1. Molecular taxonomy of recently described
Phytophthora and Pythium
Dr. Lassaad Belbahri

2. Declining Polish forests
Alnus glutinosa, Kolo, Poland.

3. Direct PCR, DNA barcoding
Collaboration with FASTERIS SA (Geneva)

4. Single tube nested PCR for
sensitive detection of oomycetes

5. ARISA Analysis of oomycete infected
Rhododendron leaves

6. ITS rDNA PCR Amplification of oomycete
community from soil DNA

7. ARISA analysis of oomycete community
from soil DNA

8. Evolution rate of new species discovery during
time

9. Genomic assisted Phylogeny in the genus Phytophthora
Blair et al. (2007) Fungal Genetics and Biology 45 266-277

10. Morphological description : Ph. polonica

11. Morphological description: Py. recalcitrans
Hypha and hyphal swellings Appressoria
Antheridia Oospore

12. Phylogenetic analysis of Phytophthora spp.
Phylogenetic analysis based
on four concatenated genes
(ß-tubuline, EF1-α, NADH1
and COX1), induced by
Maximum Likelihood (PhyML)
Ph. sylvatica sp. nov
Ph. hungarica sp. nov

13. Phylogenetic analysis of Pythium spp.
Unrooted
phylogenetic analysis
Py.attra
Py.debaryanu
Py.helic
P
Py.dimorphu
Py.inte
Py.sylvaticum
Py.undulatum
Py.p
y.ma
Py
Py.a
Py.pro
ntheridiu
Py.
aro
based on Internal
cros
Py
.hy
rm
andrum
olig
ec
na nd
.am
dno
edium
poru
latum
an d
Py
an d
as
spo m
Py
m
m
.pe
m
r um
Py
cu
rum
ru m
m
.ac
.a
rip
Transcribed Spacers
rum
linu
r
an
Py
rh
loc m
en
th
.a
ue ale
Py
um
r
icu s
om rum
ist
um
.v s
re
de
os
an
an
Py .bo
rc
Py co
po
te
1 and 2 induced by
e
m
.p rp
Pythium quercum
Py
tra ilu
.q
yr
ilo ooli os ch
Py bu i y. edo
.in m P .o m on
fla ilu ph
Py
.pe tum Py s t er hy
ae
Bayesian Inference
Py riil y.
.gr u P am es
Py.
pl u
am
risp
inic m
ola Py
P
.ch
y.h
elic
oid
bita
cea
rum Pythium sterilum
Py.s oru
m cu cur
(MrBayes v.3.1.2) Py. ae
ulca ofer
tum ndig
Py.d
issim Py.i s
Py.sc ile exan
lerote Py.v
ichum nn
Py.zingib Py.nu
eris onon
Py.orthog
Py.myriotylum n
Py.acanthophoro
Py.tracheiphilum Py.perplexum
Py.salpingophorum Py.buismaniae
horum Py.polym
Py.conidiop astum
losum Py.unc
Py .follicu inulatu
m
um Py.m
rulos astop
Py.to m Py.c horu
m
u latu ylind
at en P y. rosp
Py.c atu
m irre orum
ust Py gula
ang tum .sp re
Py. olu Py i cu
.v e lum
Py ns .m
v oe Py am
.fle um .
Py kunm atum
ill
Py clin
.di m
Py .spin
Py oto
iss arin
cu
um
m
Py .ac
.h ro
os
um
ing
en
se Pythium spiculum
.d riu yp gyn
Py y.
m
ta
Py .erin sum
m
P og um
.lu
ch ratu
.e
Py
yn
.aq aule
Py
ch
um
Py
tile
o
in
ol
yc
Py
.ra
um
ul
.c
ua
Py
ac
at
phy a
Py
.vio amai
dio
Py.
rm
l
um
rae
eu
a
rico
.iw
Py.o
.p
s
Py.d
Py.o
pe
lae
Py
m
Py.ap
e r en
Py.hete
pad
Py
Py.insidios
ag
Py.glomer
Py.ultimumvar.u
Py.splendens
Py.ultimumvar.sporangiiferum
on d
os
kan
elien
dicu
on
kano
por
dha
.ch
hanide
.m
oga
P y.
Py
m
rothalli
Py.a
se
Py
gane
nen
atum
um
rmatu
seC
nseC
cum
ltimum
BS3
m
BS70
15.8
1.83
1

14. Towards a redefinition of oomycete genera

15. Ph. hedraiandra
Ph. hedraiandra, a
species with high
hybridization
potential spreading
on multiple hosts.

16. Ph. ramorum detection by real time PCR

17. Ph. alni detection by real time PCR
Ph. alni subsp. alni
Ph. alni subsp. multiformis
Ph. alni subsp. uniformis
Ph. alni subsp. alni
Ph. alni subsp. multiformis
Ph. alni subsp. alni
Ph. alni subsp. uniformis

18. Detection of Py. spp. and Ph. spp. by microarray

19. Ph. ramorum, tissue colonization
Tissue of infected Rhododendron
leaves with Ph. ramorum structures,
hyphae with zoosporangia growing out
of stomata on discolored
leaf surface (vital staining with FUN® 1,
Molecular Probes), zoospore production
is induced, when infected
P. ramorum in Rhododendron leaves were moistened and kept at low
cortex tissue, chlamydospores temperatures (7°C)
with white-yellow
autofluorescence in cortex of
necrotic Rhododendron stem

20. Ph. ramorum, Genetic transformation
Transgenic Ph. ramorum strain BBA9/95_6G:
Transgenic Ph. ramorum strain BBA26/02- CLSM micrographs showing GFP expression in
4: CLSM micrographs indicating GFP protoplasm of zoosporangia and hyphae, yellow
expression in germinating cysts. signals: natural Ph. ramorum autofluorescence
Left: transmission image in some hyphae, sporangium wall, base and
pedicel, GFP signals were amplified with Alexa
Right: GFP signals
Fluor 488®labelled anti GFP antibodies
(Molecular Probes)

21. Genomic organisation of virulence factors
in oomycete genomes

22. Evolutionary lines of oomycete virulence factors

23. Outlook
Diversity of oomycete species using non culture based techniques
Reinforcing environmental monitoring tools (Molecular toolbox)
Evolutionary history of main virulence factors
Linking genetic phylogeny to morphology

24. Special Thanks To:
Steve Woodward
Eduardo Moralejo
Enrique Descals
Jozef Bakonyi

25. Thanks for your attention