Water Air Soil Pollut
DOI 10.1007/s11270-007-9376-2
Ozone Biomonitoring with Bel-W3 Tobacco Plants
in the City of Valencia (Spain)
Vicent Calatayud & María José Sanz &
Esperanza Calvo & Júlia Cerveró &
Wolfgang Ansel & Andreas Klumpp
Received: 23 November 2006 / Accepted: 16 February 2007
# Springer Science + Business Media B.V. 2007
Abstract A biomonitoring study using the ozonesensitive bioindicator plant Nicotiana tabacum cv.
Bel-W3 was conducted in the city of Valencia (eastern
Spain) and surrounding areas in 2002. Plants were
exposed to ambient air at seven sites, including four
traffic-exposed urban sites, a large urban garden and
a suburban and a rural station, for six consecutive
2-week periods using highly standardised methods.
Foliar injury was registered at all stations in at least
one of the exposure periods. The urban stations
submitted to intense traffic showed lower ozone
injury than the less traffic-exposed stations. Strong
changes in the intensity of ozone injury were
observed for the different exposure periods. Leaf
injury was significantly related to both mean ozone
values (24 and 12 h means) and cumulative exposure
indices (AOT20, AOT40). However, correlation
strength was moderate (rs =0.39 to 0.58), suggesting
that the plant response to ozone was modified by
V. Calatayud (*) : M. J. Sanz : E. Calvo : J. Cerveró
Fundación CEAM, Parc Tecnològic,
c/ Charles Darwin 14,
46980 Paterna, Valencia, Spain
e-mail: [email protected]
W. Ansel : A. Klumpp
Institute for Landscape and Plant Ecology and Life Science
Center, University of Hohenheim,
70593 Stuttgart, Germany
environmental factors. The use of sensitive bioindicators like tobacco Bel-W3 in cities provides complementary information to that of continuously
operating air quality monitors, as the impact of
ambient ozone levels is directly measured.
Keywords Air quality . Bioindicators . Tobacco
Bel-W3 . Urban air pollution
1 Introduction
Ozone is a major component of photochemical smog
and is formed through complex photochemical reactions involving hydrocarbons and nitrogen oxides as
precursors (Chameides and Lodge 1992). The Mediterranean area is characterized by intense solar
radiation, high temperatures, and re-circulation of
polluted air masses (Millán et al. 1997, 2000; Sanz
and Millán 1998) favouring the formation of this
secondary pollutant. In some parts of this area, ozone
levels are high enough to cause phytotoxic effects
both in crops (Fumagalli et al. 2001) and in native
sensitive plant species (Sanz and Millán 2000).
Several plant species that develop visible injury have
been used as bioindicators of ozone. Among them,
ozone-sensitive tobacco cv. Bel-W3 is the most
commonly used worldwide, and its response to ozone
is the best described. In Europe, this tobacco cultivar
has been used in biomonitoring studies at different
Water Air Soil Pollut
scales, from extensive areas (e.g., covering the whole
British Isles, Ashmore et al. 1978) through more
limited regional or local surveys (e.g., Godzik 1997;
Lorenzini et al. 1995; Ribas and Peñuelas 2003), to
urban areas (Klumpp et al. 2006b).
The European bioindicator programme EuroBionet
(LIFE Environment Programme) aimed at assessing
and evaluating air quality in 12 cities of eight
countries during the period 2000–2002 by the use of
various bioindicator plant species (Klumpp et al.
2002; Klumpp et al. 2004, http://www.eurobionet.
com). The participant cities exposed, in parallel and at
established exposure periods, the following bioindicator plants: tobacco Bel-W3 and Populus nigra
‘Brandaris’ (to assess the effects of ozone), Lolium
multiflorum (for sulphurous compounds and metals),
Tradescantia sp. clone #4430 (for mutagenic substances) and Brassica oleracea acephala (for hydrocarbons). This paper deals with the results of the
tobacco exposure experiment conducted in the city
of Valencia within this programme. Valencia joined
the EuroBionet network in 2001, but a complete
series of tobacco exposures covering from May to
the end of August was only achieved in 2002;
consequently, the analyses will be restricted to this
year. Previous results of the tobacco experiments
within the whole pan-European biomonitoring network focusing on data obtained in 2000/2001 have
already been published by Klumpp et al. (2006b),
but data from the city of Valencia were not included.
Valencia is a flat coastal city, with about 1,200,000
inhabitants and 445,000 vehicles in its metropolitan
area. The urban area is characterized by relatively
high levels of primary pollutants and relatively
moderate ozone levels. Pollutant dynamics in the
coastal area are strongly influenced by the sea-breeze,
which transports pollutants inland (western direction)
along the Turia valley, so that higher ozone levels
than in the urban area are reached in suburban areas
and especially at about 30–50 km inland, downwind
from the city. In 2002, air quality in the city and
surrounding areas was continuously recorded at eight
monitoring stations (RVCCA, http://www.cma.gva.es);
all eight included ozone monitors, but only one
Table 1 Tobacco biomonitoring stations (TBS) established in the city of Valencia and surrounding areas, type of station, co-ordinates,
closest representative continuous air monitoring station (MS) and distance between TBS and representative MS
Tobacco biomonitoring stations [TBS]
Closest continuous air monitoring station [MS]
Number Tobacco stations
Type
Co-ordinates
Location of MS
Official
name
Distance between
MS and TBS
1
Avd. Aragón
Aragón
<10 m
Jardí de Vivers
Same garden as TBS
Vivers
c. 100 m
3
Prof. Beltrán Báguena
Same square as TBS
Plaza Manuel Granero
5
G.V. Fdo. Católico
Nuevo
Centro
Pista de
Silla
Linares
<10 m
4
6
Devesa de l'Albufera
–
–
7
Campus Univ.
València – Burjassot
N: 39° 28′ 37,20204″
W: 0° 21′ 17,64735″
N: 39° 28′ 54,59709″
W: 0° 22′ 00,13493″
N: 39° 28′ 51,74849″
W: 0° 23′16,04628″
N: 39° 27′ 34,87818″
W: 0° 22′ 31,00726″
N: 39° 28′ 06,75352″
W: 0° 22′ 54,57769″
N: 39° 21’ 31,46882″
W: 0° 19’ 26,268920″
N: 39° 30′ 30,82025″
W: 0° 24′ 58,568007″
Same street as TBS
2
Urban street,
traffic
Large urban
garden
Urban street,
traffic
Urban street,
traffic
Urban street,
traffic
Rural
Paterna
c. 2.5 km
Suburban
Same square as TBS
In a street near to TBSa
Representative MS not
available
In a relatively nearby
suburban townb
<10 m
c. 500 m
a
Parallel measurements (out of the time window of this study) from this MS and another MS operating in G.V. Fdo. Católico until
2001 showed that both MS had almost identical ozone values and daily cycles (CEAM, unpublished data).
b
This MS is considered representative of the ozone levels recorded c. 2.5 km away at the University of Burjassot, where the tobacco
station was installed. Both sites are placed in a suburban area W of Valencia, and existing periods of parallel measurements (out of the
time window of this study) between both sites with monitors showed very similar daily profiles and ozone levels, higher than in streets
inside the city of Valencia (CEAM, unpublished data).
Water Air Soil Pollut
recorded meteorological data. For the present study with
tobacco as a bioindicator plant, we selected seven
exposure sites designed to cover spatially different
zones of the city and to include stations with intense
traffic, gardens, a suburban site and a rural area. Six of
these stations were placed next to some of the above
mentioned air quality monitoring stations.
The objectives of the present study were to
evaluate the phytotoxic effects of ozone on bioindicator Bel-W3 tobacco plants within the city of
Valencia and its surrounding areas, to determine
which areas have the highest impact, and to compare
leaf injury with ozone measurements of the automated
monitoring network. Another important objective of
the project was to increase the environmental awareness of the citizens. To achieve this latter objective,
information on the effects of ozone and other
pollutants was provided to the general public at each
monitoring site, results from the exposure series in all
the cities of Europe were regularly posted in a stand
(a green box, see http://www.eurobionet.com) specially designed for, posters and leaflets were distributed, and several schools were involved in different
activities (Calatayud and Sanz 2004).
Fig. 1 Location of biomonitoring sites in the city of
Valencia and surrounding
towns (in grey). Details of
the biomonitoring sites are
given in Table 1
2 Materials and Methods
2.1 Distribution of Biomonitoring Stations and Air
Quality Stations
Characteristics of the seven tobacco exposure sites are
described in Table 1, and their spatial distribution is
provided in Fig. 1. A further biomonitoring site
originally included in this study was removed from
the dataset due to exposure problems.
2.2 Cultivation and Exposure of Tobacco Plants
Seeds of the ozone-sensitive tobacco cultivar Bel-W3
were obtained from the State Institute for Crop
Production (Landesanstalt für Pflanzenbau, Rheinstetten,
Germany). Plants were cultivated in a local greenhouse using a mixture of commercially available,
standardised soil type ED73 and river sand (8:1 by
volume), plastic pots (1.5 L) and a semi-automatic
watering system made of glass fibre wicks and water
containers. The procedure employed largely corresponded to a method described in a draft version of
the guideline of the German Association of Engineers
Water Air Soil Pollut
(Verein Deutscher Ingenieure 2003, Guideline 3957
Part 6). At each exposure site, six plants were
exposed to ambient air on exposure racks (exposure
height 90 cm, frame height 180 cm) as originally
described by Arndt et al. (1985). The top and three
sides of the rack were covered with green shading
fabric (50%), while the northern side was left open.
During outdoor exposure, all plants were irrigated by
the same system of suction wicks hanging into water
reservoirs. Plant exposure started on 29 May and
ended on 22 August 2002, with the following six
consecutive 2-week exposure series: 29 May–11 June,
14–28 June, 28 June–11 July, 11 July–25 July, 25
July–8 August, and 8 August–22 August. After each
exposure series, leaf injury was assessed, and all
plants were replaced by new ones.
2.3 Visible Injury Assessment
Visual assessment of foliar injuries was performed in
5% steps by means of a photo catalogue (Klumpp et
al. 2000; Verein Deutscher Ingenieure 2003) with
exemplary images of damaged leaves. Injury was
recorded on reference leaves (leaves no. 4–6) of each
plant immediately before exposure, and at the end of
the two-week exposure period. The mean percentage
of leaf injury was calculated for each of the
bioindicator stations. Initial injury values before
exposure were taken into account in such calculations.
from 8:00 CET to 20:00 CET. For correlating these
cumulative indices with foliar injury to tobacco, two
periods (series 3 and 4) from station 3 were not
considered, as the data cover was below 50%.
Commonly, the valid monitor data cover at each
station for any single 2-week exposure was over 95%,
and it was always above 75% except in the two cases
mentioned.
2.5 Statistical Analyses
A t test analysis was applied to test for differences
between the two main types of biomonitoring stations
(‘traffic’ vs. ‘non-traffic’). An ANOVA analysis and a
Tukey b post hoc test were applied after square root
data transformation to test for differences between
exposure series. If the ANOVA homogeneity of
variance pre-requisite was not fulfilled, the nonparametric Kruskal–Wallis test was used instead. A
probability level p<0.05 or smaller was considered
statistically significant. The non-parametric Spearman
Rank Correlation Coefficient (rs) was calculated to
correlate ozone-induced leaf injury with ozone-exposure parameters (mean, AOT20, AOT40). Statistical
analyses were performed using the SPSS for Windows Release 10.0 (SPSS Inc. USA) statistical
package.
3 Results
2.4 Ozone Concentrations
3.1 Foliar Injury in Tobacco Plants
Hourly ozone concentration data were obtained from
six monitoring stations in the Air Quality Network of
the Valencian Community (RVCCA, http://www.cma.
gva.es) located near the biomonitoring sites (Table 1).
For clarification purposes, the air monitoring stations
are identified in this study by the same numbers and
names given to the tobacco stations, but their official
names are provided in Table 1. Three of these
monitoring stations were adjacent to tobacco exposure
racks (distance <10 m), another was at ca. 100 m,
while the other two were farther away (Linares and
Paterna, see Table 1). Mean ozone concentrations per
day, biweekly exposure period and whole experimental season were computed, and cumulative exposure
indices (AOT20, AOT40) were calculated according
to the methods described by the EU Directive
(European Union 2002), using mean hourly values
In the city of Valencia, ozone levels in 2002 were
high enough to induce leaf injury in tobacco plants at
all the stations for at least one of the exposure series,
and frequently for most of them (data of the injury
values for each station and series not shown). Figure 2
shows the mean leaf injury for six exposure series as a
whole, with the stations ranked in ascending order.
The highest leaf injury levels were observed at the
two sites placed outside the city of Valencia (No. 6, a
rural station about 10 km south of the city and No. 7,
a University campus garden in a suburban area W of
the city) and at the biomonitoring site No. 2,
established in a large urban garden. In the latter, the
tobacco exposure site was relatively far from the
traffic of the surrounding streets. From now on, we
will refer to these three stations as ‘non-traffic’
Water Air Soil Pollut
Urban
60
Suburban
50
Rural
Leaf Injury (%)
Large Urban Garden
40
30
20
10
0
5
3
1
4
7
2
6
Site Number
Fig. 2 Average leaf injury (± standard error) for the different
biomonitoring stations ordered by ascendent mean values.
[n=6, Kruskal–Wallis test, differences not significant at a
probability level of p<0.05]
stations. As an example, in station 6, injury affected
about 85% of the leaf surface in two of the series. The
remaining four sites, urban stations submitted to
intense traffic (‘traffic’ stations) experienced lower
mean leaf injury for the whole 3-month period. The
lowest value of all sites was recorded at site 5, an
urban street in the center of the city with intense
traffic: in four of the six exposure series we did not
detect any ozone injury at this station. The other three
urban sites (1, 3, and 4) were located in more
peripheral zones: at different entrances to the city
which also sustained heavy traffic but were more
open (and thus presumably better ventilated) sites.
Despite this consistent ranking of the biomonitoring
stations, the differences between sites for the 3-month
exposure period of the experiment were not significant (analyses based on mean site values per exposure
period, Fig. 2). This is because the large variability in
leaf injury at each single site for the six exposure
periods prevented significant differences between
sites. But, if we consider individual exposure series,
in all of them there are significant differences between
two or more stations (data of the injury values for
each station and series not shown; but the value of p<
0.05 in all the series, checked by Kruskal–Wallis test,
indicates that in all the series there were at least two
stations with significantly different injury values).
Furthermore, in four of these series, there were also
significant differences between the two types of
biomonitoring stations (see below, Fig. 3).
For the whole biomonitoring network, clear differences in leaf injury were observed in the course of the
six exposure series (Fig. 3, grey bars). The highest
injury levels were observed in the series starting on 11
July, followed by the first exposure series, which
started on 29 May. The remaining four series
exhibited significantly lower injury values (ANOVA,
p<0.05), with the series at the end of July and
beginning of August showing the lowest mean values.
Comparison of traffic and non-traffic biomonitoring
stations (Fig. 3, black and white bars) at each of the
exposure series also indicate that non-traffic stations
experience significantly higher injury levels than
traffic stations during four of the six exposure series.
3.2 Comparison of Foliar Injury with Continuous
Measurements
In six out of seven tobacco exposure stations, there
were close or representative active air monitoring
stations with ozone data available for the exposure
period (Table 1). Figure 4 shows that the suburban
monitoring station closest to the biomonitoring station
7 (U. Burjassot) experiences the highest ozone levels
during the 3-month exposure period of this survey,
followed by the station placed in a large urban garden
(station 2, Vivers, note the higher levels especially in
the afternoon). Data from another suburban monitoring station (Quart monitoring station, http://www.
cma.gva.es) not included in this study also confirm
90
*
80
Leaf injury (%)
70
70
a
ns
a
All stations
Traffic stations
Non-Traffic stations
60
ns
50
40
b
30
*
b
20
b
10
**
b
**
0
29 May
14 Jun
28 Jun
11 Jul
25 Jul
8 Aug
Starting date of the 2-week series
Fig. 3 Grey bars: Average leaf injury (± standard error)
percentage for all 7 biomonitoring stations at the six consecutive 2-week exposure times; significant differences between
the series are indicated with different letters (ANOVA, Tukey b
test, after square root transformation, n=7, p<0.05). Black and
white bars: Average leaf injury (±standard error) percentage for
traffic (No. 1, 3, 4 and 5) and non-traffic (No. 2, 6 and 7)
stations; significant differences between the two types of
stations at each exposure series are indicated with asterisks on
the white bars (t test, *p<0.05, **p<0.01, ns=not significant)
1
2
50
3
4
5
7
40
30
20
10
0
1
3
5
7
9
11 13 15
local time (h)
17 19
21
23
Fig. 4 Daily profile of ozone concentrations in continuous
monitoring stations close to the tobacco stations for the period
from 29 May to 22 August 2002. The two stations with close
symbols represent a suburban station (7), and a large urban park
(2), while the rest (open symbols) are urban stations submitted
to intense traffic. These stations have been numbered with the
numbers of the closest tobacco stations in order to gain in
clarity. Further details on their official names and distance from
the tobacco exposure racks are given on Table 1
that ozone levels in suburban zones downwind of
Valencia are overall higher than in urban areas with
intense traffic (the case of the other four stations
included in this study, 1, 3, 4 and 5). In the two
stations classified as non-traffic, AOT40 for the whole
exposure period (29 May–22 August 2002) was
5,363 ppb h (suburban station 8), and 1,480 ppb
h (station 2, large urban garden), while for the rest of
monitoring stations, all of them classified as traffic
stations, AOT40 was below 356 ppb h.
Very incipient injury (1.4%) was already observed
at ozone values as low as 24 h mean=13.6 ppb,
AOT20=169.6 ppb h, and the lowest values at which
it became evident (20.7% injury in series 2 of
Avd. Aragón) was 24 h mean=20.7 ppb, AOT20=
415.3 ppb h. In both cases, the 40 ppb threshold
(hourly mean) was not reached, AOT40 being 0.
Temporal variation of the cumulative indices AOT20
and AOT40 for the different exposure series is given
in Fig. 5. By comparing profile of Fig. 5 with that of
leaf injury (Fig. 3), consistently the latest two series
had low ozone values and low ozone-induced injury,
and the series starting on 29 May and 11 July are
among those having both comparatively higher ozone
levels and foliar injury. However, series starting on 28
June and especially series starting on 14 June show a
discordant pattern between ozone injury and ozone
levels. Series starting on 14 June had the highest
ozone values while the injury levels were among the
lowest. Meteorological data from air monitoring
station 4 (the only ones available) indicate that this
2-week period is characterized by an outstanding
combination of high temperatures and low atmospheric humidity levels, leading to the highest VPD values
of all the series: 88.5% of the daily hours (8:00 to
20:00 h) had VPD values over 1.5 KPa. As injury to
the plants is not determined by ambient levels but
rather by the effective doses (i.e. the uptake of the
pollutant by the plant), high VPD may have favored
stomatal closure and therefore avoidance of ozone
uptake, explaining in part the low injury values
observed in this series.
Correlation of foliar injury both with mean (24 and
12 h means) and cumulative exposure indices (AOT20,
AOT40) of ozone concentrations was performed
using the Spearman Rank Correlation Coefficient (rs;
Table 2). If we include all the stations and exposure
series together (n=36 [6 stations×6 series]), the
correlation is significant for all the ozone descriptors
(means and cumulative indices) considered, with rs
values between 0.39 and 0.58, indicating a modest
correlation. When the same analysis was performed
for the six consecutive 2-week exposure series of each
biomonitoring station, the resulting values for rs are
in general higher, with some of them about 0.8–0.9
in Avd. Aragón and Manuel Granero stations. However, despite these moderate to strong correlations, in
most of the cases they were not significant. This is
partly due to the small number of cases (n=6), as the
2500
2000
AOT20
AOT40
VPD
100
90
80
70
60
1500
50
40
1000
30
20
500
10
0
Hours with VPD>1.5 KPa (%)
60
AOT (ppb*h)
Ozone concentration (ppb)
Water Air Soil Pollut
0
29 May 14 Jun 28 Jun 11 Jul 25 Jul 8 Aug
Fig. 5 Mean biweekly AOT20 and AOT40 (ppb h) (mean±
standard error of the six stations with monitoring stations) and
percentage of hours with VPD>1.5 KPa for the six consecutive
tobacco exposure periods. VPD during daily hours (8:00 to
20:00 h CET) values are from the only of the six monitoring
stations (station 4) with meteorological data
Water Air Soil Pollut
Table 2 Spearman Rank Correlation Coefficient (rs) of the correlations between leaf injury and different descriptors of ozone
concentrations measured with continuous monitors for each exposure station (n=6 exposure series, except +, with only 4), and for all
stations and series (n=36 for means and n=34 for cumulative indices)
Station
24 h Mean
12 h Mean
AOT20
AOT40
1 – Avd. Aragón
2 – Vivers
3 – Beltrán Báguena
4 – Manuel Granero
5 – Fdo. Católico
7 – U. Burjassot
All stations
0.60 ns
0.60 ns
0.66 ns
0.77 ns
0.51 ns
0.52 ns
0.58**
0.94**
0.49 ns
0.40 ns
0.60 ns
0.44 ns
0.66 ns
0.52**
0.89*
0.53 ns
0.74 ns+
0.43 ns
0.44 ns
0.60 ns
0.47**
0.78 ns
0.49 ns
0.74 ns+
0.12 ns
0.37 ns
0.31 ns
0.39*
Significant correlations at probability levels of p<0.05 and p<0.01, are indicated as (*) and as (**), respectively.
critical values of rs for such a small number of cases is
very high (0.89 for a level of significance of 0.05).
4 Discussion
The present study shows that in the city of Valencia,
ozone levels in 2002 were high enough to induce leaf
injury in tobacco plants in all the stations for at least
one of the series. In some of the exposure series,
biomonitoring stations submitted to intense traffic
(‘traffic stations’) showed significantly lower injury
values than stations less directly exposed to car
emissions (stations placed in suburban or rural sites,
or in a large garden, classified as ‘non-traffic
stations’). Traffic stations support relatively high
levels of SO2, CO, NO2 and especially of NO (data
available at http://www.cma.gva.es) that act as an
scavenger of ozone, contributing to locally reduce the
levels of this secondary pollutant. The lowest injury
values were recorded in biomonitoring station 5,
placed in an urban street in the centre of the city,
with intense traffic and limited air circulation. On the
contrary, the highest injury was observed in stations
under the influence of the city plume (rural station 6
and suburban station 7) and the station established in
a large urban garden (station 2). These non-traffic
stations experienced comparatively higher ozone
levels than traffic stations. A similar general pattern
with highest ozone levels and strongest ozoneinduced plant injuries in rural and suburban sites
when compared with urban streets has also been
reported in other European cities (Godzik 1997;
Klumpp et al. 2006a, Klumpp et al. 2006b; Nali
et al. 2001). However, Klumpp et al. (2006a) pointed
out that, in some periods, ozone pollution may reach
high levels even at central locations and street sites in
some European cities. In the present study, significant
differences between stations in each exposure series
were found, but especially marked are the strong
changes in injury levels between different series.
Considerable leaf injury (values over 10%) was
observed at rather low ozone concentrations (24 h
mean=20.7 ppb, AOT20=415.3 ppb h), confirming
the high sensitivity of tobacco Bel-W3 variety.
Nevertheless, correlation of visible injury with ozone
concentrations considering all the stations and series
together (n=34) was only moderately good. Cumulative indices such as AOT20 and AOT40 (rs =0.47, p<
0.01, and rs =0.39, p<0.05, respectively) do not
represent an improvement of the correlation strength
with regard to 24 and 12 h average values (rs =0.58
and 0.52, p < 0.01, respectively). Klumpp et al.
(2006b) obtained similar results from the whole
European biomonitoring network. When such correlations were calculated for the six series of each single
station, in order to overcome part of the possible
variability due to different site characteristics, rs
tended to increase in some stations (e.g. rs about
0.90 for 12 h mean and AOT20 in station 1), but the
high critical values of rs required prevented for these
correlations to be significant in most stations. As in
the present study, other authors also found a limited
explanation of the variance in leaf injury on the basis
of ambient ozone concentrations (Klumpp et al.
2006b; Krupa et al. 1993; Ribas and Peñuelas
2003). Environmental factors such as temperature,
humidity (or VPD, which combines both parameters)
and wind speed affect ozone uptake by the plants
Water Air Soil Pollut
through changes in stomatal behaviour (Ribas et al.
1998). Thus, environmental variables may have a
strong effect on the effective dose taken by the plant,
and consequently on the final intensity of injury. Such
particular environmental conditions with high VPD
(that would induce stomatal closure) might explain
the discrepancy between relatively high ozone levels
in the second exposure series and low injury observed
in the present study. The use of models taking into
account not only ozone concentrations but also
meteorological and physiological variables (e.g. stomatal conductance) may contribute to better characterize the dose–response relationships in tobacco
plants; this was however out of the scope of the
present study.
In any case, using sensitive bioindicator plants like
tobacco Bel-W3 has to be considered a complementary rather than a substitute approach to ozone
concentration monitoring by physico-chemical methods (Nali et al. 2006). With this technique, the impact
of the effective ozone dose on a living organism is
directly measured (Klumpp et al. 2006b). KostkaRick (2003) exposed tobacco Bel-W3 plants together
with horticultural and ornamental crops, and concluded that frequently the predictive power of this
sensitive indicator was better for crop leaf injury than
various ozone concentration or dose predictors.
Therefore, injury information provided by tobacco
Bel-W3 may be relevant to assess risk to other less
sensitive cultures. In this sense, it is interesting to note
that Valencia is surrounded by an area of intensive
irrigated agricultural use, poorly covered by air
monitors, and injury in crops such as potato and
watermelon has been regularly observed year after
year, including the year 2002, when the present study
was carried out (Sanz et al., unpublished). This is
consistent with the results of this study showing
episodes of intense phytotoxic effects on hypersensitive tobacco Bel-W3 especially in the surrounding
areas of the city. In these areas, the use of tobacco
plants would be potentially appropriate to identify
those zones at higher risk, and also to provide
information on the spatial and temporal variation of
phytotoxic impacts of the city plume on the crop
cultures, including the characterization of episodes
with high phytotoxic effects.
Finally, the fact that in the present study mean
ozone values and ozone cumulative indices (AOT20,
AOT40) are equally well correlated with leaf injury
suggests that assessment of ozone concentrations by
passive samplers – a much cheaper technique than
active air monitoring providing mean concentrations
over a exposure period (Krupa and Legge 2000; Sanz
et al. 2007; Sanz et al. 2001) – in combination with
meteorological data if available, would be a very
appropriate approach to characterize the response of
sensitive tobacco plants to ambient ozone concentrations. It would be especially useful in areas not or
poorly covered by air quality stations.
Acknowledgements This study was supported by the LIFE
Environment Programme of the European Commission, DG
Environment, under the grant LIFE/99/ENV/D/000453. We
thank the Ayuntamiento de Valencia, Oficina Técnica de la
Devesa-Albufera (A. Vizcaino, A. Quintana) for their valuable
help in this study. José Jaime Diéguez helped to process the air
monitoring data. Fundación CEAM members are also indebted
to Generalitat Valenciana and Fundación Bancaja for continuous support to the activities of the Foundation.
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