“Effetti dell’Ozono sulla Vegetacione”22 February, Torino

MONITORING AND RESEARCH ON OZONE EFFECTS ON VEGETATION IN SPAINWith emphasis on VegetPollOzone project

Vicent Calatayud

Ozone is the most relevant pollutant for forests

In general, there is a tendency towards an increase of background levels and climate change may produce a higher frequency of high ozone episodes

INTRODUCTION

O3

NO2

NH3

SO2

Advantages- Low cost- No power suply needed

Passive Sam plers /Spain

y = 0,9558x

R2 = 0,9095

0

10

20

30

40

50

60

70

0 10 20 30 40 50 60 70

Passive Sam pler (ppb)

Act

ive

Mon

itor

(ppb

)

Gandia

y = 0,9416xR2 = 0,8516

0

10

20

30

40

50

0 10 20 30 40 50

Pas s ive s am ple r (ppb)

Act

ive

Mo

nito

r (p

pb

)

Be n ifaió

y = 0,8969xR2 = 0,8743

0

10

20

30

40

50

0 10 20 30 40 50

Pas ive Sam pling (ppb )

Act

ive

Mon

ito

r (p

pb

)

V allibona

y = 0,9841xR2 = 0,7037

30

40

50

60

70

30 40 50 60 70

Pas s ive Sam ple r (ppb)

Act

ive

Mon

itor

(pp

b)

A

B

C D

Calibration against monitors

A

B C

PASSIVE SAMPLERS

<30 ppb

60-75 ppb75-90 ppb

30-45 ppb45-60 ppb

>=90 ppb

Year 2002April-SeptemberMean conc.

REDES

<30 ppb

60-75 ppb75-90 ppb

30-45 ppb45-60 ppb

>=90 ppb

Year 2003April-SeptemberMean conc.

REDES

<30 ppb

60-75 ppb75-90 ppb

30-45 ppb45-60 ppb

>=90 ppb

Year 2002April-SeptemberMax 4-weeks conc.

REDES

<30 ppb

60-75 ppb75-90 ppb

30-45 ppb45-60 ppb

>=90 ppb

Year 2003April-SeptemberMax 4-weeks conc.

REDES

y = 102.08e-2E-06x

R2 = 0.4039p<0.0001, N=91

0

10

20

30

40

50

60

70

80

350000 400000 450000 500000 550000 600000 650000

Latitude (ddmmss)

Mea

n oz

one

conc

entr

atio

n 20

00-2

004

(ppb

)

ICP-Forest NetworkPassive samplers

Spatial patterns, trends, risks....

PASSIVE SAMPLERS

OTHER NETWORKS

PASSIVE SAMPLERS

- 31 points with ozone measurements

-12 Non urban network of monitors

-22 points with passive samplers.

- Meteorological data available

- 3 of the points with passive + monitors

PASSIVE SAMPLERS

PassivePassive

MonitorPassive

MonitorPassive

0

10

20

30

40

50

60

S. J

ordi

S. J

ordi

Val

libon

a(T

.G.)

Val

libon

a(T

.G.)

Vill

ar A

rz.

Vill

ar A

rz.

Ozo

ne c

once

ntra

tions

(M

ean

Apr

il-Se

ptem

ber 2

007,

ppb

)

PASSIVE SAMPLERS

Type Plot MeanMonitor Borriana 20.93Passive Saler 25.33Passive Benifaió (Peira) 26.07Passive Guardamar 28.42Passive Benicarló 30.84Monitor T. Endomenech 32.80Passive Villar Arz. 33.57Passive S. Jordi 36.06Monitor Viver 36.20Monitor Cirat 36.48Monitor Onda 38.99Passive Muela Cortes 39.31Monitor Ontinyent 39.64Passive Vallivana 39.91Passive Relleu 40.13Passive Vallanca 40.52Monitor Penyeta 40.83Monitor Vilafranca 40.96Monitor Sorita 41.03Passive Agost 42.01Passive Ayora 42.12Passive Alborache 42.18Monitor Coratxar 44.60Passive Serra Bèrnia 44.86Passive Albatera 45.86Monitor C. Fuentes 46.16Passive Montdúver 47.15Passive Mariola 49.66Passive Vallibona (T.G.) 50.84Monitor Morella 53.05Passive Bartolo 56.88

Provisional data

PASSIVE SAMPLERS

y = 0.0137x + 32.407R2 = 0.5095

0

10

20

30

40

50

60

0 200 400 600 800 1000 1200 1400

Altitude (m)

Mea

n O

zone

Con

c.

(Apr

il-Se

ptem

ber 2

004,

ppb

)

y = -0.8172x + 39.46R2 = 0.0019

0

10

20

30

40

50

60

-2.0 -1.5 -1.0 -0.5 0.0 0.5 1.0

Longitude (Degrees)

Mea

n O

zone

Con

c.

(Apr

il-Se

ptem

ber 2

004,

ppb

)y = 0.3871x + 24.45

R2 = 0.0014

0

10

20

30

40

50

60

37 38 39 40 41 42

Latitude (Degrees)

Mea

n O

zone

Con

c.

(Apr

il-Se

ptem

ber 2

004,

ppb

)

Provisional data

INTRODUCCIÓN

PASSIVE SAMPLERS

0

10

20

30

40

50

60

701:

002:

003:

004:

005:

006:

007:

008:

009:

0010

:00

11:0

012

:00

13:0

014

:00

15:0

016

:00

17:0

018

:00

19:0

020

:00

21:0

022

:00

23:0

00:

00

UTC time (April-September 2007)

Ozo

ne c

once

ntra

tions

(ppb

)

Morella (1150 m)Sant Jordi (168 m)Borriana (53 m)

VISIBLE INJURY

Pinus halepensis

Sambucus ebulus

Fraxinus excelsior

Ulmus glabra

Vivurnum lantanaAcer campestre

Pinus halepensis

Sambucus ebulus

Fraxinus excelsior

Ulmus glabra

Vivurnum lantanaAcer campestre

Typically symptomatic species in Spain

VISIBLE INJURY

ICP-FOREST METHOD

-149 different species identified

1 Agost2 Albatera3 Ayora4 Bartolo5 Benicarló-Peníscola6 Borriana7 Caudete de las Fuentes8 Cirat9 Coratxar

10 Cortes de Pallás11 Guardamar12 Mariola13 Montduver14 Morella15 Onda16 Ontinient17 Relleu18 Sant Jordi19 Serra Bèrnia20 Sorita21 Torre en Domenech22 Tossal Gros23 Vallanca24 Vilafranca El Bovalar25 Villar del Arzobispo26 Viver27 Zarra

0

20

40

60

80

100

120

140

160

180

200

Ros

mar

inus

offic

inal

is

Thym

usvu

lgar

is

Pinu

sha

lepe

nsis

Ule

xpa

rviflo

rus

Que

rcus

cocc

ifera

Que

rcus

ilex

Bupl

euru

mfru

tices

cens

Gen

ista

scor

pius

Num

ber o

f Rec

tang

les

ICP-FOREST METHOD- Pinus halepensis present in 20 points, symptoms in 8 points

Number Plots Symptomatic PercentageCirat 14 4 29Cortes de Pallás 3 2 67Ontinient 12 4 33Sant Jordi 11 2 18Serra Bèrnia 6 2 33Torre en Domenech 13 2 15Vallanca 11 1 9Villar del Arzobispo 4 4 100

ICP-FOREST METHOD

- Crataegus monogyna, symptoms in 1 point, Coratxar

TRANSECTS

TRANSECTS

• Symptoms in Mediterranean sclerophylous vegetation are rare.

• This is due to limited ozone uptake of the plants under dry conditions (avoidance), but also to detoxification adapted to different types of stresses.

• In this context ozone fluxes provides a better approach to explain plant responses than ozone exposures, and detoxification capacity of the species becomes a key issue.

REDES

VEGETPOLLOZONE

Bean (Garrofó variety)

Font Roja Nursery

A- Field B- Field

C- Open Top Chambers D- Open Top Chambers

E- Microscopy F- Microscopy

Figure 5. Confirmation of ozone induced visible injury in Prunus spinosa A)Font Roja Nursery: ozone injury in August, note the age efefct. B) Font Roja Nursery: detail of ozone injury in August. C) Ozone injury induced in Open Top Chambers in Valencia. D) Ozone injury induced in Open Top Chambers in Valencia, note the age effect. E and F) Field samples collected in August 2006 in Font Roja Nursery, observed by microscopy; note that alterations do not occur in the epidermis, but are affecting the palisade parenchyma (collapsed cells), a main target of ozone.

VEGETPOLLOZONE

Bunyol Nursery

A- Field B- Field

C- Open Top Chambers

D- Open Top Chambers

E- Microscopy

F- Microscopy

Figure 3. Confirmation of ozone induced visible injury in Viburnum lantana A) Bunyol Nursery: ozone injury in August. B) Bunyol Nursery: ozone injury in August; shade effect. C) Ozone injury induced in Open Top Chambers in Valencia. D) Incipient ozone injury induced in Open Top Chambers in Valencia. E and F) Field samples collected in August 2006 in Bunyol Nursery, observed by microscopy; note that alterations do not occur in the epidermis, but are affecting the palisade parenchyma (collapsed cells), a main target of ozone.

TRANSECTS

Bean (Garrofó variety)

Watermelon (Benifaió)

Potato

TRANSECTS

Prunus spp.

PO3PLAR

Altitudinal and ozone gradient

0

10

20

30

40

50

60

70

Apr

il-1

Apr

il-2

May

-1

May

-2

May

-3

June

-1

June

-2

July

-1

July

-2

Aug

ust-1

Aug

ust-2

Sep

tem

ber-1

Sep

tem

ber-2

2-W

eek

Ozo

ne C

once

ntra

tions

(ppb

)

BenicarlóVallivanaMorella

Benicarló (30 m)

Vallivana (700 m)

Morella (1156 m)

PO3PLAR

- Clones: Lattecaldo (Switzerland) and Holland- 6 plants per clone and site

PO3PLAR

ITCs in Benifaió (Valencia)

OPEN TOP CHAMBERS

VISIBLE INJURY

3. Compare with field

MICROSCOPIA

TZ

EIEI

P P

P

Less chlorophyll content and callose accumulation in O3+Callose represents a non specific

defense mechanisms against fungi, wounding, insects... and ozone!

Lamotea dianae

MICROSCOPY

MICROSCOPY

AdE PP

C

TZ

IS

PP

IS

ACl

ACl

PWPW

PP

IS

ACl

ACl

PWPW

PP

IS

ACl

ACl

PWPW

PP

IS

ACl

ACl

PWPW

MICROSCOPY

SENESCENCE

Not visible effects:Difficult to assess in the field:Effects on growth and accelerates senescenceExample: Hibiscus syriacus

aa

b*

a

0

1020

30

40

5060

70

8090

100

Upper half of the plant Lower half of the plant

Num

ber o

f lea

ves

CF

NF+30

*(*)*

0123456789

1011121314

21 30 36 42 48 57 64 71 78 84

Days After Starting Fumigation

Num

ber o

f Flo

wer

s pe

r Pla

ntCFNF+30

EFFECTS REPRODUCTION

Lamotea dianaeLess flowers per plant

20µm

PG

ET

E

20µm

ET

E

PG

a

a

aa

b

b

0102030405060708090

100

Class 1 [<22 µm]

Class 1 [22-29 µm]

Class 1 [>=30 µm]

Per

cent

age

of c

lass

es (%

)

CFNF+30

Lamotea dianaeLess and more altered pollen

EFFECTS REPRODUCTION

MEDITERRANEAN SPECS.CORIACEOUS LEAVES

0

500

1000

1500

2000

2500

17/5

31/5

14/6

28/6

12/7

26/7 9/8

23/8 6/9

20/9

Fecha

Indi

ce D

año

A. opalusA. campestre A. pseudoplatanusA. monspessulanum

MEDITERRANEAN SPECS.EVERGREEN-DECIDUOUS

P. lentiscus

0102030405060708090

100

May June July August

Leaf

Inju

ry (%

sur

face

affe

cted

)

CFNF+30

P. terebinthus

0102030405060708090

100

May June July August

Leaf

Inju

ry (%

sur

face

affe

cted

)

CFNF+30

V. lantana

0102030405060708090

100

May June July August

Leaf

Inju

ry (%

sur

face

affe

cted

)

CFNF+30

V. tinus

0102030405060708090

100

May June July August

Leaf

Inju

ry (%

sur

face

affe

cted

)

CFNF+30

Evergreen

Deciduous

VISIBLE INJURY

0

5

10

15

20

25

0 20 40 60 80 100 120 140 160Days since staring of fumigation

year 2006

Inju

ry In

dex

Quercus fagineaQuercus ilexQuercus roburQuercus pyrenaica

0

5

10

15

20

25

0 20 40 60 80 100 120 140 160Days since starting of fumigation

year 2007

Inju

ri In

dex

Quercus faginea

Quercus ilex

Quercus robur

Quercus pyrenaica

INTERACTIONS

How ozone interacts with pathogens?Malva sylvestris and Puccinia malvacearum

*

*

***

0

2

4

6

8

10

12

14

16

18

0 7 14 21 28 35 42 48 55 63

Days after starting of fumigation

Rust

Infe

stat

ion

Inde

x (a

.u.)

CFNF+30

THANKS

We acknowledge people from Conselleria de Medi Ambient de la Comunitat Valenciana, especially Pilar Ara, Eduardo Perez Laorga, Hugo and the people from the Regional Forest Service that helped us with the passive samplers.

VegetPollOzone project

1. Passive sampler measurements- Gain knowledge on the ozone levels of the Valencian Community - Increase spatial cover of monitors

SCOPE

2. Assessment of visible injury: three approaches2.1. ICP-Forests methodology (random selection of plots)2.2. Transects, observation of visible injury both on wild plants and crops2.3. Bioindicators: poplar

3. Experimental approach: OTC experiments - 3.1. Gain knowledge on the ozone sensitivity of different types of plants (e.g. deciduous-evergreen)- 3.2. Validation of symptoms; including microscopy.

4. Diffusion of Results and environmental awareness

1. Passive samplers: measurements- We have gained knowledge on the ozone levels of the Valencian Community - Relevant for future extension of AQMN-CV- There is a good correlation with altitude, but not with latitude or longitude.- Future: Model the ozone levels measured using meteorological and physical data (e.g. altitude)

ACHIEVEMENTS

2. Assessment of visible injury: three approaches2.1. ICP-Forests methodology (random selection of plots)

- Visible injury using a random selection is restricted to a few plots, mainly of Pinus halepensis. 2.2. Transects, observation of visible injury both on wild plants and crops

- Visible injury is widespread in some crops such as potato and, occasionally, in bean and in fruit trees.

- In native vegetation, ozone produces symptoms in a series of sensitive broadleaf species, namely Sambucus nigra or Viburnum lantana.2.3. Bioindicators:

- Lattecaldo clon is sensitive to ozone, at least under irrigated conditions, at the current ozone levels in the Valencian Community. It has a great potential for environmental awareness and allows a good comparability among plots..