Volume 5 . - 2012
Scientic Bulletin of ESCORENA
Vol.5, July 2012
Prof. Dr. Lizica Mihut, Rector of “Aurel Vlaicu” University of Arad.
2. Introductory remarks
Prof. Dr. Ryszard Michal Kozlowski, FAO/ESCORENA Focal Point Coordinator.
3. Current status of ESCORENA Network development and the new strategy and
emerging activities: Prof. Dr. Ryszard KOZŁOWSKI, Ass. Prof. Dr. Cecilia Sirghie, M.Sc. Maria
4. Emerging role of ESCORENA Network: Michal Demes, Maria Mackiewicz-Talarczyk.
5. Medicinal and Aromatic Plants Network (MAP) New Network of ESCORENA: Dr. Kirill
6. Some data about plants of the Russian Far East Flora and their using in Folk Medicine:
Dr. Kirill G. Tkachenko.
7. Possibilities of re retardants application in the protection of wooden buildings in
the open-air museums: Ryszard KOZLOWSKI, Lizica MIHUT, Maria Silvia PERNEVAN, Malgorzata
HELWIG-KUBIAK, Lidia IDZIAK.
8. Bioremediation of soil contaminated with cadmium using hemp shives. A case study
of modication of physiological parameters in triticum aestivum: Lucian Copolovici, Dana
Copolovici, Ülo Niinemets and Cecilia Sirghie.
8. BASTEURES - Project co-funded by EUROPEAN UNION trough the European Regional
Development Fund / Sectoral Operational Programme “Increase of Economic Competitiveness”
/ “Investing for your future”
Volume 5 . - 2012
BIOREMEDIATION OF SOIL CONTAMINATED WITH CADMIUM
USING HEMP SHIVES. A CASE STUDy OF MODIFICATION OF
PHySIOLOGICAL PARAMETERS IN TRITICUM AESTIVUM
Lucian Copolovici1, Dana Copolovici1, Ülo Niinemets2 and Cecilia Sirghie1
1 ”Institute of Research , Development and Innovation in Technical and Natural Sciences ”
”Aurel Vlaicu” University , Arad, Romania
2 Estonian University of Life Sciences, Institute of Agricultural and Environmental Sciences
In the present study we show the possibility to use the hemp shives as biomass
remedying of soil contaminated with cadmium. The Triticum aestivum were employed as test
plant and assimilation rates and stomatal conductance to water vapor were measured. The
hemp shives determine a remediation of the physiological parameter for plants treated with 1
mg/L of cadmium.
Cadmium is one of the toxic heavy metal for human (Jomova and Valko 2011; Luparello
et al. 2011; Nzengue et al. 2011), animals (Thevenod 2010, 2009) and plants (Jahangir et al. 2009;
Verbruggen et al. 2009). It was shown that an expose to cadmium determine lung cancer (Park
et al. 2012), kidney disfunction (Liang et al. 2012), brast cancer (Julin et al. 2012) and testicular
injury (Wong and Cheng 2012; Siu et al. 2009). The source of cadmium contamination include
the rubber tires, industrial water cooling, plastics, pigments, plated ware, alloys, insecticides
(reviewed in (Reeves and Chaney 2008; Bertin and Averbeck 2006). The soil contamination
with cadmium can occur by direct inltration of contaminants from solid wastes, sewage, or
The removal of cadmium (and other heavy metals) from contaminated soil can been
done by transfer of contaminated soil to landlls. This procedure is expensive and determines
a risk sources for the whole ecosystem (Oberg and Bergback 2005). Due to this reason, the
bioremediation become low-cost and eco-friendly alternatives for cadmium removal from soil.
Bioremediation, as was shown in Mohanty and Patra (2011), typically use living organisms, to
SCIENTIFIC BULLETIN OF ESCORENA
remove toxic elements from the environment. There are many studies of dierent bacteria,
microbes and plants used for cadmium removal. For example metabolic active cells of
Saccharomyces cerevisiae have a potential application in cadmium removal (Wang et al.
2012). The recent study have been showing that Spinacia oleracea plants are not accumulate
cadmium so it can be used for phytoremediation of contaminated soils (Salaskar et al. 2011).
Other monitored natural attenuation (MNA) of contaminated soils (as is described in (Declercq
et al. 2012)) include in situ techniques which use the microbial biomass and bioaccumulators
(see (Mohanty and Patra 2011) for removal of pollutants.
It was shown that hemp plants (Cannabis sativa L.) accumulated heavy metals using
cellular mechanisms which allowing it to cope with high metal concentrations (Linger et al.
2005; Citterio et al. 2003; Gasiorek and Kozlowski 2003). Even more, the previous studies reveal
that short hemp bers have a good sorption potential (at a level of mmols) for Cd2+ ions
(Pejic et al. 2009). Based on this feature of Cannabis sativa plants, we used the hemp shives
as biomass remediation of soil contaminated with cadmium. We used Triticum aestivum as
monitoring plants to test the bioremediation capacity of hemp shives.
Materials and methods
Wheat (Triticum aestivum L.) seeds (cv. Lovrin, source: Fundulea, Romania) were used
for the experiment. 40 seeds of T. aestivum were sown in plastic pots (5×5×5 cm) lled with
commercial garden soil including slow release NPK fertilizer with microelements (Biolan,
Finland). The sowing depth was 1 cm. The plants were grown in a growth chamber (Percival,
IA, USA) under a light intensity of 1000 µmol m-2 s-1 provided for a 12 h light period and day/
night temperatures of 25°C/18°C.
The soil was articial contaminated with cadmium at a level of 1 mg/L. For
bioremediation in the soil was added between 1 and 5 % of hemp shives. The hemp shives
were obtained from dew-retted bers. The measurements were done at three Zadoks growth
stage: 1.0 (6 days), 1.1 (9 days) and 1.2 (12 days).
The photosynthetic parameters of the plants were monitored using the GFS 3000
Portable Gas Exchange System (Walz, Eeltrich, Germany) as was described in previous studies
(Niinemets et al. 2010; Niinemets et al. 2011). The measurements were performed at a chamber
CO2 concentration of 385 μmol mol-1, photosynthetic quantum ux density was kept at 1000
μmol m-2 s-1, leaf temperature at 25ºC and chamber relative humidity at 70%. The air ow rate
was 750 μmol s-1.
The rates of net assimilation (A) and stomatal conductance to water vapor (gs) were
calculated from these measurements according to von Caemmerer and Farquhar (1981).
Results and discussions
In previous studies were shown that T. aestivum exposed to high concentration of
cadmium leads to depresses growth rate, reduction of photosynthesis, decreased chlorophyll
content, changing in phenols and enzymes activities (Lakhdar et al. 2012; Wang et al. 2011; Ci
et al. 2010; Duan et al. 2010; Khan et al. 2008; Samiullah et al. 2007; Ouzounidou et al. 1997). In
our case, the assimilation rate decreased with more than 10 % for plants growth in soil treated
Volume 5 . - 2012
with cadmium. The inhibition of photosynthesis is the result of damage to the PSII reaction
center in the leaf (see (Duan et al. 2010)). The plants growth in the soil treated with hemp
shives have the same level of photosynthesis as the control plants.
Figure 1. Changes in net assimilation rate (A) with the remediation agent (hemp shives) concentration
Even more, for the mature plants (12 days) the assimilation rates are higher than for the control
plants in case of hemp shives in soil of 1.5% (Figure 1). This trend can be explicate by the
possibility that the hemp shives to act as chelator of the cadmium ions as was shown for C.
sativa chestnuts shell extract (Stingu et al. 2012).
The values of stomata conductance to water vapor (gs) are as well inuence by the cadmium
Figure 2. Changes in stomatal conductance to water vapor (gs) with the remediation agent (hemp shives)
SCIENTIFIC BULLETIN OF ESCORENA
From gure 2 can be seen that in plants treated with cadmium the gs decrease in
average with 20 %. Even that the mechanism of stomata closure is not totally understand
(Sha et al. 2011), we can speculate that the negative eect of Cd2+ to gs are relating with the
inhibition of primary carbon metabolism (Vassilev et al. 1997). Using even a small concentration
of remediation agent (hemp shives – 1%) the gs values are comparative with control plants
(Figure 2). Interesting, the values of gs increased with the increasing of the hemp shives
concentration until more than 30% comparative with control plant at remediation agent
concentration of 2%. After that, the values of gs decreased slowly.
In the present study we have shown that hemp shives act as a soil remediation
agent against cadmium soil pollution. The treatment of the soil contaminate with cadmium
even with a small concentration of hemp shives determine the recover of the physiological
parameters in the Triticum aestivum L. plants. More work is necessary in order to understand of
the mechanism of the eect of the cadmium and hemp shives on the plant-soil interactions.
This work was supported by project co-funded by European Union through European
Regional Development Fund Structural Operational Program “Increasing of Economic
Competitiveness” Priority axis 2, operation 2.1.2. ID project 679, cod SMIS CNSR 12638.
Bertin, G., & Averbeck, D. (2006). Cadmium: cellular eects, modications of biomolecules,
modulation of DNA repair and genotoxic consequences (a review). Biochimie, 88, 1549-1559.
Ci, D., Jiang, D., Wollenweber, B., Dai, T., Jing, Q., & Cao, W. (2010). Cadmium stress in wheat
seedlings: growth, cadmium accumulation and photosynthesis. Acta Physiologiae Plantarum, 32,
Citterio, S., Santagostino, A., Fumagalli, P., Prato, N., Ranalli, P., & Sgorbati, S. (2003). Heavy
metal tolerance and accumulation of Cd, Cr and Ni by Cannabis sativa L. Plant and Soil, 256, 243-
Declercq, I., Cappuyns, V., & Duclos, Y. (2012). Monitored natural attenuation (MNA) of
contaminated soils: State of the art in Europe-A critical evaluation. Science of the Total Environment,
Duan, Y.-P., Yuan, S., Tu, S.-H., Feng, W.-Q., Xu, F., Zhang, Z.-W., Chen, Y.-E., Wang, X., Shang,
J., & Lin, H.-H. (2010). Eects of cadmium stress on alternative oxidase and photosystem II in three
wheat cultivars. Zeitschrift Fur Naturforschung Section C-a Journal of Biosciences, 65, 87-94.
Gasiorek, J., & Kozlowski, R. 2003. Removal of heavy metals from solid waste and the
application of the latter to agriculture. In Proceedings of the 8th International Conference on
Environmental Science and Technology, Vol B, Poster Presentations, 230-239.
Jahangir, M., Abdel-Farid, I. B., Kim, H. K., Choi, Y. H., & Verpoorte, R. (2009). Healthy and
unhealthy plants: the eect of stress on the metabolism of Brassicaceae. Environmental and
Volume 5 . - 2012
Experimental Botany, 67, 23-33.
Jomova, K., & Valko, M. (2011). Advances in metal-induced oxidative stress and human
disease. Toxicology, 283, 65-87.
Julin, B., Wolk, A., Bergkvist, L., Bottai, M., & Akesson, A. (2012). Dietary cadmium exposure
and risk of postmenopausal breast cancer: a population-based prospective cohort study. Cancer
Research, 72, 1459-1466.
Khan, N. A., Singh, S., Anjum, N. A., & Nazar, R. (2008). Cadmium eects on carbonic
anhydrase, photosynthesis, dry mass and antioxidative enzymes in wheat (Triticum aestivum)
under low and sucient zinc. Journal of Plant Interactions, 3, 31-37.
Lakhdar, A., Slatni, T., Iannelli, M. A., Debez, A., Pietrini, F., Jedidi, N., Massacci, A., & Abdelly,
C. (2012). Risk of municipal solid waste compost and sewage sludge use on photosynthetic
performance in common crop (Triticum durum). Acta Physiologiae Plantarum, 34, 1017-1026.
Liang, Y., Lei, L., Nilsson, J., Li, H., Nordberg, M., Bernard, A., Nordberg, G. F., Bergdahl, I. A., &
Jin, T. (2012). Renal function after reduction in cadmium exposure: an 8-year follow-up of residents
in cadmium-polluted areas. Environmental Health Perspectives, 120, 223-228.
Linger, P., Ostwald, A., & Haensler, J. (2005). Cannabis sativa L. growing on heavy metal
contaminated soil: growth, cadmium uptake and photosynthesis. Biologia Plantarum, 49, 567-
Luparello, C., Sirchia, R., & Longo, A. (2011). Cadmium as a transcriptional modulator in
human cells. Critical Reviews in Toxicology, 41, 73-80.
Mohanty, M., & Patra, H. K. 2011. Attenuation of Chromium Toxicity by Bioremediation
Technology. In Reviews of Environmental Contamination and Toxicology, Vol 210, 1-34.
Niinemets, Ü., Copolovici, L., & Hüve, K. (2010). High within-canopy variation in isoprene
emission potentials in temperate trees: implications for predicting canopy-scale isoprene uxes.
Journal of Geophysical Research - Biogeosciences, 115, G04029.
Niinemets, U., Kuhn, U., Harley, P. C., Staudt, M., Arneth, A., Cescatti, A., Ciccioli, P.,
Copolovici, L., Geron, C., Guenther, A., Kesselmeier, J., Lerdau, M. T., Monson, R. K., & Penuelas, J.
(2011). Estimations of isoprenoid emission capacity from enclosure studies: measurements, data
processing, quality and standardized measurement protocols. Biogeosciences, 8, 2209-2246.
Nzengue, Y., Candeias, S. M., Sauvaigo, S., Douki, T., Favier, A., Rachidi, W., & Guiraud,
P. (2011). The toxicity redox mechanisms of cadmium alone or together with copper and zinc
homeostasis alteration: Its redox biomarkers. Journal of Trace Elements in Medicine and Biology,
Oberg, T., & Bergback, B. (2005). A review of probabilistic risk assessment of contaminated
land. Journal of Soils and Sediments, 5, 213-224.
Ouzounidou, G., Moustakas, M., & Eleftheriou, E. P. (1997). Physiological and ultrastructural
eects of cadmium on wheat (Triticum aestivum L) leaves. Archives of Environmental Contamination
and Toxicology, 32, 154-160.
Park, R. M., Stayner, L. T., Petersen, M. R., Finley-Couch, M., Hornung, R., & Rice, C. (2012).
Cadmium and lung cancer mortality accounting for simultaneous arsenic exposure. Occupational
and Environmental Medicine, 69, 303-309.
Pejic, B., Vukcevic, M., Kostic, M., & Skundric, P. (2009). Biosorption of heavy metal ions from
SCIENTIFIC BULLETIN OF ESCORENA
aqueous solutions by short hemp bers: eect of chemical composition. Journal of Hazardous
Materials, 164, 146-153.
Reeves, P. G., & Chaney, R. L. (2008). Bioavailability as an issue in risk assessment and
management of food cadmium: A review. Science of the Total Environment, 398, 13-19.
Salaskar, D., Shrivastava, M., & Kale, S. P. (2011). Bioremediation potential of spinach
(Spinacia oleracea L.) for decontamination of cadmium in soil. Current Science, 101, 1359-1363.
Samiullah, Khan, N. A., Nazar, R., & Ahmad, I. (2007). Physiological basis for reduced
photosynthesis and growth of cadmium-treated wheat cultivars diering in yield potential. Journal
of Food Agriculture & Environment, 5, 375-377.
Sha, M., Bakht, J., Razuddin, Hayat, Y., & Zhang, G. P. (2011). Genotypic dierence in the
inhibition of photosynthesis and chlorophyll uorescence by salinity and cadmium stresses in
wheat. Journal of Plant Nutrition, 34, 315-323.
Siu, E. R., Mruk, D. D., Porto, C. S., & Cheng, C. Y. (2009). Cadmium-induced testicular injury.
Toxicology and Applied Pharmacology, 238, 240-249.
Stingu, A., Volf, I., Popa, V. I., & Gostin, I. (2012). New approaches concerning the utilization
of natural amendments in cadmium phytoremediation. Industrial Crops and Products, 35, 53-60.
Thevenod, F. (2009). Cadmium and cellular signaling cascades: To be or not to be? Toxicology
and Applied Pharmacology, 238, 221-239.
Thevenod, F. (2010). Catch me if you can! Novel aspects of cadmium transport in mammalian
cells. Biometals, 23, 857-875.
Vassilev, A., Yordanov, I., & Tsonev, T. (1997). Eects of Cd2+ on the physiological state and
photosynthetic activity of young barley plants. Photosynthetica, 34, 293-302.
Verbruggen, N., Hermans, C., & Schat, H. (2009). Mechanisms to cope with arsenic or
cadmium excess in plants. Current Opinion in Plant Biology, 12, 364-372.
von Caemmerer, S., & Farquhar, G. D. (1981). Some relationships between the biochemistry
of photosynthesis and the gas exchange of leaves. Planta, 153, 376-387.
Wang, H., McCarthney, A., Qiu, X., & Zhao, R. (2012). Cd2+ impact on metabolic cells
of Saccharomyces cerevisiae over an extended period and implications for bioremediation.
Geomicrobiology Journal, 29, 199-205.
Wang, Y., Hu, H., Xu, Y., Li, X. X., & Zhang, H. J. (2011). Dierential proteomic analysis of
cadmium-responsive proteins in wheat leaves. Biologia Plantarum, 55, 586-590.
Wong, E. W. P., & Cheng, C. Y. (2012). Impacts of environmental toxicants on male
reproductive dysfunction. Trends in Pharmacological Sciences, 32, 290-299.