# Foliar surface cleaning techniques for analysis of particulate chemical composition in aire pollutants monitoring

Xavier Laffray, Matylda Domanski, Jean-François Castell

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Xavier Laffray, Matylda Domanski, Jean-François Castell. Foliar surface cleaning techniques for analysis of particulate chemical composition in aire pollutants monitoring. 27th Task Force Meeting of the ICP Vegetation, Jan 2014, Paris, France. hal-01622985

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27TH TASK FORCE MEETING OF THE ICP VEGETATION
IN FRANCE

Support for the International Cooperative Programme on Effects of Air Pollution on Natural Vegetation and Crops (UN-EEC/WGE/ICP-Veg), and participation in the international workshop on the estimation of the effects of the ozone on vegetation by relations dose-impact (Paris, 2014)

Juillet 2014

Étude réalisée pour le compte de l'ADEME par : INRA
N° de contrat : 1362c0039

Coordination technique ADEME : GALSOMIES Laurence - Direction\Service : VANVES DVTD SEQA

<figure><img src="image_2.png" /></figure>

RAPPORT D'ETUDE

En partenariat avec :
REMERCIEMENTS

Les membres du comité d'organisation sont remerciés :

Jean-François CASTELL (AgroParisTech - INRA)

Olivier BETHENOD (INRA)

Sebastien LEBLOND (MNHN)

Anne REPELLIN (UPEC)

Laurence GALSOMIÈS (ADEME)

Sophie FORMISANO (INRA)

CITATION DE CE RAPPORT

ADEME. Castell J-F. 2015. Rencontre 2014 du PIC-Végétation en France (27th Task Force Meeting of the ICP-Végétation). Rapport final, convention n°1362c0039, 77 pages.

Cet ouvrage est disponible en ligne [www.ademe.fr/en](http://www.ademe.fr/en), rubrique Médiathèque (URL)

Any representation or reproduction of the contents herein, in whole or in part, without the consent of the author(s) or their assignees or successors, is illicit under the French Intellectual Property Code (article L 122-4) and constitutes an infringement of copyright subject to penal sanctions. Authorised copying (article 122-5) is restricted to copies or reproductions for private use by the copier alone, excluding collective or group use, and to short citations and analyses integrated into works of a critical, pedagogical or informational nature, subject to compliance with the stipulations of articles L 122-10 – L 122-12 incl. of the Intellectual Property Code as regards reproduction by reprographic means.
TABLE DES MATIERES

<table><tr><td>1. Contexte</td><td>5</td></tr><tr><td>1.1. International</td><td>5</td></tr><tr><td>1.2. National</td><td>6</td></tr><tr><td>2. Bilan / Principaux résultats obtenus</td><td>7</td></tr><tr><td>2.1. Participants</td><td>7</td></tr><tr><td>2.2. Organisation</td><td>7</td></tr><tr><td>2.3. Principaux résultats</td><td>8</td></tr><tr><td>2.3.1. Session Ozone</td><td>8</td></tr><tr><td>2.3.2. Session Métaux-Azote-POPs</td><td>10</td></tr><tr><td>3. Conclusions / Perspectives</td><td>11</td></tr><tr><td>Références bibliographiques</td><td>13</td></tr><tr><td>Sigles et acronymes</td><td>14</td></tr><tr><td>ANNEXE 1 – Participants/27<sup>ème</sup> édition du PIC-Végétation</td><td>15</td></tr><tr><td>ANNEXE 2 – Comité d'organisation</td><td>17</td></tr><tr><td>ANNEXE 3 – Programmes (Séminaire ozone et 27<sup>ème</sup> édition du PIC-Végétation)</td><td>18</td></tr><tr><td>ANNEXE 4 – Poster de travaux français (Session poster du PIC-Végétation)</td><td>22</td></tr><tr><td>ANNEXE 5 – Résumés des travaux</td><td>23</td></tr></table>
## RESUME

Le projet a consisté à organiser en France la 27ᵉ réunion du Programme International Coopératif dédiés aux impacts de la pollution atmosphérique sur les cultures et la végétation semi-naturelle (PIC-Végétation) dans le cadre du Groupe de Travail sur les Effets issu de la commission économique pour l'Europe des Nations Unies (CEE-NU). Cet événement international s'est tenu à Paris du 29 au 30 janvier 2014. Il a été organisé de manière conjointe avec la journée internationale du 28 janvier 2014 à Paris sur de nouveaux développements de méthodes d'estimation des impacts de l'ozone sur les végétaux à partir de l'estimation des flux absorbés.

Ces deux journées ont rassemblé jusqu'à 84 participants provenant de 22 pays (dont 8 pays hors de l'union européenne). Plusieurs représentants de l'Afrique du nord (Algérie et Egypte) ont notamment participé ce qui est un point important à souligner. En effet, une des problématiques actuelles du PIC-Végétation est de mieux renseigner les effets de l'ozone sur la végétation méditerranéenne. Une autre activité du PIC-Végétation est l'estimation grâce à la biosurveillance des dépôts atmosphériques métalliques accumulés par les mousses.

Des résultats intéressants ont permis des avancées significatives pour améliorer la quantification des effets de l'ozone sur les végétaux. Ainsi, des fonctions doses-réponses ont pu être proposées pour de nouvelles espèces, forestières, prairiales ou de grandes cultures comme le maïs, pour lesquelles il n'existait pas encore de fonctions définies. Ces nouveaux résultats scientifiques seront utiles pour actualiser le chapitre 3 du manuel de cartographie et de modélisation des niveaux critiques pour la végétation. C'est un outil mis en commun pour les pays signataires de la convention de Genève sur la pollution atmosphérique transfrontière longue distance pour évaluer les impacts de l'ozone sur les végétaux en Europe.

En ce qui concerne la biosurveillance des dépôts atmosphériques par les mousses, le PIC-Végétation a validé le transfert de coordination, pour l'organisation de la prochaine campagne quinquennale in 2015, du Centre pour l'écologie et l'hydrologie (à Bangor, Royaume-Uni) vers l'institut de recherche nucléaire (à Doubna, Fédération de Russie). Il est ainsi souhaité une meilleure participation des pays d'Europe de l'est et de l'Asie, et la mise en place d'un groupe de discussion sur les techniques d'analyse des mousses. Des résultats intéressants présentés d'ores et déjà suggèrent que les polluants organiques persistants (POPS) seront à considérer lors des prochaines campagnes de biosurveillance par les mousses, dès 2015 notamment.

## ABSTRACT

The project was to organize in France the 27th meeting of the International Cooperative Programme on effects of air pollution on natural vegetation and crops (ICP Vegetation) on 29 and 30 January 2014. This meeting was coupled with an international workshop hold at Paris on the flux-based quantification of ozone impacts on vegetation.

The both meeting was attended by 84 experts from 22 countries, of which 8 are outside the European Union. The participation of representatives from North Africa (Algeria and Egypt) was an important point, because one of the issues facing the PIC vegetation is to improve the estimations of the impacts of ozone on Mediterranean vegetation.

Significant progress in impact functions of ozone on plant production was presented. Dose-impact functions are now available for new species of trees, grassland and crops. These advances will be used to update Chapter 3 of "modeling and mapping manual" which is the common tool of the signatories of the LRTAP Convention countries to assess the impacts of ozone on plants in Europe.

Regarding the monitoring of atmospheric deposition on mosses, the Task Force approved the transfer of the coordination of the program to the Nuclear Research Institute in Dubna (Russian Federation), with the aim to enhance the participation of Eastern Europe and Asian countries for the next survey 2015. The Task Force approved the establishment of a group discussion on moss analysis techniques. The results presented suggest that the inclusion of persistent organic pollutants (POPs) can now be considered for future monitoring campaigns.
# 1. Contexte

## 1.1. International

La Convention sur la pollution atmosphérique transfrontière à longue distance (dite convention de Genève ou CLRATP en anglais : *Convention on Long-range Transboundary Air Pollution*) est le premier instrument multilatéral conçu pour protéger l'environnement, à l'origine contre la menace des pluies acides et des brouillards photo-oxydants. La France fait partie des Etats membres qui ont adopté les objectifs de la CLRTAP pour lutter contre la pollution atmosphérique. Plusieurs protocoles ont été élaborés et ratifiés au fil des années suivant les polluants pour mieux cadrer les engagements pris par les Etats membres pour réduire leurs émissions de polluants.

La contribution d'origine transfrontière des polluants atmosphériques est aujourd'hui mieux caractérisée grâce aux observatoires de la pollution de fond et à longue distance. Cet apport transfrontalier des polluants est souvent non négligeable en milieu urbain, jusqu'à 40 à 70% des imports sont dus à des transferts de polluants en provenance de sources d'autres régions ou pays. C'est ce qui a pu être estimé récemment pour la région Île-de-France.

Pour une amélioration efficace de la qualité de l'air, il est nécessaire d'agir à différents niveaux, du local/régional à l'international, et pour ce faire de mieux connaître la pollution d'origine transfrontière.

Les directives et les protocoles européens fixant les plafonds nationaux d'émissions de polluants dans l'air à respecter s'appuient notamment sur des informations scientifiques fournies par la convention de Genève. La CLRTAP est de fait un élément important de la stratégie de réduction des émissions de polluants dans l'air.

Une cinquantaine de pays, dont la France, ont ratifié la convention de Genève mise en place sous l'égide de la Commission économique pour l'Europe des Nations Unies (CEE-ONU). La convention a élaboré une stratégie de surveillance et d'évaluation du transport longue distance de polluants atmosphériques (Programme coopératif EMEP, 1984) et des stratégies de réduction des émissions de polluants dans l'air (6 protocoles sur le soufre, l'azote, les COV, les métaux, les POPs : entre 1985 et 1998). Depuis le protocole de Göteborg multi-polluants et multi-effets (1999), les stratégies prennent aussi en compte les effets indésirables de la pollution sur la santé, la végétation et les matériaux, notamment par le concept des charges critiques qui permet de quantifier les dommages irréversibles dus aux effets des dépôts atmosphériques.

Les trois instances de la CLRTAP sont :

* WGE (GTE en français) – Groupe de Travails sur les Effets ;

* EMEP – Programme de Surveillance et d'Evaluation de polluants atmosphériques en Europe ;

* WGSR (GTSR en français) – Groupe de Travail sur les Stratégies et Révision.

Les parties à la Convention ont, dès le départ, souhaité fonder les décisions politiques pour lutter contre la pollution atmosphérique sur des bases scientifiques solides. Le Groupe de Travail sur les Effets de la convention (en anglais : *Working Group on Effects*, WGE), se décline en programmes internationaux coopératifs (PICs, en anglais : *ICPs International Cooperative Programmes*) et Tasks Force et concerne des activités scientifiques internationales qui produisent des données fiables (validées scientifiquement) et utiles dans les prises de décisions sur la réduction des émissions de polluants atmosphériques. Le PIC-Végétation créé en 1987 est l'un de ces programmes. Depuis plus de 30 ans, les experts de ce PIC collaborent pour établir des relations dose-effet en appuis aux réglementations de protection de l'environnement.

Les objectifs du PIC-Vegetation sur l'ozone sont :

* établir des relations dose-réponse réalistes pour les cultures et les végétations semi-naturelles

* définir les niveaux critiques d'ozone (tant en termes de concentrations que de flux) pour les cultures et les végétations semi-naturelles

* établir les cartes de dépassement de ces niveaux d'ozone pour l'Europe

* réaliser des synthèses thématiques explicitant les coûts et bénéfices à attendre du développement de stratégies de réduction de la pollution atmosphérique
A partir de 2001, les activités du PIC-Végétation ont aussi porté sur l'estimation des dépôts atmosphériques métalliques par les végétaux (notamment par les mousses). Le programme a ainsi pris la responsabilité de la coordination du réseau européen de biosurveillance des retombées en métaux par les mousses (en anglais *European Heavy Metals in Mosses Survey*), qui avait été initié et développé par le Nordic Council of Ministers à partir de 1985.

Dans ce cadre, des observations sont réalisées en Europe tous les cinq ans. Des cartes de concentrations en métaux accumulés dans les mousses sont régulièrement publiées à l'échelle européenne. Une trentaine de pays participent en moyenne à chaque campagne européenne du PIC-Végétation (2000/1, 2005/6, 2010/11). Les résultats cartographiés sont utilisés pour mieux connaître la variabilité spatiale et temporelle des dépôts estimés pour ces polluants (10 métaux). Ces données sont aussi utiles pour améliorer les dépôts calculées par le modèle EMEP à partir des concentrations d'émissions. Récemment, le PIC-Végétation a également intégré le suivi dans les mousses des concentrations accumulées en azote et des polluants organiques persistants (POPs) pour établir une estimation de ces dépôts atmosphériques.

Plus de 200 scientifiques en moyenne, provenant de 35 pays, participent au Programme du PIC-Végétation. Ils se réunissent, chaque année, au cours de journées d'échanges : séminaire scientifique (en anglais *Workshop*) et réunion stratégique d'orientation des activités (en anglais *Task Force Meeting*). Ils présentent et discutent des récents résultats acquis (volet ozone, volet métaux, azote) et des futurs développements du programme.

Ces journées annuelles scientifiques et stratégiques sont organisées, à tour de rôle, dans un pays. La France a l'occasion en 2014, pour la première fois, d'accueillir les journées du PIC-Végétation pour sa 27ème édition depuis 1987. La France avait en revanche déjà accueilli et organisé les journées d'échanges annuelles pour d'autres PICs du GTE : En 2005 à Paris le PIC-Matériau, et en 2009 à Paris également le PIC-Cartographie et Modélisation. Ces événements en France ont été possibles grâce au soutien de l'Etat aux travaux français du GTE, notamment le soutien de l'ADEME pour la coordination et le financement des points nationaux focaux (PFN français : mousse, charges critiques écosystèmes et matériaux) qui produisent les données françaises transmises à la convention de Genève au sein des programmes du GTE.

## 1.2. National

La France participe au PIC-Végétation depuis l'année 2000. La délégation française du PIC-Végétation était composée, au départ, de plusieurs personnes issues de l'ADEME et du MNHN de Paris. Grâce au travail des scientifiques français impliqués, la France a transmis principalement des données pour l'estimation au niveau national des dépôts métalliques atmosphériques (via le dispositif BRAMM). Par la suite, et surtout ces dernières années, la délégation française a pu s'agrandir avec des représentants de l'UMR AgroParisTech-INRA : sa première participation s'inscrit dans le cadre des programmes de recherche sur l'ozone soutenu notamment par le programme PRIMEQUAL (avec le projet BIOPOLLATM démarré au début des années 2000 et mené par l'unité EGC «Environnement et Grandes cultures »). Ensuite, des collaborations se sont développées grâce à l'unité EGC de l'UMR AgroParisTech-INRA avec l'UMR Bioemco (une unité mixte UPMC-UPEC-ENS-AgroParisTech-CNRS-INRA-IRD) notamment dans le cadre du projet IFLOZ puis du projet ANR VULNOZ, ce qui a conduit à renforcer la participation française sur le volet «ozone» des travaux du PIC-Végétation.

En revanche, pour le volet « Métaux » du PIC-Végétation, la participation française s'appuie sur l'expérience acquise en 1996 lors de la première contribution de la France aux activités de biosurveillance des dépôts métalliques dans le cadre de la coordination européenne assurée par le Conseil Nordique. Depuis 2000, l'UMR 7205 «OSEB» du MNHN, coordonne et met en œuvre dans le cadre du PIC-Végétation le dispositif français BRAMM (*Biosurveillance des Retombées Atmosphériques Métalliques par les Mousses*). Ce dispositif national de biosurveillance des dépôts atmosphériques cartographie en situation de pollution de fond les niveaux de concentration de polluants (métaux, azote et POP) accumulés dans les mousses. Quatre campagnes sont réalisées en France à ce jour (1996, 2000, 2006 et 2011). Pour chaque campagne, des espèces de mousses sont collectées sur 500 sites environ en moyenne. L'ADEME soutient le fonctionnement et le développement du dispositif depuis 1996.

L'implication de l'ADEME dans les travaux de la Convention consiste à :

* Développer les connaissances scientifiques nécessaires, être acteur du réseau de compétence sur le sujet, partager les expertises et les retours d'expérience, et ainsi disposer d'éléments de positionnement à la fois sur les polluants, les impacts environnementaux et les technologies de réduction des émissions ;
* Contribuer à fournir des données françaises pertinentes et de qualité (notamment à la CLRTAP/GTE) et disposer de projections de l'évolution des impacts environnementaux de la pollution atmosphérique longue distance (ces données alimentent les travaux de négociations et de suivi des protocoles de la Convention) ;

* Partager les connaissances scientifiques et technico-économiques produites par la Convention.

Il est apparu important de pouvoir aider la participation française aux travaux du PIC-Végétation et de renforcer l'implication des experts scientifiques, en proposant d'organiser en France pour la première fois les journées d'échanges annuelles du PIC-Végétation, et en les couplant aussi à une journée de séminaire international dédiée à l'amélioration des fonctions d'impact de l'ozone (relations dose-réponse) puisque la contribution sur ce thème de la part des experts français est importante, notamment avec la valorisation des résultats du programme «VULNOZ».

Enfin, soutenir les journées annuelles du PIC-Végétation est un moyen pour valoriser des travaux de la recherche française et pour contribuer à faire évoluer la réglementation. Ce soutien couplé à une manifestation scientifique internationale peut permettre de favoriser de nouveaux projets et collaborations.

## 2. Bilan / Principaux résultats obtenus

### 2.1. Participants

Les journées 2014 ont réuni 84 participants (88 inscriptions, Annexe 1), mais auxquels s'ajoutent une dizaine d'auditeurs supplémentaires (étudiants et doctorants français) qui ont pu assister à certaines sessions. Au total, la participation française d'une vingtaine de personnes est considérée importante.

Les journées annuelles de la 27ème édition du PIC-Végétation ont rassemblé 22 nationalités différentes, principalement des pays de l'Union Européenne ou d'autres pays signataires de la convention de Genève (17 pays représentés : Albanie, Allemagne, Belgique, Croatie, Espagne, Etats-Unis, Fédération de Russie, Finlande, France, Grande Bretagne, Grèce, Italie, Norvège, Pays-Bas, Pologne, Slovaquie, Slovénie, Suède, Suisse). Le Japon et l'Afrique du Nord (Algérie et Egypte) étaient également représentés.

### 2.2. Organisation

Deux événements étaient couplés. Le séminaire international sur l'ozone s'est déroulé à Paris le 28 janvier 2014 à l'AgroParisTech. La rencontre annuelle du PIC-Végétation a eu lieu les 29 et 30 janvier 2014 à l'auditorium de la grande galerie de l'Evolution du Museum National d'Histoire Naturelle de Paris.

Le comité local organisateur des journées, représenté par plusieurs organismes (dont l'ADEME), s'est réuni en 2013 pour mener à bien toute l'organisation sur les différents à Paris (Annexe 2).

En annexe 3, le détail des programmes des deux évènements (Séminaire ozone et rencontre annuelle 2014 du PIC-Végétation-2ème édition).

Le séminaire international sur l'ozone est organisé en deux sessions successives sur une journée :

* Quantification des impacts de l'ozone sur les arbres (8 présentations)

* Quantification des impacts de l'ozone sur les végétations semi-naturelles et les cultures (7 présentations, parmi lesquelles deux contributions françaises)

Le programme des deux journées d'échanges du PIC-Végétation a été organisé selon les recommandations du centre européen de coordination (CEH, Banghor). Le programme en annexe 4 est dédié aux connaissances sur les effets de la pollution atmosphérique sur les écosystèmes semi-naturels et les grandes cultures (PIC-Végétation) :
* Une session plénière d'introduction (discours d'accueil de l'ADEME par L. Galsomiès et du Muséum National d'Histoire Naturelle de Paris par J-P. Siblet, puis présentation des objectifs de la 27ème édition du PIC-Végétation)

* Une session plénière consacrée aux deux volets « ozone » et « Métaux-Azote-POPs » (2 contributions françaises)

* Quatre sessions organisées en parallèle pour présenter des communications orales sur le volet « Métaux-Azote-POPs » d'une part, et le volet s« Ozone » d'autre part.

* Une session finale pour la synthèse des résultats présentés et pour rendre compte des décisions et des recommandations adoptées pour le programme de travail à venir du PIC-Végétation.

Au total, 34 communications scientifiques sont présentées à l'oral et une session «poster» a aussi été organisée (avec 10 posters sur l'ozone, 13 posters sur les dépôts de métaux, 4 posters sur les dépôts azotés et un poster sur les dépôts particulaires présenté par une équipe française (annexe 4)).

## 2.3. Principaux résultats

Les résumés des présentations présentés au cours des 2 journées du PIC-Végétation sont rappelés plus loin. En revanche concernant le séminaire international dédié aux questions spécifiques d'impacts de l'ozone sur la végétation, les résumés des présentations ne sont pas présentés dans ce rapport car l'organisation conjointe avec cette journée n'a pas fait l'objet d'une aide de l'ADEME en particulier.

Le compte-rendu complet des journées 2014 du PIC-Végétation est disponible en anglais à l'adresse suivante :
http://icpvegetation.ceh.ac.uk/publications/documents/Minutes27thICPVegetationTFM.pdf

### 2.3.1. Session Ozone

L'étude présentée par Pleijel et al (M9) indique que les teneurs en ozone mesurées en Europe du nord restent élevées et susceptibles de provoquer des dégâts sur les végétaux. Elles diminuent néanmoins quand on remonte vers le nord. Les pics de concentration en ozone les plus élevés ont lieu en été dans la partie sud de la zone étudiée (45° de latitude) et sont de plus en plus précoces (printemps) quand on remonte vers le nord. Pour l'ensemble de la zone étudiée, les périodes pendant lesquels les niveaux d'ozone sont les plus élevées tendent à être de plus en plus précoces. Il semble bien que ce soit une conséquence du dérèglement climatique.

Une des activités « historiques » du PIC-Végétation est de produire chaque année des preuves des impacts visuels de l'ozone sur les végétaux (dégâts foliaires). Pour faciliter la participation des membres du PIC à cette partie du programme, Sharps (M8) a présenté une application pour smartphone en cours de développement. Celle-ci permet à un observateur qui aurait identifié des dégâts foliaires dus à l'ozone sur des végétations naturelles d'en prendre une photo avec un smartphone et de la transmettre directement avec les éléments de géolocalisation du lieu d'observation au centre de coordination du PIC.

Sur le plan de l'estimation des impacts, des travaux de plus en plus nombreux permettent maintenant de s'affranchir des limites des approches dites « de niveau I » (basées sur la notion d'AOT40).

La plupart des présentations du meeting, comme celles du workshop, dont les « minutes » sont en libre accès sur le site du PIC-Végétation [1], ont confirmé que les estimations basées sur les relations « flux-impact », réalisées à partir d'une estimation du flux d'ozone entrant dans les feuilles par voie stomatique permettent de meilleures estimations que les relations « dose-impact ». Cependant, l'inconvénient majeur de ces relations « flux-impact » réside dans la complexité des modèles qui permettent de calculer la conductance stomatique qui contrôle ce flux d'ozone pénétrant dans la feuille, puis de savoir estimer à partir de quelle valeur seuil de ce flux l'ozone devient toxique pour les plantes.

De nouvelles relations dose-impact ont été présentées pour plusieurs essences forestières : *Pinus cembra* et *Pinus halepensis* (de Marco et al, W1), *Quercus robur* et *Quercus petrea* (Braun W6). Ces relations s'avèrent assez variables d'une espèce à l'autre ce qui rend plutôt délicate l'estimation des impacts à l'échelle de la région ou d'une forêt composée d'un mélange hétérogène d'essences.

Une approche semi-empirique, dite « S-POD » proposée par une équipe suédoise (Karlsson et al, W5) peut permettre de résoudre cette question en paramétrant une fonction empirique de conductance stomatique qui ne dépendrait que
du climat et qui serait valable pour l'ensemble des espèces présentes. Les résultats présentés pour les forêts suédoises sont prometteurs, car ils montrent un assez bon compromis entre la précision requise pour estimer les impacts à l'échelle d'une essence et la précision requise pour estimer des impacts régionaux concernant des forêts plurispécifiques. Il sera intéressant de tester cette approche sur l'ensemble de l'Europe, mais pour l'adapter aux forêts méditerranéennes, il faudra la paramétrer pour les essences des forêts du sud et sans doute la compléter pour qu'elle prenne en compte le déficit hydrique du sol.

Une approche plus générale a été présentée par Büker *et al* (W7) Elle ne s'appuie plus sur des relations empiriques entre conductance et facteurs climatiques, mais sur le modèle DO3SE [2], qui est une modélisation plus mécaniste du fonctionnement biophysique de la forêt (couplage de modèles plus ou moins complexes de bilan énergétique, hydrique et photosynthétique des plantes). Pour l'année 2000 et à l'échelle européenne, ce modèle indique que l'ozone serait responsable d'une réduction de production de biomasse forestière d'environ 19% sur l'ensemble de l'Europe, l'impact étant plus fort sur les forêts bien alimentées en eau (France, Allemagne, Pologne, Finlande et Suède) que sur les forêts du sud (Büker *et al*, W4).

Cependant, ce type de modèle est beaucoup plus lourd à utiliser qu'une approche semi-empirique. Notamment pour la paramétrisation de la conductance stomatique, grandeur qui détermine directement la valeur du flux d'ozone qui entre dans la plante. Les versions disponibles de DO3SE dans le Mapping manual reposent sur une approche empirique de la conductance, basée sur un concept assez ancien, dit « approche de Jarvis » [3], et que l'on utilise aussi actuellement dans le « mapping manual » [4], outil commun d'estimation des impacts de l'ozone à l'échelle européenne. Dans cette approche, la conductance est considérée comme dépendant directement des facteurs environnementaux (température, rayonnement, vent, humidité de l'air et état hydrique de la plante). Depuis quelques années, une autre approche se développe, dite de « Ball-Berry » [5], dans laquelle c'est la photosynthèse qui peut être calculée assez directement en fonction des facteurs environnementaux et de lois biochimiques, la conductance étant ensuite déduite indirectement de cette valeur de photosynthèse. Les présentations de Fares *et al* (W2), Buker *et al* (W7) et les discussions qui ont suivi, ont montré que chaque approche présente autant d'avantages et d'inconvénients que l'autre. Il a donc été décidé de présenter les deux méthodes d'estimation de la conductance dans le mapping manual sans indiquer de préférence.

En ce qui concerne les plantes de grandes cultures et les prairies, des résultats analogues ont été présentés. Les contributions françaises ont permis de présenter un nouveau dispositif d'exposition des plantes à l'ozone en plein champ (Castell *et al*, M4 ; Bethenod *et al*, W9) et de mettre à la disposition de la communauté scientifique une paramétrisation simple de type Jarvis de la conductance stomatique pour le Maïs (Bethenod *et al*, W9).

Stella *et al* (W10) ont présenté un modèle français (SURFATM-O3) permettant de calculer le flux d'ozone absorbé par les feuilles d'une façon voisine du modèle DO3SE. Il semble donner d'une façon plus précise que DO3SE la partition entre le dépôt « non stomatique » (ozone qui se dépose ailleurs qu'à l'intérieur des feuilles, sur les épidermes et le sol principalement) et le flux stomatique (ozone qui entre directement dans les feuilles et que va avoir un effet toxique sur les plantes). Des comparaisons entre ces modèles (voire d'autres) sont envisagées dans l'avenir afin de vérifier la solidité et la validité du modèle DO3SE, voire pour le remplacer. Sur le plan expérimental, la présentation de Gerosa *et al* (M19) montre l'importance des dépôts non stomatiques à l'échelle d'une forêt.

Les travaux français ont également montré que les fonctions dose-réponses moyennes que l'on utilise actuellement pour le blé à l'échelle européenne (présentés par Mills *et al*, W13) peuvent être assez sensiblement différentes de celles que l'on peut calculer pour deux espèces de blé en Ile-de-France (Castell *et al*, M4). Des résultats analogues ont été obtenus sur des variétés de blé dur Italiennes et Espagnoles (Gerosa *et al*, W11).

En ce qui concerne les espèces prairiales, les travaux de F. Hayes *et al* (W12) ont permis d'établir des fonctions dose-réponse pour une quinzaine d'espèces. La difficulté principale de l'estimation des impacts de l'ozone sur les prairies vient du fait que celles-ci sont constituées de mélanges complexes d'espèces pour lesquels les modèles comme DO3SE ou SURFATM permettent difficilement d'estimer correctement des flux absorbés par chaque espèce présente. La solution pourrait être de ne pas considérer chaque espèce dans le peuplement, mais d'établir directement des fonctions dose-réponse par type de peuplement, défini en termes d'association d'espèces.

Plutôt que de multiplier les expérimentations et de calculer de façon empiriques des seuils de toxicité et des fonctions dose-réponse, il peut être intéressant de chercher les raisons physiologiques des différences observées. Pour cela, l'analyse des réponses en situation de stress multiples peut être utile.

Ainsi, Cotrozzi et al (M30) reviennent sur l'impact combiné de l'ozone et de la sécheresse sur les arbres (*Quercus Cerris*) en milieu méditerranéen.
Un aspect assez nouveau pour les membres du PIC-Végétation est l'analyse du lien entre l'ozone et l'état de nutrition minérale des plantes. L'étude présentée par Volk, Bassin et al (M10) indique ainsi qu'il existe des relations complexes entre les facteurs climatiques, l'ozone et le niveau de nutrition azotée sur la production des prairies alpines. De la même façon, la présentation de Hewitt et al (M11) montre des interactions entre l'ozone et la capacité des légumineuses à fixer l'azote. De même, Hayes et al (M18) ont montré que le niveau de nutrition azoté des arbres (Betula pendula) influe à la fois sur le flux stomatique et sur la fonction dose-réponse.

Ce lien entre azote et ozone est relativement nouveau, et doit être mis en lien avec l'accroissement des dépôts azotés sur les végétations. L'objectif conjoint des PICs « Modelling and Mapping » et « Végétation » est précisément de développer la prise en compte de ces deux aspects des dépôts atmosphériques (Hettelingh et al, M3). L'approche de type « Ball-Berry » est prometteuse sur ce point, car les modèles de photosynthèse auxquels elle fait appel peuvent être couplés avec la prise en compte de l'état azoté de la plante. La présentation de Verbeeke et al (M5) a montré qu'une première tentative de ce couplage a été faite avec la prise en compte de l'ozone dans le modèle français ORCHIDEE, qui simule la production primaire à l'échelle du globe.

Les approches nouvelles sur les relations dose-réponse sont donc moins empiriques que par le passé et s'appuient donc de plus en plus sur des considérations écophysiologiques. Pour être pertinentes et efficaces, elles doivent tenir compte des résultats obtenus au niveau cellulaire. Ainsi, la présentation de Havé et al (M23) donne des éléments pour mieux comprendre les différences entre les fonctions d'impact observées chez le blé (Castell et al, M4) en comparant l'expression des gènes et la mise en place des processus d'autophagie chez ces génotypes. La présentation de Madkour et al (M24) revient sur le rôle des régulateurs de croissance (Jasmonate et Acide Abscissique) dans l'expression de la réponse à l'ozone chez la tomate. Cependant, Maia et al (M26) ont montré chez Vigna unguiculata, que beaucoup reste à faire pour bien intégrer ces aspects biochimiques dans les modèles de végétation, car les mécanismes physiologiques de détoxication qui contrôlent la valeur seuil du flux d'ozone considérée comme toxique sont modifiés quand l'effet de l'ozone est couplé à celui de la sécheresse.

Sur le plan opérationnel, les informations apportées par les participants seront utilisées pour mettre à jour de manière consensuelle les fonctions d'impact de l'ozone du « Mapping manual » [4], d'une part en fournissant des fonctions dose-réponse pour de nouvelles espèces cultivées ou essences, d'autre part en améliorant les fonctions existantes. Un point important, longuement discuté est de parvenir à une meilleure prise en compte du couplage ozone-sécheresse dans les fonctions existantes, notamment dans le contexte méditerranéen.

### 2.3.2. Session Métaux-Azote-POPs

Cette session a rassemblé un peu moins de participants que la session «ozone». Les présentations ont été aussi moins nombreuses (une dizaine de présentations contre plus de trente pour la session «ozone», y compris à la journée du séminaire international).

La coordination du programme « mousses » sera désormais assurée par Marina Frontasyeva de la Fédération de Russie (au Joint Institute for Nuclear Research, Doubna). Dans la présentation qu'elle a donné (M7), elle explique que cette nouvelle coordination peut permettre à davantage de pays d'Europe de l'Est, du Caucase et de l'Asie Centrale (EECCA) de se joindre aux activités du PIC-Végétation et ainsi de mieux répondre à la volonté forte d'étendre la liste des parties signataires de la convention de Genève (CLRTAP). D

En ce qui concerne les résultats de dépôts azotés, la présentation de Harmens et al (M14) montre qu'il est possible d'établir une relation entre ces dépôts ou les concentrations en azote dans les précipitations et la concentration en azote des mousses. Cette relation est néanmoins complexe, et non- univoque à l'échelle européenne. Par ailleurs, les données présentées par Skudnick et al (M16) rendent compte d'une relation qui n'est valable qu'au pas de temps annuel, et non valide pour des valeurs moyennes calculées sur 3 ans. Néanmoins, le suivi des concentrations d'azote dans les mousses peut être utilisé pour identifier les zones à risque. Dans une analyse assez comparable, mais prenant en compte les dépôts et concentrations par unité paysagère, Schröder et al (M15) montrent que dans certaines unités paysagères, la corrélation est plutôt car le dépôt est statistiquement comme étant le principal facteur. Dans d'autres unités paysagères, la corrélation est moins bonne car d'autres facteurs comme le type de mousse récoltée est un facteur de variabilité important. Kluge et al (M16) ont présenté des résultats indiquant que ces différences peuvent en partie s'expliquer par des différences de couverture végétale au-dessus des mousses récoltées (collecte en sous-bois ou en zone ouverte). Cependant, Skudnick et al (M16) ne constatent pas de différence nette de teneur en azote entre mousses de sous-bois et mousses de zone ouverte en raison de la forte variabilité induite par l'hétérogénéité locale de la densité du feuillage et des arbres.
La présentation de M. Frontasyeva (M20) a montré que les concentrations de métaux et d'éléments radioactifs mesurées dans les mousses peuvent être utiles pour identifier l'origine des polluants dans les régions industrielles extrêmement polluées. Au niveau Européen, Ilyin (M27), montre qu'une bonne corrélation des concentrations dans les mousses peut être établie avec les dépôts estimés par des modèles (EMEP), mais ces relations varient selon les régions. Pour la première fois, une relation a été établie entre les dépôts et les concentrations de benzo[a]pyrène.

Dans le but d'identifier les écosystèmes à risques, Schaap et al (M28) ont présenté une méthodologie pour estimer les dépôts totaux de métaux sur les écosystèmes, basée à la fois sur l'analyse des données existantes et les sorties d'un modèle de transport (LOTO-EUROS).

Plusieurs études de cas ont également été présentées : Lazo et al (M29) ont présenté les résultats d'une étude de biosurveillance active (moss bags) menée autour de la baie de Vlora (Albanie) pour interpréter les fortes teneurs en métaux des eaux de mer, et Dunaev et al (M34) l'utilisation des techniques de biosurveillance par les mousses pour le suivi des politiques environnementales dans la région de Ivanov (Fédération de Russie). Enfin, l'analyse des données issues des campagnes 2004 et 2010 pour la Croatie montre que le niveau de pollution accumulé dans les mousses a peu varié en 5 ans, bien que les émissions soient en diminution (Spiric et al, M31).

Une étude originale (Esposito et al, M33) montre qu'il est possible de compléter les données classiques de bioaccumulation passive (accumulation de métaux dans les mousses) avec des informations relevant plutôt de la bioindication (mise en évidence d'effets biologiques des polluants) : en effet, la toxicité induite par l'accumulation de métaux se traduit par l'augmentation d'une protéine « heat shock » (Hsp70) dans les tissus des mousses collectées.

Suite aux échanges entre les participants, la décision a été prise de créer un groupe de discussion sur les techniques d'analyse des mousses. Les résultats présentés suggèrent que la prise en compte des polluants organiques persistants (POPS) peut être maintenant envisagée pour la prochaine campagne de surveillance qui sera lancée en 2015.

## 3. Conclusions / Perspectives

Les objectifs ont été atteints de façon spectaculaire pour organiser deux événements scientifiques en France sur les questions, d'une part, d'impacts de l'ozone sur la végétation, et d'autre part, de biosurveillance des dépôts atmosphériques. Le comité français d'organisation a réussi à accueillir à Paris un nombre important de participants, près de 84 personnes, pour suivre les journées annuelles du PIC-Végétation notamment et a permis d'inscrire au programme plusieurs présentations orales de résultats français : 6 présentations (contre zéro à deux habituellement), principalement dans la session «ozone». Un grand nombre de travaux réalisés dans le cadre du programme national « Vulnoz » financé par l'ANR ont été valorisés par des posters.

Les discussions menées au cours des journées ont donné lieu à des contacts intéressants entre des équipes françaises (notamment, INRA et UPEC) et des équipes étrangères, en vue d'éventuelles collaborations et pour mieux préparer les réponses communes aux appels d'offre européens, comme par exemple dans le cadre de la modélisation agrométéorologique des effets de l'ozone sur les cultures.

Malgré un attrait certain pour Paris de la part des chercheurs étrangers, l'objectif pour la 27ème édition du PIC-Végétation d'accroître fortement le nombre de participants par rapport aux années précédentes a été atteint partiellement (70-80 participants/an en moyenne). Ceci peut s'expliquer, d'une part par la date choisie (fin janvier) qui était probablement assez peu intéressante pour les obligations de service de certains enseignants-chercheurs, et d'autre part, la possible concurrence avec une conférence internationale en Chine sur les effets de l'ozone sur la végétation qui était prévue en mai 2014.

Les principales conclusions à retenir sur les activités 2014 du PIC-Végétation présentées à Paris lors de la 27ème édition concernent :

* La création d'un groupe de discussion entre experts dans le domaine des techniques d'analyse pour estimer les dépôts atmosphériques par les mousses

* La prise en compte dans l'agenda de travail 2015 d'un besoin de données sur les polluants organiques persistants (POPs)
* Un transfert de coordination à la Fédération de Russie pour l'organisation, dès 2015, des campagnes européennes de biosurveillance des dépôts atmosphériques par les mousses (métaux, azote et POPs)

* Une participation à l'actualisation du chapitre 3 du Manuel de cartographie des impacts de l'ozone (ou en anglais « *Mapping Manual* ») utilisé par le programme international concerté « modélisation et cartographie » pour compléter avec des fonctions dose-réponse établies pour des espèces qui n'étaient pas encore représentées, notamment le maïs et quelques essences forestières [4].

* Une proposition de fonctions dose-réponse spécifiques pour les cultures en climat méditerranéen, où la sécheresse est un paramètre indispensable à prendre en compte.
Références bibliographiques

[1] Workshop « ozone critical levels » - ICP vegetation Informal document. Paris, 28 January 2014.
http://icpvegetation.ceh.ac.uk/publications/documents/Minutesozoneworkshop.pdf

[2] ASHMORE, M.R. et al, 2007. Modelling stomatal ozone flux and deposition to grassland communities across Europe. Environmental Pollution 146 : 659-670.

[3] JARVIS, P.G. 1976. The interpretation of the variations in leaf water potential and stomatal conductance found in canopies in the field. Philosophical Transactions of the Royal Society of London (B), 273:593-610

[4] UNECE, 2010. Mapping Critical Levels for vegetation. Revised Chapter 3 of the UNECE Manual on methodologies and criteria for modelling and mapping Critical loads and Levels.
http://www.icpmapping.org/Mapping_Manual

[5] BALL, J.T. et al 1987. A model predicting stomatal conductance and its contribution to the control of photosynthesis under different environmental conditions. Progress in Photosynthesis research IV, Dordrecht : 221-225.
Sigles et acronymes

<table><tr><td>ADEME</td><td>Agence de l'environnement et de la maîtrise de l'énergie</td></tr><tr><td>AgroParisTech</td><td>Institut des sciences et industries du vivant et de l'environnement</td></tr><tr><td>ANR</td><td>Agence nationale de la recherche</td></tr><tr><td>BIOEMCO</td><td>Biogéochimie et écologie des milieux continentaux</td></tr><tr><td>BIOPOLLATM</td><td>Biosphère et pollution atmosphérique en zone rurale et périurbaine</td></tr><tr><td>BRAMM</td><td>Biosurveillance des retombées atmosphériques métalliques par les mousses</td></tr><tr><td>CEH</td><td>Centre for Ecology and Hydrology</td></tr><tr><td>CNRS</td><td>Centre national de la recherche scientifique</td></tr><tr><td>ENS</td><td>Ecole Normale Supérieure</td></tr><tr><td>IFLOZ</td><td>Impact des flux d'ozone sur les couverts végétaux</td></tr><tr><td>INRA</td><td>Institut national de la recherche agronomique</td></tr><tr><td>IRD</td><td>Institut de recherche pour le développement</td></tr><tr><td>MNHN</td><td>Museum national d'histoire naturelle</td></tr><tr><td>OSEB</td><td>Origine structure et évolution de la biodiversité</td></tr><tr><td>PRIMEQUAL</td><td>Programme de recherche inter organismes pour une meilleure qualité de l'air à l'échelle locale</td></tr><tr><td>UPEC</td><td>Université Paris-est Créteil</td></tr><tr><td>UPMC</td><td>Université Pierre et Marie Curie</td></tr><tr><td>VULNOZ</td><td>Vulnérabilité des agro-écosystèmes à l'ozone. Quels risques à l'horizon 2020-2030 ?</td></tr></table>
Annexe 1 – Participants/27ème édition du PIC-Végétation

(en grisé, les participants français)

<table><tr><td>Achermann</td><td>Beat</td><td>Federal Office for the Environment</td><td>Switzerland</td></tr><tr><td>Ballarin-Denti</td><td>Antonio</td><td>UNECE-CLRTAP-Working Group on Effects</td><td>Italy</td></tr><tr><td>Bancal</td><td>Marie-Odile</td><td>AgroParisTech - INRA</td><td>France</td></tr><tr><td>Bassin</td><td>Seraina</td><td>Agroscope Research Station ART</td><td>Switzerland</td></tr><tr><td>Bender</td><td>Jürgen</td><td>Thünen-Institute of Biodiversity</td><td>Germany</td></tr><tr><td>Bermejo</td><td>Victoria</td><td>CIEMAT</td><td>Spain</td></tr><tr><td>Bethenod</td><td>Olivier</td><td>INRA - EGC</td><td>France</td></tr><tr><td>Boltersdorf</td><td>Stefanie</td><td>University of Trier</td><td>Germany</td></tr><tr><td>Borowiak</td><td>Klaudia</td><td>Poznan University of Life Sciences</td><td>Poland</td></tr><tr><td>Bouchenafa</td><td>Nadia</td><td>Université Ibn Khaldoun Tiaret</td><td>Algeria</td></tr><tr><td>Braun</td><td>Sabine</td><td>Institute for Applied Plant Biology</td><td>Switzerland</td></tr><tr><td>Büker</td><td>Patrick</td><td>SEI/University of York</td><td>U.K.</td></tr><tr><td>Burkey</td><td>Kent</td><td>USDA-ARS</td><td>USA</td></tr><tr><td>Castell</td><td>Jean-Francois</td><td>AgroParisTech - INRA</td><td>France</td></tr><tr><td>Cieslik</td><td>Stanislaw</td><td>Joint Research Centre</td><td>Italy</td></tr><tr><td>Cotrozzi</td><td>Lorenzo</td><td>University of Pisa</td><td>Italy</td></tr><tr><td>Danielsson</td><td>Helena</td><td>Swedish Environmental Research Institute</td><td>Sweden</td></tr><tr><td>De Marco</td><td>Alessandra</td><td>ENEA</td><td>Italy</td></tr><tr><td>De Temmerman</td><td>Ludwig</td><td>CODA-CERVA</td><td>Belgium</td></tr><tr><td>Dellal</td><td>Abdelkader</td><td>University of Tiaret</td><td>Algeria</td></tr><tr><td>Dunaev</td><td>Anatoliy</td><td>Ivanovo State University of Chemistry and Technology</td><td>Russian Federation</td></tr><tr><td>Esposito</td><td>Sergio</td><td>Università di Napoli "Federico II"</td><td>Italy</td></tr><tr><td>Falk</td><td>Richard</td><td>University of Exeter/Centre for Ecology and Hydrology</td><td>U.K.</td></tr><tr><td>Fares</td><td>Silvano</td><td>Agricultural Research Council</td><td>Italy</td></tr><tr><td>Frontasyeva</td><td>Marina</td><td>Joint Institute for Nuclear Research</td><td>Russian Federation</td></tr><tr><td>Fumagalli</td><td>Ivan</td><td>Joint Research Centre</td><td>Italy</td></tr><tr><td>Fusaro</td><td>Lina</td><td>"Sapienza" University of Rome</td><td>Italy</td></tr><tr><td>Galsomiès</td><td>Laurence</td><td>ADEME</td><td>France</td></tr><tr><td>Gerosa</td><td>Giacomo</td><td>Universita' Cattolica del Sacro Cuore</td><td>Italy</td></tr><tr><td>Gorelova</td><td>Svetlana</td><td>L.N. Tolstoy Tula State Pedagogical University</td><td>Russian Federation</td></tr><tr><td>Grünhage</td><td>Ludger</td><td>Justus-Liebig University Giessen</td><td>Germany</td></tr><tr><td>Harmens</td><td>Harry</td><td>Centre for Ecology and Hydrology</td><td>U.K.</td></tr><tr><td>Havé</td><td>Marien</td><td>Université Paris-Est Créteil</td><td>France</td></tr><tr><td>Hayes</td><td>Felicity</td><td>Centre for Ecology and Hydrology</td><td>U.K.</td></tr><tr><td>Hettelingh</td><td>Jean-Paul</td><td>ICP M&M - Coordination Centre for Effects (CCE)</td><td>The Netherlands</td></tr><tr><td>Hewitt</td><td>Dan</td><td>Centre for Ecology and Hydrology</td><td>U.K.</td></tr><tr><td>Ilyin</td><td>Ilia</td><td>EMEP/MSC-E</td><td>Russian Federation</td></tr><tr><td>Karlsson</td><td>Per Erik</td><td>Swedish Environmental Research Institute</td><td>Sweden</td></tr><tr><td>Kluge</td><td>Michaela</td><td>Chair of Landscape Ecology, University of Vechta</td><td>Germany</td></tr><tr><td>Koroleva</td><td>Yuliya</td><td>Immanuel Kant Baltic Federal University, Kaliningrad</td><td>Russian Federation</td></tr></table>
<table><tr><td>Kubin</td><td>Eero</td><td>Finnish Forest Research Institute</td><td>Finland</td></tr><tr><td>Laffray</td><td>Daniel</td><td>Université Paris-Est Créteil</td><td>France</td></tr><tr><td>Laureau</td><td>Constance</td><td>Université Paris-Est Créteil</td><td>France</td></tr><tr><td>Lazo</td><td>Pranvera</td><td>University of Tirana</td><td>Albania</td></tr><tr><td>Le Gall</td><td>Anne Christine</td><td>ICP M&amp;M – INERIS</td><td>France</td></tr><tr><td>Leblond</td><td>Sébastien</td><td>Muséum National d'Histoire Naturelle</td><td>France</td></tr><tr><td>Leitao</td><td>Luis</td><td>Université Paris-Est Créteil</td><td>France</td></tr><tr><td>Madkour</td><td>Samia</td><td>Damanhour University</td><td>Egypt</td></tr><tr><td>Maia</td><td>Yuri</td><td>Université Paris-Est Créteil</td><td>France</td></tr><tr><td>Maňkovská</td><td>Blanka</td><td>ILE SAS Bratislava</td><td>Slovakia</td></tr><tr><td>Marzuoli</td><td>Riccardo</td><td>Universita' Cattolica del Sacro Cuore</td><td>Italy</td></tr><tr><td>Mills</td><td>Gina</td><td>Centre for Ecology and Hydrology</td><td>U.K.</td></tr><tr><td>Nali</td><td>Cristina</td><td>University of Pisa</td><td>Italy</td></tr><tr><td>Olendrzynski</td><td>Krzysztof</td><td>UNECE LRTAP Secretariat</td><td>Switzerland</td></tr><tr><td>Pellegrini</td><td>Elisa</td><td>University of Pisa</td><td>Italy</td></tr><tr><td>Pihl Karlsson</td><td>Gunilla</td><td>IVL Swedish Environmental Research Institute</td><td>Sweden</td></tr><tr><td>Pleijel</td><td>Håkan</td><td>Biological and Environmental Sciences, University of Gothenburg</td><td>Sweden</td></tr><tr><td>Poikolainen</td><td>Jarmo</td><td>Finnish Forest Research Institute (METLA)</td><td>Finland</td></tr><tr><td>Puga Freitas</td><td>Ruben</td><td>Université Paris-Est Créteil</td><td>France</td></tr><tr><td>Qarri</td><td>Flora</td><td>University of Vlora</td><td>Albania</td></tr><tr><td>Rakmani</td><td>Ahmed</td><td>Université Paris-Est Créteil</td><td>France</td></tr><tr><td>Rihm</td><td>Beat</td><td>Meteotest</td><td>Switzerland</td></tr><tr><td>Saitanis</td><td>Costas</td><td>Agricultural University of Athens</td><td>Greece</td></tr><tr><td>Schaap</td><td>Martijn</td><td>TNO</td><td>The Netherlands</td></tr><tr><td>Schröder</td><td>Matthias</td><td>Justus-Liebig University Gießen</td><td>Germany</td></tr><tr><td>Schröder</td><td>Winfried</td><td>Chair of Landscape Ecology, University of Vechta</td><td>Germany</td></tr><tr><td>Schuetze</td><td>Gudrun</td><td>Federal Environment Agency</td><td>Germany</td></tr><tr><td>Sharps</td><td>Katrina</td><td>Centre for Ecology and Hydrology</td><td>U.K.</td></tr><tr><td>Sicard</td><td>Pierre Louis</td><td>ACRI-ST</td><td>France</td></tr><tr><td>Sitch</td><td>Stephen</td><td>University of Exeter</td><td>U.K.</td></tr><tr><td>Skudnik</td><td>Mitja</td><td>Slovenian Forestry Institute</td><td>Slovenia</td></tr><tr><td>Spiric</td><td>Zdravko</td><td>OIKON Ltd. - Institute for Applied Ecology</td><td>Croatia</td></tr><tr><td>Stella</td><td>Patrick</td><td>UMR SAD-APT AgroParisTech/INRA</td><td>France</td></tr><tr><td>Thöni</td><td>Lotti</td><td>FUB - Research Group for Environmental Monitoring</td><td>Switzerland</td></tr><tr><td>Turk</td><td>Boris</td><td>University of Ljubljana, Biotechnical Faculty, Agronomy Dept.</td><td>Slovenia</td></tr><tr><td>Tuzet</td><td>Andrée</td><td>INRA – EGC</td><td>France</td></tr><tr><td>Uggerud</td><td>Hilde</td><td>NILU</td><td>Norway</td></tr><tr><td>Urbaniak</td><td>Justyna</td><td>Poznan University of Life Sciences</td><td>Poland</td></tr><tr><td>Velissariou</td><td>Dimitris</td><td>Technological Educational Institute of Peloponnese</td><td>Greece</td></tr><tr><td>Verbeke</td><td>Thomas</td><td>LSCE</td><td>France</td></tr><tr><td>Volk</td><td>Matthias</td><td>Agroscope, Zürich</td><td>Switzerland</td></tr><tr><td>Werner</td><td>Willy</td><td>University of Trier</td><td>Germany</td></tr><tr><td>Yamamoto</td><td>Hidemasa</td><td>Ministry of the Environment</td><td>Japan</td></tr></table>
Annexe 2 – Comité d'organisation

Les membres du comité d'organisation local sont :

<table><tr><td>Jean-François CASTELL (AgroParisTech - INRA)</td></tr><tr><td>Olivier BETHENOD (INRA)</td></tr><tr><td>Sebastien LEBLOND (MNHN)</td></tr><tr><td>Anne REPELLIN (UPEC)</td></tr><tr><td>Laurence GALSOMIÈS (ADEME)</td></tr></table>

Les réunions du comité d'organisation se sont tenues à l'AgroParisTech aux dates suivantes (1 demi-journée en moyenne) :

<table><tr><td>13 juin 2013</td></tr><tr><td>23 Juillet 2013</td></tr><tr><td>5 septembre 2013</td></tr><tr><td>30 septembre 2013</td></tr><tr><td>17 octobre 2013</td></tr><tr><td>19 novembre 2013</td></tr></table>
# Annexe 3 – Programmes (Séminaire ozone et 27ème édition du PIC-Végétation)

Version en anglais

Tuesday 28th January, 2014

WORKSHOP OZONE : Quantifying ozone impacts on vegetation

08:45 Harry Harmens – Introduction to the workshop

**Theme 1: Quantifying ozone impacts on trees. Chair: Harry Harmens, Rapporteur: Felicity Hayes**

W1 Alexandra De Marco et al. – Ozone stomatal flux-based critical levels translated into real-world forest impacts.

W2 Silvano Fares et al – Ball-Berry vs Jarvis approaches to model stomatal ozone deposition in Mediterranean forests: validation using the test sites of Castelporziano and San Rossore forests.

W3 Beat Rihm et al. – Mapping of ozone flux in Switzerland with high spatial resolution for epidemiological analysis.

W4 Patrick Büker et al – The effect of ozone on the C sequestration potentials of European forests.

General discussion.

10:30 – 11:00 Coffee/tea

W5 Per Erik Karlsson et al – Further developments of the concept of the Simplified Phytotoxic Ozone Dose, S-POD, to assess the risk for ozone impacts on trees on the regional scale.

W6 Sabine Braun – Validation of stomatal flux in Quercus species.

W7 Patrick Büker et al – DO3SE parameterisation for new forest tree species and revision of flux-response relationships for eight European trees species (note: 15 min presentation).

W8 Harry Harmens – Structured discussion on questions, conclusions & recommendations on trees.

12:30 – 14:00 Lunch

**Theme 2: Quantifying ozone impacts on crops and (semi-)natural vegetation. Chair: Håkan Pleijel, Rapporteur: Patrick Büker**

W9 Olivier Bethenod et al – Ozone impact on maize.

W10 Patrick Stella et al – Two-year total, stomatal, cuticular, and soil ozone budgets of an agricultural field with winter wheat and maize crops near Paris, France.

W11 Giacomo Gerosa et al – Yield response of some Italian and Spanish cultivars of durum wheat to elevated ozone: a varietal screening.

W12 Felicity Hayes et al – Dose-response relationships for grassland species.

General discussion.

15:30 – 16:00 Coffee/tea

W13 Gina Mills, Håkan Pleijel et al. – Application of a generic crop flux-effect relationship in integrated assessment modelling in Europe.

W14 Patrick Büker et al – Overview of the DO3SE parameterisation for new crop species.
W15 Häkan Pleijel – Structured discussion on questions, conclusions & recommendations on crop and grassland species; amendments to chapter 3 of the Modelling and Mapping Manual.

17:30 End of workshop

Tuesday 28th January: 18:30-20:30

(Amphithéâtre Tisserand, AgroParisTech)

**27th MEETING of the ICP VEGETATION**

Welcome buffet/reception and registration Task Force Meeting

Wednesday 29th January, 2014

(Auditorium de la Grande Galerie de l'Evolution, MNHN)

**27th MEETING of the ICP VEGETATION**

**Session 1: 09:15 – 10:30 Plenary session – Chair : Jean-François Castell**

**Welcome addresses:**

Laurence Galsomiès (ADEME)

Jean-Philippe Siblet (MNHN)

M1 Krzysztof Olendrzynski (Secretariat LRTAP Convention) – Update on the activities under the LRTAP Convention.

M2 Harry Harmens et al – Overview of the achievements of the ICP Vegetation in 2013 and future workplan (2014-2016).

M3 Jean-Paul Hettelingh, Anne Christine Le Gall et al. – Overview of the activities of ICP Modelling and Mapping and plans for the future.

10:30 – 11:00 Coffee/tea

**Session 2: 11:00 – 12:30 Plenary session Chair: Jürgen Bender**

M4 Jean-François Castell et al. – Validity of usual dose-response functions in the case of wheat exposed to free-air fumigation in the Paris region.

M5 Thomas Verbeke et al. – Incorporating a more mechanistic ozone impact parameterisation in the ORCHIDEE global vegetation model.

M6 Beat Achermann et al. – Effects of nitrogen deposition on species richness and diversity in Switzerland.

M7 Marina Frontasyeva – Transfer of the coordination of the European moss survey to the Russian Federation: plans for the future.

M8 Katrina Sharps – New ICP Vegetation smart-phone App for recording incidences of ozone injury on vegetation.

General discussion.

12:30 – 14:00 Lunch and putting up posters - Caves Esclangon - University of Paris VI

**Session 3: 14:00 – 15:30 Two parallel sessions: Ozone – Moss survey**

**Session 3a: Ozone - Chair: Alessandra De Marco**

M9 Häkan Pleijel et al. – Latitude dependence and temporal trend of the yearly peak in surface ozone in middle and northern Europe.

M10 Matthias Volk, Seraina Bassin et al. – Seven years of N- and O3 pollution × climate interaction in subalpine grassland: ecosystem carbon budget, nitrogen pools and O3 -flux modelling (POD).

M11 Dan Hewitt et al. – Is ozone reducing the N fixation capacity of current managed pasture?

M12 Demonstration and discussion on plans for use of ICP Vegetation smart-phone App for recording ozone damage to vegetation.

**Session 3b: Moss survey - Chair: Gunilla Pihl-Karlsson**
M13 Harry Harmens, Lotti Thöni et al. – Relationship between site-specific nitrogen concentrations in mosses and bulk atmospheric nitrogen deposition.

M14 Winfried Schröder et al – Correlations between nitrogen concentrations in atmospheric deposition and mosses mapped for natural landscapes in Europe.

M15 Michaela Kluge et al – Differences in concentration of nitrogen in mosses due to canopy drip effects - case study Germany.

M16 Mitja Skudnik et al – Indicative content of N, S and δ15N in the moss Hypnum cupressiforme in the southern part of Central Europe.

General discussion.

15:30 - 16:00 Refreshments

## Session 4: 16:00-17:15 Two parallel sessions Ozone and Moss survey

### Session 4a: Ozone - Chair: Silvano Fares

M17 Gina Mills et al – Overview of progress for the EU FP7 ECLAIRE project.

M18 Felicity Hayes et al – Impacts of ozone and nitrogen on silver birch.

M19 Giacomo Gerosa et al – Stomatal uptake and non-stomatal ozone removal by a mixed oak-hornbeam mature forest in the Po Valley: Results of the ECLAIRE long-term campaign.

General discussion.

### Session 4b: Moss survey - Chair: Harry Harmens

M20 Marina Frontasyeva – Moss biomonitoring of trace elements and radionuclides in rural and urban areas experiencing environmental stress.

M21 Harry Harmens and Marina Frontasyeva – Preparations for the European moss survey in 2015: monitoring manual, coordination, participation, stimulating participation in more EECCA countries and Asia.

18:00- 19:30 Poster session and buffet – Caves Esclangon – University of Paris VI

Thursday 30th January, 2014

27th MEETING of the ICP VEGETATION

### Session 5: 9:00 – 10:30 Two parallel sessions: Ozone and Moss survey

#### Session 5a: Ozone - Chair: Kent Burkey

M22 Muhammad Adrees et al. – Ambient air ozone impact on growth, yield and nutritional quality of wheat, maize and mung bean at two sites of Faisalabad, Pakistan.

M23 Marien Havé et al. – Vacuolar endoproteolytic activities and protein carbonylation levels in leaf as a model for depicting ozone tolerance and sensitivity in winter wheat (Triticum aestivum L.).

M24 Samia Madkour et al – The role of jasmonate and salicylic acid in conferring tolerance against ozone injury in tomato plants.

M25 Elisa Pellegrini et al. – Circadian profiles of photosynthetic parameters and primary metabolites in grapevine varieties exposed to ozone.

M26 Yuri Maia et al. – Involvement of plastidial terminal oxidase (PTOX) and mitochondrial proteins (AOX and pUCP) in the response of Vigna unguiculata (cowpea) to combined drought stress and ozone exposure.

General discussion

#### Session 5b: Moss survey - Chair: Marina Frontasyeva

M27 Ilia Ilyin et al – Evaluation of heavy metal and POP pollution levels in the EMEP region using atmospheric deposition modeling and measurements in mosses.
**M28** Martijn Schaap *et al* – Atmospheric deposition of heavy metals to terrestrial ecosystems in Germany.

**M29** Pranvera Lazo *et al* – The survey of atmospheric deposition of Al, Cr, Fe, Ni, V and Zn in Albania by using moss biomonitoring and ICP-AES.
General discussion, recommendations and decisions.

10:30 – 11:00 Refreshments

**Session 6: 11:00-12:30 Two parallel sessions: Ozone and Moss survey**

**Session 6 a: Ozone - Chair: Håkan Pleijel**

**M30** Lorenzo Cotrozzi *et al* – Response of Quercus cerris to combined ozone and drought stress.
General discussion, recommendations and decisions (including from the ozone workshop).

**Session 6b: Moss survey - Chair: Sébastien Leblond**

**M31** Zdravko Spiric *et al* – Croatia participated in the ICP Vegetation HM survey since 2005.

**M32** Flora Qarri *et al* – Assessment of trace elements pollution around Vlora area, Albania.

**M33** Sergio Esposito *et al* – A possible role of Heat Shock Proteins as sensor of heavy metal pollution in Bryophyta.

**M34** Anatoliy Dunaev *et al* – Biomonitoring of trace elements in Ivanovo region (Central Russia).
General discussion.

12:30 – 14:00 Lunch - Restaurant of the Great Mosque of Paris

**Session 7: 14:00 – 16:00 Plenary session - Chair: Harry Harmens**

- Reporting decisions and recommendations.

- ICP Vegetation work programme 2014 – 2016.

- Collaboration with other relevant bodies/organizations.

- Patrick Büker *et al*. – Outreach activities: Raising awareness of air pollution impacts on vegetation in South Asia.

- Conclusions and review of the 27th Task Force Meeting.

- Next Task Force Meeting.

- Any other business.

Excursion: 16:00 – 18:00 Grande Galerie de l'Evolution

20:00 Conference dinner: dinner-cruise on the Seine River
Annexe 4 – Poster de travaux français (Session poster du PIC-Végétation)

<figure><img src="image_3.png" /></figure>

Le résumé du poster est en annexe 5 (point 5.3) du rapport.
# Annexe 5 – Résumés des travaux

## 5.1 SÉMINAIRE INTERNATIONAL SUR l'OZONE

### W9 OZONE IMPACT ON MAIZE

<sup>a</sup>Bethenod O., <sup>c</sup>Leitao L., <sup>b</sup>Castell J.F., <sup>a</sup>Massad R., <sup>a</sup>Tuzet A., <sup>d</sup>Le Thiec D., <sup>c</sup>Repellin A., <sup>c</sup>Bagard M.

Ozone (O₃) is the air pollutant that currently represents one of the greatest environmental concerns in the troposphere, with particles and nitrogen oxides. Ground levels of O₃ have increased continuously over the three decades before 2000, at a rate of 0.5-2% per year¹. Although a stagnation or slight decrease over Europe and North America was observed since 2000, O₃ concentration is still increasing in Asia and others world parts². Based on dose-effect curves, Mills et al. (2007)³ identified three crops groups according to ozone sensibility: ozone sensitive crops (such as wheat), moderately sensitive crops (such as maize) and ozone resistant crops (such as barley)⁴.

For a given AOT40, the decrease in leaf photosynthesis (established using the A/Ci curves) is similar in maize and wheat whereas the decrease in grain dry matter is about doubled in wheat compared to maize. To understand this discrepancy, we investigated changes in maize (Zea mays L.) Leaf Area Index (LAI), leaf gas exchanges and harvest index, due to ozone using a new linear field fumigation device.

When fumigation occurred before flowering, leaf area decreased up to 15%⁴. If the LAI remained over 4, the intercepted radiation by the maize plants was maximal. Therefore, in this case, ozone impact on maize intercepted radiation was limited. When fumigation occurred after flowering, the decrease in leaf photosynthesis in the upper leaves of the plants negatively impacted their dry matter production. In response to an AOT40 of 22 mmol mol⁻¹h, leaf photosynthesis decreased by 30% whereas the grain dry matter was reduced by 5% only, as previously observed by Mills et al. (2007)³. In addition, the shoot dry matter production was reduced by more than 11% and the harvest indexes were 51.6 % in control plants and 59 % in fumigated plants. Moreover, ozone-induced moderation of stomatal conductance resulted in a limitation of gas fluxes (CO₂, O₃) entering in the leaves. This moderation of stomatal conductance was similar to that observed for leaf photosynthesis. Since the stomatal conductance of maize leaves was approximately half that of wheat leaves, the phytotoxic ozone dose (POD) ⁵ in maize was half that of wheat POD, for a given AOT40.

In conclusion, maize grain production was less impacted by ozone than wheat one due in part to lower maize gs and a grain sink priority.

### References

¹ Vingarzan, 2004, Atmospheric Environment, 38, 3431-442.

² Oltman et al., 2013, Atmospheric Environment, 67, 331-351.

³ Mills et al. 2007, Atmospheric Environment, 41, 2630-2643.

⁴ Leitao et al., 2007, Plant Biol. (Stuttg). 9, 478-88.

⁵ Grunhage et al., 2012, Environmental Pollution, 165, 147-157.

<footnote><sup>a</sup>INRA, UMR 1091 Environnement et Grandes Cultures, F-78850 Thiverval-Grignon, <sup>b</sup>AgroParisTech, UMR 1091 Environnement et Grandes Cultures, F-78850 Thiverval-Grignon</footnote>

<footnote><sup>c</sup>UMR 7618 IEES Paris, équipe Intéraction Plante-Environnement, Université Paris Est Créteil, 61 Avenue du Général De Gaulle F-94010 Créteil</footnote>

<footnote><sup>d</sup>UMR 1137, Ecologie et Ecophysiologie Forestières, Université de Lorraine, Vandœuvre-lès-Nancy F-54506, France</footnote>
## W6 VALIDATION OF STOMATAL FLUX IN QUERCUS SP.

Braun,S.

IAP, Sandgrubenstrasse 25, CH-4124 Schönenbuch, sabine.braun@iap.ch

Oak species (mainly *Quercus petraea* and *Q. robur*) are important in Switzerland, and it is expected that their significance in forestry will increase in future due to climate change. Ozone effects on *Quercus* are therefore an important issue, and their quantification needs estimates of ozone uptake. The recommendations for flux calculations of oaks in the Mapping Manual (UNECE, 2010) are based for several important parameters on the generic deciduous tree parameterisation. The aim of the present investigation was (1) to validate the proposed parameterisation using sap flow and (2) to test if *Q. petraea* and *Q. robur* have to be treated separately. Sap flow has been used successfully to validate stomatal uptake of ozone in *Fagus sylvatica* (Braun et al., 2010).

Three forest plots in Switzerland including *Fagus sylvatica*, *Quercus petraea* and/or *Q. robur* were equipped with sap flow sensors in the years 2012 and 2013. Analysis of the oak sap flow data was performed in a similar way as described earlier for beech. The multivariate nonlinear regression approach was applied successfully but some questions remained open. Some of the *Q. petraea* showed a strange sapflow pattern in the outer sapwood, and it seems that the optimum temperature may vary from year to year depending on the climatic conditions. The first results suggest that *Q. petraea* and *Q. robur* need different parameterisations for calculating ozone uptake.

### References

* Braun, S., Schindler, C., Leuzinger, S., 2010. Use of sap flow measurements to validate stomatal functions for mature beech (*Fagus sylvatica*) in view of ozone uptake calculations. Environmental Pollution 158, 2954-2963.

* UNECE, 2010. Mapping Critical Levels for Vegetation. Revised Chapter 3 of the UNECE Manual on methodologies and criteria for Modelling and Mapping Critical Loads & Levels and Air Pollution Effects, Risks and Trends. Prepared under the Convention on Long-range Transboundary Air Pollution (UNECE) by the International Cooperative Programme on Effects of Air Pollution on Natural Vegetation and Crops (ICP Vegetation, Centre for Ecology & Hydrology (CEH), Bangor, UK.

## W4 CURRENT DAY TROPOSPHERIC OZONE CONCENTRATIONS SUBSTANTIALLY REDUCE THE CARBON SEQUESTRATION POTENTIAL OF LIVING TREE BIOMASS ACROSS EUROPE

Patrick Büker<fnref n="1" />, Harry Harmens², Gina Mills², Lisa Emberson<fnref n="1" />

Tropospheric ('ground level') ozone (O3) is not only the third most important greenhouse gas, it is also an important air pollutant that reduces plant growth and crop yield. This damaging impact of O3 on vegetation reduces the amount of carbon (C) sequestered in living biomass and therefore may enhance climate change by increasing atmospheric CO2 concentrations. Trees have the greatest C sink capacity of any vegetation and represent the largest stock of stored biomass C.

In this study we estimate the impacts of O3 on sequestered C - expressed as net annual increment (NAI) - of European forest trees using re-analysed UNECE LRTAP (Convention of Long-range Transboundary Air Pollution) flux-based dose-response relationships. The methodology is applied for the year 2000 using EMEP (European

<footnote n="1">¹ Stockholm Environment Institute, Environment Department, University of York, York, UK.
patrick.bueker@york.ac.uk; l.emberson@york.ac.uk
² Centre for Ecology and Hydrology, Bangor, UK. hh@ceh.ac.uk; gmi@ceh.ac.uk</footnote>
Monitoring and Evaluation Programme) meteorology and O3 data and the effect of soil water conditions and species' sensitivity (distinguishing only conifer and deciduous vs. specific parameterisation for six tree species) on O3 flux and C sequestration response is investigated.

Ambient stomatal O3 fluxes in 2000 reduced C storage in the living biomass of trees on average by approximately 19% across Europe, with the highest and lowest relative reductions in Central/Western Europe and Southern Europe, respectively. The highest absolute reductions generally occurred in forest-rich countries such as France, Germany, Poland, Finland and Sweden. The inclusion of the potential for soil water to limit O3 uptake reduced the O3 fluxes and hence C sequestration losses especially in the Mediterranean countries. The species-specific parameterisation led to generally slightly higher fluxes, suggesting that a parameterisation only accounting for deciduous and coniferous trees might underestimate the risk O3 poses to C sequestration.

## Acknowledgement

We thank the UK Department for Environment, Food and Rural Affairs (Defra) for funding this work. Further financial support was provided to the ICP Vegetation Programme Coordination Centre by the UNECE and the UK Natural Environment Research Council (NERC).

# W1 OZONE STOMATAL FLUX-BASED CRITICAL LEVELS TRANSLATED INTO REAL-WORLD FOREST IMPACTS

De Marco A. <fnref n="1" />, Sicard P. <fnref n="2" />, Dalstein-Richier L. <fnref n="3" />, Paoletti E. <fnref n="4" />

alessandra.demarco@enea.it & pierre.sicard@acri-st.fr

Ground-level ozone (O3) is an important atmospheric pollutant, a pressing sanitary problem for human and ecosystem health, and a serious climate forcer. The European region at highest O3 risk is the Mediterranean area. Indeed, the climate change is expected to be more pronounced in the Mediterranean Basin than in most other regions of the world. In the Western European Mediterranean area, we demonstrated that the ozone pollution is a major air quality issue in both remote and urban atmospheres.

To date, most experiments to establish biologically relevant plant responses to ozone have been performed under controlled conditions, not representative of actual field conditions, and the results may not provide realistic results for developing standards. An assessment based on phytotoxic ozone dose (PODY) and on real plant damage is more appropriate. The LIFE+ FO3REST project allows testing the current exposure-based critical levels (AOT40) and suggesting new ozone flux-based critical levels for Mediterranean forest protection against ozone pollution.

A field survey of ozone-induced symptoms was carried out (ICP-Forests protocol) in 54 plots in South-eastern France and North-western Italy. Stomatal ozone fluxes were modelled and correlated to real-world forest impacts in terms of visible injury (stippling/mottling, crown discoloration and leaf loss) in 2012 and 2013. The indicators POD0 and POD1 were calculated thanks to the DO3SE model using the available parameterization for Mediterranean and continental deciduous broadleaf forests and conifers. Spearman test was carried out to understand the relative contribution of O3 (concentrations or stomatal uptake) to visible injury occurrence. From the flux-effect relationships we derived and proposed species-specific flux-based critical levels CLef for forest protection. In a climate change

<footnote n="1">¹ENEA - 76, Lungotevere Thaon de Revel, Roma, Italy.</footnote>

<footnote n="2">²ACRI-ST, 260 route du Pin Montard, 06904 Sophia Antipolis, France.</footnote>

<footnote n="3">³GIEFS - 60, Avenue des Hespérides, 06300 Nice, France.</footnote>

<footnote n="4">⁴ IPP-CNR - Via Madonna del Piano 10, 50019 Sesto Fiorentino, Italy.</footnote>
context, with information in hand, policymakers can make decisions about new legislation for air pollution control strategies in Europe.

## Acknowledgment

This work was made possible with the contribution of the LIFE financial instrument of the European Union (LIFE10 ENV/FR/208) for the FO3REST project.

# W2 BALL-BERRY VS JARVIS APPROACHES TO MODEL STOMATAL OZONE DEPOSITION IN MEDITERRANEAN FORESTS: VALIDATION USING THE TEST SITES OF CASTELPORZIANO AND SAN ROSSORE FORESTS

Fares S<fnref n="1" />, Savi F<fnref n="1" />,<fnref n="2" />, Hoshika<fnref n="3" />, Y., De Marco A<fnref n="4" />, Cieslik S<fnref n="5" />, Gruening C<fnref n="5" />, Matteucci M.<fnref n="6" />, Goded I.<fnref n="5" />, Sicard P. <fnref n="7" />, Paoletti E.<fnref n="3" />

Models to predict stomatal uptake require a proper parameterization. Field sites where ozone fluxes are measured continuously with Eddy Covariance are ideal to extract fundamental information on stomatal ozone fluxes to validate models. In this study, we collected ozone flux measurements from two representative Mediterranean forest ecosystems in Italy: A Holm Oak forest (Quercus ilex) located inside the Presidential Estate of Castelporziano, Rome, and a stone pine (Pinus pinea) forest in San Rossore, Pisa. Measurements were carried out during all seasons in order to capture ecophysiological responses to seasonal changes in meteorology and stomatal fluxes were calculated using the Monteith evaporative/resistive methods.

Two modelling approaches were used: the first is the empirical Jarvis approach based on multiplicative algorithm. We used the most recent literature to parameterize reducing functions in the algorithm reported in the UNECE Mapping Manual (release 2010). A second and process-based approach involved use of Ball-Berry equation to estimate stomatal conductance as a function of carbon assimilation, testing the equation with classical parameterization retrieved from literature and with data directly measured in the field using the eddy covariance technique. Results show that the Jarvis approach better predicts absolute magnitude of stomatal ozone fluxes, while the Ball-Berry approach better reproduced the dynamics of stomatal ozone fluxes during the day but it diverged from the measured fluxes. Current research is aimed at refining parameterization of the Ball-Berry approach in order to obtain a good agreement with measurements.

<footnote n="1">¹Consiglio per la ricerca e la sperimentazione in agricoltura - Research Center for the Soil- Plant System, Rome, Italy.
silvano.fares@entecra.it</footnote>

<footnote n="2">²Department for Innovation in Biological, Agro-Food and Forest Systems (DIBAF), University of Tuscia, via S. Camillo de Lellis, 01100 Viterbo, Italy.</footnote>

<footnote n="3">³National Research Council, Institute for Plant Protection, Firenze, Italy.</footnote>

<footnote n="4">⁴ENEA, Rome, Italy.</footnote>

<footnote n="5">⁵European Commission – Joint Research Centre, IES, Ispra, Italy.</footnote>

<footnote n="6">⁶Department of Agriculture, Food and Environment, The University of Pisa, Pisa, Italy</footnote>

<footnote n="7">⁷ACRI-ST, Sophia-Antipolis, France.</footnote>
W11 YIELD RESPONSE OF SOME ITALIAN AND SPANISH CULTIVARS OF DURUM WHEAT TO ELEVATED OZONE: A VARIETAL SCREENING

Gerosa G.<fnref n="1" />, Marzuoli R.<fnref n="1" />, Monga R.<fnref n="2" />, Faoro F.<fnref n="2" />, Gonzalez I.<fnref n="3" />, Alonso R.<fnref n="3" />, Bermejo V.<fnref n="3" />

Durum wheat is the main wheat species grown in Southern Europe countries. Hence, the evaluation of the response of the main cultivars of durum wheat to tropospheric ozone is a priority for Southern Europe. The present work investigated the O3 effects on five cultivars of durum wheat, three of them from Italy (Colombo, Sculptur, Faraon) and two from Spain (Gallareta and Vitron) in terms of yield and related physiological drivers.

The experiment was performed in the open-top chambers (OTC) facility of C.R.IN.E.S at Curno, Italy, from 09th March to 15th June 2013. Each cv was sown in 12 pots (three plants per pot) and pots were randomly assigned to 4 OTC (three pots per OTC for each cv), two of them supplied with charcoal filtered air (-50% of ambient air ozone = CF) and two with ozone enriched air (+50% of ambient air ozone = EN). During the experiment the ozonated plants were exposed to a daylight AOT40 value of 11.561 ppm•h, well above the UN/ECE critical level of 3 ppmh, while the control (CF) plants experienced an exposure of 0.432 ppm h.

Periodical measurements of stomatal conductance, chlorophyll-a fluorescence and growth were performed during the growing season. Furthermore, macroscopic (chlorotic/necrotic spots) and microscopic (cell death and H2O2 deposits in mesophyll tissue) symptoms development were followed on the flag leaves. At the end of the season plants were

harvested and the stems and the ears of each plant were separately dried and weighted. Then the grain yield was assessed per each OTC as total grain weight and hectoliter weight.

The Italian cv Sculptur, the most productive one without ozone, together with the cv Colombo resulted the most ozone sensitive cvs, with a grain yield decrease of 16% (p<0.05) and 10% (ns) respectively in the EN treatment. Nevertheless very small differences were observed in the hectoliter weight between the two ozone treatments. The remaining cvs did not show any negative effect of ozone but rather a slight yield stimulation (between 1 and 5%, ns). The Spanish Gallareta resulted the best performing cv when both CF and EN yield are considered.

Ozone affected the earing and the plants growth: the number of ears was quite uniformly reduced by ozone, with a remarkable low for Sculptur, and the losses of the aboveground biomass confirm the ozone sensitivity of Sculpur and Colombo (-20% and -25% respectively, both p<0.01).

The stomatal conductance was greatly decreased by ozone in all cvs (between 11% and 37%, p<0.01), but it is notable that the most sensitive cvs showed the highest gs values, even under the ozone treatment, which implies a highest ozone dose. This would confirm that the stomatal dose is an important driver of the plants' response to this pollutant.

On the contrary the overall performance of the photosynthesis (PI) - assessed by the OJIP test on the chlorophyll fluorescence measurements - was not affected by ozone. Thus, the observed reduction of productivity in the sensitive cvs is not to be ascribed to a damage of the photosystems but rather to a dissipation of the available energy for the metabolic defense against the oxidative stress.

Finally, the observation of the cv Sculpure behaviour suggests that the breeding strategy of selecting a greater gs to increase the yield may not be appropriate under increasing ozone conditions.

<footnote n="1">¹ Università Cattolica del S. C., via Musei 41, Brescia (I), giacomo.gerosa@unicatt.it</footnote>

<footnote n="2">² Università degli Studi, via Celoria 2, Milano (I), franco.faoro@unimi.it</footnote>

<footnote n="3">³ CIEMAT, Av. Complutense 40, Madrid (E), victoria.bermejo@ciemat.es</footnote>
W5 FURTHER DEVELOPMENTS OF THE CONCEPT OF THE SIMPLIFIED PHYTOTOXIC OZONE DOSE, S-POD, TO ASSESS THE RISK FOR OZONE IMPACTS ON TREES ON THE REGIONAL SCALE

Karlsson, P.E¹., Danielsson, H¹., Pleijel, H²., Simpson, D.³, Uddling, J²., Pihl Karlsson, G¹.

¹IVL Swedish Environmental Research Institute Inc., P.O. Box 53021, 40014 Göteborg, Sweden.
pererik.karlsson@ivl.se

²University of Gothenburg, Biological and Environmental Sciences, P.O. Box 461, 40530 Göteborg,
Sweden,

³Meteorologisk Institututt, Meteorologisk institutt, Postboks 43 Blindern, 0313 OSLO, Norway

A new concept was developed, based on the “Simplified Phytotoxic Ozone Dose”, S-POD. The exposure index in intended to be used to assess ozone impacts on vegetation on the regional scale in northern Europe and it should be possible to estimate the ozone exposure based on relatively few, low cost, in situ measurements.

The S-POD is estimated from measured hourly ozone concentrations according to:

$$S-POD_x = \sum \text{daylight} (([O_3] * f) >x); \quad f = \max[f_{\text{min}}, (f_{\text{phen}} * f_{\text{temp}} * f_{\text{VPD}} * f_{\text{gmax}})).$$

The functions used are as far as possible similar to what is stated in the LRTAP Mapping Manual. The fgmax is introduced as a possibility to normalize between different plant species based on the combination of gmax and gb. The S-POD concept was tested on experimental data from Sweden and Finland on young Norway spruce and Silver birch and compared with POD and AOT40. The strongest dose-response relationships (D-R) were still obtained with POD (with a threshold >2). However, the D-R derived from S-POD was almost as good as for POD and far better than for AOT40. It is concluded that the S-POD concept is well suited to be used e.g. as an indicator in the Swedish Environmental Quality Objectives.

<figure><img src="image_4.png" /><figcaption>Figure 1. Dose-responserelationships for ozone impacts on the biomass of young Norway spruce (Picea abies) and Silver birch (Betula pendula) in multi-year exposure experiments in Sweden (SE) and Finland (FI). In one dataset with P. abies ozone exposure was combined with a drought stress treatment. Exposure is expressed as yearly exposure and response as a yearly decrease (1.0 represents no effect) in total (SE) or above ground (FI) biomass.</figcaption></figure>

■ SE P. abies

▲ SE P. abies + drought

■ SE B. pendula

▲ FI B. pendula

<figure><img src="image_5.png" /></figure>

<figure><img src="image_6.png" /></figure>
# W13 APPLICATION OF A GENERIC CROP FLUX-EFFECT RELATIONSHIP IN INTEGRATED ASSESSMENT MODELLING IN EUROPE

Mills, G.¹, Pleijel, H.², Danielsson, H.³, Norris, D.¹, Harmens, H.¹, Simpson, D.⁴ et al.

¹ ICP Vegetation, Centre for Ecology and Hydrology, Bangor, UK. gmi@ceh.ac.uk,

² University of Gothenburg, P.O. Box 461, 40530 Göteborg, Sweden,

³ IVL Swedish Environmental Research Institute, P.O. Box 53021, 40014 Göteborg, Sweden.

⁴ Chalmers University of Technology, SE-412 96 Gothenburg, Sweden.

The full parameterisation of the DO₃SE flux model for ozone is data intensive, including a requirement for hourly resolution climate and soil moisture data on a 50 x 50 km (or smaller) grid. At the request of integrated assessment modellers within the LRTAP Convention, participants at the most recent critical levels workshop (Ispra, October, 2009) and follow-on ICP Vegetation Task Force Meeting in Tervuren (Belgium, January, 2010) discussed a simplified modelling approach for a generic crop and tree species that is suitable for application in scenario analysis within GAINS. Parameterisations were derived that excluded soil moisture, a particularly problematic variable for modelling, and were included in the LRTAP Convention's Modelling and Mapping Manual (LRTAP Convention, 2010). Guidance was provided that these models should be used to identify areas at risk of damage rather than to quantify the economic impacts of such damage.

More recently, the Task Force on Integrated Assessment Modelling have requested a mechanism for quantifying the extent of risk within GAINS using a dose-response relationship for the generic flux models. Hakan Pleijel and Helena Danielsson re-visited the wheat database used to derive the full flux model and have calculated a yield-effect relationship for the generic crop flux model which is based on wheat. This relationship (Figure 1a) can be applied to assess risk to crops on the assumption that soil moisture is not limiting. To meet the needs of the LRTAP Convention to quantify effects and benefits for the various scenarios discussed during the recent revision of the Gothenburg Protocol, the response function was applied spatially to the wheat-growing areas of EU27+CH+NO. The larger countries with the highest potential percentage effects were Hungary, Italy and Poland, with the mean percentage yield loss for EU27+CH+NO being 12.4% for GP2005, the baseline scenario for 2005 and 10.3% for GP CLE2020, the Gothenburg Protocol current legislation scenario for 2020.

## Acknowledgement

We thank the UK Department for Environment, Food and Rural Affairs (Defra) for funding the ICP Vegetation Programme Coordination Centre. Further financial support was provided by the UNECE and the UK Natural Environment Research Council (NERC).

<figure><img src="image_7.png" /><figcaption>Figure 1(a) New response function for integrated assessment modelling based on wheat and accumulated over 45 days; (b) Percentage yield loss for GP2005 for wheat-growing grid squares, calculated using the equation in (a).</figcaption></figure>
# W3 MAPPING OF OZONE FLUX IN SWITZERLAND WITH HIGH SPATIAL RESOLUTION FOR EPIDEMIOLOGICAL ANALYSIS

Rihm B.<fnref n="1" />, Braun S.<fnref n="2" />

This study aims for maps of ozone flux that (1) can be used for epidemiological analysis of ozone effects on tree growth, i.e. have a sufficient spatial resolution (250 m raster) to reflect local meteorological conditions and pollutant levels, (2) cover the whole country allowing for national assessments of ozone-effects and (3) show possibilities of simplifying the flux calculation in order to reduce data demand and to make it more manageable in the practice of air pollution control.

In a first step, the DO3SE model was run at 24 rural monitoring stations in the period 1991-2006 and at 37 stations in the period 2007-2011 to calculate ozone flux doses POD1 for beech and spruce. The required hourly input-data (ozone and climatic parameters) were measured on-site. Medium water storage capacity was used uniformly for all calculations as flux differences due to soil water variation were small.

For producing maps two approaches are pursued. In the first approach, the fluxes calculated at the monitoring stations were spatially interpolated: The average fluxes of each period were related to maps of predicting parameters (long-term ozone concentration, global radiation and wind speed) by regression analysis. The mean stomatal flux in the resulting map for deci-duous forest was 20.5, for the coniferous forest 27.8 mmol m⁻<fnref n="2" /> year⁻<fnref n="1" />. The area-weighted average growth reduction for all forests was 11% (Figure 1). In a further step, provisional yearly POD1 maps were produced by spatially interpolating POD ratios (POD1 of the specific year divided by the average POD1) of the monitoring stations using an inverse-distance weighting (IDW) method. The yearly fluxes were successfully used for provisional epidemiological analysis of stem increment data.

<figure><img src="image_8.png" /><figcaption>**Figure 1:** Growth reduction, average 1991-2011.</figcaption></figure>

A drawback of that approach is that the yearly flux maps attained by the IDW method do not include further local information. Therefore, existing monthly meteorological maps were included in the second mapping approach: The calculated hourly stomatal functions (for temperature, VPD and light) and fluxes at the monitoring stations as well as the hourly input data were aggregated to monthly values. The relation between monthly averages of the stomatal functions and the input parameters was analyzed using nonlinear regression. The analysis showed that the fluxes can be estimated based on monthly means of radiation, temperature, dew point and ozone concentrations. The monthly ozone maps are produced by geo-statistical interpolation of monitoring data.

**Acknowledgments:** This work was financed by the Federal Office for the Environment (FOEN), Air Pollution Control and Chemicals Division.

<footnote n="1">¹ Meteotest, Fabrikstrasse 14, CH-3012 Bern, [beat.rihm@meteotest.ch](mailto:beat.rihm@meteotest.ch)</footnote>

<footnote n="2">² IAP, Sandgrubenstrasse 25, CH-4124 Schönenbuch, [sabine.braun@iap.ch](mailto:sabine.braun@iap.ch)</footnote>
# W10 TWO-YEAR TOTAL, STOMATAL, CUTICULAR, AND SOIL OZONE BUDGETS OF AN AGRICULTURAL FIELD WITH WINTER-WHEAT AND MAIZE CROPS NEAR PARIS, FRANCE

Stella P., Personne E., Lamaud E., Loubet B., Cellier P.

AgroParisTech, UMR INRA/AgroParisTech SADAPT, Paris, France. [patrick.stella@agroparistech.fr](mailto:patrick.stella@agroparistech.fr)

In this study, we evaluated ozone ($O_3$) deposition to an agricultural field with winter-wheat and maize crops over a period of two years. The Surfatm-$O_3$ model (Personne et al., 2009, Stella et al., 2011), a two-layer soil-vegetation-atmosphere-transfer model, was used to partition the $O_3$ flux between the soil, the cuticular and the stomatal pathways.

The comparison between measured and modeled $O_3$ fluxes exhibited a good agreement, independently of the canopy structure and coverage and the climatic conditions. This statement implicitly validates the $O_3$ flux partitioning. Indeed, sensitivity analysis using other parameterization for soil and cuticular resistances induced underestimations of the total $O_3$ deposition ranging between 0% to around 20%. Therefore, the Surfatm-$O_3$ model is used to establish the total, soil, cuticular and stomatal $O_3$ budgets.

Over the two-year period, total ecosystem $O_3$ deposition was 87.5 kg ha$^{-1}$. Non-stomatal deposition, i.e., soil and cuticular pathways, dominated the deposition budget. In particular, soil deposition accounted for up to 50% of the total deposition. However, phenological and physiological differences between maize and winter-wheat induced large differences in the stomatal deposition budgets of these two crops: while it represented 5.4 kg ha$^{-1}$ (22% of the total deposition) for the maize crop, stomatal sink accounted for 10.5 kg ha$^{-1}$ (36% of the total deposition) for the winter-wheat from sowing to harvest.

Finally, crop yield losses were estimated for winter-wheat and maize crops using the AOT40 and POD6 approaches. According to these methods, crop yield loss was 1.5-4.2% for the winter-wheat whereas maize was not affected by $O_3$.

## References

Personne, E., Loubet, B., Herrmann, B., Mattsson, M., Schjoerring, J.K., Nemitz, E., Sutton, M.A., and Cellier, P. (2009), Surfatm-NH3: a model combining the surface energy balance and bi-directional exchanges of ammonia applied at the field scale, Biogeosci., 6, 1371-1388.

Stella, P., Personne, E., Loubet, B., Lamaud, E., Ceschia, E., Béziat, P., Bonnefond, J.M., Irvine, M., Keravec, P., Mascher, N., and Cellier, P. (2011a), Predicting and partitioning ozone fluxes to maize crops from sowing to harvest: the Surfatm-$O_3$ model, Biogeosci., 8, 2869-2886.
## 5.2 JOURNEES D'ECHANGES DU PIC VEGETATION

### M6 EFFECTS OF NITROGEN DEPOSITION ON SPECIES RICHNESS AND DIVERSITY IN SWITZERLAND

Achermann B.<sup>1</sup>, Roth T.<sup>2</sup>, Kohli L.<sup>3</sup>, Rihm B.<sup>4</sup>.

Relationships between atmospheric nitrogen deposition and biodiversity monitored at the habitat (plots of 10 m<fnref n="2" />) and landscape (plots of 1 km<fnref n="2" />) level in Switzerland were analysed on the basis of vascular plants and bryophytes data from the Swiss Biodiversity Monitoring (BDM, www.biodiversitymonitoring.ch) and nitrogen deposition data at a high spatial resolution. Higher mean nutrient values (according to Landolt et al. 2010) of recorded plant species at the habitat level are observed with increasing nitrogen deposition in alpine pastures, mountain habitats, forests, meadows and pastures. In mountain hay meadows (EUNIS code E2.3) the analysis showed a decrease of oligotrophic vascular plant and bryophyte species richness with increasing nitrogen deposition and a decrease of community uniqueness of vascular plant species (Roth et al. 2013). Increasing nitrogen deposition was also negatively related to species richness of target plant species for the agricultural sector. On the other hand an increase in species richness and a decrease in community uniqueness was detected for eutrophic bryophyte species with increasing nitrogen deposition. Overall the results indicate a trend towards floristic homogenization with increasing nitrogen deposition mainly due to the decrease and loss of oligotrophic plant species. On the basis of the results the Critical Load for Nitrogen for mountain hay meadows should not be set higher than 10-15 kg N ha⁻¹yr⁻<fnref n="1" />. First results from investigations at the landscape level indicate as well a decrease of the species richness of oligotrophic plant species with increasing nitrogen deposition.

### References

Landolt E. et al., 2010. Flora indicative, Ecological indicator values and biological attributes of the Flora of Switzerland and the Alps. Haupt Verlag Bern-Stuttgart-Wien. ISBN 978-3-258-07461-0.

Roth T., Kohli L., Rihm B., Achermann B., 2013. Nitrogen deposition is negatively related to species richness and species composition of vascular plants and bryophytes in Swiss mountain grassland. Agriculture, Ecosystems and Environment 178, 121-126.

### M22 AMBIENT AIR OZONE IMPACT ON GROWTH, YIELD AND NUTRITIONAL QUALITY OF WHEAT, MAIZE AND MUNG BEAN AT TWO SITES OF FAISALABAD-PAKISTAN

Adrees, M.*<fnref n="1" />, Khalid, S.<fnref n="2" />, Ibrahim, M.<fnref n="1" />, Jabeen, F.<fnref n="1" />, Saleem, F.<fnref n="1" /> & Tanvir, M.<fnref n="1" />

Ground level ozone (O₃) is the most important atmospheric pollutant that likely to become a threat to global food production by damaging crops because of its high level of toxicity. Ambient O₃ concentrations are reducing the quality and total production of crops in many countries worldwide including Pakistan, East Asia, India and Europe.

<footnote n="1">¹ Swiss Federal Office for the Environment, CH-3003 Bern, [beat.achermann@bafu.admin.ch](mailto:beat.achermann@bafu.admin.ch)</footnote>

<footnote n="2">² Hintermann & Weber AG, CH-4153 Reinach, [roth@hintermannweber.ch](mailto:roth@hintermannweber.ch)</footnote>

<footnote n="3">³ Hintermann & Weber AG, CH-3011 Bern, [kohli@hintermannweber.ch](mailto:kohli@hintermannweber.ch)</footnote>

<footnote n="4">⁴ Meteotest, CH-3012 Bern, [beat.rihm@meteotest.ch](mailto:beat.rihm@meteotest.ch)</footnote>

<footnote n="1">¹Department of Environmental Sciences, Government College University, Faisalabad, 38000, Pakistan.
[madrees@gcuf.edu.pk](mailto:madrees@gcuf.edu.pk)</footnote>

<footnote n="2">²Department of Environmental Sciences, Fatima Jinnah Women University, Rawalpindi, Pakistan</footnote>
Rolling industrialization and economic growth over much of Asia has resulted in increased level of O3 emission precursor pollutants and consequently elevated concentrations of O3. The concentration of ambient O3 is high enough in Faisalabad city to pose serious threat to the agricultural crop growth and yield.

A project was designed to study the impact of ambient air ozone on the yield and quality of wheat, maize and mung bean at two sites of Faisalabad-Pakistan. Suitable varieties were selected and sown in pots at research area of Ayub Agricultural Research Institute, Faisalabad. Half of the pots were exposed to elevated ambient ozone levels at GTS Chowk near Faisalabad Railway Station (referred to as high ozone treatment), while the other half were kept at Farm Area of Ayub Agricultural Research Institute, Faisalabad (referred as control). During the whole exposure period, ambient air O3 was recorded. Data regarding O3 visible injury, stomatal conductance, photosynthetic rate, and total chlorophyll contents were collected at different stages of crop growth. At the time of harvest, the data of plant yield and its attributes were recorded for further analyses.

Results revealed that exposure of plants of wheat, maize and mung bean plants at elevated pollution site caused significant decreases in stomatal conductance, photosynthetic rate, total chlorophyll content and yield of these crops. The grain protein content of these crops showed significant reduction in response to elevated O3 exposure. These results suggest that O3 could be a hidden threat to global food security.

**Keywords:** ozone pollution, food security, mung bean, wheat

## M4 VALIDITY OF USUAL DOSE-RESPONSE FUNCTIONS IN THE CASE OF WHEAT EXPOSED TO FREE-AIR FUMIGATON IN THE PARIS REGION

Castell, J-F.<sup>a</sup>, Bethenod, O.<sup>b</sup> and Laffray, D.<sup>c</sup>

Two wheat varieties (Premio and Soissons) were grown in the field in the Paris region and exposed to moderate levels of ozone through a 50m long linear free-air fumigation device. Plants were ozone-fumigated daily for 56 days (from 12 May to 6 July 2009). O3 exposures at 3 different distances from the fumigation source: 2 m, 3.5 m and 5 m downwind of the source, were compared to control located 15m upwind of the linear source. Hourly average concentrations and cumulative exposure index (AOT40) were calculated. Stomatal conductance measurements and modelling allowed the calculation of the Phytotoxic Ozone Dose (POD06) absorbed by the upper leaves of the canopy.

The results show strong correlations between grain yield and both ozone exposure (AOT40 or POD06) indicators. However, there are differences in the dose-response relationship obtained for each variety. Moreover, these relationships are also quite different from the dose-response relationships found in the literature.

This can be partly explained by the fact that the use of free-air fumigation systems does not allow to achieve a true control treatment (plants exposed to ozone-free air). The highest production values that could be achieved in an atmosphere without ozone must therefore be extrapolated from experimental data, and this may increase the inaccuracy of calculations of relative yield losses.

<footnote><sup>a</sup> AgroParisTech, UMR EGC, 78850 Thiverval-Grignon, France - [castell@grignon.inra.fr](mailto:castell@grignon.inra.fr)</footnote>

<footnote><sup>b</sup> INRA, UMR 1091 EGC, 78850 Thiverval-Grignon, France - [bethenod@grignon.inra.fr](mailto:bethenod@grignon.inra.fr)</footnote>

<footnote><sup>c</sup> UPEC, UMR Bioemco, 61 Avenue du Général de Gaulle, 94010 Créteil, France - [laffray@u-pec.fr](mailto:laffray@u-pec.fr)</footnote>
M30 RESPONSE OF QUERCUS CERRIS TO COMBINED OZONE AND DROUGHT STRESS

Cotrozzi L., Remorini D., Pellegrini E., Lorenzini G., Massai R., Nali C.

Department of Agriculture, Food and Environment, University of Pisa - Via del Borghetto 80
56124 Pisa Italy - lorenzo.cotrozzi@for.unipi.it

Urban plants can help to improve air quality, and consequently human health. Due to the negative future prospects in 2050 for the urban environment caused by global climatic change [such as elevated ozone (O3) concentration and drought]. There is a need to monitor and proactively manage urban forests, adjusting them to change and using them to help cities to adapt to change. Special attention must be paid to the species that populate environments characterized by above-optimal temperatures in summer, such as the Mediterranean ones. Under field conditions, the study of the response to environmental stresses is difficult because environmental factors can be synergistically or antagonistically modified. For these reasons, the aim of this work is the analysis of the impact of a combined stress (O3 and drought) to simulate under controlled conditions the impact of a 2050 environmental scenario on the physiological performance of the Mediterranean *Quercus cerris*. Profiles related to leaf gas exchange, chlorophyll a fluorescence and leaf water potential were analyzed in 2-years old *Q.cerris* saplings exposed to O3 (80-100 ppb, 4 h day⁻¹ for 12 consecutive weeks), to drought (daily irrigated with 30% of effective evapotranspiration) and to O3 combined with drought in order to improve the response to oxidative stress. At the end of the exposure, plants do not exhibit any foliar symptoms. Photosynthetic processes, however, were significantly changed. Since the first week of treatment, oxidative stress induced decrease in net photosynthesis (A), above all in drought-stressed plants (about 2-fold), in comparison to controls. This trend unvaried until the end of the exposure due to a reduction of the stomatal conductance (gs). gs was lower in drought and combined-stressed plants during the whole exposure period (until a minimum about 5-fold after 5 weeks when compared to controls). In O3-treated plants, intermediate A [8 vs 3-11 (other three thesis' range) µmol CO₂ m⁻² s⁻¹] and gs [0.10 vs 0.07-0.22 (other three thesis' range) mol H₂O m⁻² s⁻¹, after 4 weeks] values were observed in comparison to the other three thesis explained by a gas avoidance mechanism activated by a stomata regulation causing a medium decrease of the net photosynthesis. Combined stress-treated plants did not show significant changes in comparison to drought stressed individuals in terms of photosynthetic status. Although intercellular CO₂ concentration (Ci) significantly changed after every treatment. Although the variable and maximal fluorescence ratio (Fv/Fm) did not show significant change during the exposure, all the plants showed values inside the optimal range. A regulatory adjustment of photosynthetic processes was highlighted during the exposure by the higher values of no-photochemical quenching (qNP) of all plants in comparison to controls and therefore suggests a tendency to increase the efficiency of thermal energy dissipation within PSII. Measurements of predawn leaf water potential revealed a leaf drought stress (around -2.0 vs -0.5 MPa in controls) in plants grown under water deficit conditions especially in combined stress plants suggesting a synergistic effect on the hydric status. On this basis drought stress (single and combined) should be considered more harmful to *Q. cerris* than O₃.

Acknowledgement

This research was supported by a grant from MIUR (PRIN 2010-2011).
## M34 BIOMONITORING OF TRACE ELEMENTS IN IVANOVO REGION, CENTRAL RUSSIA

Dunaev A.M.<fnref n="1" />, Frontasyeva M.V.<fnref n="2" />, Grinevich V.I.<fnref n="1" />

The results on biomonitoring of atmospheric deposition of trace elements in Ivanovo Region, Central Russia, in moss survey 2010/2011 are reported. An attempt to assess air quality in the large cities of the Ivanovo Region and wildlife preserve "Klyazminskiy" was made. A total of 47 elements were determined in 96 moss samples, collected evenly over the study area, by two complementary analytical techniques, neutron activation analysis performed at JINR, Dubna, and atomic absorption spectrometry in ISUCT, Ivanovo. GIS distribution maps of element-pollutants and some statistical data treatment are presented.

In autumn 2013 the Ivanovo State University of Chemistry and Technology initiated the development of the environmental protection strategy for the Ivanovo Region till 2020. It includes creation of an open-access database comprising data both from the state monitoring system and scientific research based on biomonitoring. Such policy will result in reliable assessment of air quality and will allow conversion of trace element concentrations in moss to absolute deposition values for trace elements from precipitation collected near the same moss sampling sites. Thus, interpreting data for use in baseline human health *risk assessments* will be possible.

## M33 A POSSIBLE ROLE OF HEAT SHOCK PROTEINS AS SENSORS OF HEAVY METAL POLLUTION IN BRYOPHYTA

S. Esposito<fnref n="1" />*, M. Cardi<fnref n="1" />, M. Lentini<fnref n="1" />, A. De Lillo<fnref n="1" />, B. Conte<fnref n="1" />, S. Sorbo<fnref n="2" />, A. Basile<fnref n="1" />

Bio-monitoring represents a highly precise and low-cost strategy method(s) to achieve rapid and affordable results in the examining of the enrichment of pollutants in the environment; in this aim, Bryophyta show a high capability to accumulate metals because of the high surface/volume ratio and the presence of a thin cuticle. Bryophytes do not have roots, and their rhizoids do not primarily contribute to the uptake of substances; therefore, most of the elements measured in mosses are due to atmospheric depositions: thus, elements concentrations in epiphytic bryophytes reflect the amount of atmospheric depositions, and mosses are used as air pollution indicator. Therefore, different Bryophyta are currently studied in relation to uptake, and subcellular localisation, of heavy metals.

Heat Shock Proteins 70 (Hsp70) represent a group of chaperones involved in protein folding, but it has been assessed that they play a central role in counteracting the toxic effects of heavy metals on proteins and enzymes, protecting them from misfolding and proteolysis. Even if the understanding of roles and functions of Hsp70 are yet to be completely described, recent studies evidenced their role(s) in the response to abiotic stress.

The aim of this work is to indicate Hsp70 as simple and powerful indicators of heavy metal pollution in Briophyta and other aquatic plants. The subcellular localisation of pollutants can be investigated using X-ray microanalysis, associate to transmission electron microscopy (TEM). The damages caused by pollution were observed using TEM and SEM

<footnote n="1">¹Ivanovo State University of Chemistry and Technology (ISUCT), F.Engels pr. 7, Ivanovo, Russian Federation, amdunaev@ro.ru</footnote>

<footnote n="2">²Joint Institute of Nuclear Research (JINR), Joliot-Curie 6, Dubna, Russian Federation</footnote>

<footnote n="1">¹Dipartimento di Biologia</footnote>

<footnote n="2">²Centro Interdipartimentale per la Microscopia Elettronica (C.I.S.M.E.)</footnote>

<footnote>Università di Napoli "Federico II" - Naples, Italy. sergio.esposito@unina.it</footnote>
microscopy, in specimens collected in undisturbed and polluted areas, and compared to changes observed in HSP70 levels and occurrence measured using western blotting. The results were compared and related with the effects observed after *in vitro* exposition of specimens to different heavy metals (Cd, Pb, Cu, Zn).

The results will be discussed in the aim to indicate Hsp70 as a possible bioindicator able to measure the biological effects of atmospheric pollution in plants.

## M20 MOSS BIOMONITORING OF TRACE ELEMENTS AND RADIONUCLIDES IN RURAL AND URBAN AREAS EXPERIENCING ENVIRONMENTAL STRESS

Frontasyeva, M.

Joint Institute for Nuclear Research, 14180 Dubna, Moscow Region, Russian Federation. marina@nf.jinr.ru

The application of mosses as biomonitors of trace elements and radionuclides in selected rural and urban areas affected by intense anthropogenic activity is reviewed. These include the western part of the Kola Peninsula (NW Russia), the most heavily industrialized area in the entire Arctic, and the South Urals (Karabash) considered amongst the most polluted areas in the world where human impact on the environment is largely irreversible. The major pollutants are heavy metals and long-lived radionuclides from full-scale activities and accidents at the radiochemical "Mayak" Production Association (PA). Moss was successfully used to study spatial deposition of ¹³⁷Cs and ²¹⁰Pb in Belarus and Slovakia 20 years after the Chernobyl accident. A combination of analytical data (NAA and AAS in our case) with principle component analysis and GIS technologies allowed pollution source characterization and apportioning in the sampled areas around (1) copper mines in Karabash (RF), Bor (Serbia), South of Poland; (2) Zn-Pb smelters in Baia Mare (Romania), Krdjali (Bulgaria) and Veles (Macedonia); (3) Fe-V plant in Tula (RF); (4) Fe-Cr industry in Tikhvin (RF) and Mo-i-Rana (Norway); (5) oil refinery industry in Yaroslavl (RF), and (6) thermal power plant in a Moscow district. Besides passive (terrestrial) moss biomonitoring, active moss biomonitoring (moss bags technique) proved effective in biomonitoring air pollution in large cities (street canyons of Belgrade and Moscow). The results obtained at a local scale in the areas experiencing environmental stress can be used to establish emission levels of pollutants and to provide information for health-related institutions.

## M19 STOMATAL UPTAKE AND NON-STOMATAL OZONE REMOVAL BY A MIXED OAK-HORNBEAM MATURE FOREST IN THE PO VALLEY: RESULTS OF THE ECLAIRE LONG-TERM CAMPAIGN

Gerosa G.<fnref n="1" />, Marzuoli R.<fnref n="1" />, Monga<fnref n="1" /> R., Hardersen S.<fnref n="2" />, Gorian F.<fnref n="2" />, Minari E.<fnref n="2" /> and Finco A<fnref n="1" />.

In the framework of the ÉCLAIRE Project of the 7th EU framework programme a joint field campaign and a following long-term one had been conducted at an Oak-Hornbeam mixed mature forest in the Po Valley (Bosco Fontana, Mantova, Italy). Inside the forest, a 42 m tall micrometeorological tower was installed to measure the mass (O₃, CO₂, H₂O) and energy exchange between the ecosystem and the atmosphere. The focus of this work will be on the ozone flux measurements which started in June 2012 and are currently running.

<footnote n="1">¹ Catholic University, via dei Musei 41, 25121 Brescia (I), angelo.finco@unicatt.it</footnote>

<footnote n="2">² CFS-MIPAAF, CNRBF "Bosco Fontana e Peri", Via Ederle 16, 37126 Verona (I)</footnote>
The ecosystem behaved as a relevant sink for tropospheric ozone, both in summer and in winter, with an average monthly ozone removal of 5.8 kg ha⁻<fnref n="1" /> and a total annual deposition of more than 70 kg ha⁻<fnref n="1" />. The stomatal flux, estimated by deriving the bulk stomatal resistance through the inversion of the Monteith equation. The maximum stomatal fraction was observed in July 2013 (55%) and the minimum in October 2012 (13%). A strong interannual variability about the stomatal deposition between summer 2012 and 2013 was observed. The phytotoxic ozone doses taken up by the plants were respectively 13.5 mmol m⁻² in 2012 and 29 mmol m⁻² in 2013.

The non-stomatal deposition, obtained as a residual between the total and the stomatal ozone fluxes, was investigated too. In the joint field campaign Neimitz et al. (2013) found that half of the non-stomatal deposition, was due to NO soil emission, which were on average above 100mg N m⁻² s⁻<fnref n="1" />. A minor part of the non-stomatal fluxes (6%) was attributed to isoprene reactions with ozone. A large part of the non-stomatal deposition (44%) was not explained. An investigation on this part, with particular attention on the deposition on non-transpiring surfaces, will be showed.

Additional measurements of O₃, NO and NO₂ concentrations along the tower profile, allowed to estimate the ozone storage inside the trunk space. The ozone storage can be seen as a part of the deposition flux which is temporarily stored into the trunk space. Taking it into account, the daily course of the non-stomatal deposition significantly changed, with important consequences for non-stomatal interpretation and for proper modelling of the non-stomatal processes.

An estimation of the ozone impact on the whole ecosystem was assessed by looking at the departure from the carbon dioxide balance closure. First results of CO₂ exchange seem to indicate a net emission of CO₂ from the ecosystem, as it should not be expected from a healthy mature forest ecosystem where, typically, the carbon balance is zero. Again, a general limitation of the net carbon uptake has been observed at high ozone stomatal fluxes.

Finally, the great differences observed in the two years highlight the need for long time series of flux measurements. Moreover, this kind of studies will help in raising the awareness of the policy makers on the ecosystem services offered by the vegetal ecosystems.

## Reference

E. Nemitz, B. Langford, C.F. Di Marco, M. Coyle, C. Braban, M. Twigg, G. Gerosa, A. Finco, A. Valach, J.Acton, B. Loubet, S. Schallart, R. Gasche, E. Diaz-Pines, S. Fares, J. Westerlund, Å. Hallquist, C. Gritsch, S. Zechmeister-Boltenstern and M.A. Sutton, 2013. Quantifying Chemical Interactions in a Forest Canopy – First Results from the ÉCLAIRE Campaign at Bosco Fontana, Po Valley. ACCENT-Plus Symposium, Urbino (Italy), 17-20 September 2013.

# M2 OVERVIEW OF THE ACHIEVEMENTS OF THE ICP VEGETATION IN 2013 AND FUTURE WORKPLAN (2014 – 2016)

Harmens, H.<fnref n="1" />, Mills, G.<fnref n="1" />, Hayes, F.<fnref n="1" />, Norris D.A.<fnref n="1" />, and the participants of the ICP Vegetation

The ICP Vegetation is an international programme that reports on the effects of air pollutants on natural vegetation and crops [1]. It reports to the Working Group on Effects (WGE) of the UNECE Convention on Long-range Transboundary Air Pollution (LRTAP). In particular, the ICP Vegetation focuses on the following air pollution problems: quantifying the risks to vegetation posed by ozone pollution and the atmospheric deposition of heavy metals, nitrogen and persistent organic pollutants (POPs) to vegetation. The ICP Vegetation also studies the impacts of pollutant mixtures (e.g. ozone and nitrogen), consequences for ecosystem services and biodiversity, and interactions between air pollutants and climate change. When possible, economic evaluations of the impacts of ozone are included.

<footnote n="1">¹ICP Vegetation Programme Coordination Centre, Centre for Ecology and Hydrology, Bangor, Gwynedd LL57 2UW, UK.
hh@ceh.ac.uk</footnote>
At the 27th Task Force Meeting we will report on the achievements of the ICP Vegetation in 2013 [1], including: Impacts of ozone pollution on ecosystem services and biodiversity [2]; Outcome of the European heavy metals and nitrogen in mosses survey 2010/11 [3] and the pilot study on POPs; In addition, we will report on the contribution of ICP Vegetation to the common work plan items of the WGE for 2013, such as: Report [4] and brochure [5] on benefits of air pollution control for biodiversity and ecosystem services; Guidance document on health and environmental improvements [6].

We will also discuss the future workplan, including progress made with items to be reported to the LRTAP Convention in 2014 [7], such as: Report on air pollution deposition to, and impacts on vegetation, in EECCA/SEE countries and South-East Asia; Update chapter 3 of the Modelling and Mapping Manual of the LRTAP Convention.

## Acknowledgement

We thank the UK Department for Environment, Food and Rural Affairs (Defra) for funding the ICP Vegetation Programme Coordination Centre. Further financial support was provided by the UNECE and the UK Natural Environment Research Council (NERC).

# M13 RELATIONSHIP BETWEEN SITE-SPECIFIC NITROGEN CONCENTRATIONS IN MOSSES AND BULK ATMOSPHERIC NITROGEN DEPOSITION

Harmens H<sup>a</sup>, Schnyder E<sup>b</sup>, Thöni L<sup>b</sup>, Cooper D<sup>a</sup>, Mills G<sup>a</sup>, Leblond S<sup>c</sup>, Mohr K<sup>d</sup>, Poikolainen J<sup>e</sup>, Santamaria J<sup>f</sup>, Skudnik M<sup>g</sup>, Zechmeister H<sup>h</sup>, Lindroos A-J<sup>e</sup>, Hanus-Ilnar A<sup>i</sup>

To assess the relationship between nitrogen concentrations in mosses and bulk nitrogen deposition or concentrations in precipitation, both moss tissue and precipitation were sampled within a distance of 1 km of each other at sites in seven countries across Europe. As previously described for modelled nitrogen deposition [1], the relationship between nitrogen concentration in mosses and measured (total) bulk deposition or concentration in precipitation across Europe is best described by an asymptotic relationship. The asymptotic relationship is much stronger for ammonia-N than for nitrate-N in bulk deposition or precipitation, with high ammonium deposition rates at German sites being most influential in providing evidence of the asymptotic behaviour. Saturation appears to occur at bulk nitrogen deposition rates of ca. 15 – 20 kg ha<sup>-1</sup> y<sup>-1</sup>. Up to such deposition rates, linear relationships have been observed in some countries (Finland and Switzerland) but not in others. Considerable scatter was observed in the relationship at the European level, although less than previously found with modelled deposition [1]. Nitrogen concentration in mosses can be applied to identify areas at risk of high nitrogen deposition at the European scale.

## Acknowledgement

We thank the UK Department for Environment, Food and Rural Affairs (Defra) for funding the ICP Vegetation Programme Coordination Centre. Further financial support was provided by the UNECE and the UK Natural

<footnote><sup>a</sup> Centre for Ecology and Hydrology, Bangor, UK. hh@ceh.ac.uk</footnote>

<footnote><sup>b</sup> FUB-Research Group for Environ. Monitoring, Rapperswil, Switzerland. fub@fub-ag.ch</footnote>

<footnote><sup>c</sup> Muséum National d'Histoire Naturelle, Paris, France. sleblond@mnhn.fr</footnote>

<footnote><sup>d</sup> Landwirtschaftskammer Niedersachsen, Germany. Karsten.Mohr@LWK-Niedersachsen.de</footnote>

<footnote><sup>e</sup> Finnish Forest Research Institute Oulu, Finland. Jarmo.Poikolainen@metla.fi;</footnote>

<footnote><sup>f</sup> University of Navarra, Pamplona, Spain. chusmi@unav.es</footnote>

<footnote><sup>g</sup> Slovenian Forestry Institute, Ljubljana, Slovenia. mitja.skudnik@gozdis.si.</footnote>

<footnote><sup>h</sup> University of Vienna, Vienna, Austria. Harald.Zechmeister@univie.ac.at</footnote>

<footnote><sup>i</sup> Umweltbundesamt, Vienna, Austria.</footnote>
Environment Research Council (NERC). We also thank the funding bodies in the various countries for their financial support.

## References

[1] Harmens et al. (2011). Nitrogen concentrations in mosses indicate the spatial distribution of atmospheric nitrogen deposition in Europe. Environmental Pollution 159: 2852-2860.

# M23 VACUOLAR ENDOPROTEOLYTIC ACTIVITIES AND PROTEIN CARBONYLATION LEVELS IN LEAF AS A MODEL FOR DEPICTING OZONE TOLERANCE AND SENSITIVITY IN WINTER WHEAT (TRITICUM AESTIVUM L.)

Havé M.ª, Leitao L.ª, Bagard M.ª, Laffray D.ª, Bethenod O.ᵇ, Castell J. F.ᵇ, Repellin A.ª

Current global background concentrations in tropospheric ozone (O₃) have been shown to have significant adverse effects on crop yield, including wheat. As selection of O₃ resistant cultivars (cvs) could help reduce O₃-induced agricultural losses (Avnery et al., 2013), we investigated whether protein oxidation (carbonylation) and vacuolar endoproteolytic activities measured in the flag leaf could differentiate between ozone sensitive and tolerant wheat cvs. Two winter wheat (Triticum aestivum L.) cvs released in 1986 (Soissons) and 2006 (Premio) were grown in the field and exposed to ambient and semi-controlled chronic O₃ concentrations, from pre-anthesis to harvest, using a new linear O₃ fumigation device that generates gradients of the pollutant. Determination of resistance to O₃ from grain yield and quality showed that Soissons was the most sensitive cv. For both cvs, an important decline in total chlorophyll contents was observed in response to O₃ suggesting it induced a premature development of leaf senescence, causing the shortening of the assimilation and grain-filling periods. O₃ induced an increase in protein carbonyl groups (PCO) for both cvs. However, Soissons was characterized by a higher intrinsic level of protein carbonyls. Increased PCO levels and losses in total chlorophyll contents were concurrent, suggesting a link between the extent of oxidative stress and senescence development. Consistent with a premature induction of leaf senescence, O₃ induced a stimulation of endoproteolytic activities that resulted mostly from increases in cysteine protease (CP) activities, for both cvs. However, CP stimulation was more important in the resistant cv. Increased endoproteolytic activities were associated with increased PCO and with a decline in total soluble protein contents. In conclusion, intrinsic PCO levels and induction of CP in response to O₃ could help distinguish between resistant and sensitive wheat cvs to O₃.

## References

Avnery S, Mauzerall DL, Fiore AM (2013) Increasing global agricultural production by reducing ozone damage via methane emission controls and ozone-resistant cultivar selection. Global Change Biology 19: 1285-1299.

# M18 IMPACTS OF OZONE AND NITROGEN ON SILVER BIRCH

Hayes, F., Harmens, H., Mills, G.

Centre for Ecology and Hydrology, Bangor, UK. fhay@ceh.ac.uk

Birch trees (Betula pendula) were exposed to factorial combinations of seven ozone and four nitrogen regimes for five months in solardomes at CEH Bangor in 2012 and 2013. The ozone regime was designed to investigate the benefits of changes in air quality policy that are anticipated to reduce both background and peak ozone concentrations, but with a

<footnote>ª Equipe IBIOS, UMR 7618 Bioemco, Université Paris Est-Créteil, 61 Avenue du Général de Gaulle, 94010 Créteil Cedex, France. marien.av@hotmail.fr</footnote>

<footnote>ᵇ UMR 1091 Environnement et Grandes Cultures, INRA, 78850 Thiverval-Grignon, France</footnote>
larger reduction for the peaks. The range of ozone exposure seasonal means was 35 ppb to 70 ppb (24h mean) and the nitrogen treatments were applied weekly as ammonium nitrate to give treatments equating to 10, 30, 50 and 70 kg $ha^{-1}yr^{-1}$. Measurements were made at both the leaf level and at whole-tree level to investigate whether:

- nitrogen modifies the response to ozone;

- nitrogen and/or ozone treatment alter the $DO_3SE$ parameterisations for birch;

- fluxes of ozone and carbon become uncoupled;

- whole-tree alterations in fluxes are a consequence of individual leaf physiological responses or via alterations in tree biomass.

The main findings in the second season of ozone exposure where:

- Reduced tree size and leaf number;

- Premature leaf loss

- Tendency to reduced leaf photosynthetic capacity ($V_c,max$, $J_{max}$) later in the season, linked to reduction chlorophyll index due to early leaf die-back.

In the first year of exposure we had shown that whole tree carbon fluxes were reduced with increasing ozone treatment.

Effects of nitrogen included:

- Increased tree size, leaf number and bud length;

- Increased stomatal conductance;

- Increased chlorophyll content and photosynthetic capacity, particularly early in the season.

Potential interactions between ozone and nitrogen were also observed, including:

- Impacts on tree size and woody biomass;

- High nitrogen seemed to delay early leaf die-back at high ozone exposure later in the season, delaying the reduction in photosynthetic capacity due to ozone.

## Conclusions

- Ozone pollution decreases growth and therefore carbon sequestration of birch trees;

- Nitrogen treatment affects stomatal fluxes of birch and therefore nitrogen deposition should be accounted for when calculating ozone fluxes to inform assessments of vegetation at risk of ozone pollution;

- The cumulative effects of ozone and nitrogen pollution on trees require further study over several years as leaf-level measurements indicate that effects and interactions may occur over longer timescales.

## Acknowledgement

This work was funded by the EU ECLAIRE project (FP 7, grant agreement no. 282910) and the Natural Environment Research Council, UK.
M3 OVERVIEW OF THE ACTIVITIES OF ICP MODELLING AND MAPPING AND PLANS FOR THE FUTURE

Hettelingh J-P<fnref n="1" />, Le Gall A-C<fnref n="2" />, Posch M<fnref n="1" />, Slootweg J<fnref n="1" />, Harmens H<fnref n="3" />, Mills G<fnref n="3" />

The Coordination Centre for Effects (CCE) is the programme centre of the ICP Modelling and Mapping (ICP-M&M; www.icpmapping.org) and its network of National Focal Centres developing methods and databases on critical thresholds. These thresholds are used in the **Greenhouse Gas and Air Pollution Interactions and Synergies (GAINS)**-model in support of abatement policies that aim to protect European ecosystems against adverse effects of atmospheric deposition. In recent protocol negotiations, the focus of effects-based knowledge by the ICP-M&M and ICP-V as used in GAINS, was on geo-chemical modelling of exceedances of critical loads for eutrophication and acidification (Figure 1) and ozone critical levels respectively.

<figure><img src="image_9.png" /><figcaption>**Figure 1:** The temporal development since 1880 of the area at risk (in %, left) and magnitude (in eq ha⁻¹yr⁻<fnref n="1" />, right) of exceedance (AAE) of acidification (red) and eutrophication (green), using Gothenburg Protocol – Current Legislation (GP-CLE) scenario depositions from 2010 onwards (Source: Hettelingh et al. EEA technical report XX/2013 in press)</figcaption></figure>

Critical thresholds are embedded in the “GAINS-model” to interactively optimize emission reduction patterns to regionally diminish adverse ecosystem effects. Alternatively, critical thresholds can be used outside of the model, in a “GAINS-system” mode to post-analyse adverse ecosystem effects of air pollution for selected emission scenario-outputs of the GAINS-model. This approach resulted in collaboration between most ICPs to produce values for a range of indicators in two collaborative reports entitled “Guidance document on health and environmental improvements”<fnref n="1" /> and “Benefits of Air Pollution control for benefits and ecosystem services”<fnref n="2" />. This collaboration between ICPs in general and with ICP-V in particular is further strengthened under the LRTAP Convention workplan for 2014-2015.

Near future work of the ICP-M&M and ICP-V focusses on the identification and application of novel indicators and thresholds within the EU Framework VII project *Effects of Climate Change on Air Pollution and Response Strategies for European Ecosystems* (ECLAIRE: http://www.eclaire-fp7.eu). Vegetation change dynamics are coupled to soil chemistry to model changes of plant species diversity using selected indicators that capture adverse effects of air pollution under climate change. The challenge for the ICP-M&M and ICP-V is to address the trade-offs between (a) the nitrogen induced increase, and ozone induced decrease, of plant productivity in the short term and (b) plant productivity in the short term and the N induced impoverishment of soil chemistry in the long term.

<footnote n="1">¹ICP-M&M/CCE at RIVM, P.O.Box 1, NL-3729 BA Bilthoven,
jean-paul.hettelingh@rivm.nl</footnote>

<footnote n="2">²Chair, TF-ICP/M&M at INERIS, P.O.Box 2, FR-60550, Verneuil-en-Halatte,</footnote>

<footnote n="3">³TF-ICP/Vegetation at CEH, Bangor, Gwynedd LL57 2UW, UK</footnote>

<footnote n="1">¹ http://www.unece.org/fileadmin/DAM/env/documents/2013/air/eb/ECE_EB.AIR_2013_8_E.pdf</footnote>

<footnote n="2">² http://www.unece.org/fileadmin/DAM/env/documents/2013/air/wge/ECE_EB.AIR_WG.1_2013_14_ENG_01.pdf</footnote>
## M11 IS OZONE REDUCING THE N FIXATION CAPACITY OF CURRENT MANAGED PASTURE?

Hewitt, D<sup>1,2*</sup>, Mills, G<sup>1</sup>, Hayes, F<sup>1</sup> & Davies, W<sup>2</sup>.

<sup>1</sup> CEH Bangor, Environment Centre Wales, Deiniol Rd, Bangor, Gwynedd, UK, LL57 2UW. [danhew@ceh.ac.uk](mailto:danhew@ceh.ac.uk)

<sup>2</sup> Lancaster Environment Centre, Lancaster University, Lancaster, UK, LA1 4YQ.

Clovers (*Trifolium* spp.) are an important component of temperate pasture, providing a source of fixed nitrogen (N) to soil and improving forage quality and productivity. Although known to be highly sensitive to ozone pollution, surprisingly little is known about the effects of ozone on the modern cultivars of clover sewn by farmers for use in intensively grazed pasture. In 2012, we exposed monocultures of white clover (*T. repens* cv. Crusader) and red clover (*T. pratense* cv. Merviot) to a range of ozone profiles for 12 weeks. We found reductions in root biomass and root nodule number in response to ozone in both cultivars, with Crusader also displaying reductions in the size and mass of nodules and N-fixation rates. Applying the AOT40-based response function spatially to the UK, we found that there was potential for a median 9% reduction in clover root nodule mass (mean for all 10 x 10km grid squares containing pasture) in a high ozone spring such as the year 2006.

In 2013, we exposed Crusader, in mixture with a current cultivar of high-sugar ryegrass (*L. perenne* cv. AberMagic), to current and near-future ozone scenarios for a period of 16 weeks. We investigated effects on N-fixation, and associated physiology and growth throughout the growing season. The results are discussed in relation to potential mechanisms and impacts on agricultural sustainability.

## M15 DIFFERENCES IN CONCENTRATION OF NITROGEN IN MOSSES DUE TO CANOPY DRIP EFFECTS - CASE STUDY GERMANY

Kluge, M.<sup>a</sup>, Pesch, R.<sup>a</sup>, Schröder W.<sup>a</sup>, Hoffmann, A.<sup>b</sup>

**Background.** Atmospheric nitrogen (N) deposition into terrestrial ecosystems is frequently considered as a threat to phyto-diversity. In Germany, N concentrations in mosses were recorded in 2004 as part of a regional investigation at 54 locations in the North-West (Schröder et al. 2007) and in 2005 at 726 locations across the whole territory. Following the methodology presented by Schröder et al. (2013), differences of correlations between N concentrations in atmospheric deposition and in mosses between landscapes across Europe could be corroborated (Schröder et al. 2014). These investigations (Schröder et al. 2007, 2013, 2014) were accomplished by studies focusing on spatial differences of N concentrations in mosses at the site level.

**Methods.** To this end, N concentrations were determined in mosses sampled at sites with and without canopy drip effects in several European states (see *Discussion*), amongst them Germany. This article deals with that study conducted in 2012 comparing N concentrations in mosses sampled within 30 forest stands and in 26 adjacent open fields in Northwestern Germany (Kluge et al. 2013). The N concentrations were converted to N atmospheric deposition values by use of a regression model derived from Schröder et al. (2014). These deposition estimations enabled to calculate N critical loads exceedances.

<footnote><sup>a</sup> Chair of Landscape Ecology, University of Vechta, POB 15 53, 49377 Vechta, [mkluge@iuw.uni-vechta.de](mailto:mkluge@iuw.uni-vechta.de), [rpesch@iuw.uni-vechta.de](mailto:rpesch@iuw.uni-vechta.de), [wschroeder@iuw.uni-vechta.de](mailto:wschroeder@iuw.uni-vechta.de)</footnote>

<footnote><sup>b</sup> LUFA Nord-West, Institute for Fertilizers and Seeds, 31787 Hameln [andreas.hoffmann@lufa-nord-west.de](mailto:andreas.hoffmann@lufa-nord-west.de)</footnote>
**Results.** Compared to the average N concentration in mosses sampled in open fields 2012 (7.4 kg / ha*a), the average N concentrations in mosses within adjacent forests were almost four times higher (26.6 kg / ha*a), and the maximum within the stands accounted for ~ 56 kg / ha*a. Compared to 2005,

there was a slight decline of the average N deposition by 2.4 kg / ha*a in open fields. However, the average N concentrations in mosses within forests stands in 2012 remained nearly the same since 2004 (29 kg / ha*a). The atmospheric N deposition as estimated from the N concentration in mosses ranged between the minimum and maximum N critical load at 71 % of the 56 sites investigated. At 14 % of the sites the N deposition was close to the maximum N critical load value which was exceeded in 11%.

**Discussion.** The results presented could be compared to concentrations of N in mosses sampled at both, canopy drip influenced sites and open areas, in several European countries (Austria, Estonia, France, Finland, Germany, Switzerland, The Netherlands, and Slovenia). In addition, results of a re-sampling in 2013 in Northwestern Germany and of mosses collected at two German ICP Integrated Monitoring and EMEP deposition monitoring sites (Neuglobsow, Bayerischer Wald) could be presented and discussed.

**Conclusions.** The study at hand revealed statistically significant differences between N concentrations measured in mosses sampled within forests and in open fields (Kluge et al. 2013). The presented findings should be accounted for both modelling and mapping atmospheric N deposition into terrestrial ecosystems on the one hand and related estimations of N critical load exceedances on the other hand.

**References.**

* Kluge M. et al. 2013. Accounting for canopy drip effects of spatiotemporal trends of the concentrations of N in mosses, atmospheric N depositions and critical load exceedances: a case study from North-Western Germany. Environ Sci Europe 2013, 25:26 (13 pp + 4 suppl files).

* Schröder W. et al. 2007. Nitrogen and metals in two regions in Central Europe: Significant differences in accumulation in mosses due to land use? *Environ Monit Assess* 133:495-505

* Schröder W. et al. 2013. Correlation between atmospheric deposition of Cd, Hg and Pb and their concentrations in mosses specified for ecological land classes covering Europe. *AtmosPollut Res* 4:267-274

* Schröder W. et al. 2014. Mapping correlations between nitrogen concentrations in atmospheric deposition and mosses for natural landscapes in Europe. *EcolInd*36:563-571.

M29 THE SURVEY OF ATMOSPHERIC DEPOSITION OF AL, CR, FE, NI, V AND ZN IN ALBANIA
BY USING MOSS BIOMONITORING AND ICP-AES

Pranvera Lazo<fnref n="1" />, Flora Qarri<fnref n="2" />, Trajce Stafilov<fnref n="3" />, Marina Frontasyeva<fnref n="4" />, Lirim Bekteshi<fnref n="1" />, Katerina Baceva<fnref n="3" />, Jani Marka<fnref n="1" />

The atmospheric deposition of Al, Cr, Fe, Ni, V and Zn in Albania was investigated by using carpet-forming-moss species (*Hypnum cupressiforme*) as bioindicators. This research is a part of the international programme (ICP Vegetation Programme, UNECE) carried out in most of European countries since 1987 investigating the impacts of air pollutants on crops and natural vegetation. Sampling was done in dry seasons of autumn 2010 and summer 2011 at 62 sites distributed over Albania. Unwashed, dried samples were totally digested by using microwave digestion and the concentrations of metal elements were determined by ICP-AES. The results reflect local emission points. The elements with high values of concentration compare with other European countries, such as Al, Cr, Fe, Ni, V and Zn, were separately treated in this paper. The median values of chromium (4.75 mg/kg, DW), iron (1618 mg/kg, DW), nickel (5.85 mg/kg, DW), vanadium (3.51 mg/kg, DW), zinc (13.77 mg/kg, DW) and aluminum (6974 mg/kg, DW) are similar to those of neighboring countries, but higher to those of European countries. EWMA and univariate control chart was used to investigate the moving range of successive observations and to estimate the variability of the data.

<footnote n="1">¹ Department of Chemistry, Faculty of Natural Sciences, University of Tirana, Albania [pranveralazo@gmail.com](mailto:pranveralazo@gmail.com)</footnote>

<footnote n="2">² Department of Chemistry, University of Vlora, Albania</footnote>

<footnote n="3">³ Institute of Chemistry, Faculty of Science, Sts. Cyril and Methodius University, Skopje, Macedonia</footnote>

<footnote n="4">⁴ Joint Institute for Nuclear Research, Dubna, Russian Federation</footnote>
Certain local emitters were identified like, iron-chromium metallurgy and cement industry, oil refinery, mining industry, and transport. In addition, the natural sources, from the accumulation of these metals in mosses caused by metal-enriched soils, associated with wind blowing soils particularly in south-east direction of the country, was pointed as another possibility of local emitting factors.

**Keywords:** air pollution, metals; moss biomonitoring; ICP-AES, EWMA, multivariate analysis, Albania.

<figure><img src="image_10.png" /></figure>

# THE ROLE OF JASMONATE AND SALICYLIC ACID IN CONFERRING TOLERANCE AGAINST OZONE INJURY IN TOMATO PLANTS

Madkour, Samia and Abou-Salem, Amal

Faculty of Agriculture, Damanhour University, Damanhour, Egypt.
agric@damanhour.edu.eg

Enhancing stress tolerance in plants has major implications in agriculture. However, a simple method for inducing tolerance in plants without undesirable side effects has not been available till now. Both Salicylic acid (SA) and Jasmonic acid (JA) signaling pathways were found to be activated in response to O3 exposure. The endogenous accumulation of SA and JA during O3 – fumigation was reported repeatedly in the literature. In the present study exogenous applications of SA and JA prior to O3 exposure were used to elucidate their effect in altering the response of tomato plants to O3-induced oxidative stress, to investigate whether plants exhibiting different levels of O3-tolerance exhibited different responses to signal transduction; and to test the hypothesis that treatment with SA and JA is able to alter or reduce O3 injury symptoms. Eight different tomato cultivars were tested for O3 – sensitivity. Tests were undertaken with different O3 concentrations (100-150ppb and 150-200ppb) at different stages of vegetative growth (4-5 leaves and 7-8 leaves). Data collected from those screening tests yielded two sensitive cultivars, Strain B and Peto86. There were no significantly tolerant species in the group, rather different degrees of intermediate responses. Chico (intermediate/sensitive), Ace 55 and Castle rock (intermediate/tolerant) were selected on the basis of their consistent responses in the repeated experiments. Electrolyte leakage which is an early event leading to cell death and lesion formation was investigated. Dramatic increases in electrolyte

leakage were recorded on both the O3 – sensitive cultivars (Strain B and Peto86), while significant but less striking enhancement of e-leakage and cell death were the results of O3 exposure of the intermediate cultivars (Chico, Ace 55 and Castle Rock). Ethylene (ET) emission has been shown to correlate to O3 sensitivity in several plant species. Induction of ET synthesis in O3 – sensitive tomato plants occurred within 0.5-2hr after the onset of ozone exposure and reached its peak value at 6h from the start of O3 fumigation. Data collected 18hr afterwards (24hr sampling) were dramatically lower but remained stable in the subsequent sampling time (48h). The amount of ET produced generally correlated well with the degree of visible lesion formation that developed at least 18-24hr from the end of O3 exposure. Exogenous SA induced a clear and significant increase in O3 – sensitivity in all the tomato cultivars studied. Visible symptoms were more severe and spread over larger leaf areas. The increase in foliar symptoms was supported by the enhancement of ET emission from SA – treated plants, especially in O3 – sensitive cultivars. A general increase in loss of cell integrity and cell death was also observed. On the other hand, pre-treatment with exogenous JA dramatically reduced the magnitude of visible foliar injury caused by O3 and foliar lesions were significantly attenuated on all the cultivars investigated. Electrolyte leakage decreased significantly pointing to a lower rate of cell death. Cell death was almost completely abolished in all the cultivars except for Strain B, the most sensitive cultivar. ET levels in JA – treated plants declined sharply, but the general ET emission trend was maintained. We concluded that SA is most probably involved in lesion propagation and would thus promote cell death, while JA influences lesion attenuation and containment, therefore, would play a role in enhancing stress tolerance.
# NEW ICP VEGETATION SMART-PHONE APP FOR RECORDING INCIDENCES OF OZONE INJURY ON VEGETATION

Mills, G.<fnref n="1" />, Bacon, J.<fnref n="2" />, Harmens, H.<fnref n="1" />, Hayes, F.<fnref n="1" /> et al.

In 2007, the ICP Vegetation published a synthesis report documenting over 500 incidences of ozone injury on crops, grassland species and shrubs growing in the field under ambient air conditions in 17 countries of Europe (Hayes et al., 2007). We plan to revisit this study by compiling new spatial data on current incidences of ozone injury. Using smart-phone technology for i-phones and android phones, and web-based recording methodology we are developing a new way of recording incidences of ozone injury in the field. A new App is nearing completion that will allow participants to upload photographs of ozone injury direct from the field together with the coordinates for the location where the injury was detected (derived from a zoom-able Google map). Participants will be taken through a series of questions designed to assist with quality assurance, including being asked if they have previous experience of identifying ozone damage or plant diseases and recent weather conditions. The App could be used anywhere in the world and thus is suitable for use in the outreach activities as well as the European activities of the ICP Vegetation. The aim is to finalise the App before the forthcoming growing season allowing it to be rolled out to participants in March/April. We would like to encourage as many people as possible to participate in using the App to record ozone injury in 2014 allowing us to fully test the App by developing a large database for analysis.

The web-based recording facility will be available at our webpage: http://icpvegetation.ceh.ac.uk. News of the completion and launch of the web-based and smart-phone App will be provided by email and on the web.

## Acknowledgement

We thank the UK Department for Environment, Food and Rural Affairs (Defra) and the UK Natural Environment Research Council (NERC) for funding this project.

## References

Hayes, F., Mills, G., Harmens, H., Norris, D. (2007) Evidence of widespread ozone damage to vegetation in Europe (1990 – 2006). Programme Coordination Centre of the ICP Vegetation, Centre for Ecology and Hydrology, Bangor, UK. ISBN 978-0-9557672-1-0.

http://icpvegetation.ceh.ac.uk/publications/documents/EvidenceReportFINALPRINTEDVERSIONlow-res.pdf

## M17 OVERVIEW OF PROGRESS FOR THE EU FP7 ECLAIRE* PROJECT

Mills, G.<fnref n="1" />, Sutton, M.<fnref n="2" />, and many colleagues from the ECLAIRE project

<footnote n="1">¹ ICP Vegetation Programme Coordination Centre, Centre for Ecology and Hydrology, Bangor, Gwynedd LL57 2UW, UK.
gmi@ceh.ac.uk; hh@ceh.ac.uk; fhay@ceh.ac.uk</footnote>

<footnote n="2">² Jim Bacon, Centre for Ecology and Hydrology, Wallingford, Oxfordshire OX10 8BB.
jame2@ceh.ac.uk</footnote>

<footnote n="1">¹ ICP Vegetation Programme Coordination Centre, Centre for Ecology and Hydrology, Bangor, Gwynedd LL57 2UW, UK.
gmi@ceh.ac.uk</footnote>

<footnote n="2">² Centre for Ecology and Hydrology, Penicuik, Midlothian EH26 0QB. ms@ceh.ac.uk</footnote>
Running for just over two years, the EU FP7 project ECLAIRE brings together expertise from many specialists to develop new ways of modelling the risks posed to European ecosystems from the combined effects of air pollution and climate change. This unique combination of scientists covering

effects at the leaf scale through to modelling impacts on European ecosystem services and climate, and including experts on ozone, nitrogen, aerosols and climate change met recently in Zagreb, Croatia to discuss progress at the half-way stage of the project. This presentation will provide a brief overview of progress with the project, with particular emphasis on aspects of relevance to ICP Vegetation and policy makers at the LRTAP Convention.

The project is split into five main scientific components: (1) Emissions and Exchange processes; (2) Emissions and exchanges at local to EU and global scales; (3) Ecological response processes and thresholds; (4) Ecological responses at regional and European scales; and (5) Integrated risk assessment and policy tools. Many representatives of bodies of the LRTAP Convention are participating in this project, e.g. ICP Vegetation, CCE (ICP Modelling and Mapping), ICP Forests, Task Force on & Centre for Integrated Assessment Modelling, NEBEI, EMEP/MSC-West, Task Force on Reactive Nitrogen.

ICP Vegetation participants have greatest involvement in component 3, including analysis of data mined from the published literature to derive dose-response relationships for physiological and growth responses, experimental exposure of crops, grassland and tree species to combinations of ozone and nitrogen pollutants, investigations of leaf-scale processes and model development including photosynthesis-based ozone uptake and growth models. MADOC for species changes and JULES (a land-surface climate model). As well as providing an overview, this paper will also include results from the data mining activities showing how cross-species response functions are being derived from a data extraction exercise involving over 300 papers. The following two papers will describe results from tree O3 and N exposure experiments at Curno, Italy (Gerosa et al.) and Bangor, UK (Hayes et al.).

* Effects of climate change on air pollution impacts and response strategies for European ecosystems.

## Acknowledgement

We thank the EU for funding this project under the Seventh Framework Programme, grant agreement no. 282910.

# M25 CIRCADIAN PROFILES OF PHOTOSYNTHETIC PARAMETERS AND PRIMARY METABOLITES IN GRAPEVINE VARIETIES EXPOSED TO OZONE

Pellegrini E.<fnref n="1" />, Campanella A.<fnref n="1" />, Paolocci M.<fnref n="2" />, Lorenzini G.<fnref n="1" />, Nali C.<fnref n="1" />, Muganu M.<fnref n="2" />

A comparative study on the diurnal courses of photosynthetic gas exchange parameters, chlorophyll a fluorescence characteristics and photoprotection mechanisms has been conducted on two *Vitis vinifera* varieties [black-berried cv. Aleatico (ALE) and white-berried cv. Trebbiano giallo (TRE)] exposed to chronic ozone (O3) fumigation simulating the typical circadian profile of this pollutant in Central Italy. Experiments started in early spring before budbreak. Two-year-old plants grown in five liter pots were acclimated inside a greenhouse ventilated with charcoal filtered air and treated for 28 days with a pulse of O3 concentration between 8:00 a.m. and 1:00 p.m., with a target concentration of 80 ppb and between 1:00 p.m. and 6:00 p.m., at 40 ppb. At the constitutive level, TRE showed both higher average value

<footnote n="1">¹ Department of Agriculture, Food and Environment, University of Pisa - Via del Borghetto
80 56124 Pisa Italy - elisa.pellegrini@for.unipi.it</footnote>

<footnote n="2">² Department of Agriculture, Forests, Nature and Energy, University of Tuscia - Via S. Camillo de Lellis snc 01100 Viterbo, Italy</footnote>
of whole leaf thickness (157 ±3.5 TRE, vs 139 ±2.3 µm ALE) and higher stomatal conductance to water vapour (Gw) levels (+29% in term to daily values) in comparison to ALE. On the other hand, stomatal density, violaxanthin content and abscisic acid concentration were lower in TRE compared to ALE (143 ±30.8 stomata mm⁻<fnref n="2" /> TRE, vs 249 ±31.8 ALE, -75 and -37%, respectively), suggesting a different stomatal behaviour of the varieties. At the end of the treatment, both varieties showed visible spots (Ø 1-2 mm) of yellowing tissue located in the interveinal area of fully expanded leaves. SEM observations of the abaxial surface of treated samples showed an irregular presence of deformed swelling areas between veins. These modifications were more frequent and developed in ALE. Oxidative stress modified the diurnal pattern of leaf CO₂ assimilation rate (A): in both varieties, A values were significantly reduced during all day and a single-peak was obtained at 4:00 p.m. In ALE, this reduction was mainly due to a reduced mesophyllic functioning, as evidenced by the marked increase in intercellular CO₂ concentration (+33% in terms of daily values). On the contrary, in TRE, insufficient stomatal conductance was found to be the main reason for the alteration of photosynthesis, as confirmed by the decrease of Gw levels (-29% in terms of daily values). In both varieties, the alteration of photosynthetic performance observed at the end of the exposure was accompanied by PSII damage. The decrease of maximum PSII photochemical efficiency (Fv/Fm ratio) recorded in the early afternoon (at 4:00 p.m. and between 2:00 and 4:00 p.m., respectively in ALE and TRE) is indicative of photoinhibition. The reversible change of this parameter found during the day (in both varieties, Fv/Fm levels recovered at the constitutive values in the noon) suggests that photoprotection rather than photodamage occurred. At the end of the exposure, both varieties displayed mechanisms that play a role in dissipating the excess energy, as evidenced by: (i) increased content of total carotenoids (+64 and +30% in term to daily values, respectively in ALE and TRE); (ii) enhanced efficiency of thermal energy dissipation within PSII (+32 and +20%) that closely correlated with the increased de-epoxidation index (+26 and +22%), and (iii) partitioning of carbon to total carbohydrates (+14 and +9%) in comparison to controls. These results indicate a different efficiency of the thermal dissipation mechanisms and photorespiratory pathway among varieties and a major need for carotenoid-mediated photoprotection in ALE.

## M9 LATITUDE DEPENDENCE AND TEMPORAL TREND OF THE YEARLY PEAK IN SURFACE OZONE IN MIDDLE AND NORTHERN EUROPE

Pleijel H.<fnref n="1" />, Klingberg J.<fnref n="1" />, Karlsson P.E.<fnref n="2" />, Engardt M.<fnref n="3" />

The yearly dynamics of the surface ozone concentration [O₃] was studied for 25 monitoring stations ranging from Austria and Switzerland in the south, Ireland in the west to the Arctic Circle in Sweden and Finland. The aim was to investigate the level and timing of the annual peak of [O₃] in relation to latitude and growing season. Further, the change in the timing of the annual O₃ peak was studied, since this may affect risk assessment for O₃. The [O₃] yearly maximum and maximum 8h-mean significantly declined with increasing latitude as did the number of days with 8h-mean [O₃]>120 µg m⁻<fnref n="3" />. However, the yearly minimum in [O₃] (lowest running two-week average) correlated positively with latitude, thus being higher in the north. The day number of maximum [O₃] declined strongly and significantly with increasing latitude. This means that the annual peak in [O₃] is shifted from the summer in the southern part of the region to spring in the north (Figure 1). This is of particular importance in the north, since the yearly O₃ peak today occurs partly before the onset of the growing season. Climate change is likely to lead to an earlier onset of the growing season, for which there is already evidence in the last few decades e.g. in northern Sweden. Thus, the growing season may increasingly overlap with the O₃ peak in the boreal/subarctic region in the

<footnote n="1">¹University of Gothenburg, Biological and Environmental Sciences, P.O. Box 461, 40530 Göteborg, Sweden, hakan.pleijel@bioenv.gu.se</footnote>

<footnote n="2">²IVL Swedish Environmental Research Institute Inc., P.O. Box 53021, 40014 Göteborg, Sweden</footnote>

<footnote n="3">³Swedish Meteorological and Hydrological Institute, Folkborgsvägen 1, 60176 Norrköping, Sweden</footnote>
future. It has, however, been suggested, that the pronounced spring peak in [O3] in the north is largely a result of low
deposition to snow, ice and vegetation which is not physiologically active and

the spring peak would be reduced or occur earlier if the growing season starts earlier. Our study shows that there is
currently a time trend for an earlier yearly maximum in [O3] in the whole region covered by the study with no latitude
dependence. Preliminary results indicate that the rate of change in the timing of the yearly [O3] peak is smaller than
that in onset of the growing season. This suggests a development towards increasing risk for O3 damage to
vegetation in northern Scandinavia with respect to phenology and timing of the yearly [O3] peak, the reasons for
which will be further discussed in the presentation.

<figure><img src="image_11.png" /><figcaption>Figure 1. Day number of maximum daytime mean and 90-percentile of the ozone concentration in relation to latitude. For both ozone variables running two-week means were used. Both relationships are strongly significant (p<0.001).</figcaption></figure>

50

M32 ASSESSMENT OF TRACE ELEMENT POLLUTION AROUND
VLORA AREA, ALBANIA

Flora Qarri<fnref n="1" />, Sonila Kane<fnref n="1" />, Pranvera Lazo<fnref n="2" />

The environmental assessment of air and seawater pollution was carried out on coastals part of Vlora Bay, Albania. The air quality survey was carried out by using the moss Biomonitoring technique, where the moss Hypnum cupressiforme was employed as a long- term biomonitoring system. The urban area of Vlora city was investigated by using the active biomonitoring technique. The moss bags were exposed parallel with ditches for six months at nine sites in Vlora, along the main streets. Moss Biomonitoring at coastal and inland sites was used for investigation of the distribution of heavy metals in the air. The air deposition of mobilized traces metals was investigated by using ICP/AES analysis of moss samples. The seawater quality survey was also carried out from April-May 2011 and in February 2012 and heavy metals content in water samples from 15 sites in Narta and Orikumi lagoons and Vlora Bay were determined.

Heavy metals (Cu, Pb, Zn, Mn, Fe and Cd) in seawater and moss bag samples were determined via Atomic Absorption Spectrometry (flame, electrothermal systems). The area studied is moderately polluted as a result of high vehicular emissions and use of adulterated fuel in vehicles. Locations in the city were categorized on the basis of metal concentrations in the mosses and data statistical treatment. Comparison of exposed and unexposed moss helped assess the factors that adulterated the exposed moss samples. Correlation analysis helped determine the geochemically mobile elements and geochemically bound elements.

<footnote n="1">¹ Dep. of Chemistry, Faculty of Technical Sciences, Ismail Qemali University, Vlora, Albania [flora.qarri@gmail.com](mailto:flora.qarri@gmail.com)</footnote>

<footnote n="2">² Dep. of Chemistry, Faculty of Natural Sciences, University of Tirana, Tirana, Albania</footnote>
The heavy metal concentrations in seawater samples (Cd, Cr, Cu, Fe, Hg, Mn, Ni, Pb and Zn) were quite similar with the values reported for other regions in the European coastal waters. This study proved that the multivariate analysis is a powerful tool in environmental study. It was found that the Bay receives particulate matter, nutrient and dissolved element inputs from Vjosa and Shushica Rivers, as well from industrial, urban and agricultural activities along this river and the estuary around the Vlora Bay, that caused an increase of the concentration of Fe, Mn and Zn in the vicinity of the delta of the Vjosa River.

Harbors activities, such as dredging and cargo handling, the dumping of ship waste and other coastal activities are reflected on the concentrations of Fe, Cd, Cr, Cu and other metals in seawater of Vlora Bay, that cause anthropogenic pollution of the bay.

Some other anthropogenic factors such as atmospheric sources of dust generated by the erosion of land surfaces, wind transported dusts and by direct emission from anthropogenic activities, such as industry, agriculture and transportation, were found as less important factors for dissolved metals content in seawater of the Vlora Bay.

The dissolved form of the metals in the upper layer of seawater depended on weather conditions and red/ox process in seawater that may cause the variations in the concentration of dissolved form of heavy metals in seawater.

**Keywords:** active biomonitoring, urban area, seawater, heavy metals, accumulation factor, multivariate analysis, moderately polluted.

## M28 ATMOSPHERIC DEPOSITION OF HEAVY METALS TO TERRESTRIAL ECOSYSTEMS IN GERMANY

Schaap M.<fnref n="1" />, Wichink Kruit R.<fnref n="1" />, Builtjes P.<fnref n="1" />, Nickel. S<fnref n="2" />, Pesch R.<fnref n="2" />, Schröder W<fnref n="2" />., Nagel H-D<fnref n="3" />

Quantifying the atmospheric deposition of heavy metals to ecosystems remains a challenge as experimental data for many metals are few and, if present, emission estimates are highly uncertain. University of Vechta, TNO, and ÖKO-DATA started a project to establish the atmospheric deposition of heavy metals to terrestrial ecosystems in Germany and to identify ecosystems at risk. In total, 17 metals are in focus. As the information availability on the different metals differs substantially, the metals have been divided in three groups, indicating the effort put into their assessment from high to low.

Group A
Cd, Pb, Hg

Group B
As, Cr, Cu, Ni, Zn

Group C
V, Mn, Sb, Ti, Th, Co, Mo, Pt

The first step in the project is to quantify the atmospheric input to ecosystems. For this purpose two methodologies will be used. For all metals a first order assessment will be made based on experimental data. Available air concentration and wet deposition data will be classified as function of station type. Through combination of air concentrations with effective dry deposition velocities typical ranges for dry deposition fluxes can be estimated for each land use class for rural, urban and industrial areas. Combination with wet deposition data will provide first order estimates for the total deposition.

<footnote n="1">¹TNO, P.O. Box 80015, 3508 TA, Utrecht, the Netherlands, martijn.schaap@tno.nl</footnote>

<footnote n="2">²Chair of Landscape Ecology, University of Vechta, P.O. Box 1553, 49364, Vechta, Germany,</footnote>

<footnote n="3">³ÖKO-DATA GmbH, Hegermühlenstraße 58, 15344 Strausberg, Germany</footnote>
In the second methodology the chemical transport model LOTOS-EUROS will be used to calculate land use dependent dry deposition distributions across Germany for the metals that allow explicit modeling. Calculations will be performed using dedicated existing emission databases or through the modeling of PM in combination with metal contents. An example for the latter is the estimation of Vanadium levels based on modeled primary PM from heavy oil combustion. The observations from methodology 1 will be used for validation. As systematic biases are anticipated between model and observations the modeled fields may be scaled up or down towards the measured concentrations provided that the spatial gradients are resolved well. Wet deposition fluxes will be determined through *Residual Kriging* of observed fluxes.

The second step in the project will entail a comparison to other studies, e.g. EMEP, and an exploration to the use of
moss monitoring data to improve the deposition estimates. At the meeting we will present the set-up of the study and
illustrate the first results for deposition of Cadmium, Lead and Vanadium to German ecosystems.

<figure><img src="image_12.png" /><figcaption>Figure 1. Left: Modelled Cd concentrations (ng/m³) across Europe for the year 2010, Right: Observed Cd concentrations across Germany as function of station type for 3 years.</figcaption></figure>
M14 CORRELATIONS BETWEEN NITROGEN CONCENTRATIONS IN ATMOSPHERIC DEPOSITION AND MOSSES MAPPED FOR NATURAL LANDSCAPES IN EUROPE

Schröder W.<sup>a</sup>, Pesch R.<sup>a</sup>, Simon Schönrock S.<sup>a</sup>, Harmens H.<sup>b</sup>, Mills G.<sup>b</sup>, Fagerli H.<sup>c</sup>

<sup>a</sup> Chair of Landscape Ecology, University of Vechta, PO Box 1553, D-49356 Vechta, Germany, rpesch@iuw.uni-vechta.de, simon.schoenrock@uni-vechta.de, wschroeder@iuw.uni-vechta.de

<sup>b</sup> Centre for Ecology and Hydrology, Bangor, UK, hh@ceh.ac.uk; gmi@ceh.ac.uk

<sup>c</sup> Meteorological Synthesizing Centre-West of EMEP, The Norwegian Meteorological Institute, P.O.Box 43-Blindern, N-0313 Oslo, Norway, Hilde.Fagerli@nilu.no

Nitrogen (N) concentrations in mosses are primarily determined by atmospheric deposition. The correlations are country- and N compound-specific and agree well with spatial patterns and temporal trends across Europe as a whole and in single European countries. Following the methodology presented by Schröder et al. (2013) this study investigates whether correlations between the concentration of N in atmospheric deposition and mosses within the units of an ecological land classification of Europe can be established (Schröder et al. 2014). To this end, N measurements from the 2005 European moss survey and modelled N atmospheric deposition in 2005 were intersected with a map of European landscapes. Then, considering minimum numbers of sampling sites required across Europe, in single European countries and within the landscapes of Europe and accounting for spatial auto-correlation, the correlations between the N concentration in mosses and corresponding deposition were calculated and mapped for each of those landscape units containing moss sampling sites. Using an example of one landscape with positive correlation and one landscape with no correlation between N concentrations in deposition and in mosses, influencing factors were ranked based on investigating the multivariate interactions between moss concentrations and, amongst others, atmospheric deposition, land use, elevation or moss species by classification and regression trees. From this study it could be concluded that the numbers of sampling sites within Europe and most participating countries as well as within most of the landscapes covering Europe are sufficient for reliable statistics. Spatial patterns of correlations between the atmospheric N deposition and N concentration in mosses proven to vary across the landscapes of Europe. Where clear positive correlations between N concentrations in deposition and mosses exist in landscapes, multivariate ranking identifies the deposition as main influencing factor. In cases with no correlation between deposition and N concentrations in mosses, other factors such as e.g. moss species collected may be of importance. Therefore, mosses were proven to serve as biological indicators for atmospheric depositions and ecologically defined land classes could be identified as complex indicators which allow relating exposure monitoring with effects assessment. This investigation, enabling to quantitatively describe differences of correlations between N concentrations in atmospheric deposition and in mosses between landscapes, was accomplished by studies concentrating on spatial differences of N concentration in mosses at the site level. To this end the N concentration in mosses sampled at sites with and without canopy drip effects in several European states, amongst them Germany (Kluge et al. 2013). The respective results are presented in the talk given by Michaela Kluge.

## References

**Kluge M. et al. 2013.** Accounting for canopy drip effects of spatiotemporal trends of the concentrations of N in mosses, atmospheric N depositions and critical load exceedances: a case study from North-Western Germany. Environ Sci Europe 2013, 25:26 (13 pp + 4 suppl files) **Schröder W. et al. 2013.** Correlation between atmospheric deposition of Cd, Hg and Pb and their concentrations in mosses specified for ecological land classes covering Europe. *Atmosph Pollut Res* 4:267-274

**Schröder W. et al. 2014.** Mapping correlations between nitrogen concentrations in atmospheric deposition and mosses for natural landscapes in Europe. *Ecological Indicators* 36:563-571.
M16 INDICATIVE CONTENT OF N, S AND δ¹⁵N IN THE MOSS HYPNUM CUPRESSIFORME IN THE SOUTHERN PART OF CENTRAL EUROPE

Skudnik M. ¹,*, Kastelec D.², Jeran Z.³, Simončič P.⁴ & Batič F.²

¹ Department of Forest and Landscape Planning and Monitoring, Slovenian Forestry Institute, Večna pot 2, 1000 Ljubljana, Slovenia. mitja.skudnik@gozdis.si

² Department of Agronomy, Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, 1000 Ljubljana, Slovenia

³ Department of Environmental Sciences, Jožef Stefan Institute, Jamova 39, 1000 Ljubljana, Slovenia

⁴ Department of Forest Ecology, Slovenian Forestry Institute, Večna pot 2, 1000 Ljubljana, Slovenia

In years 2005/6 almost 3000 moss samples were collected in 16 European countries and used for N analysis (Harmens et al., 2008). With the goal of testing if mosses could be appropriately used to assess atmospheric N deposition, Harmens et al. (2011) compared the N content in mosses and modelled atmospheric N deposition (EMEP model). For countries in the south-eastern part of Europe, they found a weak relationship between EMEP modelled N deposition and N in mosses.

With the aim of evaluating the monitoring programme in the southern part of central Europe in 2010, we collected samples of Hypnum cupressiforme Hedw. at 18 sites near locations where the quality of deposition (bulk and throughfall) are also assessed. From 18 sites, 4 were near locations of Slovenian national quality of deposition sites and 14 near locations of UN-ECE ICP-Forest Level II plots in Slovenia and neighbouring countries (Austria, Italy and Croatia). To test the influence of site characteristics, especially of the canopy on element content in moss samples, mosses were additionally collected at 91 sites at intersections of the regular 8x16 km grid throughout the country. At each sampling site two types of samples were taken, namely below the canopy and in the open and analysed for N and S content and δ¹⁵N signature. Additionally, in the field the distances between moss collecting locations and the nearest tree crown were assessed.

Exploring the influence of atmospheric element deposition measured in bulk deposition on the element content in mosses showed that mosses collected in the open reflect atmospheric bulk N deposition, but only for cumulative one year deposition and not for the three year average. No influence of N in throughfall deposition on N in moss sampled under the canopy, nor influence of S deposition on S in moss (open and under canopy) was found. All sites showed no dependence between the δ¹⁵N values of the mosses and the NH4+/NO3 ratio in the deposition. However, if 4 of the 18 sites with a cumulative yearly precipitation of less than 1000 mm were excluded, the dependence was significant for the δ¹⁵N values in mosses collected in the open as well as under the canopy.

Evaluation of data for systematic sampling of mosses below the canopy and as far away as possible from the nearest tree from all 91 sites showed that canopy drip and litterfall bias the relationship for N and S, but not for δ¹⁵N. Significantly higher values of N and S were found in mosses under the canopy in comparison to those at least 1 m away from the nearest tree. Namely, the moss samples collected at a least 3 m distance from the nearest tree crown have on average 41% and 34% lower values of N and S, respectively, in comparison to those sampled below the canopy.

## References

Harmens, H., Norris, D., Cooper, D., Hall, J., 2008. Spatial trends in nitrogen concentrations in mosses across Europe in 2005/2006, Report on Nitrogen in European Mosses Work package 4. ICP Vegetation Programme Coordination Centre, Gwynedd, p.18.

Harmens, H. et al. 2011. Nitrogen concentrations in mosses indicate the spatial distribution of atmospheric nitrogen deposition in Europe. Environmental Pollution 159, 2852-2860.
M31 CROATIA PARTICIPATED IN THE ICP VEGETATION SURVEY SINCE 2005

Špirić Z. <fnref n="1" />, Stafilov T. <fnref n="2" />, Vučković I. <fnref n="2" />, Kušan V. <fnref n="1" />,
Barišić D, <fnref n="3" /> Vekić B. <fnref n="3" />, Šmit Z <fnref n="4" /> Glad M. <fnref n="5" />, Frontasyeva M. <fnref n="6" />

**Key words:** biomonitoring, moss, air, Croatia

Croatia participated 2006 and 2010 in moss survey in the framework of the International Cooperative Programme on Effects of Air Pollution on Natural Vegetation and Crops heavy metals in Europe. http://icpvegetation.ceh.ac.uk

Moss samples were collected during the summer/autumn of 2006 and 2010, from locations evenly distributed over the country. The most dominant moss species in this study area were *Hypnum cupressiforme*, *Pleurozium schreberi*, *Brachythecium rutabulum* and *Homalothecium sericeum*.

In addition to the comprehensive qualitative and quantitative chemical analysis of all samples collected (Špirić et al., 2012 and Špirić et al., 2013), laboratory research by using the Kjeldahl analytical method was conducted in order to determine nitrogen concentration in all collected moss samples. In addition to that, 22 out of 161 moss samples from 2010 were subjected to gamma-spectrometric analyses for assessing activity of the naturally occurring radionuclides. Also, some selected samples were analysed for PAHs, PCBs, PCDDs and PCDFs.

According to this research and in comparison of the results in 2006 with 2010, it is obvious that the state of anthropogenic pollution in the last five years in Croatia has not changed significantly, although it is obvious that the anthropogenic influence in decreasing.

**References**

Špirić, Z., Frontasyeva, M., Steinnes, E., & Stafilov, T. (2012). Multi-element atmospheric deposition study in Croatia. *International Journal of Environmental Analytical Chemistry*, *92*(10), 1200-1214.

Špirić, Z., Vučković, I., Stafilov, T., Kušan, V., & Frontasyeva, M. (2013). Air Pollution Study in Croatia Using Moss Biomonitoring and ICP-AES and AAS Analytical Techniques. *Archives of Environmental Contamination and Toxicology*, *65*:33-46.

<footnote n="1">¹ OIKON Ltd. – Institute for Applied Ecology, Trg senjskih uskoka 1-2, 10020 Zagreb, Croatia</footnote>

<footnote n="2">² Institute of Chemistry, Faculty of Natural Sciences and Mathematics, Ss. Cyril and Methodius University, Skopje, Macedonia</footnote>

<footnote n="3">³ Ruđer Bošković Institute, Zagreb</footnote>

<footnote n="4">⁴ Public Health Institute „Dr. Andrija Štampar“ Zagreb</footnote>

<footnote n="5">⁵ Teaching Institute of Public Health, Primorsko-Goranska County, Rijeka, Croatia</footnote>

<footnote n="6">⁶ Frank Laboratory of Neutron Physics, Joint Institute for Nuclear Research, Moscow, Russia</footnote>
M5 IMPACT OF O3 ON TERRESTRIAL ECOSYSTEMS: A MORE MECHANISTIC PARAMETRIZATION IN THE GLOBAL VEGETATION MODEL ORCHIDEE

T. Verbeke<sup>a</sup>, N. Viovy<sup>a</sup>, J-F. Castell<sup>b</sup>, J. Lathière<sup>a</sup>, S. Szopa<sup>a</sup>, D. Hauglustaine<sup>a</sup>

Tropospheric ozone is a major phytotoxic agent which can severely damage vegetation functioning. We propose a semi-mechanistic parameterization to integrate the impact of ozone on ecosystems in the dynamic global vegetation model ORCHIDEE. This study promotes an approach of ozone-induced effect on photosynthesis using flux-response relationships, in replacement to the former empirical function based on the external concentration AOT40 (Accumulated exposure Over a Threshold of 40 ppb). The new concept links a direct response of the maximum rate of photosynthesis Vcmax to the phytotoxic ozone dose (POD), also influenced by the accelerated senescence and countered by detoxification processes. Calibration of the parameters for C3 crops, grassland and deciduous forest was performed with sites measurements provided by field fumigation experiments. Furthermore, the correlation between observed and modelled relative reduction of Vcmax is higher basing the damage function on POD than on the ozone exposure indicator AOT40. As our photosynthesis scheme is coupled to a stomatal conductance module, we also compared the indirect effect on conductance with the impact simulated by the UNECE multiplicative conductance model, updated by the LRTAP 2010 convention. We found similar level of performance supporting the integration of the process-based flux-response module of impact in ORCHIDEE.

M10 SEVEN YEARS OF N- AND O3 POLLUTION × CLIMATE INTERACTION IN SUBALPINE GRASSLAND: ECOSYSTEM CARBON BUDGET, NITROGEN POOLS AND O3-FLUX MODELLING (POD)

Volk, M.*, Bassin, S.*, Wolff, V.*, Lehmann, M.**, Widmer, F.* and Fuhrer, J.*

We tested the effects of increased N and O3 deposition on subalpine grassland and the interacting effects of climate on the response to the above pollutants. The fully factorial field experiment had five levels of N deposition (0, 5, 10, 20 und 50 kg ha⁻¹ a⁻¹) and three levels of [O3] (ambient, amb.×1.4 and amb.×1.7) in a free air fumigation system.

**Carbon budget:** Climatic conditions and management change have led to increased productivity and increased soil C content in the Alp Flix grassland. The N-treatment increased productivity, while O3-treatment did not affect plant growth. Net ecosystem exchange measurements, covering two complete years of contrasting climate, detected extreme short-term effects on the CO₂ C-balance, with antagonistic climate effects on ecosystem respiration and gross primary productivity. Carbon isotope analysis revealed that O3- and N-treatment made soil air, root material and bulk soil δ¹³C less negative, but independent of the pollutant deposition treatment bulk soil became more negative over the years. While the bulk soil overall C content increased over time, soil density fractionation revealed a decreased C content in the oldest soil fraction.

<footnote><sup>a</sup>Laboratoire des Sciences du Climat et de l'Environnement, Gif-sur-Yvette, France. [thomas.verbeke@lsce.ipsl.fr](mailto:thomas.verbeke@lsce.ipsl.fr)</footnote>

<footnote><sup>b</sup>AgroParisTech, UMR 1091 EGC, F-78850 Thiverval-Grignon, France.</footnote>

<footnote>* Agroscope Zürich, Air Pollution/Climate Group, Zurich, Switzerland [matthias.volk@agroscope.admin.ch](mailto:matthias.volk@agroscope.admin.ch)</footnote>

<footnote>**Institute for Environmental Geoscience, University of Basel, Basel, Switzerland</footnote>
In summary, single parameters of climate and atmospheric pollution are not sufficient to predict aboveground yield responses, and yield responses are not sufficient proxies for the ecosystem C-budget. Instead, the interaction of multiple parameters needs to be evaluated.

**Nitrogen pools:** N contents were calculated for green phytomass, necromass, roots, microbial biomass, soluble soil N, extractable soil N, and immobilised soil N. In addition, soil microbial communities were analysed by parallel 454 pyrosequencing of bacterial and fungal ribosomal sequences tags. N deposition increased all plant N pools: +38% N in green phytomass, +73% in necromass and +33% in root biomass (growing season average). Thus, added N was primarily stored in phytomass. In contrast, most of the N pools were unaffected by O₃ exposure; only soil microbial biomass N was slightly increased by 0.85 g m⁻² (+8%). However, a highly significant O₃ x N interaction was found for root N pool, revealing a negative effect of O₃ in unfertilized plots and a positive effect of O₃ in fertilized plots. A similar response pattern was observed in microbial biomass and soluble soil N, potentially related to changes in nutrient allocation and/or altered root and litter decomposition. However, the first reason can be excluded since the belowground responses were not accompanied by reciprocal changes in aboveground N pools. Rather, decomposition processes were affected, as indicated by the strong separation of fungal and bacterial communities among the four treatment combinations. However, a straightforward explanation for the interactive O₃ x N effect is missing yet.

**Ozone flux modelling:** On average, POD0 and POD1 were increased by a factor of 1.33 and 1.37 (O₃+) and 1.83 and 1.91 (O₃++), respectively, compared to the O₃ control treatment. Both maximum (2006) and minimum (2009) POD values occurred in the two warmest and driest years, respectively. In high O₃, POD reached values found to be detrimental to plant growth elsewhere. Thus, based on our data, the high O₃ tolerance of the investigated vegetation cannot be explained by limited stomatal O₃ uptake.
## 5.3 POSTERS

### EVALUATION OF PINOLENE AS PROTECTANT AGAINST OZONE PHYTOTOXICITY

Evgenios Agathokleous<fnref n="1" />, Costas J. Saitanis<fnref n="1" />, Yolanda Papatheohari<fnref n="2" />

**Introduction** : Tropospheric ozone (O3), a widespread air pollutant, constitutes a real menace to plants, threatening food security. A major still unsolved issue is the lack of some appropriate (in terms of effectiveness and cost) protectant of plants against ozone that would be widely used in the agricultural practice. EDU is an effective protectant but is not commercially available, and it is quite expensive to be produced for laboratory use. Pinolene (Di-1-p-menthene) has been suggested as a potential protectant.

**Purpose** : In the framework of this research we evaluated the efficacy of EDU and Vapor Gard (containing 96% pinolene) to protect plants of the sensitive to O3 Bel-W3 tobacco (*Nicotiana tabacum* L.) plants against ozone.

**Materials and methods** : Two months old plants were exposed either to ozone ([O3+]: 90 ppb x 12 days x 8 hours/day) or to ozone free air ([O3-]) and treated either by Water (control group) or by EDU (at 10, 100 and 500 mg/lit) or by Vapor (at 1, 5, 10 and 50 ml/lit). The measured parameters were: leaf area (LA), foliar visible injury (VI), plant height (Height) & dry weight (DW), total chlorophylls (ChlT) & total carotenoids (Car), ChlT/Car, Chlα/Chlβ ratio, quantum yield of photosynthesis (QY), leaf greenness (SPAD) and stomatal resistance (SR).

**Results and discussion** : EDU, when applied at low concentration (10 mg/lit) did not protect the plants against O3. However, the plants treated by the higher EDU concentrations (100 and 500 mg/lit) exhibited much fewer or not at all symptoms: plants were protected in all the measured parameters. In contrast, pinolene only at 1ml/lit prevented ozone-induced reduction of the parameters LA, DW, ChlT and Car. The ChlT did not differ between [O3+] and [O3-] groups. Pinolene, at 50 ml/lit, caused reduction of height and ChlT in both [O3+] and [O3-] groups. In [O3+] group, all of the subgroups of plants treated by substances (except those treated by EDU at 500mg/lit) the Chl/Car ratio was reduced when compared pairwise with that of the corresponding subgroups in the [O3-], suggesting a higher reduction of carotenoids in comparison with chlorophylls; however, this difference was statistically significant only in plants treated by pinolene at 10 ml/lit. Chlorophylls α/β ratio did not differ neither between [O3+] and [O3-] groups nor among the substances subgroups. As to the QY, Car and SR, pinolene did not exhibit any effect. Concerning SPAD within the [O3+] group, only those plants treated by pinolene 1 ml/lit had statistically significantly higher SPAD value in comparison with those sprayed only with water. However, pinolene, when applied at 10 and 50 ml/lit caused phytotoxic symptoms - necrotic spots very similar to those caused by the O3; these phytotoxic symptoms were developed only a few hours after the pinolene application. At the end of the experiment (after 12 days) the plants injured by the highest concentration of pinolene were shorter and stunted with distorted leaves.

**Conclusions** : EDU remains the best available protectant against ozone; pinolene may has some protective role on Bel-W3 plants at low concentrations (1 ml/lit) that should be further investigated for several plant species and ozone levels. At higher concentrations (10 and 50 ml/lit) pinolene is phytotoxic, at least to Bel-W3 tobacco plants.

<footnote n="1">¹Lab of Ecology and Environmental Science, Agricultural Univ. of Athens, Iera Odos 75, 11855, Athens, Greece</footnote>

<footnote n="2">²Lab of Agronomy, Agricultural Univ. of Athens, Iera Odos 75, 11855, Athens, Greece e-mail address: evgenios_ag@hotmail.com</footnote>
# AIR POLLUTION STUDY IN MACEDONIA BY USING MOSS BIOMONITORING TECHNIQUE, NAA, ICP-AES, AND AAS

Barandovski L.<fnref n="1" />, Frontasyeva M.V.<fnref n="2" />, Stafilov T.<fnref n="3" />, Šajn R.<fnref n="4" />, Bačeva K.<fnref n="3" />, Dmitriev A.Yu<fnref n="2" />

The third moss survey in the Republic of Macedonia took place in August and September 2010 when 72 samples of the terrestrial mosses *Homalothecium lutescens* and *Hypnum cupressiforme* were collected over the whole territory of the country using the same sampling network grid as for the previous surveys in 2002 and 2005. Using neutron activation analysis (NAA), inductively coupled plasma-atomic emission spectrometry (ICP-AES), and atomic absorption spectrometry (AAS), a total of 47 elements (Al, As, Au, Ba, Br, Ca, Cd, Ce, Cl, Co, Cr, Cs, Cu, Dy, Eu, Fe, Hf, Hg, I, In, K, La, Li, Mg, Mn, Mo, Na, Nd, Ni, P, Pb, Rb, Sb, Sc, Se, Sm, Sr, Ta, Tb, Th, Ti, U, V, W, Yb, Zn, and Zr) were determined. Distribution maps were prepared to point out the regions most affected by pollution and related to known sources of contamination. To reveal hidden multivariate data structures and to identify and to characterize different pollution sources Principal Component Analysis (PCA) was used. A comparison of the results obtained in 2010 and 2005 [1, 2] surveys was carried out to examine temporal trends in the elemental depositions.

## References:

1. L. Barandovski, M. V. Frontasyeva, T. Stafilov, R. Šajn, S. Pavlov, V. Enimiteva, Trends of atmospheric deposition of trace elements in Macedonia studied by the moss biomonitoring technique, *Journal of Environmental Science and Health, Part A*, **47**(13), 2000-2015 (2012).

2. L. Barandovski, T. Stafilov, R. Šajn, M.V. Frontasyeva, K. Bačeva, Air pollution study in Macedonia by using moss biomonitoring technique, ICP-AES and AAS, *Macedonian Journal of Chemistry and Chemical Engineering*, **32**(1), 89-107 (2013).

# ATMOSPHERIC DEPOSITION OF HEAVY METAL IN EASTERN ALBANIA BY MOSS BIOMONITORING TECHNIQUE

L. Bekteshi<fnref n="1" />, F. Qari<fnref n="2" />, P. Lazo<fnref n="3" />, T. Stafilov<fnref n="4" />

The moss biomonitoring technique was applied for assessing environmental situation of air pollution in eastern Albania. The carpet-forming-mosses have some particular properties that make them suitable for monitoring air pollutants. Sampling was performed in accordance with the LRTAP convention-ICP protocol and sampling strategy of the European Program on Biomonitoring of HM Atmospheric Deposition. The *Hypnum cupressiforme* moss species were used as bioindicator for air pollution, that were collected during June-July, 2011 at 19 sites distributed over the eastern

<footnote n="1">¹ Institute of Physics, Faculty of Natural Sciences and Mathematics, Ss Cyril and Methodius University, POB 162, 1000 Skopje, Macedonia</footnote>

<footnote n="2">² Frank Laboratory of Neutron Physics, Joint Institute for Nuclear Research, 141980 Dubna, Moscow Region, Russia; e-mail: marina@nf.jinr.ru</footnote>

<footnote n="3">³ Institute of Chemistry, Faculty of Natural Sciences and Mathematics, Ss Cyril and Methodius University, POB 162, 1000 Skopje, Macedonia; e-mail: trajcest@pmf.ukim.mk</footnote>

<footnote n="4">⁴ Geological Survey of Slovenia, Dimičeva ul. 14, 1000 Ljubljana, Slovenia</footnote>

<footnote n="1">¹ University of Elbasan. lirimbekteshi@ymail.com</footnote>

<footnote n="2">² University of Vlora</footnote>

<footnote n="3">³ University of TiranaTirana, Albania</footnote>

<footnote n="4">⁴ Institute of Chemistry, Faculty of Science, Sts. Cyril and Methodius University, Skopje, Macedonia</footnote>
part of Albania. A total of 19 elements (Al, As, Cd, Cr, Cu, Fe, Ni, Pb, V and Zn) were determined by ICP-AES technique. The results reflect local emission point.

<figure><img src="image_13.png" /><figcaption>Figure 1 Normal distribution of Al, As, Cd, Cr, Cu, Fe, Ni, Pb, V and Zn</figcaption></figure>

<figure><img src="image_14.png" /></figure>

These elements show high variability compare to their mean values. High values of skewness and curtosis are also evident that mean the data are strongly skewed, are not normal distributed, except Al (Fig. 1, P<0.05). The data matrix was treated statistically by using Minitab 15 software. Factor analysis with VARIMX rotation was used to identify and characterize different pollution sources and to identify the areas receiving the highest metal pollution load. The data of the factor analysis were applied to distinguish elements mainly of anthropogenic origin from those predominantly originating from natural sources.

Geogenic factor, wind soil dust and weather conditions were identified as main factors of contamination of this area.

**Key word:** mosses, biomonitoring, heavy metal, ICP/AES, factor analysis, Albania
# PHYSIOLOGICAL RESPONSES: DO LICHENS, MOSSES AND TREE BARK REACT WITH DIFFERENT CARBON AND NITROGEN ISOTOPE PATTERNS ALONG A NITROGEN DEPOSITION GRADIENT?

Boltersdorf S. H.¹, Pesch R.², Schröder W.², Werner, W.¹

¹ University of Trier, Department of Geobotany, Behringstr. 21, 54296 Trier, Germany.
stefanie.boltersdorf@gmx.de, werner@uni-trier.de

² University of Vechta, Chair of Landscape Ecology, PO Box 1553, 49364 Vechta, Germany. wschroeder@iuw.uni-vechta.de, rpesch@iuw.uni-vechta.de

Mankind has dramatically influenced the N fluxes between soil, vegetation, water and atmosphere, the global N cycle. Many problems are caused by the human-accelerated environmental change. Long-term ecological monitoring programs are fundamental to evidence-based environmental decision-making. Biomonitoring is essential, particularly where less deposition measurement sites exist. In Germany still key societal threats of excess reactive nitrogen (N<sub>r</sub>) are on hand.

An overview is given of the physiological response of different bioindicators towards different anthropogenic N impacts in the environment. Abundance of ¹³C and ¹⁵N contents and total C and total N of lichens, mosses and tree bark, were reported.

Lichen, moss and bark samples were collected at 16 deposition measurement sites of the Air Monitoring Network of the Federal Environment Agency of Germany (UBA) in September and October 2008 (lichens and tree bark samples) and from September to October 2006 (moss samples). The investigation has tested 442 lichen samples, 203 tree bark samples and

16 moss samples and considered the epiphytic lichen species *Hypogymnia physodes*, *Parmelia sulcata*, *Xanthoria parietina*, *Physcia tenella* and *Physcia adscendens* (pooled *Physcia* sp.), the moss species *Hypnum cupressiforme*, *Pleurozium schreberi* and *Pseudoscleropodium purum*, and the tree species *Acer platanoides*, *Acer pseudoplatanus*, *Alnus glutinosa*, *Betula pendula*, *Carpinus betulus*, *Corylus avellana*, *Fraxinus excelsior*, *Malus domestica*, *Pinus sylvestris*, *Populus × canadensis* agg., *Prunus avium*, *Prunus domestica*, *Quercus petraea*, *Quercus robur*, *Salix* sp., *Sorbus torminalis*, *Tilia cordata* and *Tilia platyphyllos*. The various tree species were not taken into account separately.

Isotope specificity of species and of taxonomic groups relating to δ¹⁵N and δ¹³C values were indicated, probably due to differences in plant physiology and biochemistry.

All bioindicators have shown an increase of the N content and simultaneously a decrease of the C/N ratio while enhanced agriculture-related N impact exist. C pool turnover rates greatly increased in response to N impact which point to N deposition as a stress factor.

Furthermore, lichens and bark samples were characterized by an increase of less depleted δ¹⁵N values in highly agricultural areas which proves the existence of particulate ammonium, originating from agricultural immissions.
RELATIONS BETWEEN ACCUMULATION OF TRACE ELEMENTS, PHOTOSYNTHETIC ACTIVITY, WATER STATUS AND GROWTH PARAMETERS OF ITALIAN RYEGRASS EXPOSED IN AMBIENT AIR CONDITIONS

Borowiak K.<fnref n="1" />, Zbierska J.<fnref n="1" />, Barałkiewicz D.<fnref n="2" />, Hanć A.<fnref n="2" />, Kayzer D.<fnref n="3" />, Budka A.<fnref n="3" />

The aim of the study was to evaluate an effect of heavy metals to Italian ryegrass (Lolium multiflorum L.) exposed in ambient air conditions. Plants were cultivated for six weeks in the greenhouse conditions and afterwards were transported to five exposure sites located in Wielkopolska region. Two sites were located in the Poznan city, one in suburban area, one in Landscape Park and the last one in the rural area. Plants were exposed for four weeks and four exposure series were performed in 2012 growing season, beginning in the middle of May. Selected trace elements (Ni, Cr, As, Cd, Pb) were analysed before and after exposure, as well as relative water content (RWC) and specific leaf area (SLA). While photosynthesis activity parameters were measured before, in the middle and at the end of every exposure series.

The experiment revealed an increase of leaf thickness due to increased heavy metal concentrations in ambient air, while higher trace element concentrations caused a decrease of relative water content of exposed plants. Hence, based on our results, we can conclude that trace element concentrations in ambient air were high enough to affect *Lolium multiflorum* L. water relations, but too low to influence leaf blade thickness.

Net photosynthesis rate and stomatal conductance were mostly negatively related with all analysed trace elements, while intercellular CO2 concentrations revealed opposite tendency. However, there were some exceptions, such as for arsenic in two exposure series, when lack or low relations with PN and gs were observed. The lowest levels of PN and gs were mostly noted at two sites located in the city, where high concentrations of Cd and Pb was recorded in L. multiflorum leaves. Moreover, the lowest levels of PN were noted at all sites during third exposure series, when the highest Cr and Ni levels were recorded.

Acknowledgement

Presented investigations were supported by project grant No.: N 0540/B/P01/2011/40 from the National Science Centre.

<footnote n="1">¹Department of Ecology and Environmental Protection, Poznan University of Life Sciences,
ul. Piątkowska 94C, 60-649 Poznań, Poland</footnote>

<footnote n="2">²Department of Trace Element Analysis by Spectroscopy Method, Adam Mickiewicz
University, ul. Grunwaldzka 6, 60-780 Poznań, Poland</footnote>

<footnote n="3">³Department of Mathematical and Statistical Methods, Poznan University of Life Sciences, Wojska Polskiego 28, 60-637 Poznań, Poland</footnote>
# MAPPING OF AIR POLLUTION ROADSIDE ORIGIN BY SOME TRACE METALS (CD AND CU) USING THE TRANSPLANTATION OF A BIO ACCUMULATOR LICHEN XANTHORIA PARIETINA IN THE CITY OF TIARET (ALGERIA)

Bouchenafa N¹, Mazouz S² Dellal A¹

¹ Laboratoire Agro-Biotechnologie- Université Ibn khaldoun - Tiaret

² Direction de l'Environnement - Tiaret

This study seeks to quantify air pollution by two trace metals Cd and Cu picked by lichen Xanthoria parietina bio
accumulator in the city of Tiaret (Algeria). This measure of pollution is by using the method of atomic absorption
spectrophotometry provides results highlighting low levels of air pollution by the two elements mentioned above. This
does not mean that air pollution is absent but that these two trace elements are spread by wind over the rugged
terrain of the region.

**Keywords** : pollution, trafic, Cu, Cd, lichen, Xanthoria parietina, bio accumulation, Tiaret, Algérie

TEMPERATURE EFFECTS ON YIELD OF R123 AND S156 SNAP BEANS – ESTABLISHING
CRITERIA FOR EFFECTIVE USE AS AN OZONE BIO-INDICATOR SYSTEM

Kent O. Burkey

USDA-ARS Plant Science Research Unit, 3127 Ligon Street, Raleigh, NC, 27607 USA
[Kent.Burkey@ars.usda.gov](mailto:Kent.Burkey@ars.usda.gov)

Snap bean recombinant inbred lines derived from a cross between ozone (O3)-sensitive and tolerant parents (Reinert and Eason, 2000) are being tested as an O3 bio-indicator system. The S156 (O3-sensitive) and R123 (O3-tolerant) snap bean genotypes were originally identified based on similar yield under clean air conditions and differential yield responses reflected in declining S156/R123 yield ratios when grown in elevated O3 environments. The S156/R123 pair has been tested previously in open-top chambers (Burkey et al., 2000) and is currently being utilized in the ICP-Vegetation snap bean project.

While S156 and R123 consistently exhibit differential foliar injury in response to O3 exposure, differential yield responses to assess O3 impacts appear to be more complex. At the 2013 Task Force meeting, we reported observations that suggested high temperature may limit mature pod formation in a manner that could complicate interpretation of O3 impacts in terms of S156/R123 yield ratios. To directly test this hypothesis, mature pod yield in the absence of ozone was measured over a range of growth temperatures. Mature pod yield declined significantly at a growth temperature of 32°C and was practically zero at a growth temperature of 36°C (Table 1). The decline in mature pod yield at increasing growth temperature was associated with large increases in small undeveloped pods (Table 1), supporting the hypothesis that high temperatures can inhibit pollination and limit pod yield potential. Given that ambient O3 is highest in the summer months, the use of the S156/R123 pod yield ratio as an O3 bio-indicator may not be appropriate in regions with average temperatures high enough to impact mature pod development. Foliar injury ratings may still be useful in these situations.
<table><thead><tr><th rowspan="2">Target T (D/N) (°C)</th><th rowspan="2">Actual T (D/N) (°C)</th><th colspan="2">Pod Yield (g/plant)</th><th rowspan="2">Yield ratio</th><th colspan="2">Undeveloped pods/plant</th></tr><tr><th>S156</th><th>R123</th><th>S156/R123</th><th>S156</th><th>R123</th></tr></thead><tbody><tr><td>24/20</td><td>24/21</td><td>68±10</td><td>79±6</td><td>0.87±0.20</td><td>13±1</td><td>9±2</td></tr><tr><td>28/24</td><td>27/25</td><td>48±8</td><td>64±13</td><td>0.80±0.29</td><td>20±4</td><td>32±4</td></tr><tr><td>32/28</td><td>32/28</td><td>23±6</td><td>18±1</td><td>1.30±0.25</td><td>28±4</td><td>115±63</td></tr><tr><td>36/32</td><td>34/31</td><td>3±3</td><td>1±1</td><td>----</td><td>73±7</td><td>71±12</td></tr></tbody></table>

Table 1. Snap bean genotypes S156 and R123 were grown from planting through harvest in charcoal-filtered air (≤ 2 ppb O3) under 4 temperature regimes during the fall of 2013. Plants were grown in 15-liter pots of Sunshine MVP mix with Osmocote Plus slow-release fertilizer using Outdoor Plant Environment Chambers (OPECs) to control day (D)-night (N) temperature (T). The experimental design (4 T x 2 blocks) considered each of the eight OPECs as an experimental plot with 4 plants per genotype randomly assigned within each plot. Pod yield (mature pods with seeds) and counts of undeveloped pods were measured at 82 days after planting. Values are chamber means ± standard error (n = 2).

## References

Burkey, K.O., J.E. Miller, and E.L. Fiscus (2005) J Environmental Quality 34: 1081-1086
Reinert, R.A. and G. Eason (2000) J American Society of Horticultural Science 125: 222-2

# OZONE: AN ELICITOR OF SECONDARY COMPOUNDS IN HYPERICUM PERFORATUM?

Campanella A., Lorenzini G., Nali C.

Department of Agriculture, Food and Environment, University of Pisa, Via del Borghetto 80,
56124 Pisa, Italy; e-mail: cristina.nali@unipi.it

Ozone (O3) is considered the most ubiquitous and phytotoxic air pollutant in industrialized and in developing countries. Many secondary metabolites are active compounds of medicinal plants and show a wide range of biological activities. They play important role in the adaptation of the plant to the environment, contributing to its resistance to biotic and abiotic stresses. It is interesting to study the interaction of O3 with wild medicinal plants, with special regard to the effect on the secondary metabolism. In this work, attention has been given to a species of great scientific and economic interest, *Hypericum perforatum* L.. Commonly called St. John's wort, it is an herbaceous perennial plant, long known for its putative medicinal properties. Its macerate in oil has been traditionally used as a vulnerary to treat minor burns, wounds, skin inflammation and nerve pain, but its alcoholic extracts are gaining increasing popularity as an alternative treatment for mild to moderate depression. Considering O3 as elicitor of secondary metabolites, the study evaluated the change of bioactive compounds (flavonoids, the naphthodianthrone hypericin and the acylphloroglucinol hyperforin) of plants obtained from local germplasm in vegetative stage. Six-month old plants were exposed under controlled environmental conditions to O3 (100 ppb, 5 h); control plants were maintained in charcoal filtered air. Samples were taken immediately at the end of treatment and after 24 h. In all cases, there were no visible symptoms. The search of the metabolites was performed by HPLC on methanolic extracts obtained from apical shoots. Twenty-four h after the end of the exposure, it was observed a significant increase of hyperforin and