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Two aquatic macrophytes as bioindicators for medium-high copper
concentrations in freshwaters
Paolo Zuccarinia
; Saša Kampušb
a
Dipartimento di Biologia delle Piante Agrarie, Università di Pisa, Viale delle Piagge 23, Pisa, Italy b
Fakulteta za družbene vede, Univerza v ljubljani, Kardeljeva ploščad 5, Ljubljana, Slovenija
First published on: 26 May 2011
To cite this Article Zuccarini, Paolo and Kampuš, Saša(2011) 'Two aquatic macrophytes as bioindicators for medium-high
copper concentrations in freshwaters', Plant Biosystems - An International Journal Dealing with all Aspects of Plant
Biology,, First published on: 26 May 2011 (iFirst)
To link to this Article: DOI: 10.1080/11263504.2010.547677
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2. Two aquatic macrophytes as bioindicators for medium-high
copper concentrations in freshwaters
PAOLO ZUCCARINI1
& SASˇA KAMPUSˇ2
1
Dipartimento di Biologia delle Piante Agrarie, Universita` di Pisa, Viale delle Piagge 23, 56124, Pisa, Italy and
2
Fakulteta za druzˇbene vede, Univerza v ljubljani, Kardeljeva plosˇcad 5, 1000, Ljubljana, Slovenija
Abstract
Plant scions of Ceratophyllum demersum L. and Potamogeton natans L. were exposed in controlled conditions to different
concentrations of copper during approximately 2 weeks; Fv/Fm was monitored at regular intervals and relative growth rate
(RGR) was calculated at the end of the trial. P. natans was affected by Cu concentrations starting from 2 mM; C. demersum
started to show significant reductions in growth and photosynthetic efficiency from 4 mM Cu. As it results from the observed
data, the two aquatic macrophytes can be used as valid bioindicators for medium-high copper concentrations in freshwaters.
Keywords: Ceratophyllum demersum, Potamogeton natans, copper, bioindicators, freshwaters
Introduction
Despite of its relative mildness of character, copper
can be highly toxic to plants even at micromolar
concentrations (Cabral 2003). It is an important
pollutant for aquatic biota (Granmo et al. 2002;
Katranitsas et al. 2003), in which environments
(rivers, shallow lakes) tend to accumulate for several
reasons, one of the most relevant being its release
from antifouling paints (Nichols 1988; Smith 1996;
Lambert et al. 2006).
Several kinds of aquatic organisms have been
studied as potential biological indicators for copper
(Monnet et al. 2006; Nicolau 2007) exhibiting a
wide range of sensitivity, lower on average for algae,
and higher for higher aquatic plants (Nor 1987).
Submerged macrophytes proposed as potential
biomonitors have been, among the others, Hydrilla
verticillata Royle (Gupta et al. 1996), Typha latifolia
L. (Muller et al. 2001), Elodea canadensis Michaux.
(Mal et al. 2002).
The aim of the present work was to test resistance
to copper of Potamogeton natans L. and Ceratophyllum
demersum L., characterized by good adaptability and
proved resistance to several abiotic stresses (Rama
Devi and Prasad 1998; Bernez et al. 2004), in the
perspective of their use as potential bioindicators for
higher levels of copper toxicity than the ones applied
on average to the previously mentioned plants.
Methods
Ceratophyllum demersum was chosen among free-
floating aquatic macrophytes in order to avoid
additional stress due to lack of substrate, and because
of the average higher tolerance to copper of pleusto-
phytes rather than hydrophytes, as shown by Maleva
et al. (2004). Potamogeton natans was chosen because,
in virtue of its rhizophytic nature, it was supposed to
show slightly lower tolerance than C. demersum, being
therefore a suitable bioindicator for medium copper
concentrations. Apical parts of plants of C. demersum
and P. natans were collected, and pre-cultivated
under continuous light in aquaria containing tap
water (total copper 51 mg lv1
). Ten-centimeter plant
scions were cut, tied with raffia to wire net, and
placed into 2000 ml beakers containing Hoagland’s
medium diluted to 1/100 strength (Eliasson 1978) at
a pH of 5.5 + 0.3 with different concentrations of
copper (supplied as CuSO4): 0, 1, 2, 4 and 6 mM,
corresponding to 0, 65, 130, 260 and 390 mg l71
.
Six repetitions were done for each species and for
each copper concentration. Plants were incubated at
Correspondence: Paolo Zuccarini, Dipartimento di Biologia delle Piante Agrarie – Sez., Universita` degli Studi di Pisa, Fisiologia Vegetale – Via Mariscoglio 34,
56127, Pisa, Italy. Tel: þ39-349-1298437. Fax: þ39-050-2216532. Email: p.zuccarini@virgilio.it
Plant Biosystems, 2011; 1–4, iFirst article
ISSN 1126-3504 print/ISSN 1724-5575 online ª 2011 Societa` Botanica Italiana
DOI: 10.1080/11263504.2010.547677
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3. 228C under 12 h of light period (450 mmol m72
s71
)
during 12 days. Photosynthetic efficiency was mea-
sured at regular intervals as Fv/Fm (ratio between
variable and maximum fluorescence) with a Plant
Efficiency Analyzer (PEA, Hansatech); for the calcu-
lation of relative growth rate (RGR) fresh weights were
measured at the end of the trial, and initial fresh
weights were estimated as average values out of 30
scions, 10 cm long, of the examined species. Statis-
tical analysis of the data was performed by using the
SAS statistical software program (SAS Institute 1990).
Fv/Fm and growth data, relative to the two plants
taken separately and at the different levels of copper,
were subjected to analysis of variance and the signi-
ficance of difference between means was then deter-
mined with Duncan’s multiple range test; paired t-test
was used to compare differences in growth between
the two species at different copper concentrations.
Results and discussion
Photosynthetic efficiency of C. demersum was not
affected by copper concentrations up to 2 mM, while
P. natans showed a significant drop (p 5 0.05), at the
end of the trial, already for 2 mM (Figure 1).
This is only partially in accordance with Rama Devi
and Prasad (1998), who showed how Cu accumula-
tion in C. demersum already started to induce
oxidative stress from concentrations of 2 mM. The
reason of this difference can be that at 2 mM, even if
clorophyll content starts to decrease (Rama Devi and
Prasad 1998), its efficiency is still intact, providing
optimal values of Fv/Fm. This interpretation is
supported by the work of other authors, who observed
a temporally delayed decrease of chlorophyll content
and photosynthetic efficiency in plants undergoing
stress conditions (Liu and Huang 2000; Bounfour
et al. 2002), so that in some situations chlorophyll
content is considered to be a better indicator of stress
than the Fv/Fm ratio (Rong-hua et al. 2006).
Both P. natans and C. demersum were significantly
affected by concentrations of copper of 4 and 6 mM,
but the decrease of Fv/Fm for P. natans was more
marked, reaching respectively values of 0.525
(p 5 0.01) and 0.397 (p 5 0.01) on the last day of
trial. The different tolerance to copper appears to be
species-specific, since works conducted on P. pecti-
natus showed it to have a high Cu absorption capacity
(Samecka-Cymerman and Kempers 2004), even
higher than C. demersum (Gavrilenko and Zolotu-
khina 1989). C. demersum appears to be strongly
affected too by the highest copper concentrations,
but its final values of photosynthetic efficiency are
not so critical, reaching the minimum value of 0.514
(p 5 0.01) at day 12 when exposed to 6 mM copper,
indicating still the presence of a remarkable photo-
synthetic activity even at the most extreme condi-
tions. The resistance showed by C. demersum can be
put in correlation with its properties of absorbtion,
accumulation and tolerance of heavy metals
(Keskinkan et al. 2004), that made this aquatic
macrophyte be suggested for heavy metal detoxifica-
tion (Mishra et al. 2006). P. natans, which performs
Figure 1. Trends of Fv/Fm of C. demersum and P. natans exposed to different levels of copper during 12 days of trial.
Figure 2. Relative Growth Rate of C. demersum and P. natans
during the trial. Values marked with the same letter are not
statistically different at p 5 0.05, according to Duncan’s multiple
range test. Normal letters refer to the left columns, relative to C.
demersum; letters in italic refer to the right columns, relative to P.
natans. The two groups have to be considered separately.
2 P. Zuccarini and S. Kampusˇ
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4. heavy metal uptake through roots, shoots, and leaves
(Fritioff and Greger 2006) is not affected by copper
only at concentrations of up to 1 mM. RGR of P.
natans was significantly lower than C. demersum at Cu
concentrations of 2 mM and more (Figure 2) (t-test,
p 5 0.05), confirming the observation that the
former plant is the one which undergoes the most
severe stress for medium-high copper concentra-
tions. The higher RGR of P. natans in non-stress
conditions (0 mM and 1 mM) confirms its naturally
high vigor in growth; the observation of growth data
also confirms the tolerance thresholds deduced with
Fv/Fm, since C. demersum showed significant growth
reduction from 4 mM copper (p 5 0.05), while P.
natans already from 2 mM copper (p 5 0.05).
In conclusion, these data show C. demersum and P.
natans to be good bioindicators for higher copper
concentrations than the ones commonly associated to
other species of macrophytes (Gupta et al. 1996;
Muller et al. 2001; Mal et al. 2002), and point out C.
demersum as the one with the highest tolerance.
Figure 3 reports a list of species of aquatic macro-
phytes investigated for their tolerance to copper with
the relative thresholds: as it can be seen H. verticillata
and Potamogeton pusillus are the only ones which
performances are comparable with the ones of the two
species object of this work. Further research could be
focused on investigating the performances of copper
absorption and tolerance by rooted specimens of
P. natans, as it occurs in nature, in order to compare
them with the ones of the experimental floating scions.
Acknowledgment
The authors sincerely thank Dr. Cecilia Viegi for
valuable help in revision of the manuscript.
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