Tetracycline accumulates in Iberis sempervirens L. through apoplastic transport inducing oxidative stress and growth inhibition.

Environmental antibiotic contamination is due mainly to improper and illegal disposal of these molecules that, yet pharmacologically active, are excreted by humans and animals. These compounds contaminate soil, water and plants. Many studies have reported the bioaccumulation of antibiotics in plants and their negative effects on photosynthesis, cell growth and oxidative balance. Therefore, the principal objective of this paper was the study of antibiotic accumulation sites in plants and its uptake modality. Iberis sempervirens L., grown in soil and in agar in the presence or absence of tetracycline, were used as a model system. Using confocal and transmission electron microscopy, we demonstrated that tetracycline was absorbed and propagated in plants through apoplastic transport and also accumulated in intercellular spaces. Tetracycline was rarely detected inside cells (in cytoplasm and mitochondria where, coherent to its pharmacological activity, it probably affected ribosomes), except in stomata. Moreover, we verified and clarified further the phytotoxic effects of tetracycline on plants. We observed that the antibiotic induced a large reduction in plant growth and development and inhibition of photosynthetic activity. As tetracycline may lead to oxidative stress in plants, plant cells tried to balance this disequilibrium by increasing the amount and activity of some endogenous enzyme antioxidant agents (superoxide dismutase 1 and catalase) and levels of antiradical secondary metabolites.

[1]  Michael Martin,et al.  Distribution, fate and risk assessment of antibiotics in sewage treatment plants in Hong Kong, South China. , 2012, Environment international.

[2]  A. Gismondi,et al.  Biochemical, Antioxidant and Antineoplastic Properties of Italian Saffron (Crocus sativus L.) , 2012 .

[3]  P. Taylor,et al.  Localization of the two major allergens in rye-grass pollen using specific monoclonal antibodies and quantitative analysis of immunogold labelling , 1994, The Histochemical Journal.

[4]  A. Masia Superoxide dismutase and catalase activities in apple fruit during ripening and post‐harvest and with special reference to ethylene , 1998 .

[5]  D. Hawker,et al.  Investigation of the mechanism of uptake and accumulation of zwitterionic tetracyclines by rice (Oryza sativa L.). , 2012, Ecotoxicology and environmental safety.

[6]  L. Migliore,et al.  Effect of sulphadimethoxine contamination on barley (Hordeum distichum L., Poaceae, Liliposida) , 1996 .

[7]  J Zhang,et al.  Uptake of oxytetracycline and its phytotoxicity to alfalfa (Medicago sativa L.). , 2007, Environmental pollution.

[8]  Salvatore Cozzolino,et al.  Phytotoxicity to and uptake of enrofloxacin in crop plants. , 2003, Chemosphere.

[9]  Gerard D. Wright,et al.  Intrinsic antibiotic resistance: mechanisms, origins, challenges and solutions. , 2013, International journal of medical microbiology : IJMM.

[10]  Samir Samman,et al.  Flavonoids—Chemistry, metabolism, cardioprotective effects, and dietary sources , 1996 .

[11]  A. Polle,et al.  Responses of antioxidative enzymes to elevated CO2 in leaves of beech (Fagus sylvatica L.) seedlings grown under a range of nutrient regimes , 1997 .

[12]  I. Huseynova Photosynthetic characteristics and enzymatic antioxidant capacity of leaves from wheat cultivars exposed to drought. , 2012, Biochimica et biophysica acta.

[13]  D. Calamari,et al.  Pharmaceuticals in the Environment in Italy: Causes, Occurrence, Effects and Control , 2006, Environmental science and pollution research international.

[14]  M. Tamoi,et al.  Regulation and function of ascorbate peroxidase isoenzymes. , 2002, Journal of experimental botany.

[15]  D. Kolpin,et al.  Transport of chemical and microbial compounds from known wastewater discharges: potential for use as indicators of human fecal contamination. , 2005, Environmental science & technology.

[16]  A. Polle,et al.  Differential stress responses of antioxidative systems to drought in pendunculate oak (Quercus robur) and maritime pine (Pinus pinaster) grown under high CO(2) concentrations. , 2001, Journal of experimental botany.

[17]  Ashutosh Kumar Singh,et al.  Antibiotic Use in Agriculture and Its Impact on the Terrestrial Environment , 2005 .

[18]  S. Jørgensen,et al.  Occurrence, fate and effects of pharmaceutical substances in the environment--a review. , 1998, Chemosphere.

[19]  Satish C. Gupta,et al.  Sulfamethazine uptake by plants from manure-amended soil. , 2007, Journal of environmental quality.

[20]  Wei Wang,et al.  A universal and rapid protocol for protein extraction from recalcitrant plant tissues for proteomic analysis , 2006, Electrophoresis.

[21]  R Hirsch,et al.  Occurrence of antibiotics in the aquatic environment. , 1999, The Science of the total environment.

[22]  D. Inzé,et al.  Signal transduction during oxidative stress. , 2002, Journal of experimental botany.

[23]  Marilena E. Dasenaki,et al.  Multi-residue determination of seventeen sulfonamides and five tetracyclines in fish tissue using a multi-stage LC-ESI-MS/MS approach based on advanced mass spectrometric techniques. , 2010, Analytica chimica acta.

[24]  R. Gryglewski,et al.  On the mechanism of antithrombotic action of flavonoids. , 1987, Biochemical pharmacology.

[25]  Diana S. Aga,et al.  Potential Ecological and Human Health Impacts of Antibiotics and Antibiotic-Resistant Bacteria from Wastewater Treatment Plants , 2007, Journal of toxicology and environmental health. Part B, Critical reviews.

[26]  H. Lichtenthaler CHLOROPHYLL AND CAROTENOIDS: PIGMENTS OF PHOTOSYNTHETIC BIOMEMBRANES , 1987 .

[27]  H. Jeske,et al.  Remission of the free-branching pattern of Euphorbia pulcherrima by tetracycline treatment. , 2000 .

[28]  K. Wakasa,et al.  Guanosine tetra- and pentaphosphate synthase activity in chloroplasts of a higher plant: association with 70S ribosomes and inhibition by tetracycline. , 2004, Nucleic acids research.

[29]  A. Gómez-Cadenas,et al.  Metabolomics as a Tool to Investigate Abiotic Stress Tolerance in Plants , 2013, International journal of molecular sciences.

[30]  C. Pini,et al.  Localisation of a carbohydrate epitope recognised by human IgE in pollen of Cupressaceae , 2004, Journal of Plant Research.

[31]  L. Kapustka Selection of phytotoxicity tests for use in ecological risk assessments , 1997 .

[32]  A Yonath,et al.  Crystal structures of complexes of the small ribosomal subunit with tetracycline, edeine and IF3 , 2001, The EMBO journal.

[33]  S. Allakhverdiev,et al.  Photoinhibition of photosystem II under environmental stress. , 2007, Biochimica et biophysica acta.

[34]  Marta Carballa,et al.  Behavior of pharmaceuticals, cosmetics and hormones in a sewage treatment plant. , 2004, Water research.

[35]  G. Rotilio,et al.  Purification of iron superoxide dismutase from the cyanobacterium Anabaena cylindrica Lemm. and localization of the enzyme in heterocysts by immunogold labeling , 1992, Planta.

[36]  S. Hosseinimehr,et al.  Antioxidant activity, phenol and flavonoid contents of some selected Iranian medicinal plants , 2006 .

[37]  Yaqi Cai,et al.  Occurrence of antibiotics in eight sewage treatment plants in Beijing, China. , 2012, Chemosphere.

[38]  W. Saenger,et al.  Structural basis of gene regulation by the tetracycline inducible Tet repressor–operator system , 2000, Nature Structural Biology.

[39]  A. Canini,et al.  Nutraceutical properties of honey and pollen produced in a natural park. , 2012 .

[40]  H. Aebi,et al.  Catalase in vitro. , 1984, Methods in enzymology.

[41]  D. Kolpin,et al.  Pharmaceuticals, Hormones, and Other Organic Wastewater Contaminants in U.S. Streams , 2005 .

[42]  M. M. Bradford A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[43]  L. Migliore,et al.  Oxytetracycline as environmental contaminant in arable lands. , 2007, Analytica chimica acta.

[44]  S. Milz,et al.  Characterization of eight different tetracyclines: advances in fluorescence bone labeling , 2010, Journal of anatomy.

[45]  P. Prenzler,et al.  Phenolic content and antioxidant activity of olive extracts , 2001 .

[46]  S. Jørgensen,et al.  Drugs in the environment. , 2000, Chemosphere.

[47]  W. Hillen,et al.  Tetracyclines in Biology, Chemistry and Medicine , 2001, Birkhäuser Basel.

[48]  John Jensen,et al.  Veterinary Medicines and Soil Quality: The Danish Situation as an Example , 2001 .

[49]  E. Frankel Nutritional Benefits of Flavonoids , 1997 .

[50]  E. Thurman,et al.  Pharmaceuticals, hormones, and other organic wastewater contaminants in U.S. streams, 1999-2000: a national reconnaissance. , 2002 .

[51]  C. Rosen,et al.  Antibiotic uptake by plants from soil fertilized with animal manure. , 2005, Journal of environmental quality.

[52]  Jun Yu Li,et al.  Antibiotic contamination in a typical developing city in south China: occurrence and ecological risks in the Yongjiang River impacted by tributary discharge and anthropogenic activities. , 2013, Ecotoxicology and environmental safety.

[53]  James Fletcher,et al.  Effects of Ten Antibiotics on Seed Germination and Root Elongation in Three Plant Species , 2011, Archives of environmental contamination and toxicology.

[54]  Gianfranco Brambilla,et al.  Hormetic effect(s) of tetracyclines as environmental contaminant on Zea mays. , 2010, Environmental pollution.

[55]  P. Krogh,et al.  Effects of the antibiotics oxytetracycline and tylosin on soil fauna. , 2000, Chemosphere.

[56]  M. Oturan,et al.  Study of the toxicity of sulfamethoxazole and its degradation products in water by a bioluminescence method during application of the electro-Fenton treatment , 2011, Analytical and bioanalytical chemistry.