exposed
e
to Cr
C (III) ( 20, 50, 150 and
d 200 mg/L). All indepenndent experiments
show
w a hormesiss effect (up to
o 20 mg/L), with plants showing
s
an increase of llengh and growth.
This preliminary result
r
led us to raise the hypothesis of Cr(III) being a beneficcial nutrient to
t this
crop. For that we
e analyzed some
s
physio
ological param
meters name
ely photosynnthesis. Curiously,
all Crr doses decrreased A (ne
et photosynth
hetic rate), and led to a stomatal
s
clossure. These Cr(III)
dosess did not sig
gnificantly afffect the Ci ((internal CO2
2 availability). Also, conssidering thatt plant
growtth is a resultt of cell division and exte
ension, the im
mpact of thes
se doses on mitotic inde
ex (MI)
and m
mitosis catasstrophe (micrronuclei) in tthe root apex
x is under discussion. Thhese data su
upport
that C
Cr(III) hampe
ers some photosynthetic parameters, and the pa
athways invoolved in the growth
g
stimu
ulation at low
wer doses de
eserve furthe
er studies. The growth sttimulation att lower doses also
suppo
ort that, conttrarily to Cr(V
VI), Cr(III) ha
as potential to
o be conside
ered a benefficial nutrient when
at verry low dosess, despite furtther studies are ongoing.
www.mapp.edu.pt/bioplant 28 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 5 Chemica
al profile and
a anti-a llergic scrreening off Fucus sp
piralis
P15
Linnaeus
s and Pad
dina pavon
nica (Linna
aeus) Thiv
vy
Mariana Barbosa, Joana
J
Costa, Tiago Pereira,, Patrícia Vale
então, Paula B.
B Andrade, Brrígida R. Pinho
o
REQU
UIMTE/Labora
atório de Fa
armacognosia,, Departamento de Quím
mica, Faculddade de Farrmácia,
Unive
ersidade do Po
orto, R. Jorge Viterbo Ferreiira n.º 228, 40
050-313 Porto,, Portugal.
Prese
ently, the ressearch on marine
m
produ
ucts has sho
own that seaweeds are a rich source
e of a
wide variety of natural com
mpounds be
elonging to different metabolic patthways, and
d with
nume
erous interessting properties. In fact, m
more than 15
5000 primary and second ary metaboliites
have already bee
en reported in macroalga
ae and differrent applications are asssigned to the
em [1].
From
m a human he
ealth point of
o view, both primary and
d secondary metabolites exhibit num
merous
biolog
gical activitie
es, like antiox
xidant, antitu
umor and antti-inflammato
ory [1].
Macroalgae can be classifie
ed into three
e classes ba
ased on the
eir pigmentaation: green algae
(Chlo
orophyta), bro
own algae (P
Phaeophyta)) and red algae (Rhodoph
hyta). There is a wide div
versity
of brrown algae that are important mem
mbers of many marine communitiees with significant
econo
omic importa
ance [2]. In th
his work, eth
hanolic extrac
cts of two bro
own seaweeeds species [F
Fucus
spirallis Linnaeus and Padina
a pavonica ((Linnaeus) Thivy],
T
collec
cted in differrent places of the
west coast of Portugal were chemically ccharacterized
d and their anti-allergic
a
pproperties studied.
Eleve
en compound
ds were iden
ntified by GC
C–MS (Table 1), F. spiraliis presentingg a great variety of
comp
pounds. The
e anti-allergic screening
g was asses
ssed by inh
hibition of ccalcium iono
ophore
stimu
ulated RBL-2
2H3 cells degranulation
d
n by non-to
oxic concenttrations of tthe two ethanolic
extracts. Prelimin
nary data point P. pavoniica as the mo
ost promising species, bbecause it red
duced
51.5 ± 8.56 % (m
mean ± SEM) the cells d
degranulation
n at 12.5 mg/mL, while F
F. spiralis red
duced
the degranulation
n in 40.0 ± 4.5
59%, for the same conce
entration.
In co
onclusion, in spite of F. spiralis
s
to be
e more chem
mically rich than P. pavoonica, the las
st one
show
wed better an
nti-allergic ac
ctivity, which
h could be re
elated, at lea
ast partially, w
with the presence
of α-linoleic acid (C18:3)
(
and of phlorogluccinol.
Tablle 1 - Metabo
olic composittion of F. spir
iralis and P. pavonica
p
sam
mples (mg/1000 g dry alga
ae)a
Co
ompounds
RIcalcb
RIlitc
F. spiralis
P. pavonnica
Proline
Phloroglucinol
C14:0
Mannitol
C16:0
C18:3
C18:1
C18:0
C20:4
C20:5
F
Fucosterol
1286
1631
1837
1896
2043
2225
2238
2253
2389
2398
3361
12
299 [3]
16
647 [4]
18
843 [4]
19
912 [4]
20
040 [4]
22
218 [4]
22
248 [4]
22
234 [5]
23
373 [4]
23
380 [4]
33
330 [5]
39.68
3
(9.68)
< LOQ
6.98
6
(0.27)
12
27.38 (3.48)
60.60
6
(2.06)
216
69.06 (166.76))
20.84
2
(2.14)
46.56
4
(2.90)
21.70
2
(1.14)
31.76
3
(0.94)
13.48 (2. 78)
21.02 (0. 06)
38.32 (1. 58)
12.16 (3. 38)
186.02 (22.20)
146.92 (44.88)
14.04 (2. 42)
16.48 (0. 56)
a
Valu
ues are exprressed as mean
m
(SD) off three deterrminations; LOQ
L
– limit oof quantification. b
c
RIcalc: Retention in
ndex obtaine
ed in the exp
periment. RIIlit: Retention index descrribed in litera
ature.
Acknowledgemen
nts: This wo
ork was supp
ported by Fundação
F
pa
ara a Ciênciia e a Tecno
ologia
(FCT
T): (PEst-C//EQB/LA0006/2011). B.. R. Pinho
o is indebtted to FCT
T for the grant
(SFR
RH/BD/63852
2/2009).
Referrences: [1] Andrade
A
et al.
a (2013) Valluable compounds in macroalgae exttracts, Food Chem
138:1
1819-1828; [2]
[ Amsler et
e al. (2005) Defensive and
a
Sensory
y Chemical E
Ecology of Brown
B
Algae
e. In: Callow
w JA, editor. Advances
A
in
n Botanical Research,
R
Vo
olume 43: Accademic Pre
ess; p.
1-91. [3] Meyer et al. (200
09) Remode
eled respirattion in nduffs4 with low
w phosphory
ylation
ency suppresses Arabidopsis germin
nation and growth
g
and alters
a
controll of metaboliism at
efficie
Plant
Physiol
night,
151:603
3–619.
[4]
NIST
T
chemi stry
webbook
(http://webbook.nist.gov/chem
mistry/name-sser.html). [5]] Isidorov et al. (2003) G
GC–MS analy
ysis of
comp
pounds extra
acted from buds
b
of Popu
ulus balsamifera and Po
opulus nigra , Z Naturforsch C
58:35
55–360.
www.mapp.edu.pt/bioplant 29 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 P16
6 Carga orrgânica ad
dicionada a FitoETA
AR por colmos de P
Phragmite
es
australis
s mantidos
s nos leito
os durante
e o outono
o/inverno
Luis M
Manuel Oliveirra1, António Mário
M
Almeida2 e Isabel A-P. Mina1, 3
1
Depa
artamento de Biologia, Esco
ola de Ciência
as da Universiidade do Minh
ho (DB-ECUM)
M)
Centtro de Física, Escola de Ciê
ências da Univversidade do Minho
M
(CF-EC
CUM)
3
Centtro de Investig
gação em Tec
cnologia Agro-A
-Ambiental e Ciências
C
Biológ
gicas (CITAB))
2
As E
Estações de Tratamento de Águas Residuais (ETAR) têm por objetivoo reduzir a carga
orgân
nica e aumen
ntar a qualidade sanitária
a das águas afluentes.
ara o tratamento secundário de águaas residuais sendo
As FitoETARs utilizam leitos de
d plantas pa
s
nto de águ
sobre
etudo adequ
uadas para o tratamen
uas residuaiis doméstica
cas de pequenas
comu
unidades. É já considerá
ável o núme
ero de FitoET
TARs existentes em Porrtugal e a maioria
m
utiliza
a leitos plan
ntados com Phragmites australis (c
caniço). Entre setembro e fevereiro, esta
Poacceae cosmop
polita seca, e os seus co
olmos são, em muitas FittoETARs, maantidos nos leitos.
Neste
e período do
d ano, frio e geralmen
nte caracterrizado por fortes aguacceiros, os colmos
permanecem ime
ersos durantte largos pe
eríodos.Para
a estimar a carga orgânnica adiciona
ada a
leitoss de FitoETA
ARs pelo encharcamento
o causado pelas
p
chuvas
s invernais, realizaram-s
se em
laborratório ensaio
os em microcosmos.Os ensaios prelliminares até
é agora realizzados evidenciam
que, colmos de caniços
c
imerrsos em águ
ua da torneirra durante uma semana , conduzem a um
consiiderável teorr de carga orgânica
o
na ““água da ch
huva experim
mental”. Ao loongo do tem
mpo, a
carga
a orgânica da
d “água da chuva” estim
mada pela Carência
C
Bio
oquímica de Oxigénio (C
CBO5)
dimin
nui, enquantto que o pH e a tenssão superfic
cial aumenta
a, mostrandoo tendência para
estab
bilizar ao fim de algumas semanas.
P17
7 Halotole
erant Plantts for Phy
ytoremedia
ation in Sa
aline Envi ronments
s
Jesussa, J., Gonçalvvesb, Ana T., Mina
M c, Isabel, B
Borgesb,d, Maria-Teresa,
a – FE
EUP – CIGAR
R - Centro de Investigação
I
e
em Geo-Ambiiente e Recurs
sos, Faculdadde de Engenha
aria da
Unive
ersidade do Po
orto, Rua Dr. Roberto
R
Frias,, s/n 4200-465
5 Porto, Portug
gal
b - FC
CUP – Departa
amento de Bio
ologia, Faculd
dade de Ciênciias da Univers
sidade do Port
rto,
Rua C
Campo Alegre
e s/n, 4169-007 Porto, Portu
ugal.
c – U
UM - Departa
amento de Biiologia - Univversidade do Minho, Camp
pus de Gualttar 4710-057 Braga
Portug
gal
d - CIIIMAR – Centro Interdiscip
plinar de Invesstigação Mariinha e Ambien
ntal, Universiddade do Porto
o, Rua
dos B
Bragas 289, 40
050-123 Porto, Portugal.
Halottolerant and halophytic plants have
e been the subject of intense reseearch due to
o their
capaccity to withsstand high sa
alinity which
h opens up new
n
opportu
unities for phhytoremediattion in
extreme condition
ns. This study focus on T
Typha latifolia
a and Arund
do donax abillity to tolerate
e high
salinity and on methods
m
to successfully propagate and acclima
atize these sspecies aiming at
gathe
ering practica
al information
n for in situ re
remediation applications
a
like saline efffluents treatment.
Typha latifolia was
w
collected and prop
pagated with
h intact rhiz
zomes. It reevealed ade
equate
adapttation to the
e tested subs
strate, expan
nded clay, after
a
2 weeks
s. However, survival rate
e was
dependent on sa
ampling seas
son, with hig
gher values in autumn (7
75 to 90% suurvival) and lower
value
es in late spring (11%), probably du e to a lowerr carbohydra
ate concentraation in rhizo
omes.
Furth
hermore, it was
w
able to withstand ssalinity value
es of up to 2.4% with 994% survival rate,
despiite some gro
owth inhibition. Arundo do
onax was collected and propagated
p
tthrough seco
ondary
shootts. The best results were
e obtained in
n the spring. Expanded clay
c
utilizatioon increased
d plant
growtth and survivval. With proper acclimattion, survival rate was hig
gh (=100%) uup to 3.5% salinity
s
level.. Furthermore, Typha latiifolia was alsso able to withstand and treat highly saline waste
ewater
from a recirculating aquacultu
ure farm. Ho
owever, it re
emains uncle
ear if these pplants specie
es are
able to uptake dissolved
d
salts. Althoug
gh expanded
d clay itself is able to adsorb significant
amou
unts of salts some
s
salinity
y reduction ccannot be explained by substrate adssorption alone.
Furth
her studies, therefore, will
w be aimed
d at clarifying the pote
ential use off these plan
nts for
reme
ediation of sa
aline environm
ments.
www.mapp.edu.pt/bioplant 30 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 P18
8 Monogalactosyl diacylglyce
d
erols from
m the ediblle brown aalgae Fucus
spiralis Linnaeus:
L
Anti-infla
ammatory potential on RAW 2264.7
macroph
hage cell line
G. Lop
pes1, G. Dalettos2, P. Proksch2, P. B. And
drade1 and P. Valentão1
1
Department of
REQ
QUIMTE/Laborratory of Pharrmacognosy, D
o Chemistry, University off Porto, R. de Jorge
Viterb
bo Ferreira, 22
28, 4050-313 Porto,
P
Portuga
al
2
Intitu
ute of Pharma
aceutical Biolo
ogy and Biote
echnology, Heinrich-Heine
H
University, U
Universitätstra
asse 1,
40225
5 Düsseldorf, Germany
As the majority off marine orga
anisms, seaw
weeds have developed physical
p
and chemical de
efence
mech
hanisms in order to survive in a competitive environmen
nt, with extrreme atmospheric
condiitions. These
e defence strategies
s
re
esulted in the production
n of a signiificant divers
sity of
secon
ndary metab
bolites, which
h have attractted scientists
s’ attention in
n the past few
w years.
The e
edible brown
n seaweed Fucus
F
spiraliis Linnaeus is widely dis
stributed oveer the Portug
guese
west coast. This species is rich in second
dary metabolites, among which fatty acids constiitute a
very special group. Fatty acid
ds are one o
of the most re
epresentative
e classes off compounds
s, both
in divversity and contents,
c
and
d are associa
ated with div
verse biological activitiess. F. spiralis has a
diversse lipid composition, dom
minated by m
mono and po
olyunsaturate
ed long chainn fatty acids, oleic
acid being the major one [1]. A purified ffraction containing two monogalactos
m
syl diacylglycerols
(MGD
DGs) and a monoglycerid
m
de (Figure 1
1) were isolatted from the non-polar frraction of the
e ethyl
aceta
ate extract off this seawee
ed by using reversed phase silica ch
hromatographhy. Their stru
ucture
was ascertained by 1H NM
MR and MS spectroscop
py. The antti-inflammatoory activity of
o the
comp
pounds was assessed
a
on
n RAW 264.7
7 macrophage cell line.
Gly
ycerol
C188:1 Ω9
Figurre 1. Monoglyceride isolated from th
he non-polar fraction of th
he ethyl aceetate extract of the
n algae F. sp
brown
piralis.
The MGDGs fra
action and th
he monoglyyceride displayed anti-in
nflammatory activity with
h IC50
value
es of 85.67 and
a
65.96 µg/mL,
µ
respe
ectively. These results highlight the potential of these
comp
pounds as an
nti-inflammattory agents.
Acknowledgemen
nts:The autho
ors are grate
eful to Funda
ação para a Ciência e a Tecnologia (FCT)
through grant no. PEst-C/EQ
QB/LA0006/2 011. G. Lop
pes (SFRH/B
BD/61565/20009) is indebted to
FCT, FSE and PO
OPH for the grant.
g
[1] Foo
od Chemistry
y, 2013, 138
8, 1819-18288.
www.mapp.edu.pt/bioplant 31 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 P19
9 Neuroprrotective effect
e
of stteroidal allkaloids frrom Lycop
persicon
esculenttum Mill. on
o glutama
ate-induce
ed toxicity
y in SH-SY
Y5Y cells
aveira, M. Macciejewska, M. Costa,
C
I. Costta, P. Valentão
o, P. B. Andrade
M. Ta
UIMTE/Labora
atório de Fa
armacognosia,, Departamento de Quím
mica, Faculddade de Farrmácia,
REQU
Unive
ersidade do Po
orto, Rua de Jorge
J
Viterbo F
Ferreira, nº. 22
28, 4050-313 Porto, Portuga
gal
Lycop
persicon escculentum Milll. (tomato pla
ant) is one off the most im
mportant cropp in the world
d, with
econo
omic and health impact. Neverthelesss, there are several pestts that can deestroy its cultures,
such as Spodop
ptera littoralis
s (Bois.). Th
he losses ca
aused by this pest justiified the che
emical
chara
acterization and
a the evalu
uation of the
e biological potential
p
having in mind itts possible use
u as
sourcce of bioactive compounds, taking profit from the infestation. The meetabolic profile of
severral S. littoraliis materials (larvae,
(
pupa
ae, adults an
nd excrements) and the hhost plant ma
aterial
(leave
es) were ch
haracterized in terms o f phenolics, alkaloids, amino
a
acidss, fatty acids
s and
sterols. These me
etabolites we
ere determin
ned by HPLC
C-DAD and GC-MS
G
techhniques. Phe
enolics
were identified only in L. esc
culentum lea
aves, being absent in S.
S littoralis m
materials. Ste
eroidal
alkalo
oids were ide
entified in all matrices, e
especially in leaves and S.
S littoralis exxcrements. Amino
A
acidss and fatty accids seem to be bioaccum
mulated by S.
S littoralis larrvae.
The antioxidant capacity of these matr ices was ch
hecked in chemical miccroassays against
severral reactive species (Fig
gure 1). The
e ethanolic extract
e
from tomato leavves was the most
active
e antioxidantt, followed by
y that of S. litttoralis excre
ements.
Figurre 1. Effect of
o ethanolic extracts aga
ainst DPPH●, O2●- and ●NO.
N Values show mean ± SE
from three experiments perforrmed in tripliccate.
The a
acetyl and bu
utyrylcholinesterase inhib
bitory capaciity was also evaluated ussing a colorimetric
micro
oassay. As verified for the antioxiidant potenttial, the tom
mato leavess and S. litttoralis
excre
ements were
e the most active
a
matrice
es. Their ac
ctivity can be
e associatedd to the conttent in
steroidal alkaloids. By evalua
ating the bio
ological fate of these compounds in the S. littorralis/L.
escullentum syste
em we can discover the sstrategies us
sed by the plant to overcoome the pes
st, and
also those develo
oped by the insect to fe
eed and surv
vive on the plant. Basedd on the obttained
inform
mation, a pra
actical applic
cation would be in the bre
eeding of cro
op plants moore resistant to the
attackk by the inse
ect.
Acknowledgemen
nts: The auth
hors are gratteful to Fundação para a Ciência e a Tecnologia (FCT)
for Grant No. PEsst-C/EQB/LA
A0006/2011. M. Taveira (SFRH/BD/62
2662/2009) iis indebted to
o FCT
for the Grant.
www.mapp.edu.pt/bioplant 32 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 0 Grindelia
a robusta Nutt. : Ch
haracteriza
ation of th
he phenoliic profile and
a
P20
evaluatio
on of its in
nhibitory p
potential against
a
brrain enzym
mes and
oxidative
e species
C. Gro
ossoa, C. Azevedoa, F. Ferrreresb, A. Gil-IIzquierdob, P. Valentãoa, P. B. Andradea
a
REQ
QUIMTE/Laboratory of Pha
armacognosy, Chemistry De
epartment, Fa
aculty of Pharrmacy, Univerrsity of
Porto,, Portugal
b
Rese
earch Group on
o Quality, Safety
S
and Bio
oactivity of Pllant Foods, Department
D
off Food Scienc
ce and
Techn
nology, CEBAS (CSIC), P.O
O. Box 164, 30
0100 Campus University Espinardo, Murccia, Spain
Oxida
ative stress is considered to be one of the cause
es of the dev
velopment oof neuropsychiatric
disord
ders, such as Alzheim
mer’s disea se (AD) an
nd depression. Moreovver, inhibito
ors of
acetyylcholinestera
ase (AChE) and butyryl cholinestera
ase (BuChE) ameliorate the symptoms of
AD, while mono
oamine oxida
ase A (MAO
O-A) inhibito
ors are use
ed to treat tthe symptom
ms of
depre
ession. Thesse last also prevent the prroduction of H2O2, a reactive oxygen sspecies (RO
OS).
This study reportts the biological potentia
al of two extracts (aqueo
ous and metthanol/water) from
Grind
delia robusta
a Nutt. as RO
OS (O2●− an
nd H2O2) and
d reactive nitrogen speccies [RNS, (●NO)]
scave
engers and as
a AChE, Bu
uChE and MA
AO-A inhibito
ors. The phenolic profile w
was characterized
by HP
PLC-DAD-ES
SI-MSn, both
h extracts beiing dominate
ed by the pairs diosmetinn-7-O-glucosy
yl-3´O-pentoside+apig
genin-7-O-glu
ucosyl-4´-O--pentoside and
a
apigenin-7-O-glucooside+diosme
etin-7O-glu
ucoside (Fig. 1). Concerning the an
ntioxidant ac
ctivity, the extracts
e
dispplayed betterr O2●−
scave
enging capacity than ascorbic acid, but they we
ere weaker than
t
the refeerence comp
pound
again
nst the otherr oxidative species.
s
The
e ability of th
he extracts to
t inhibit choolinesterases
s was
weakk, but the aqu
ueous extrac
ct proved to be a good MAO-A
M
inhibitor, which caan be attribu
uted to
its hig
gher content of hidroxycinnamic acidss in comparis
son with the hydromethaanolic one.
Figurre 1 - HPLC
C-UV chroma
atogram (34
40 nm) of th
he aqueous (A) and hyddromethanollic (B)
extracts of Grind
delia robusta
a. 1 – 5-ca
affeoylquinic acid; 2 – 3-feruloylqu inic acid; 3 – 4oylquinic aciid; 4 – quercetin-3-O-he
exosyl-7-O-glucoside; 5 – 5-feruloylqquinic acid; 6 – 4caffeo
ferulo
oylquinic acid; 7 – 3,5-di-feruloylqu
uinic acid; 8 – 3,5-di-ca
affeoylquinic acid; 9 – 3,4-di3
caffeo
oylquinic acid; 10 – querrcetin-7-O-gl ucoside; 11 – quercetin--3-O-hexosidde; 12 – luteo
olin-7O-glu
ucosyl-3´/4´-O
O-pentoside;; 13 – 4,5-dii-caffeoylquin
nic acid; 14 – luteolin-7--O-glucoside; 15 –
diosm
metin-7-O-glu
ucosyl-3´-O-p
pentoside; 1
16 – apigen
nin-7-O-gluco
osyl-4´-O-peentoside; 17 – 3methyyl-quercetin glucoside; 18 – 3-metthyl-quercetin glucoside isomer; 199 – apigenin
n-7-Ogluco
oside; 20 – diosmetin-7-O
d
O-glucoside;; 21 – querc
cetin; 22 – lu
uteolin-3´/4´-O
O-pentoside; 23 –
3-me
ethyl-quercetiin pentoside; 24 – luteoliin; 25 – diosmetin-3´-O-p
pentoside; 266 – apigenin-4´-Opento
oside; 27 – 3-methyl-quercetin; 28 – dimethyl-q
quercetin pentoside; 29 – apigenin; 30 –
diosm
metin; 31 – quercetageti
q
n-3,6,3´/4'-trrimethyletherr; 32 – querrcetin-3,3´-di methylether;; 33 –
querccetin-3,7- dim
methylether; 34 – kaem
mpferol-3-me
ethylether; 35
5 – quercettagetin-3,6,7,3´/4´tetram
methylether; 36 – 6-hydroxy-kaempfe
erol-3,6,7/4´--trimethylethe
er; A1 – hyd roxycinnamic acid
1; A2
2 – hydroxycinnamic acid
d 2; A3 – hyd
droxycinnam
mic acid 3; F1
1 – flavonoidd 1; F2 – flav
vonoid
2.
Acknowledgemen
nts: The auth
hors are gratteful to Fundação para a Ciência e a Tecnologia (FCT)
for grant no. PE
Est-C/EQB/LA
A0006/2011,, to "Operaç
ção NORTE-07-0124-FE
EDER-00006
69", to
CYTE
ED Program
mme (Ref. 112RT0460
0) CORNUC
COPIA Thematic Netw
work and project
p
AGL2
2011-23690 (CICYT). Clara Gro
osso thanks
s FCT for the Posst-Doc fello
owship
(SFR
RH/BPD/6392
22/2009).
www.mapp.edu.pt/bioplant 33 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 1 Enzymes
s inhibitio
on by Pipe
er betle L. leaves
eia P. Oliveira
a1, Joana Ferrreira1, Sofia Riboira1, Fed
derico Ferreres2, Angel Gil-l-Izquierdo2, Patrícia
P
1
1
ntão , Paula B.. Andrade
QUIMTE/Laborratório de Fa
armacognosia
a, Departame
ento de Quím
mica, Faculddade de Farrmácia,
ersidade do Po
orto, Rua de Jorge
J
Viterbo F
Ferreira, nº 22
28, 4050-313 Porto,
P
Portugaal
earch Group on
o Quality, Safety
S
and Bio
oactivity of Pllant Foods, Department
D
off Food Scienc
ce and
nology, CEBAS (CSIC), P.O
O. Box 164, 30
0100 Campus University Espinardo, Murccia, Spain
r betle L., a member of
o Piperacea
ae, is a wid
dely distributted plant in the tropica
al and
ropical region
ns of the wo
orld, recognizzed as a trad
ditional herbal remedy foor many dise
eases.
leaves act as breath-fres
shener and a
are used in the Indian system of meedicine and health
heir medicinal propertiies, such a
hepatoprote
as anti-inflammatory, anti-allergic,
a
ective,
xidant, antifu
ungic, antihelmintic, xanth
hine oxidase
e inhibitor and
d chemoprevventive [1, 2,, 3]. In
work, aqueou
us lyophilized and ethan olic extracts of P. betle were analyzzed regarding
g their
mical compossition and bio
ological pote
ential. Amino
o acids, fatty acids, sterools and triterpenes
determined by GC-MS.B
Both extractss showed a good
g
potentia
al against accetylcholinestterase
E) and butyrylcholineste
erase (BuChE
E). In a general way, the
e aqueous eextract revea
aled to
ore active ag
gainst BuChE (IC50 = 0.6
601 mg/mL) while
w
the eth
hanolic one w
was more efffective
nst AChE (IC
C50 = 0.264 mg/mL)
m
(Fig. 1A and 1B). Furthermorre, the α-gluccosidase inhibitory
ty was also evaluated, being
b
ethano
olic extract the
t most inte
eresting (IC550 = 0.069 mg/mL)
m
1C).
C
% α -Glucosidase inhibition
B
00
80
60
40
20
0
0
1
2
100
Aqueous lyopphilized extract
Ethanolic exttract
80
60
40
20
0
3
[Piper betlee] (mg/mL)
0
1
2
3
[
[Piper
betle] (mg/mL)
a
of aq
queous lyoph
hilized and ethanolic exxtracts of P. betle
. Enzymatic inhibitory activity
es: (A) AChE
E, (B) BuCh
hE and (C) α-glucosidas
se. Values show
s
mean + SE from three
riments, perfformed in trip
plicate.
results sugg
gest that P. betle leavess could be useful
u
to the
e prevention and treatment of
eimer´s dise
ease and diabetes,
d
du
ue to their effects aga
ainst cholineesterases and αosidase, resp
pectively.
rences:
anguly, M. et al., 2007. The
T Journal o
of Pharmacy and Pharma
acology, 59, 711-718
ajumdar, B. et al., 2002. Indian Journ
nal of Clinica
al Biochemisttry, 17, 49-577.
urata, K. et al.,
a 2009. Jou
urnal of Natu
ural Medicine
es, 63, 355-3
359.
nowledgmen
nts: The authors are gra
ateful to Fun
ndação para a Ciência e a Tecnolog
gia for
t no. PEst-C//EQB/LA0006/2011.
www.mapp.edu.pt/bioplant 34 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 2 Glandora diffusa (Lag.)
(
D. C
C. Thomas
s: metabolic profile in
P22
authentic
city contro
ol and the
e key of bioactivity
2
3
F. Ferrnandes1, P. Almeida
A
, F. Ferreres
F
, A. G
Gil-Izquierdo3, P. Valentão2, P. B. Andradee
1
Interrdisciplinary Centre
C
for Marine and Envi
vironmental Re
esearch (CIIM
MAR/CIMAR), Rua dos Bragas nº
289, 4
4050-123 Portto, Portugal.
2
REQ
QUIMTE/Labo
oratório de Farmacognosi
F
ia, Departam
mento de Quíímica, Faculddade de Farrmácia,
Unive
ersidade do Po
orto, Rua de Jorge
J
Viterbo F
Ferreira, nº 22
28, 4050-313 Porto,
P
Portugaal
3
Ressearch Group on Quality, Safety
S
and Bio
ioactivity of Plant Foods, Department
D
off Food Scienc
ce and
Techn
nology, CEBAS (CSIC), P.O
O. Box 164, 30
0100 Campus University Espinardo, Murccia, Spain.
Gland
dora genus belongs
b
to Boraginaceae
e family, in which Glandorra diffusa (Laag.) D. C. Th
homas
is inccluded. Usua
ally known as
a scramblin
ng-gromwell or shrubby-g
gromwell, thhe consumpttion of
this sspecies hass been asso
ociated with diuretic, de
epurative, an
ntihypertensivve [1] and, more
recen
ntly, antidiab
betic and antioxidant pro
operties [2].T
This work aimed the usee of the che
emical
profile
e as a tool for
f authenticity control, a
and the asse
essment of some
s
biologiccal activities of G.
diffussa, purchased in the local market, from
m three diffe
erent medicin
nal plants disstributors.
Amino acids, fattty acids, ste
erols and tritterpenes we
ere determin
ned by GC-M
MS. The phenolic
profile
e was chara
acterized by HPLC-DAD
D. The studie
ed samples revealed a similar qualitative
comp
position. Nevvertheless, different quan
ntitative proffiles were ob
bserved.The three comm
mercial
samp
ples showed
d a potent dose-depend
d
dent respons
se against free radicals and a mod
derate
inhibiitory effect on
o the enzymes acetylccholinesteras
se and butyrrylcholinesterrase. Additio
onally,
extracts were cyto
otoxic to botth human co lorectal aden
nocarcinoma
a cell line (Caaco-2) and human
h
gastrric adenocarrcinoma celll line (AGS)). Sample B revealed to the bestt anticarcino
ogenic
prope
erties (Fig. 1).
5
50
100
2.
0
1.
0
0.
5
0.
25
0
0.
12
0
0.
06
2.
0
1.
0
0.
5
0
0.
01
0.
02
0.
03
0.
06
0.
12
0.
25
0
200
0.
03
10
00
300
10
00
0.
01
50
% Cell Viability
20
00
Caco-2 cells
c
% LDH leakage
100
15
50
0.
02
30
00
AGS
% LDH leakage
% Cell Viability
150
Fig.1.
Cyttotoxic
effectt of an extracct from G. diiffusa (samp le B) on AGS
S and Caco--2 cells. Resuults are pres
sented
as me
ean ± SEM of
o 4 independ
dent experim
ments perform
med in triplica
ate.
Sam
mple B (mg/mL))
Sa
ample B (mg/m
mL)
phenolic com
mpounds qua
alitative proffile of the three samples
s revealed too be similar to the
The p
one p
previously re
eported [2], confirming
c
th
heir authentic
city. Additionally, the diveersity of desc
cribed
comp
pounds, toge
ether with itts biologic p
potential, su
uggests G. diffusa
d
as a good sourrce of
bioacctive compou
unds, and points its consu
umption as a probable su
urplus to hum
man’s health.
[1] Crrellin, J. K., ett al. (1990). Herbal
H
medicin
ne past and present:
p
a refe
erence guide to medicinal plants:
Duke University Pre
ess.
erreres, F., et al.
a Food Chem
m., 2013, 136, 1390-1398.
[2] Fe
Acknowledgmentts: The authors are gratteful to Fund
dação para a Ciência e a Tecnolog
gia for
grantt no. PEst-C//EQB/LA0006/2011 and tto "Operação
o NORTE-07
7-0124-FEDE
ER-000069".
www.mapp.edu.pt/bioplant 35 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 3 New insights on SIZ1
S
function in the Arabidop
psis thalian
na stress
P23
response
e
Autho
ors: Freitas S, Castro PH, Couto
C
D, Ruiz--Albert J, Tava
ares RM, Beja
arano ER, Azeevedo H
Affiliattion: Center for
f Biodiversity
y, Functional & Integrative Genomics (B
BioFIG), Plantt Functional Biology
B
Cente
er, University of Minho, Cam
mpus de Gua
altar, 4710-05
57 Braga, Porttugal (P.H.C., D.C., S.F., R.M.T.,
R
H.A.)
uto de Hortofr
fruticultura Su
ubtropical y M
Mediterránea “La
“
Mayora”, Universidad de Málaga-C
Consejo
Institu
Superrior de Investiigaciones Cien
ntíficas (IHSM
M-UMA-CSIC),, Dept. Biolog
gía Celular, Geenética y Fisiiología,
Unive
ersidad de Mállaga, Campus
s Teatinos, 290
071 Málaga, Spain
S
(P.H.C., J.R.-A., E.R.B
B.)
Rapid
d and reverssible posttran
nslational mo
odifications (PTMs)
(
are essential
e
for the function
ning of
a livin
ng organism
m, particularly
y in response
e to a constantly changing environm
ment. One cla
ass of
PTMss subject to increasing fo
ocus of rese
earch employ
ys small mod
difying peptiddes like the Small
Ubiqu
uitin-related Modifier (SU
UMO). Modiffication by SUMO
S
may produce
p
diffeerent effects
s on a
targe
et protein, such as conforrmational con
ntrol, and cre
eation or blocking of inteeracting interfaces.
n many aspe
ects of plant growth and developmennt, but also acts
a
in
SUMO is not onlyy involved in
the re
esponse to abiotic stres
ss, including extreme tem
mperatures, drought, sallinity, and nu
utrient
availa
ability stressses. SUMO-c
conjugates a
accumulate quickly
q
upon several stim
muli and gra
adually
disap
ppear in a re
ecovery phase. SUMO rregulates nu
umerous nuc
cleus-associaated mechan
nisms,
name
ely by modullating transcrription and cchromatin rem
modeling fac
ctors for a raapid transcrip
ptional
repro
ogramming. Recent res
search effo rts have been
b
employed on deetermining plant’s
p
sumo
oylation targets, particularly in wha
at concerns the plant re
esponse to stress. We have
comp
piled this info
ormation and performed
d a gene on
ntology analy
ysis, highlighhting that SUMOconju
ugates inclu
ude chloropllast-targeted
d proteins. Moreover, our microarrray analyses of
Arabiidopsis T-DN
NA insertion
n mutants in
n the sumoy
ylation pathw
way show a de-regulation of
chloro
oplast-assocciated genes
s. Taking the
ese observa
ations into co
onsideration,, we perform
med a
functiional characcterization of the major A
Arabidopsis SUMO
S
E3 lig
gase siz1 m
mutant. Indee
ed, the
siz1 mutant dissplayed alte
ered pigmen
nt content, de-regulate
ed reactive oxygen sp
pecies
home
eostasis and changes in chloroplast ultrastructurre. Overall re
esults establlish a new ro
ole for
sumo
oylation in the
e control of chloroplast
c
fu
unctioning.
Workk was suppo
orted by FCT
T/FEDER (re
efs. PTDC/B
BIA-PLA/3850
0/2012 and FCOMP-01--0124FEDE
ER-028459).
www.mapp.edu.pt/bioplant 36 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 4 Distributtion of ara
abinogalac
ctan proteins and pectins in
n Quercus
s
P24
suber fem
male flower
M.I.Am
morim, a, b C.S
Sousa,a M.Cos
sta,a,b and S.C
Coimbra,a,b
a
b
Dep
partment of Biiology, Faculty
y of Science, U
University of Porto,
P
Portugall.
Centter for Biodive
ersity, Function
nal & Integratiive Genomics – BioFIG, Porrto, Portugal
Coarkk oak (Querrcus suber) is a domina
ant Fagacea
a tree specie
es in the fo rests of Sou
uthern
Iberia
an Peninsula
a. This is a monoecious
m
ttree species with a long progamic
p
phaase that provide a
comp
prehensive system
s
for comparative
c
d
and sexuall reproductio
on [1].
studies in development
Arabiinogalactan proteins (AG
GPs) and pe
ectins belong to a supe
erfamily of h ighly glycosylated
hydro
oxyproline-ricch glycoprote
eins cell com
mponents fou
und in the entire plant kkingdom, in almost
a
all pla
ant organs and
a cell types
s from root to
o flowers. Att the subcellular level, AG
GPs can be found
in the
e cell wall, in the apoplas
st or anchore
ed to the plas
sma membra
ane via a GP
PI anchor atta
ached
to the
e C-terminal domain of th
he AGP backkbone. Pectins, together with other ppolymers, are
e plant
cell w
wall compone
ents that give
e physical sttrength to the plant body
y and providee a barrier against
the o
outside enviro
onment. In
n reproductivve tissues, th
he expressio
on of AGPs iis associated
d with
the sporophyte–g
gametophyte transition, a
as our own previous
p
worrk has show
wn, a specific
c AGP
ession pattern during plant gameto
ogenesis in Arabidopsis thaliana [2]] and in Tri
rithuria
expre
subm
mersa [3]. A set
s of monoc
clonal antibo
odies (mAbs)) directed ag
gainst the carrbohydrate moiety
m
of ce
ell wall polysaccharides were
w
used fo
or the immunolocalizatio
on of AGPs aand pectins,, such
as: JIM 8, JIM13, JIM17 and LM7. The l abeling obta
ained with an
nti-AGP antibbodies in t fe
emale
ers show a dynamic distrribution of th ose sugar epitopes in re
eproductive ttissues. More
eover,
flowe
these
e labeling ma
ay function as
a a molecullar marker fo
or the tissue specific exppression patttern in
pistil tissues. Th
he changes in AGP orr pectins sp
pecific mAbs
s distributionn reveal tha
at the
ession of these is spatially regulated iin cork oak fe
emale flowerr.
expre
The authors are
e grateful forr the financiial support through
t
FCT
T for the Prooject PTDC//AGR0
GPL//118508/2010
[1] Bo
oavida L., Varela
V
C. and
d Feijo J.A.,, 1999 Sexu
ual reproducttion in the ccork oak (Qu
uercus
suberr L.).I.The prrogamic phas
se Sex Plantt Reprod 11:3
347–353
[2] Coimbra S., Almeida
A
J., Junqueira
J
V.., Costa, M. and Pereira L.G. 2007. Arabinogalactan
eins as mo
olecular ma
arkers in Arrabidopsis thaliana
t
sexu
ual reproduuction. Journ
nal of
prote
Botany
58
8:
40274035.
[3]Costa,M., Pereira,A.M
M., Rudall,P.J
J. and
Experimental
mbra,S. 2013.. Immunoloc
calization off arabinoga
alactan prote
eins (AGPs)) in reproductive
Coim
structtures of an early-diverge
e
ent angiosperrm, Trituria (Hydatellace
(
a), Annals oof Botany 111
1:183190
www.mapp.edu.pt/bioplant 37 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 Listta de Parrticipanttes / Listt of Partiicipants
___
______
_______
______
______
_______
______
_______
__
Albe
erto Pesso
oa
Protein T
Trafficking and Develo
opment/Bio
oFIG/FCUP –
pessoa.a
[email protected]
pt
o
Ana Assunção
PlantEvo
ol/ CIBIO-UP – agla@c
cibio.up.pt
Ana Clara Gro
osso
Farmaco
ognosia/ RE
EQUIMTE/ FFUP
F
–
claragrossso@hotma
ail.com
Ana Cláudia Oliveira
O
CIBIO - up2009060
[email protected]
Ana Cunha
DB - UM – accunha@b
a
bio.uminho..pt
CITAB, D
Ana Luísa Silv
va
Bioactive
e Natural Products /IB
BMC
analfgsilv
[email protected]
Ana Paula Porrtela
CIBIO/U P - up2009
[email protected]
Ana Raquel Allmeida
ologia e Citó
ómica/UA, almeidaraq
[email protected]
Biotecno
Ana Rita Silva
a
BIOCON//Biodiversitty and Conservation E
Ecology/ CIBIO
rita.silva
[email protected]
Ana Sofia Vaz
z
Biodiverssity and Co
onservation Ecology/ C
CIBIO
sofia.lino
[email protected]
Andreia Patríc
cia Oliveira Farmaco gnosia/ REQUIMTE/ FFUP
[email protected]
andreiap
ónio Teixe
eira
Antó
Departam
mento de Biologia
B
/CITAB/UM
[email protected] Artu
ur Conde
CITAB, D
DB-UM, artu
urconde@b
bio.uminho..pt
Brígida Pinho
Farmaco
ognosia/ RE
EQUIMTE/ FFUP
F
brigidarp
pinho@gma
ail.com
no Peixoto
o
Brun
Protein T
Trafficking and Develo
opment/Bio
oFIG/FCUP bpeixoto
[email protected]
m
Când
dida Learm
month
cmmc.le
[email protected]
Carla Patrícia Azevedo
Biotecno
ologia e Citó
ómica/Biolo
ogia do stre
ess/UA carlapsazzevedo@ua
a.pt
olina Azev
vedo
Caro
Farmaco
ognosia/ RE
EQUIMTE/ FFUP
F
carolina..azevedo@o
outlook.com
m
Conc
ceição San
ntos
nology and cytomics/C
CESAM/UA, csantos@u
ua.pt
Biotechn
Cristtiana Maia
a Alves
CIBIO/U P - up2009
[email protected]
Cristtiano Soarres
FCUP - [email protected]
BioFIG/F
Cristtina Maris
sa Almeida
a CIIMAR – [email protected]
p.pt
Fern
nanda Fida
algo
Plant Mo
olecular Biology and Biotechnolog
gy/ Plant
Stress/ B
BioFIG/ FCU
UP – ffidalg
[email protected]
Fran
ncisca Oliv
veira
BioFIG/F
FCUP - francisca.n.oliv
[email protected]
Fran
ncisco Lim
ma
Bioactive
e Natural Products/IBM
MC –
francisco
o.lima278@
@gmail.com
Fran
ncisco Peix
xoto
CITAB/U
UTAD - fpeix
[email protected]
Grac
ciliana Lop
pes
ognosia/ RE
EQUIMTE/ FFUP
F
Farmaco
gracilian [email protected]
www.mapp.edu.pt/bioplant 38 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 Héld
der Gomes
s
Universid
dade de Av
veiro – helder.alexand [email protected]
Henrique Noro
onha
CITAB, D
DB-UM, aluno Bioplantt henrique
[email protected]
o.pt
evedo
Herllânder Aze
BioFIG/U
UM – hazev
[email protected]
Hern
nâni Gerós
s
CITAB, D
DB-UM – ge
[email protected]
Inês
s Carqueijeiro
Bioactive
e Natural Products/IBM
MC –
[email protected]
Isab
bel Amorim
m
BioFIG/F
FCUP - mpa
[email protected]
Isab
bel Mina
CITAB/U
UM - [email protected]
minho.pt
Joan
na Almeida
a
oanaalexan
ndragar@gm
mail.com
FCUP - jo
Joan
na Gonçalv
ves
Empresa
a ecoinside – joanagon
[email protected]
Joan
na Ferreira
a
Farmaco
ognosia/ RE
EQUIMTE/ FFUP
F
–
joanague
erraferreira
[email protected]
João Jesus
CIGAR /F
FEUP - joao
ojesuscta@
@hotmail.co m
ge Teixeira
a
Jorg
BioFIG / FCUP – [email protected]
José
é António Macedo
Biodiverssity and Co
onservation Ecology/C
CIBIO jos.med..mac@gma
ail.com
José
é Feijó
IGC/Gulb
benkian - [email protected]. pt
José
é Pissarra
BioFIG/F
FCUP - jpiss
[email protected]
José
é Tiago Mo
oreira
BioFIG/U
UM - josetm
moreira@gm
mail.com
Julia
ana Oliveira
Secretarriado Biopla
ant – juolive
[email protected]
Lore
ena Romerro
Bioactive
e Natural Products/IBM
MC –
lorena.allmagro@um
m.es
Luís
s Carlos An
ndrade
[email protected]
com.pt
Luís
s Manuel Oliveira
O
CITAB/U
UM - luismco
[email protected]
Man
nuela Costa
a
UM – [email protected]
o.pt
BioFIG/U
Marc
cos André
é Monteiro
Farmaco
ognosia/ RE
EQUIMTE/ FFUP
F
taveira.m
marcos@gm
mail.com
Mariia da Costta
Biotechn
nology and Cytomics/U
UA/aluna B ioplant –
costama [email protected]
Fátim
ma Fernan
ndes
m
[email protected]
CIIMAR//CIMAR - mfgfernande
om
Mariia Teresa Borges
Biodiverssidade Costteira (LBC)- CIIMAR //FCUP mtborge
[email protected]
Mariia Teresa Braga
[email protected]
up.pt
Plant Strress/ FCUP – up20080
Mariia Teresa Lino Neto
BioFIG/U
UM – [email protected]
Mariiana Nune
es Barbosa
a FFUP - m
mariana.nun
[email protected]
om
Mariiana Sotto
omayor
FCUP/IB MC/Diretorra Bioplant – msottom
[email protected]
p.pt
Mars
slin Grego
ory
DB-UM/aluno Bioplant –
CITAB/D
marsling
gregory@gm
mail.com
Migu
uel Ângelo
o Faria
Plant Evo
olution Gro
oup/CIBIO - mfaria@c ibio.up.pt
Patrrícia Duartte
Bioactive
e Natural Products/IBM
MC – [email protected]
up.pt
Paulla Melo
BioFIG / FCUP /CC Bioplant – [email protected]
Pedrro Humberto Castro
o UM/BioF IG – pedro1berto@bio
o.uminho.p
pt
www.mapp.edu.pt/bioplant 39 4ª Workkshop Anual Biioplant 18 / 19 Julho 2013 Pedrro Albuque
erque
MDE - M icrobial Div
versity and Evolution/ CIBIO [email protected]
palbuque
Pedrro Miguel Almeida
Farmaco
ognosia/ RE
EQUIMTE/ FFUP
F
–
mfgferna
[email protected]
Rais
ssa Cristina Faria
Fisiologia
a vegetal/C
CESAM/UA ra_issa_
[email protected]
m
Rich
hard Maykel Breia
CITAB, D
DB-UM – ric
chardgonca
[email protected]
uminho.pt
Rui T
Tavares
UM/CC B
Bioplant – tavares@bio
t
o.uminho.p
pt
Sara
a Bettenco
ourt Ramo
os Bioactive
e Natural Products/IBM
MC –
[email protected]
Sara
a Freitas
BioFIG/U
UM - [email protected]
minho.pt
Sílviia Coimbra
a
BioFIG/F
FCUP – [email protected]
up.pt
Simã
ão Pedro Neves
N
BioFIG/F
FCUP - sbra
ancon@gma
ail.com
Sofia
a Barroso Riboira
Farmaco
ognosia/ RE
EQUIMTE/ FFUP
F
–
sofiaribo
oira92@gma
ail.com
Valé
érie Pantan
no
FCUP – v
[email protected]
Vane
essa Vieira
Protein T
Trafficking and Develo
opment/Bio
oFIG/FCUP –
[email protected]
ana Martin
ns
Vivia
CITAB, D
DB-UM, aluna Bioplantt –
[email protected]
nho.pt
Williian Giuseb
bepe Bedo
o Fisiologia
a vegetal/C
CESAM/UA - will_bedo
[email protected]
com
www.mapp.edu.pt/bioplant 40