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Antioxidant activities and reactive oxygen species in
flag leaves of Fusarium-inoculated wheat genotypes Szeged – Timişoara axis for the safe food and feed
SZETISA1HURO/0901/147/2.2.2
Ágnes GalléDepartment of Plant Biology
University of Szeged
2nd Progress Report MeetingSzeged, 22-23 September, 2011
Reporting period: 01.06. 2011 – 30.09. 2011
www.huro-cbc.euwww.hungary-romania-cbc.eu
DisclaimerThe content of this page does not necessarily represent the official position of the European Union.
Two countries, one goal, joint success!
Fusarium head blight (FHB) is an important disease of wheat and has worldwide distribution, which results in yield loss and mycotoxin contamination of grain when the disease is caused by a mycotoxin-producing Fusarium species.
Fusarium graminearum and F. culmorum are the Fusarium species that most commonly cause FHB of wheat and both can produce trichothecene mycotoxins, predominantly deoxynivalenol (DON, also known as vomitoxin), zearalenone, nivalenol, dideoxynivalenol, 3-acetyldeoxynivalenol
Fusarium graminearum Fusarium culmorum
Vomitoxin, also known as deoxynivalenol (DON), is a type B trichothecene, an epoxy-sesquiterpeoid.
When compared to other trichothecene mycotoxins which can form in grains and forages, vomitoxin is relatively mild.
Vomitoxin belongs to a class of mycotoxins which are strong protein inhibitors. Inhibition of protein synthesis following exposure to vomitoxin causes the brain to increase its uptake of the amino acid tryptophan and, in turn, its synthesis of serotonin. Increased levels of serotonin are believed to be responsible for the anorexic effects of DON and other tricothecenes.
Irritation of the gastrointestinal tract may also play a role in reducing feed intake.
Zearalenone is also a trichothecene mycotoxin, and has estrogen effect, and can cause inflame in the uterus in swine.Nivalenol was shown to inhibit protein synthesis in rabbit reticulocytes in vitro, with an ID50 of 2.5 mg/ml, probably by impairment of the ribosomal function.
Nivalenol inhibited the synthesis of nucleic acids in vitro (Ueno and Fukushima, 1968). This response appeared at doses much higher than the inhibition of protein synthesis. The mechanism of nucleic acid inhibition is not known.
Before inoculation sampling (ROS and total antioxidant capacity)
anthesis
inoculation
SAMPLING
Dough stagePremilk stage Milk stage
GK Mini Manó/Nobeokabozu genetic mapping population (Prof. Mesterházy Ákos )
Nobeokabozu: FHB resistant lineA recombinant line from China: ‘WSY’, Wangshubai/Sumai3/Yangangfanzhou
GK Mini ManóArthur 71 / Sava // Rusalka / NS 171.2 /3/ F 30.74Resistant line to e.g. leaf rust
Plant material
Total intracellular ROS before inoculation
0
100
200
300
400
500
600
700
800
900
474 r 501 r 483 s 488 s
DC
F fl
uo
resc
ence
Total intracellular ROS after inoculation with F.c.
0
100
200
300
400
500
600
700
800
900
561 r 474 r 501 r 483 s 488 s
DFC
fluo
resc
ence
Total intracellular ROS after inoculation with F.g.
0
100
200
300
400
500
600
700
800
900
561 r 474 r 501 r 483 s 488 s
DFC
fluo
resc
ence
Total intracellular ROS after inoculation
0
100
200
300
400
500
600
700
800
900
561 r 474 r 501 r 483 s 488 s
DCF
fluo
resc
ence
Antioxidant activity (FRAP)
0
1000
2000
3000
4000
5000
6000
7000
8000
561 r 474 r 501 r 483 s 488 s
Ant
ioxi
dant
act
ivity
(m
mol
FeS
O4/
g F
W)
Antioxidant activity (FRAP)After inoculation with F. c.
0
1000
2000
3000
4000
5000
6000
7000
8000
561 r 474 r 501 r 483 s 488 sA
ntio
xida
nt a
ctiv
ity
(mm
ol F
eSO
4/ g
FW
)
Antioxidant activity (FRAP)After inoculation with F. g.
0
1000
2000
3000
4000
5000
6000
7000
8000
561 r 474 r 501 r 483 s 488 s
Ant
ioxi
dant
act
ivity
(m
mol
FeS
O4/
g F
W)
Antioxidant activity (FRAP)Before inoculation
0
1000
2000
3000
4000
5000
6000
474 r 501 r 483 s 488 s
An
tio
xid
an
t a
cti
vit
y
(mm
ol F
eS
O4/
g F
W)
Enzymatic antioxidants
Peroxidases (POX; EC 1.11.1.7) are oxido-reductive enzymes that participate in the wall-building processes such as oxidation of phenols, suberization, and lignification of host plant cells during the defense reaction against pathogenic agents.
Accumulation of lignin and phenolic compounds have been correlated with disease resistance in a number of plant–pathogen interactions. These include wheat, tomato and rice.
Enhanced POX activity has been correlated with resistance in rice, wheat, barley and sugarcane following the inoculation with phytopathogens.
POX activity was rather delayed or remained unchanged during the compatible interaction in susceptible plants. .
Peroxidase specific activity in heads of wheat cultivars. C, non-inoculated control; I, inoculated plants. (F. graminearum)
POD activity of the selected wheat lines
0
500
1000
1500
2000
2500
3000
561 r 474 r 501 r 483 s 488 sSp
ecif
ic P
OD
act
ivit
y (U
/ mg
p
rote
in)
POD activity of the selected wheat lines after F. culmorum infection
0
500
1000
1500
2000
2500
3000
561 r 474 r 501 r 483 s 488 sSp
ecif
ic P
OD
act
ivit
y (U
/ mg
p
rote
in)
POD activity of the selected wheat lines after F. graminearum infection
0
500
1000
1500
2000
2500
3000
561 r 474 r 501 r 483 s 488 sSp
ecif
ic P
OD
act
ivit
y (U
/ mg
p
rote
in)
SOD activity of the selected wheat lines
0
2
4
6
8
10
12
14
561 r 474 r 501 r 483 s 488 sSp
ecif
ic S
OD
act
ivit
y (U
/ mg
p
rote
in)
SOD activity of the selected wheat lines after F. culmorum infection
0246
8101214
561 r 474 r 501 r 483 s 488 sSp
ecif
ic S
OD
act
ivit
y (U
/ mg
p
rote
in)
SOD activity of the selected wheat lines after F. graminearum infection
02
468
10
1214
561 r 474 r 501 r 483 s 488 sSp
ecif
ic S
OD
act
ivit
y (U
/ mg
p
rote
in)
KAT activity of the selected wheat lines
0
100
200
300
400
500
600
561 r 474 r 501 r 483 s 488 sSp
ecif
ic K
AT
act
ivit
y (U
/ mg
p
rote
in)
KAT activity of the selected wheat lines after F. culmorum infection
0
100
200
300
400
500
600
561 r 474 r 501 r 483 s 488 sSp
ecif
ic K
AT
act
ivit
y (U
/ mg
p
rote
in)
KAT activity of the selected wheat lines after F. graminearum infection
0
100
200
300
400
500
600
561 r 474 r 501 r 483 s 488 sSp
ecif
ic K
AT
act
ivit
y (U
/ mg
p
rote
in)
GR activity of the selected wheat lines
0
100
200
300
400
500
600
561 r 474 r 501 r 483 s 488 s
Sp
ecif
ic G
R a
ctiv
ity
(U/ m
g
pro
tein
)
GR activity of the selected wheat lines after F. culmorum infection
0
100
200
300
400
500
600
561 r 474 r 501 r 483 s 488 sSp
ecif
ic G
R a
ctiv
ity
(U/ m
g
pro
tein
)
GR activity of the selected wheat lines after F. graminearum infection
0
100
200
300
400
500
600
561 r 474 r 501 r 483 s 488 sSp
ecif
ic G
R a
ctiv
ity
(U/ m
g
pro
tein
)
Some protein families (examples) which were induced during Fusarium infection : Zhou et al 2005
• Proteins with an antioxidant function: superoxide dismutase, dehydroascorbate reductase, and glutathione S-transferases (GSTs)
• Expression of proteins with highest similarity to dehydroascorbate reductase and TaGSTF5 (a glutathione S-transferase) differed following FHB infection in susceptible and resistant cultivars.
GST activity of the selected wheat lines
0
10
20
30
40
50
561 r 474 r 501 r 483 s 488 sSp
ecif
ic G
ST
act
ivit
y (U
/ mg
p
rote
in)
GST activity of the selected wheat lines after F. culmorum infection
0
10
20
30
40
50
561 r 474 r 501 r 483 s 488 sSp
ecif
ic G
ST
act
ivit
y (U
/ mg
p
rote
in)
GST activity of the selected wheat lines after F. graminearum infection
0
10
20
30
40
50
561 r 474 r 501 r 483 s 488 sSp
ecif
ic G
ST
act
ivit
y (U
/ mg
p
rote
in)
Parameter 561 474 501 483 488
POD
SOD
KAT
GR
GST
Total ROS
Total antioxidant capacity
FUSARIUM CULMORUM
Parameter 561 474 501 483 488
POD
SOD
KAT
GR
GST
Total ROS
Total antioxidant capacity
FUSARIUM GRAMINEARUM
Parameter 561 474 501 483 488
POD
SOD
KAT
GR
GST
Total ROS
Total antioxidant capacity
CONTROL
Conclusions:
The Fusarium inoculation on the ear influenced the hole antioxidant system in the flag leaves of wheat in dough stage!
There were differences between the resistant and the sensitive cultivars in POD activity inductions.
BUT according our measurements differences were detecable in control circumstances.
And the amount of ROS, antioxidant capacity and the GST activity correlated with the resistance.
Thank you for your attention!