Post on 18-Jan-2016
Participant 8
Assistant Contractor to Participant 7
Michael Tausz, Dieter Grill
Karin Herbinger, Klaus Remele, Maria Müller
Institute of Plantphysiology, University of Graz, Schubertstraße 51, A-8010 Graz, Austria
Investigations
• Biochemical defense– Ascorbate system– Glutathione system– Tocopherols– Chloroplast pigments
• Chromosomal damages– Aberration rates
Experiments
Container exposed young beech trees
Branch cuvette exposures
Mature beech trees
sampling 2003
Container exposed young (3-yrs-old) trees
exposed at 2 canopy levels: sun and shade
- Pigments and tocopherols
June, July, September
- Ascorbate and glutathione
due to insufficient material only September
Leaves:
Roots:- Ascorbate, glutathione and tocopherols
due to insufficient material only September
- Chromosomal aberration analysis
September
interface to partner 7
Branch cuvette exposures
- Pigments, tocopherols, ascorbate and glutathione
September
in leave samples included in branch cuvette fumigation systems
interface to partner 7
Mature trees
- tocopherols
May, June, July, September, Oktober
- Chromosomal aberration analysisMay, June, July, September, Oktober
interface to partner 4 who contributes ascorbate and glutathione
Leaves:
- Pigments and tocopherolsMay, June, July, September, Oktober
Roots:
Chromosomal aberrations
- are approved indicators for genetic damages and
- could be defined as changes in the structure of chromosoms
Chromosomal aberration analysis
Material: root tips of young and old beech trees
We adjusted the method
- in connection with the staining procedure (double staining: Schiff reagent, Giemsa, or Schiff reagent and carmine acetic acid)
- in connection with the identification of chromosomal aberrations in meta- and anaphases
(bridge, sticky chromosomes, vagrant, fragment etc)
Aberration types- sticky chromosomes - anaphase bridges
- vagrant chromosomes anaphasemetaphase
Problems:
The cell walls of most tips were lignified
and/or mycorrhized –
but:
enough root tips for the classification of
chromosomal aberrations
viol
axan
thin
neox
anth
in
zeax
anth
in
anth
erax
anth
in
chlo
roph
yll a
-c
arot
ene
Pigments: Chromatogram
chlo
roph
yll b
lute
in
-ca
rote
ne
0
2
4
6
8
10
12
14
A A
b
July 2003
young treesmature trees
SHADESHADE SUNSUN
total chlorophyll [mg g
-1 dry weight]
1 x ozone 2 x ozone
tota
l chl
orop
hyll
[mg
g-1 d
ry w
eigh
t]0
2
4
6
8
10
12
14
B
C
aa
b
sig. diff. mature/young:Sun 1 x ozoneSun 2 x ozoneShade 1 x ozone
Total chlorophyll
0
2
4
6
8
10
12
14
AAbc
June 2003
young treesmature trees
SHADESHADE SUNSUN
total chlorophyll [mg g
-1 dry weight]
1 x ozone 2 x ozone
tota
l chl
orop
hyll
[mg
g-1 d
ry w
eigh
t]
0
2
4
6
8
10
12
14B
B
a
cab
sig. diff. mature/young:Shade 1 x ozoneShade 2 x ozone
0
2
4
6
8
10
12
14
Mai
Juni
Juli
OktoberSeptemberJulyJuneMay
0
2
4
6
8
10
12
14
16
Juni Ju
li
young trees
mature trees
September OktoberJulyJuneMay
Sun 1 x ozone Sun 2 x ozone Sha 1 x ozone Sha 2 x ozone
tota
l chl
orop
hyll
[mg
g-1 d
ry w
eig
ht]
Total chlorophyll
?Containers removed
-Carotene
0
50
100
150
200
250
300
350
400
450
sig. diff. mature/young:Sun 1 x ozoneSun 2 x ozoneShade 1 x ozoneShade 2 x ozone
AB
AA
c
July 2003
young treesmature trees
SHADESHADE SUNSUN
-carotene [µ
g g-1 dry w
eight]
1 x ozone 2 x ozone
-ca
rote
ne [µ
g g-1
dry
wei
ght]
0
50
100
150
200
250
300
350
400
450
ab
bc
a
0
50
100
150
200
250
300
350
400
450
sig. diff. mature/young:Sun 1 x ozoneSun 2 x ozone
C
A
B
A
c
June 2003
young treesmature trees
SHADESHADE SUNSUN
-carotene [µ
g g-1 dry w
eight]
1 x ozone 2 x ozone
-ca
rote
ne [µ
g g-1
dry
wei
ght]
0
50
100
150
200
250
300
350
400
450
a
b
a
0
50
100
150
200
250
300
350
Mai Ju
ni
Juli
OktoberSeptemberJulyJuneMay
0
50
100
150
200
250
300
350
400
Juni
Juli
young trees
mature trees
September OktoberJulyJuneMay
Sun 1 x ozone Sun 2 x ozone Sha 1 x ozone Sha 2 x ozone
-ca
rote
ne [
µg
g-1 d
ry w
eig
ht]
-Carotene
Containers removed
-Carotene
0
50
100
150
200
250
300
350
400
450
AA
B
AB
b
a aab
July 2003
young treesmature trees
SHADESHADE SUNSUN
-carotene [µg g
-1 dry weight]
1 x ozone 2 x ozone
-ca
rote
ne [µ
g g-1
dry
wei
ght]
0
50
100
150
200
250
300
350
400
450
sig. diff. mature/youngSun 1 x ozoneSun 2 x ozoneShade 1 x ozoneShade 2 x ozone
0
50
100
150
200
250
300
350
400
450
b
aab
ab
ba abab
June 2003
young treesmature trees
SHADESHADE SUNSUN
-carotene [µg g
-1 dry weight]
1 x ozone 2 x ozone
-ca
rote
ne [µ
g g-1
dry
wei
ght]
0
50
100
150
200
250
300
350
400
450
no sig. diff. mature/young
050
100150200250300350400450
Mai
Juni
Juli
OktoberSeptemberJulyJuneMay
050
100150200250300350400450500
Juni
Juli
young trees
mature trees
September OktoberJulyJuneMay
Sun 1 x ozone Sun 2 x ozone Sha 1 x ozone Sha 2 x ozone
-ca
rote
ne [
µg
g-1 d
ry w
eig
ht]
-Carotene
Containers removed
0
100
200
300
400
500
Mai
Juni
Juli
OktoberSeptemberJulyJuneMay
0
100
200
300
400
500
600
Juni
Juli
young trees
mature trees
September OktoberJulyJuneMay
Sun 1 x ozone Sun 2 x ozone Sha 1 x ozone Sha 2 x ozone
neox
anth
in [
µg
g-1 d
ry w
eig
ht]
Neoxanthin
Containers removed
0100200300400500600700800900
1000
Mai
Juni
Juli
OktoberSeptemberJulyJuneMay
0100200300400500600700800900
10001100
Juni
Juli
young trees
mature trees
September OktoberJulyJuneMay
Sun 1 x ozone Sun 2 x ozone Sha 1 x ozone Sha 2 x ozone
lute
in [
µg
g-1 d
ry w
eig
ht]
Lutein
Containers removed
Xanthophyll cycle
Zeaxanthin
Antheraxanthin
Violaxanthin
Light-protection
Light-harvesting
De-epoxidised
Epoxidised
Protection from excessive light energy
Violaxanthin + Antheraxanthin + Zeaxanthin
0
50
100
150
200
250
300
350
400
450
500
550July 2003
young treesmature trees
SHADESHADE SUNSUN
V+
A+
Z [µ
g g-1 dry w
eight]
1 x ozone 2 x ozone
V+
A+
Z [µ
g g-1
dry
wei
ght]
0
50
100
150
200
250
300
350
400
450
500
550
AA
A
Aaa
aa
sig. diff. mature/young:Sun 1 x ozoneShade 1 x ozoneShade 2 x ozone
0
50
100
150
200
250
300
350
400
450
500
550June 2003
young treesmature trees
SHADESHADE SUNSUN
V+
A+
Z [µ
g g-1 dry w
eight]
1 x ozone 2 x ozone
V+
A+
Z [µ
g g-1
dry
wei
ght]
0
50
100
150
200
250
300
350
400
450
500
550
B
BB
Aaaaa
sig. diff. mature/young:Shade 1 x ozoneShade 2 x ozone
0
50100150200250300350400450
Mai
Juni
Juli
OktoberSeptemberJulyJuneMay
050
100150200250300350400450500
Juni
Juli
young trees
mature trees
September OktoberJulyJuneMay
Sun 1 x ozone Sun 2 x ozone Sha 1 x ozone Sha 2 x ozone
V+
A+
Z [
µg
g-1 d
ry w
eig
ht]
Violaxanthin + Antheraxanthin + Zeaxanthin
Containers removed
Deepoxidstate
0.0
0.2
0.4
0.6
0.8
1.0July 2003
young treesmature trees
SHADESHADE SUNSUN
Deepoxidstate
1 x ozone 2 x ozone
Dee
poxi
dsta
te
0.0
0.2
0.4
0.6
0.8
1.0
no sig. diff. mature/young:
BB
A A
bb
aba
0.0
0.2
0.4
0.6
0.8
1.0June 2003
young treesmature trees
SHADESHADE SUNSUN
Deepoxidstate
1 x ozone 2 x ozone
Dee
poxi
dsta
te
0.0
0.2
0.4
0.6
0.8
1.0
sig. diff. mature/young: Sun 2 x ozone
BB
AA
c
bab
a
0.0
0.2
0.4
0.6
0.8
Mai
Juni Ju
li
OktoberSeptemberJulyJuneMay
0.0
0.2
0.4
0.6
0.8
1.0
Juni
Juli
young trees
mature trees
September OktoberJulyJuneMay
Sun 1 x ozone Sun 2 x ozone Sha 1 x ozone Sha 2 x ozone
Dee
poxi
dsta
te
Deepoxidstate
Containers removed
0
100
200
300
400
500
600
700
800
900
1000
sig. diff. mature/young:Sun 2 x ozone
BB
AA
a
bb
a
June 2003
young treesmature trees
SHADESHADE SUNSUN
-tocopherol [µ
g g-1 dry w
eight]
1 x ozone 2 x ozone
-to
coph
erol
[µg
g-1 d
ry w
eigh
t]
0
100
200
300
400
500
600
700
800
900
1000
2222
-Tocopherol
First results on pigments and -tocopherol show differences between sun and shade crown.
Shade leaves exhibited higher pigment content and lower deepoxidate state of xanthophyll cyclus.
Only marginal differences were found between ozone treatments.
Summerised
Outlook
for the next months
Investigations of
- pigments and tocopherol (remaining sampling dates)
- Ascorbate and gluthatione (in young trees, branch cuvetts)
- Chromosomal abberations
statistical evaluation
comparison with the results of other participants
and interpretation