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Max-Planck-Institute of Plant Breeding
Dept. of Plant Microbe Interactions (P. Schulze-Lefert)
Research areas:
-Recognition and Signalling in Disease Resistance (Paul Schulze-Lefert)
-Molecular Mechanisms of Defence Suppression (Ralph Panstruga)
-Signaling mechanisms in Plant Innate Immunity (Jane Parker)
-Molecular basis of Fungal Biotrophy (Richard O'Connell)
-Regulatory Protein Complexes in Plant Defense (Erich Kombrink)
-Transcriptional Regulation in Plant Defence (Imre E. Somssich)
-The Mechanism of FL2 Signaling in Plant Innate Immunity (Silke Robatzek)
-Functional Genomics in Barley (Thomas Koprek); until end of 2005
Mlo wildtype mlo mutant
Mutations in barley Mlo confer resistance to all isolates of Blumeria graminis f. sp. hordei
2 40
L
T
IM
A
V
EI
Y
K
S
K
I AV
VQA
W
L
GT
NV
P M
V LFFV
A
T
L
I
D Q
T P
I
H
G
G
I
L
VF
A
P
WT
S V
L
V IV
IFI
V
SI
T G L
W
F
Q
M
MI
IME
E
L
D
F
V
RA
G
K
I
D
V
E
D
S
S
K
R
T
M
I
A
Q
D
K
I
V
N
L
K
E
L
K
D
G
S
K
E
T
M
N
G
P
M
K
T A
S
F
F
A
T
H
S
V
R
K
SQ
N
P
K
V
L
P
T
K
L
M
D
S
R
P
VF
H
V
TK
H
F
A
R
S
FS
L
L
P
P
T
G
G
RF R
FM
A S
D
VL
H L
N S
GQ
Q
G
P
S
L
K
D
Q
D
V
S
T
V
H
S
V A
KG
NA
K
V P
H
SR
S
V
AD
DK
W N
Q DD
Q
E
K
R
Y
G
A
R
RP
F
S
T
P
R R
QI
D
M
D
PR
T
E
L
I
R
A
V
E
I
P
G
Q
R
S
F
M
S
L P
PS
V
RV
S
HR
Q
I
LQ
LY M
L
I
VLI G
AYVF IC
T
V A
A
A L
L
LA
TF
T
SLV
NM
F A
R
F S S S
HD
L
A
S
RE
R
T
KT
KE
K
S
Y
A
K
L TA
P
M
H A
K
N
QF
V A
IS
R
LA
N
K
A
A
E
T
W
A
H Q IL
G
S
E
R
4 0
H Q
TL
5 0
M
T
6 0
1 30
1 60
1 70
1 80
2 00 2 10
2 20
2 30
2 503 00
3 10
3 20
3 30
3 70
3 80
4 10
4 30
4 404 50
4 60
4 70
4 80
4 90
5 00
5 105 20
5 30
4 20
1 90 S
W
G
TP
P E
KG
R E
VP A
MSD
K
L
V
L
EL
MGIS
LL L
FL
H
W
H
F QS D
Y
RF
H
F
YR
E
F
V
IS
W
G
P
GTK
L
V
FW
F
P
LH
FN AF
FW
C
CSY T
P L
VQ
MGS
K
L
W
KA
NH2
COOH
MFAVV
LV S
V
A
L
G
F
HGL
HKL
HW
EM
FD
L
TL I
L
SE
DA
I
IC
D11 0
W
A
VM
P
PS
C K
K
GT
G R
R
E
LMA
V
ST
K
P
G
C
E
YDVYK
VV
E x trac e llu la r
In tra ce llu la r
2 40
L
T
IM
A
V
EI
Y
K
S
K
I AV
VQA
W
L
GT
NV
P M
V LFFV
A
T
L
I
D Q
T P
I
H
G
G
I
L
VF
A
P
WT
S V
L
V IV
IFI
V
SI
T G L
W
F
Q
M
MI
IME
E
L
D
F
V
RA
G
K
I
D
V
E
D
2 40
L
T
IM
A
V
EI
Y
K
S
K
I AV
VQA
W
L
GT
NV
P M
V LFFV
A
T
L
I
D Q
T P
I
H
G
G
I
L
VF
A
P
WT
S V
L
V IV
IFI
V
SI
T G L
W
F
Q
M
MI
IME
E
L
D
F
V
RA
G
K
I
D
V
E
D
S
S
K
R
T
M
I
A
Q
D
K
I
V
N
L
K
E
L
K
D
G
S
K
E
T
M
N
G
P
M
K
T A
S
F
F
A
T
H
S
V
R
K
SQ
N
P
K
V
L
P
T
K
L
M
D
S
R
P
VF
H
V
TK
H
F
A
R
S
FS
L
L
P
P
T
G
G
RF R
FM
A S
D
VL
H L
N S
GQ
Q
S
S
K
R
T
M
I
A
Q
D
K
I
V
N
L
K
E
L
K
D
G
S
K
E
T
M
N
G
P
M
K
T A
S
F
F
A
T
H
S
V
R
K
SQ
N
P
K
V
L
P
T
K
L
M
D
S
R
P
VF
H
V
TK
H
F
A
R
S
FS
L
L
P
P
T
G
G
RF R
FM
A S
D
VL
H L
N S
GQ
Q
G
P
S
L
K
D
Q
D
V
S
T
V
H
S
V A
KG
NA
K
V P
H
SR
S
V
AD
DK
W N
Q DD
Q
E
K
R
Y
G
A
R
RP
F
S
T
P
R R
QI
D
M
D
PR
T
E
L
I
R
A
V
E
I
P
G
Q
R
S
F
M
S
L P
PS
V
RV
S
HR
Q
I
LQ
LY
G
P
S
L
K
D
Q
D
V
S
T
V
H
S
V A
KG
NA
K
V P
H
SR
S
V
AD
DK
W N
Q DD
Q
E
K
R
Y
G
A
R
RP
F
S
T
P
R R
QI
D
M
D
PR
T
E
L
I
R
A
V
E
I
P
G
Q
R
S
F
M
S
L P
PS
V
RV
S
HR
Q
I
LQ
LY M
L
I
VLI G
AYVF IC
T
V A
A
A L
L
LA
TF
T
SLV
NM
F A
R
F S S S
HD
L
A
S
RE
R
T
KT
KE
K
S
Y
A
K
L TA
P
M
H A
K
N
QF
V A
IS
R
LA
N
K
A
A
E
T
W
A
H Q IL
G
S
ML
I
VLI G
AYVF IC
T
V A
A
A L
L
LA
TF
T
SLV
NM
F A
R
F S S S
HD
L
A
S
RE
R
T
KT
KE
K
S
Y
A
K
L TA
P
M
H A
K
N
QF
V A
IS
R
LA
N
K
A
A
E
T
W
A
H Q IL
G
S
E
R
4 0
H Q
TL
5 0
M
T
6 0
1 30
1 60
1 70
1 80
2 00 2 10
2 20
2 30
2 503 00
3 10
3 20
3 30
3 70
3 80
4 10
4 30
4 404 50
4 60
4 70
4 80
4 90
5 00
5 105 20
5 30
4 20
1 90 S
W
G
TP
P E
KG
R E
VP A
MSD
K
L
V
L
EL
MGIS
LL L
FL
H
W
H
F QS D
Y
RF
H
F
YR
E
F
V
IS
W
G
P
GTK
L
V
FW
F
P
LH
FN AF
FW
C
CSY T
P L
VQ
MGS
K
L
W
K
E
R
4 0
H Q
TL
5 0
M
T
6 0
1 30
1 60
1 70
1 80
2 00 2 10
2 20
2 30
2 503 00
3 10
3 20
3 30
3 70
3 80
4 10
4 30
4 404 50
4 60
4 70
4 80
4 90
5 00
5 105 20
5 30
4 20
1 90 S
W
G
TP
P E
KG
R E
VP A
MSD
K
L
V
L
EL
MGIS
LL L
FL
H
W
H
F QS D
Y
RF
H
F
YR
E
F
V
IS
W
G
P
GTK
L
V
FW
F
P
LH
FN AF
FW
C
CSY T
P L
VQ
MGS
K
L
W
KA
NH2
COOH
MFAVV
LV S
V
A
L
G
F
HGL
HKL
HW
EM
FD
L
TL I
L
SE
DA
I
IC
D11 0
W
A
VM
P
PS
C K
K
GT
G R
R
E
LMA
V
ST
K
P
G
C
E
YDVYK
VV
E x trac e llu la r
In tra ce llu la r
2 40
L
T
IM
A
V
EI
Y
K
S
K
I AV
VQA
W
L
GT
NV
P M
V LFFV
A
T
L
I
D Q
T P
I
H
G
G
I
L
VF
A
P
WT
S V
L
V IV
IFI
V
SI
T G L
W
F
Q
M
MI
IME
E
L
D
F
V
RA
G
K
I
D
V
E
D
S
S
K
R
T
M
I
A
Q
D
K
I
V
N
L
K
E
L
K
D
G
S
K
E
T
M
N
G
P
M
K
T A
S
F
F
A
T
H
S
V
R
K
SQ
N
P
K
V
L
P
T
K
L
M
D
S
R
P
VF
H
V
TK
H
F
A
R
S
FS
L
L
P
P
T
G
G
RF R
FM
A S
D
VL
H L
N S
GQ
Q
G
P
S
L
K
D
Q
D
V
S
T
V
H
S
V A
KG
NA
K
V P
H
SR
S
V
AD
DK
W N
Q DD
Q
E
K
R
Y
G
A
R
RP
F
S
T
P
R R
QI
D
M
D
PR
T
E
L
I
R
A
V
E
I
P
G
Q
R
S
F
M
S
L P
PS
V
RV
S
HR
Q
I
LQ
LY M
L
I
VLI G
AYVF IC
T
V A
A
A L
L
LA
TF
T
SLV
NM
F A
R
F S S S
HD
L
A
S
RE
R
T
KT
KE
K
S
Y
A
K
L TA
P
M
H A
K
N
QF
V A
IS
R
LA
N
K
A
A
E
T
W
A
H Q IL
G
S
E
R
4 0
H Q
TL
5 0
M
T
6 0
1 30
1 60
1 70
1 80
2 00 2 10
2 20
2 30
2 503 00
3 10
3 20
3 30
3 70
3 80
4 10
4 30
4 404 50
4 60
4 70
4 80
4 90
5 00
5 105 20
5 30
4 20
1 90 S
W
G
TP
P E
KG
R E
VP A
MSD
K
L
V
L
EL
MGIS
LL L
FL
H
W
H
F QS D
Y
RF
H
F
YR
E
F
V
IS
W
G
P
GTK
L
V
FW
F
P
LH
FN AF
FW
C
CSY T
P L
VQ
MGS
K
L
W
KA
NH2
COOH
MFAVV
LV S
V
A
L
G
F
HGL
HKL
HW
EM
FD
L
TL I
L
SE
DA
I
IC
D11 0
W
A
VM
P
PS
C K
K
GT
G R
R
E
LMA
V
ST
K
P
G
C
E
YDVYK
VV
E x trac e llu la r
In tra ce llu la r
2 40
L
T
IM
A
V
EI
Y
K
S
K
I AV
VQA
W
L
GT
NV
P M
V LFFV
A
T
L
I
D Q
T P
I
H
G
G
I
L
VF
A
P
WT
S V
L
V IV
IFI
V
SI
T G L
W
F
Q
M
MI
IME
E
L
D
F
V
RA
G
K
I
D
V
E
D
2 40
L
T
IM
A
V
EI
Y
K
S
K
I AV
VQA
W
L
GT
NV
P M
V LFFV
A
T
L
I
D Q
T P
I
H
G
G
I
L
VF
A
P
WT
S V
L
V IV
IFI
V
SI
T G L
W
F
Q
M
MI
IME
E
L
D
F
V
RA
G
K
I
D
V
E
D
S
S
K
R
T
M
I
A
Q
D
K
I
V
N
L
K
E
L
K
D
G
S
K
E
T
M
N
G
P
M
K
T A
S
F
F
A
T
H
S
V
R
K
SQ
N
P
K
V
L
P
T
K
L
M
D
S
R
P
VF
H
V
TK
H
F
A
R
S
FS
L
L
P
P
T
G
G
RF R
FM
A S
D
VL
H L
N S
GQ
Q
S
S
K
R
T
M
I
A
Q
D
K
I
V
N
L
K
E
L
K
D
G
S
K
E
T
M
N
G
P
M
K
T A
S
F
F
A
T
H
S
V
R
K
SQ
N
P
K
V
L
P
T
K
L
M
D
S
R
P
VF
H
V
TK
H
F
A
R
S
FS
L
L
P
P
T
G
G
RF R
FM
A S
D
VL
H L
N S
GQ
Q
G
P
S
L
K
D
Q
D
V
S
T
V
H
S
V A
KG
NA
K
V P
H
SR
S
V
AD
DK
W N
Q DD
Q
E
K
R
Y
G
A
R
RP
F
S
T
P
R R
QI
D
M
D
PR
T
E
L
I
R
A
V
E
I
P
G
Q
R
S
F
M
S
L P
PS
V
RV
S
HR
Q
I
LQ
LY
G
P
S
L
K
D
Q
D
V
S
T
V
H
S
V A
KG
NA
K
V P
H
SR
S
V
AD
DK
W N
Q DD
Q
E
K
R
Y
G
A
R
RP
F
S
T
P
R R
QI
D
M
D
PR
T
E
L
I
R
A
V
E
I
P
G
Q
R
S
F
M
S
L P
PS
V
RV
S
HR
Q
I
LQ
LY M
L
I
VLI G
AYVF IC
T
V A
A
A L
L
LA
TF
T
SLV
NM
F A
R
F S S S
HD
L
A
S
RE
R
T
KT
KE
K
S
Y
A
K
L TA
P
M
H A
K
N
QF
V A
IS
R
LA
N
K
A
A
E
T
W
A
H Q IL
G
S
ML
I
VLI G
AYVF IC
T
V A
A
A L
L
LA
TF
T
SLV
NM
F A
R
F S S S
HD
L
A
S
RE
R
T
KT
KE
K
S
Y
A
K
L TA
P
M
H A
K
N
QF
V A
IS
R
LA
N
K
A
A
E
T
W
A
H Q IL
G
S
E
R
4 0
H Q
TL
5 0
M
T
6 0
1 30
1 60
1 70
1 80
2 00 2 10
2 20
2 30
2 503 00
3 10
3 20
3 30
3 70
3 80
4 10
4 30
4 404 50
4 60
4 70
4 80
4 90
5 00
5 105 20
5 30
4 20
1 90 S
W
G
TP
P E
KG
R E
VP A
MSD
K
L
V
L
EL
MGIS
LL L
FL
H
W
H
F QS D
Y
RF
H
F
YR
E
F
V
IS
W
G
P
GTK
L
V
FW
F
P
LH
FN AF
FW
C
CSY T
P L
VQ
MGS
K
L
W
K
E
R
4 0
H Q
TL
5 0
M
T
6 0
1 30
1 60
1 70
1 80
2 00 2 10
2 20
2 30
2 503 00
3 10
3 20
3 30
3 70
3 80
4 10
4 30
4 404 50
4 60
4 70
4 80
4 90
5 00
5 105 20
5 30
4 20
1 90 S
W
G
TP
P E
KG
R E
VP A
MSD
K
L
V
L
EL
MGIS
LL L
FL
H
W
H
F QS D
Y
RF
H
F
YR
E
F
V
IS
W
G
P
GTK
L
V
FW
F
P
LH
FN AF
FW
C
CSY T
P L
VQ
MGS
K
L
W
KA
NH2
COOH
MFAVV
LV S
V
A
L
G
F
HGL
HKL
HW
EM
FD
L
TL I
L
SE
DA
I
IC
D11 0
W
A
VM
P
PS
C K
K
GT
G R
R
E
LMA
V
ST
K
P
G
C
E
YDVYK
VV
E x trac e llu la r
In tra ce llu la r
BUT: mlo mutants are more susceptible to certain hemibiotrophic (Magnaporthe grisea) and necrotrophic (Bipolaris sorokiniana) fungi
TPS 8, 263, 2003
Inducibility and expression kinetics of parsley genes upon Pmg elicitor treatment
A B
probe: nuclear run-on transcript
W1 box CTTACACACTTAATTTGACCGAGTAACATTCG
W2 box CAATTACAGCCATCAAAGTTGACCAATAATA
W3 box CTATTATGACTAAATAGTCAGAATTTCCAA
E17 box TGTCAATGGTCAACATTCAAC
D box TACAATTCAAACATTGTTCAAACAAGGAACC
U box TGTGCCATGAAGTTGAAATTCAATAGTGTGC
S box CAGCCACCAAAGAGGACCCAGAAT
GCC box CAGCCGCCAAAGAGGACCCAGAAT
Gst1 box TTCTAGCACCAGATTTGACCAAAC
Fungal Elicitor Responsive Cis-acting Elements
Rushton et al., Plant Cell l4, 749 (2002)
cis-element
TATA-box
4 x D box
2 x (W2/S/ D) box
4 x W1 box
4 x S box
MgCl P. syringae
B. graminisP. parasitica
The WRKY family of transcription factors
WRKY C-C-H-HWRKYC-C-H-H
Group I
WRKY C-C-H-H
HARF WRKY C-C-H-H
LZ WRKY C-C-H-H
Group II
C-C- -H-CWRKY
Group III
LZ WRKY C-C- -H-C
D---WRKYGQK-------PR-Y--C-----C---K--------------TY---H-H
WRKY domain:
The WRKY Family of Transcription Factors
• Novel Zn finger type DNA binding proteins• Translocated into the plant cell nucleus via NLS motif
• Bind to cis-regulatory element: W box (C/T TGAC T/C)
• Phosphorylation increases DNA binding affinity• Contain functional transactivation domains• Capable of activating but also repressing gene expression
AGTTGACCAA
AGATGACCAA
AGGTGACCAA
AGCTGACCAA
AGTTGACAAA
AGTTGACGAA
AGTTGACTAA
AGTAGACCAA
AGTTCACCAA
AGTTGAACAA
AGTTGACCAA
WRKY11At4g31550
WRKY genes are found outside of the plant phyla
Arabidopsis thaliana WRKY Family
AtWRKY8AtWRKY12AtWRKY13AtWRKY23AtWRKY24AtWRKY28AtWRKY43AtWRKY48AtWRKY49AtWRKY50AtWRKY51AtWRKY56AtWRKY57AtWRKY59AtWRKY68AtWRKY71AtWRKY75
c
AtWRKY18AtWRKY40AtWRKY60
a
AtWRKY1AtWRKY2AtWRKY3AtWRKY4AtWRKY10AtWRKY19AtWRKY20AtWRKY25AtWRKY26AtWRKY32AtWRKY33AtWRKY34AtWRKY44AtWRKY45AtWRKY58AtWRKY73
AtWRKY7AtWRKY11AtWRKY15AtWRKY17AtWRKY21AtWRKY39AtWRKY74
d
AtWRKY30AtWRKY38AtWRKY41AtWRKY46AtWRKY52AtWRKY53AtWRKY54AtWRKY55AtWRKY62AtWRKY63AtWRKY64AtWRKY66AtWRKY67AtWRKY70
AtWRKY14AtWRKY16AtWRKY22AtWRKY27AtWRKY29AtWRKY35AtWRKY65AtWRKY69
e
AtWRKY6AtWRKY9AtWRKY31AtWRKY36AtWRKY42AtWRKY47AtWRKY61AtWRKY72
b
group I group II group III
Response of AtWRKY KO lines to Ralstonia solanacearum
8 days post root inoculation with strain GMI1000 (virulent on wt Col-0 plants)
Wt AtWRKY27 KO mutant
Disease Index
0 : no wilt symptom
1 : 0 – 25 % wilted leaves
2 : 25 – 50 % wilted leaves
3 : 50 – 75 % wilted leaves
4 : 75 – 100 % wilted leaves
0
1
2
3
4
0 2 4 6 8 10 12 14
Dis
ease
In
dex
(50 plants from each line root inoculated)
Days post-inoculation
Control-Col-0
W27ETL-Col-1
Control-Col-1W27-SALK-Col-0
AtWRKY27 KO lines show enhanced resistance towards Rastonia solanacearum (GMI1000) infection
Col-0 WRKY53 KO
Col-0
WRKY40
bacte
ria
log
cfu
/cm
2
Days
WRKY18
0
2
4
6
8
10
12
14
0 2 4
WRKY53
WRKY40 KO
AtWRKY40 + 53 knockout lines are more susceptible towards P. syringae pv. tomato DC3000
