Abstract
The use of an adequate range of taxa in ecotoxicological studies is a key point for the achievement of ecologically relevant results. Higher plants are an essential part of a healthy and balanced ecosystem and new plant models are essential in the evaluation of potential impacts of pollutants. With more than 10,000 living species, ferns are the second-largest group of vascular plants. Fern spores, and spore-developed gametophytes, have long been recognized as useful models for plant research in important areas. One of the main advantages of this model is its naturally miniature size. Fern spores are single meiotic cells which develop into gametophytes which are miniature mature higher plants. The use of microtubes and microplates is imposed by the natural model. Chronic toxicity testing involves longer periods of exposure to toxicants (>48 h) and assesses the ability of a substance to disrupt a significant portion of an organism’s life stage. DNA content in developing gametophytes is a biomarker of the disturbance that the toxicant provokes in gametophyte growth and development. The use of the DNA fluorescent probe Hoechst is a rapid, sensitive, and reliable method frequently used in cell biology which has been successfully applied to spores and gametophytes. Chlorophyll autofluorescence can also be used as a biomarker of the physiological state. Both biomarkers can easily be measured using 96 multiwell plates and plate readers. The combined use of these biomarkers in chronic toxicity tests using developing gametophytes of the riparian Polystichum setiferum is yielding very satisfactory results and is a promising new model for Ecotoxicology.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Agati, G. 1998. Response of the in vivo chlorophyll fluorescence spectrum to environmental factors and laser excitation wavelength. Pure Appl Opt 7, 797–807.
Banks, J. A. 1999. Gametophyte development in ferns. Annu Rev Plant Phys 50, 163–186.
Benenati, F. E. 1990. Keynote address: Plants – kesystone to risk assessment. In Plants for toxicity assessment, Eds W-c Wang, J W Gorsuch and W R Lower. pp. 5–14. ASTM, Philadelphia.
Catalá, M., Esteban, M., Rodríguez-Gil, J. L., and Quintanilla, L. G. 2009. Development of a naturally miniaturised testing method based on the mitochondrial activity of fern spores: a new higher plant bioassay. Chemosphere 77, 983–988.
Czerniawska-Kusza, I., Ciesielczuk, T., Kusza, G., and Cichon, A. 2006. Comparison of the Phytotoxkit microbiotest and chemical variables for toxicity evaluation of sediments. Environ Toxicol 21, 367–372.
Daxhelet, G. A., Coene, M..M, Hoet, P. P., and Cocito, C. G. 1989. Spectrofluorometry of dyes with DNAs of different base composition and conformation. Anal Biochem 179, 401–403.
Doust, J. L., Schmidt, M., and Doust, L. L. 1994. Biological assessment of aquatic pollution – a review, with emphasis on plants as biomonitors. Biol Rev 69, 147–186.
Fent, K., Weston, A. A., and Caminada, D. 2006. Ecotoxicology of human pharmaceuticals. Aquat Toxicol 76, 122–159.
Ferrat, L., Pergent-Martini, C., and Romeo, 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.
Galbraith, D. W., Harkins, K. R., and Jefferson, R. A. 1988. Flow cytometric characterization of the chlorophyll contents and size distributions of plant protoplasts. Cytometry 9, 75–83.
Khetan, S. K., and Collins, T. J. 2007. Human pharmaceuticals in the aquatic environment: a challenge to green chemistry. Chem Rev 107, 2319–2364.
Krause, G. H., and Weis, E. 1991. Chlorophyll fluorescence and photosynthesis – the basics. Annu Rev Plant Phys 42, 313–349.
Labarca, C., and Paigen, K. 1980. Simple, rapid, and sensitive DNA assay procedure. Anal Biochem 102, 344–352.
Landis, W. G., Hughes, J. S., and Lewis, M. A. 1993. Environmental toxicology and risk assessment. ASTM, Philadelphia. 431p.
Lewis, M. A. 1995. Use of fresh-water plants for phytotoxicity testing – a review. Environ Pollut 87, 319–336.
Maiti, S., Maiti, P., Sinha, S. S., Mitra, R. K., and Pal, S. K. 2009. Molecular recognition of plant DNA: does it differ from conventional animal DNA? Int J Biol Macromol 44, 133–137.
Mocharla, R., Mocharla, H., and Hodes, M. E. 1987. A novel, sensitive fluorometric staining technique for the detection of DNA in RNA preparations. Nucleic Acids Res 15, 133–137.
Mohan, B. S., and Hosetti, B. B. 1999. Aquatic plants for toxicity assessment. Environ Res 81, 259–274.
OECD 2007 Section 2 – Effects on Biotic Systems (Draft). In Guidelines for the Testing of Chemicals. Organisation for Economic Co-Operation and Development, Paris.
Quintanilla, L. G., and Escudero, A. 2006. Spore fitness components do not differ between diploid and allotetraploid species of Dryopteris (Dryopteridaceae). Ann Bot 98, 609–618.
Rodriguez-Gil, J. L., Catala, M., Alonso, S. G., Maroto, R. R., Valcarcel, Y., Segura, Y., Molina, R., Melero, J. A., and Martinez, F. 2010. Heterogeneous photo-Fenton treatment for the reduction of pharmaceutical contamination in Madrid rivers and ecotoxicological evaluation by a miniaturized fern spores bioassay. Chemosphere 80(4), 381–388.
USEPA 1972 The Clean Water Act (33 U.S.C. 1251 et seq.) (Federal Water Pollution Control Act Amendments of 1972, 1977, and 1987). U.S. Environmental Protection Agency, Washington D.C.
USEPA 2010 Series 850 – Ecological Effects Test Guidelines (Draft). In Harmonized Test Guidelines. U.S. Environmental Protection Agency, Office of Chemical Safety and Pollution Prevention (OCSPP), Washington D.C.
USEPA 1997 Terms of Environment: Glossary, Abbreviations and Acronyms (EPA Publication No.175-B-97-001). U.S. Environmental Protection Agency, Washington D.C.
Wang, W. C. 1991. Literature-review on higher-plants for toxicity testing. Water Air Soil Poll 59, 381–400.
Acknowledgments
The authors want to thank Raquel Feito for giving permission to use unpublished results to illustrate the examples shown in this chapter.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this chapter
Cite this chapter
Catalá, M., Rodríguez-Gil, J.L. (2011). Chronic Phytotoxicity in Gametophytes: DNA as Biomarker of Growth and Chlorophyll Autofluorescence as Biomarker of Cell Function. In: Kumar, A., Fernández, H., Revilla, M. (eds) Working with Ferns. Springer, New York, NY. https://doi.org/10.1007/978-1-4419-7162-3_18
Download citation
DOI: https://doi.org/10.1007/978-1-4419-7162-3_18
Published:
Publisher Name: Springer, New York, NY
Print ISBN: 978-1-4419-7161-6
Online ISBN: 978-1-4419-7162-3
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)