Participant 8 Assistant Contractor to Participant 7 Michael Tausz, Dieter Grill Karin Herbinger, Klaus Remele, Maria Müller Institute of Plantphysiology,

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


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


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


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


0

50

100

150

200

250

300

350

400

Juni

Juli

young trees

mature trees



-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


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


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


050

100150200250300350400450500

Juni

Juli

young trees

mature trees



-ca

rote

ne [

µg

g-1 d

ry w

eig

ht]

-Carotene

Containers removed

0

100

200

300

400

500

Mai

Juni

Juli


0

100

200

300

400

500

600

Juni

Juli

young trees

mature trees



neox

anth

in [

µg

g-1 d

ry w

eig

ht]

Neoxanthin

Containers removed

0100200300400500600700800900

1000

Mai

Juni

Juli


0100200300400500600700800900

10001100

Juni

Juli

young trees

mature trees



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


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


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


050

100150200250300350400450500

Juni

Juli

young trees

mature trees



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


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


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


0.0

0.2

0.4

0.6

0.8

1.0

Juni

Juli

young trees

mature trees



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


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

Documents

Participant 8 Assistant Contractor to Participant 7 Michael Tausz, Dieter Grill Karin Herbinger, Klaus Remele, Maria Müller Institute of Plantphysiology,