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Two aquatic macrophytes as bioindicators for medium-high copperconcentrations in freshwatersPaolo 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-highcopper concentrations in freshwaters', Plant Biosystems - An International Journal Dealing with all Aspects of PlantBiology,, First published on: 26 May 2011 (iFirst)To link to this Article: DOI: 10.1080/11263504.2010.547677URL: http://dx.doi.org/10.1080/11263504.2010.547677

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Two aquatic macrophytes as bioindicators for medium-highcopper concentrations in freshwaters

PAOLO ZUCCARINI1 & SASA KAMPUS2

1Dipartimento di Biologia delle Piante Agrarie, Universita di Pisa, Viale delle Piagge 23, 56124, Pisa, Italy and2Fakulteta za druzbene vede, Univerza v ljubljani, Kardeljeva plos�cad 5, 1000, Ljubljana, Slovenija

AbstractPlant scions of Ceratophyllum demersum L. and Potamogeton natans L. were exposed in controlled conditions to differentconcentrations 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. demersumstarted to show significant reductions in growth and photosynthetic efficiency from 4 mM Cu. As it results from the observeddata, 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 copper51 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

Downloaded By: [Zuccarini, Paolo] At: 02:25 29 May 2011

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 (p5 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

(p5 0.01) and 0.397 (p5 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

(p5 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 p50.05, 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. Kampus

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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,

p5 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 (p5 0.05), while P.

natans already from 2 mM copper (p5 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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