Journal of Environment and Sociobiology

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Molecular Markers in Plant-Based Bioassays for the Detection of Molecular Endpoints to Probe of Aquatic Genotoxicity-An Overview


Affiliations
1 Plant Cytogenetics and Molecular Biology Laboratory, Post Graduate Dept of Botany, Hooghly Mohsin College, Chinsurah, Hooghly, India
2 Dept of Zoology, Hooghly Moshin College, Chinsurah, Hooghly, India
     

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Many contaminants entering the aquatic environment have been lipophilic by nature and can be readily taken up by aquatic organisms or absorbed by particulate matter. Examples of such compounds include polycyclic aromatic hydrocarbons, polychlorinated biphenyls and phthalates. Therefore, the impact of these aquatic contaminants on ecosystems largely depends upon factors including uptake and biotransformation by different species, and effects can be easily seen at the cellular level through the individual, population and ultimately the ecosystem as a whole. Research in this particular area has gained much attention and momentum in the recent past, with particular interests thrust upon the presence of different genotoxic agents surrounding in the environment. Results from the U.S. Environmental Protection Agency's (EPAs) Toxic Release Inventory (TRI) found out that several known and many dubious mutagenic and genotoxic chemicals could be readily accounted in contaminated surface waters, of which one third of these toxicants from effluent discharges are practically rodent carcinogens. Moreover, 800 metric tonnes of chemicals released into surface waters every year have been detected to be class 1, 2A or 2B carcinogens as classified by the International Agency for Research on Cancer (IARC). It is therefore clear that the assessment of complex mixtures for their genotoxic potential fits in the list of an extremely important consideration for environmental pollution monitoring and management, especially while considering the implications of these compounds in initiation, propagation and development of carcinogenesis, inherited disease and teratogenesis. Concomitantly, it is important to develop reliable and effective methods for detecting endpoints indicative of exposure to genotoxicants in particular using test methods that would be simple, rapid and cost effective.

Antioxidant enzymes, i.e., ROS scavengers, chromosomal abnormalities resultant of altered mitotic index and micronuclei formation, the alkaline COMET Assay, RAPD, AFLP coupled with RT-PCR can be employed as effective biochemical, cytogenetic and molecular markers in plants (as model systems for aquatic toxicology) which serve as best tools to detect the mode of action and levels of genotoxic endpoints upon exposure to different genotoxins in aquatic environments.


Keywords

Genotoxication, Heavy Metals, Comet, RAPD, AFLP, RT-PCR.
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  • Adams, S. M., Giesy, J. P., Tremblay, L. A. and Eason, C. T. 2001. The use of biomarkers in ecological risk assessment: Recommendations from the Christ church conference on biomarkers in ecotoxicology. Biomarkers, 6: 1– 6.

  • Aebi, C., Lafontaine, E. R., Cope, L. D., Latimer, J. L., Lumbley, S. L., McCracken, G. H., J. and Hansen, E. J. 1998. Phenotypic effect of isogenic uspA1 and uspA2 mutations on Moraxella catarrhalis O35E. Infection and Immunity, 66: 3113–3119.

  • Agarwall, M., Shrivastava, N. and Padh, H. 2008. Advances in molecular marker techniques and their applications in plant sciences. Plant Cell Reports, 27(4): 617-631.

  • Aina, R., Labra, M., Fumagalli, P., Vannini, C., Marsoni, M., Cucchi, U., Bracale, M., Sgorbati, S. and Citterio, S. 2007. Thiol-peptide level and proteomic changes in response to cadmium toxicity in Oryza sativa L. ischolar_mains. Environmental and Experimental Botany, 59 (3): 381-892.

  • Akhtar, T. A., Lampi, M. A. and Greenberg, B. M. 2005. Identification of six differentially expressed genes in response to copper exposure in the aquatic plant Lemna gibba (Duckweed). Environ. Toxicol. Chem., 24: 1705 – 1715.

  • Asada, K. 2006. Production and scavenging of reactive oxygen species in chloroplasts and their functions. Plant Physiology, 141: 391–396.

  • Atienzar, F. A. and Jha, A. N. 2006. The random amplified polymorphic DNA (RAPD) assay and related techniques applied to genotoxicity and carcinogenesis studies: a critical review. Mutation Research, 613 (2-3): 76-102.

  • Attia, H. F., Karray, N. and Lachaal, M. 2009. Light interacts with salt stress in regulating superoxide dismutase gene expression in Arabidopsis. Plant Science, 177: 161–167.

  • Aust, S. D., Morehouse, L. A. and Thomas, C. E. 1985. Role of metals in oxygen radical reactions. Free Radicals Biology and Medicine, 1: 3-25.

  • Becana, M., Dalton, D. A., Moran, J. F., Iturbe-Ormaetxe, I., Matamoros, M. A. and Rubio, M. C. 2000. Reactive oxygen species and antioxidants in legume nodules. Physiologia Plantatum, 109: 372–381.

  • Boussama, N., Ouariti, O. and Ghorbel, M. H. 1999a. Changes in growth and nitrogen assimilation in barley seedlings under cadmium stress. Journal of Plant Nutrition, 22: 731–752.

  • Boussama, N., Ouariti, O., Suzuki, A. and Ghorbal, M. H. 1999b. Cd stress on nitrogen assimilation. Journal of Plant Physiology, 155: 310–317.

  • Callaway, A. S., Abranches, R., Scroggs, J., Allen, G. C. and Thompson, W. F. 2002. High throughput transgene copy number estimation by competitive PCR. Plant Molecular Biology Reporter, 20: 265-277.

  • Chaney, R. L. 1993. Zinc phytotoxicity. In : Robson, A. D. (ed.), Zinc in soil and plants. Kluner Academic Publishers. Dovdracut, the Netherlands, pp. 135–150.

  • Chaudhry, N. and Qurat-ul-Ain, Y. 2003. Effect of growth hormones, i.e., IAA, Kinetin and Heavy metal, i.e., Lead Nitrate on the internal morphology of leaf of Phaseolus vulgaris L. Pakistan Journal of Biological Sciences, 6(2): 157-163.

  • Costa de, C. A. Casali, V. W. D., Loures, E. G., Cecon, P. R. and Jordao, C. P. 1994. Content level of heavy metals in lettuce (Lactuca sativa L.) fertilized with organic compost from urban waste. Revista Ceres, 238: 629–640. http://www.epa.gov/scipoly/sap/.

  • De Vos, C. H. R., Ten Boukum, W. M., Vooijs, R., Schar, H. and De Kok, L. J. 1993. Effect of copper on fatty acid composition and peroxidaiton of lipids in the ischolar_mains of copper tolerant and sensitive Silene cucbalus. Plant Physiology Biochemistry, 31: 151-158.

  • Doncheva, S., Nikolov, B. and Ogneva, V. 1996. Effect of excess copper on the morphology of the nucleus in maize ischolar_main meristem cells. Physiologia Plantarum, 96 (1): 118-122.

  • Dong, J., Wu, F. and Zhang, G. 2006. Influence of cadmium on antioxidant capacity and four microelement concentrations in tomato seedlings (Lycopersicon esculentum). Chemosphere, 64: 1659–1666.

  • Dorak, T. 2006. Real-Time PCR. An introduction to real-time PCR. Taylor & Francis Group. http://www.gene-quantification.de.

  • Enan, M. R. 2006. Application of random amplified polymorphic DNA (RAPD) to detect the genotoxic effect of heavy metals. Biotechnology and Applied Biochemistry, 43 (3): 147-154.

  • Erbes, M., Wessler, A., Obst, U. and Wild, A. 1997. Detection of primary DNA damage in Chlamydomonas reinhardtii by means of modified microgel electrophoresis. Environ. Mol. Mutagen, 30:448–458.

  • Ernst, W. H, O. and Peterson, P. J. 1994. The role of biomarkers in environmental assessment of terrestrial plants. Ecotoxicol., 3: 180-192.

  • Ferrat, L., Pergent-Martini, C. and Roméo, M. 2003. Assessment of the use of biomarkers in aquatic plants for the evaluation of environmental quality: Application to seagrasses . Aquat. Toxicol., 65: 187-204.

  • Fernandes, J. C. and Henriques, F. S. 1991. Biochemical, physiological and structural effect of excess copper in plants. Botanical Review, 57: 246-273.

  • Florijn, P. J, and Van Beusichem, M. L. 1993. Uptake and distribution of cadmium in maize inbred lines. Plant and Soil, 150: 25-32.

  • Forbes, V. E., Palmqvist, A. and Bach, L. 2006. The use and misuse of biomarkers in ecotoxicology. Environ. Toxicol. Chem., 25: 272-280.

  • Fridovich, I. 1995. Superoxide radical and superoxide dismutases. Annual Review Biochemistry, 64: 97–112.

  • Goupila, P., Souguira, D., Ferjanib, E., Faurec, O., Hitmid, A. and Ledoigta, G. 2009. Expression of stress-related genes in tomato plants exposed to arsenic and chromium in nutrient solution. Journal of Plant Physiology, 166(13:1): 1446-1452.

  • Grant, W. F. 1994. The present status of higher plant bioassays for the detection of environmental mutagens. Mutation Research, 310: 175-185.

  • Gupta, S. L. 1986. Copper uptake and inhibition of growth, photosynthetic pigments and macromolecules in the cyanobacterium Anacystis nidulans. Photosynthetica, 20 (4): 447-453.

  • Halliwell, B. and Gutteridge, J. M. C. 1989. Protection against oxidants in biological systems: The superoxide theory of oxygen toxicity. In: Halliwell, B., Gutteridge, J.M.C. (eds.) Free radicals in biology and medicine. Clarendon Press, Oxford.

  • Hodges, D. M., DeLong, J. M., Forney, C. F. and Prange, R. K. 1999. Improving the thiobarbituric acid reactive substances assay for estimating lipid peroxidation in plant tissues containing anthocyanin and other interfering compounds. Planta, 207: 604–611.

  • Humphrey, M. O. and Nicholls, M. K. 1984. Relationships between tolerances to heavy metals in Agrostis cupillaris L. ( A. tenuis Sibth.). New Phytology, 98: 177–190.

  • Ingham, D. J., Beer, S., Money, S. and Hansen, G. 2001. Quantitative real-time PCR assay for determining transgene copy number in transformed plants. Biotechniques, 31: 132-134, 136–140.

  • Jamers, A., Van der Ven, K. , Moens, L. , Robbens, J. , Potters, G., Guisez, Y., Blust, R. and De Coen, W. 2006. Effect of copper exposure on gene expression profiles in Chlamydomonas reinhardtii based on microarray analysis. Aquat. Toxicol., 80: 249–260.

  • Jimi, E., Aoki, K., Saito, H., D’Acquisto, F., May, M. J. and Nakamura, I. 2004. Selective inhibition of NF-kappa B blocks osteoclasto-genesis and prevents inflammatory bone destruction in vivo. Nature Medicine, 10: 617–624.

  • Joshua, S. Y., Reed, A., Chen, F. and Neal Stewart, C. 2006. Statistical analysis of realtime PCR data. BMC Bioinformatics. Methodology article. doi: 10.1186/14712105-7-85.

  • Kanazawa, T., Nakamura, S, Momoi, M., Yamaji, T., Takematsu, H., Yano, H., Sabe, H., Yamamoto, A., Kawasaki, T. and Kozutsumi, Y. 2000. Inhibition of cytokinesis by a lipid metabolite psychosine. The Journal of Cell Biology, 149: 943–950.

  • Knasmuller, S., Gottmann, E., Steinkellner, H., Fomin, A., Pickl, C., God, R. and Kundi, M. 1998. Detection of genotoxic effects of heavy metal contaminated soils with plant bioassays. Mutation Research, 420: 37–48.

  • Korpe, D. A. and Aras, S. 2011. Evaluation of copper-induced stress on egg plant (Solanum melongena L.) seedlings at the molecular and population levels by use of various biomarkers. Mutation Research, 719 (1-2): 29–34.

  • Koppen, G. and Verschaeve, L. 1996. The alkaline Comet test on plant cells: a new genotoxicity test for DNA strand breaks in Vicia faba ischolar_main cells. Mutat. Res. (Environ Mutagen RS), 360:193–200.

  • Kreuz, K. and Martinoia, E. 1999. Herbicide metabolism in plants: Integrated pathways of detoxification. In: The Proceedings of the 9th International Congress on Pesticide Chemistry: The Food-Environment Challenge. Brooks, G. T., Roberts T. R. (eds.) The Royal Society of Chemistry, London, pp: 279–287 .

  • Kuroda, S., Yano, H., Koga-Ban, Y., Tabei, Y., Takaiwa, F., Kayano, T. and Tanaka, H. 1999. Identification of DNA polymorphism induced by X-ray and UV irradiation in plant cells. Japan Agricultural Research Quarterly, 33: 223-226.

  • Labra, M., Di Fabio, T., Grassi, F., Regondi, S.M., Bracale, M., Vannini, C. and Agradi, E. 2003. AFLP analysis as biomarker of exposure to organic and inorganic genotoxic substances in plants. Chemosphere, 52(7): 1183–1188.

  • Lettieri, T. 2006. Recent applications of DNA microarray technology to toxicology and ecotoxicology. Environ. Health Perspect, 114: 4–9.

  • Li, X., Lin, H., Zhang, W., Zou, Y., Zhang, J., Tang, X. and Zhou, J. M. 2005. Flagellin induces innate immunity in nonhost interactions that is suppressed by Pseudomonas syringae effectors. Proceedings of the National Academy of Sciences of the United States of America, 102: 12990–12995.

  • Lidon, F. C. and Henriques, F. S. 1991. Effects of copper on the ascorbate, diamine and odiphenol oxidases activities of rice leaves. Oyton-International Journal of Experimental Botany, 52: 97–104.

  • Lipman, C. B. And McKinney, G. 1931. Proof of essential nature of copper for higher green plants. Plants Physiol., 6: 539–599.

  • Lin, C. C. and Kao, C. H. 1999. NaCl induced changes in ionically bound peroxidase activity in ischolar_mains of rice seedlings. Plant and Soil, 216: 147–153.

  • Lin, C. C. and Kao, C. H. 2000. Effect of NaCl stress on H2O2 metabolism in rice leaves. Plant Growth Regulation, 30: 151–155.

  • Lipman, C. B and Mekinney, G. 1931. Proof of the essential nature of copper for higher green plants. Plant Physical., 6: 539–599.

  • Liu, W., Yang, Y. S., Zhou, Q., Xie, L., Li, P. and Sun, T. 2007. Impact assessment of cadmium contamination on rice (Oryza sativa L.) seedlings at molecular and population levels using multiple biomarkers. Chemosphere, 67 (6): 1155–1163.

  • Liu, W., Li, P., Qi, X. M., Zhou, Q., Zheng, L., Sun, T. and Yang, Y. 2005. DNA changes in barley (Hordeum vulgare) seedlings induced by cadmium pollution using RAPD analysis. Chemosphere, 61: 158–167.

  • Marisa, L., Wong, J. F. and Medrano. 2005. Real-time PCR for mRNA quantitation. BioTechniques, 39: 75–85.

  • McCord, J. M. and Fridovich, I. 1969. Superoxide dismutase: an enzymic function for erythrocuprein (hemocuprein). The Journal of Biological Chemistry, 244: 6049–6055.

  • McCarty, L. S. and Munkittrick, K. R. 1996. Environmental biomarkers in aquatic toxicology: Fiction, fantasy, or functional ? Human Ecol. Risk Assess., 2: 268 –274.

  • Miller, A. F. and Sorkin, D. L. 1997. “Superoxide dismutases: A molecular perspective”. Comments on Molecular and Cellular Biophysics, 9(1): 1–48.

  • Muller, L. A. H., Lambaerts, M., Vangronsveld, J. and Colpaert, V. 2004. AFLPbased assessment of the effects of environmental heavy metal pollution on the genetic structure of pioneer populations of Suillus luteus. New Phytologist, 164: 297–303.

  • Munzuroglu, O. and Geckil, H. 2002. Effects of metals on seed germination, ischolar_main elongation and coleoptile and hypocotyl growth in Triticum aestivum and Cucumis sativus. Archives Environmental Contamination and Toxicology, 43: 203–213.

  • Nuwaysir, E. F., Bittner, M., Trent, J., Barrett, J. C. and Afshari, C. A. 1999. Microarrays and toxicology: The advent of toxicogenomics. Molecular Carcinogenesis, 24: 153–159.

  • Nedelkoska, T. V. and Doran, P. M. 2000. Characteristics of heavy metal uptake by plant species with potential for phytoremediation and phytomining. Minerals Engineering, 13: 549–561.

  • Nussbaum, S., Schmutz, D. and Brunold, C. 1988. Regulation of assimilatory sulfate reduction by cadmium in Zea mays L. Plant Physiology, 88: 1407–1410.

  • Ouzounidou, G., Eleftheriou, E. P. and Karataglis, S. 1992. Ecophysiological and ultrastructural effects of copper in Thlaspi ochroleucum (Curciferae). Canadian Journal of Botany, 70: 947–957.

  • Park, S., Polle, J. E. W., Melis, A., Lee, T. K. and Jin, E. 2006. Up-regulation of photoprotection and PSII repair gene expression by irradiance in the unicellular green alga Dunaliella salina. Marine Biotechnol., 8: 120–128.

  • Poschenrieder, C., Gunse, B. and Barcelo, J. 1989. Influence of cadmium on water relations, stomatal resitance and abscisic acid content in expanding bean leaves. Plant Physiology, 90: 1465–1371.

  • Qilin, D., D., Wang, Jin, W., Feng Bin, F., Liu Tingting, L., Chen Chen, C., Lin Honghui, L. and Shizhang, D. 2009. Molecular cloning and characterization of a new peroxidise gene (OvRCI) from Orychophragmus violaceus. African Journal of Biotechnology, 8 (23): 6511–6517.

  • Rai, L. C. and Raizada, M. 1988. Impact of chromium and lead on Nostoc muscorum: Regulation of toxicity by ascorbic acid, glutathione and sulphur-containing amino acids. Ecotoxicology and Environmental Safety, 15: 195–205.

  • Sanchez-Estudillo, L., Freile-Pelegrin, Y., Rivera-Madrid, R., Robledo, D. and Narva’ez-Zapata, J. A. 2006. Regulation of two photosynthetic pigment-related genes during stress-induced pigment formation in the green alga, Dunaliella salina. Biotechnol. Lett., 28: 787–791.

  • Sandalio, L. M., Dalurzo, H. C., Gomes, M., Romero-Puertas, M. and Del Rio, L. A. 2001. Cadmium-induced changes in the growth and oxidative metabolism of pea plants. Journal of Experimental Botany, 52: 2115–2126.

  • Sandermann, H. J. R. 1994. Higher plant metabolism of xenobiotics: the ‘green liver’ concept. Pharmacogenetics, 4(5): 225–241.

  • Santos, C., Gaspar, M., Caeiro, A., Branco-Price, C., Teixeira, A. and Ferreira, R. B. 2006. Exposure of Lemna minor to arsenite: Expression levels of the components and intermediates of the ubiquitin/ proteasome pathway. Plant Cell Physiol., 47: 1262–1273.

  • Sharma, P. and Dubey, R. S. 2005. Lead toxicity in plants. Brazilian Journal of Plant Physiology, 17: 35–52.

  • Singh, N. P., McCoy, M. T., Tice, R. R. and Schneider, E. L. 1988. A simple technique for quantitation of low levels of DNA damage in individual cells. Exp. Cell. Res., 175: 184–191.

  • Somashekaraiah, B. V., Padmaja, K. and Prasad, A. R. K. 1992. Phytotoxicity of cadmium ions on germinating seedlings of mung bean (Phaseolus vulgaris) : involvement of lipid peroxides in chlorophyll degradation. Physiologia Plantarum, 85: 85–89.

  • Sommer, A. L. 1931. Copper as an essential for plant growth. Plant Physiology, 6: 339–345.

  • Sommer, A. L. and Lipman, C. B. 1926. Evidence on the indispensable nature of zinc and boron for higher green plants. Plant Physiology, 1: 231–249.

  • Song, P., Cai, C. Q., Skokut, M., Kosegi, B. D. and Petolino, J. F. 2002. Quantitative real-time PCR as a screening tool for estimating transgene copy number in WHISKERSTM derived transgenic maize. Plant Cell Reports, 20: 948–954.

  • Soydam Aydın, S., Gokce, E., Büyük, İ. and Aras, S. 2012. Characterization of Copper and Zinc induced stress on cucumber (Cucumis sativus L.) by molecular and population parameters. Mutation Research, 746 (1): 49–55.

  • Soydam Aydın, S., Basaran, E., Cansaran-Duman, D., and Aras, S. 2013. Genotoxic effect of cadmium in okra (Abelmoschus esculantus L.) seedlings: comperative inverstigation with population parameter and molecular marker. Journal of Environmental Biology, 34: 985–990.

  • Turner, J. G., Ellis, C. H., and Devoto, A. 2002. The jasmonate signal pathway. Plant Cell, 14 (Suppl): 153–164.

  • USEPA. 2004. Potential implications of genomics for regulatory and risk assessment applications at EPA. Science Policy Council U.S. Environmental Protection Agency, Washington, DC.

  • Willekens, H., Inze , D., Van Montagu, M. and Van Camp, W. 1995. Catalases in plants. Mol. Breeding, 1: 207–228.

  • Yi, T. H. and Ching, H. K. 2003. Changes in protein and amino acid contents in two cultivars of rice seedlings with different apparent tolerance to cadmium. Plant Growth Regulation, 40: 147–155.

  • Yoshida, Y. 2002. E3 ubiquitin ligase that recognizes sugar chains. Nature, 418: 438–442.

  • Zhou, Q. X., Cheng, Y., Zhang, Q. R. and Liang, J. D. 2003. Quantitative analyses of relationships between ecotoxicological effects and combined pollution. Science in China Series, 33: 566–573.


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  • Molecular Markers in Plant-Based Bioassays for the Detection of Molecular Endpoints to Probe of Aquatic Genotoxicity-An Overview

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Authors

Dipan Adhikari
Plant Cytogenetics and Molecular Biology Laboratory, Post Graduate Dept of Botany, Hooghly Mohsin College, Chinsurah, Hooghly, India
Sarmila Pal
Dept of Zoology, Hooghly Moshin College, Chinsurah, Hooghly, India

Abstract


Many contaminants entering the aquatic environment have been lipophilic by nature and can be readily taken up by aquatic organisms or absorbed by particulate matter. Examples of such compounds include polycyclic aromatic hydrocarbons, polychlorinated biphenyls and phthalates. Therefore, the impact of these aquatic contaminants on ecosystems largely depends upon factors including uptake and biotransformation by different species, and effects can be easily seen at the cellular level through the individual, population and ultimately the ecosystem as a whole. Research in this particular area has gained much attention and momentum in the recent past, with particular interests thrust upon the presence of different genotoxic agents surrounding in the environment. Results from the U.S. Environmental Protection Agency's (EPAs) Toxic Release Inventory (TRI) found out that several known and many dubious mutagenic and genotoxic chemicals could be readily accounted in contaminated surface waters, of which one third of these toxicants from effluent discharges are practically rodent carcinogens. Moreover, 800 metric tonnes of chemicals released into surface waters every year have been detected to be class 1, 2A or 2B carcinogens as classified by the International Agency for Research on Cancer (IARC). It is therefore clear that the assessment of complex mixtures for their genotoxic potential fits in the list of an extremely important consideration for environmental pollution monitoring and management, especially while considering the implications of these compounds in initiation, propagation and development of carcinogenesis, inherited disease and teratogenesis. Concomitantly, it is important to develop reliable and effective methods for detecting endpoints indicative of exposure to genotoxicants in particular using test methods that would be simple, rapid and cost effective.

Antioxidant enzymes, i.e., ROS scavengers, chromosomal abnormalities resultant of altered mitotic index and micronuclei formation, the alkaline COMET Assay, RAPD, AFLP coupled with RT-PCR can be employed as effective biochemical, cytogenetic and molecular markers in plants (as model systems for aquatic toxicology) which serve as best tools to detect the mode of action and levels of genotoxic endpoints upon exposure to different genotoxins in aquatic environments.


Keywords


Genotoxication, Heavy Metals, Comet, RAPD, AFLP, RT-PCR.

References