Histological, Cytological and Biochemical Alterations Induced by Microcystin-LR and Cylindrospermopsin in White Mustard (Sinapis alba L.) Seedlings


This study compares the histological, cytological and biochemical effects of the cyanobacterial toxins microcystin-LR (MCY-LR) and cylindrospermopsin (CYN) in white mustard (Sinapis alba L.) seedlings, with special regard to the developing root system. Cyanotoxins induced different alterations, indicating their different specific biochemical activities. MCY-LR stimulated mitosis of root tip meristematic cells at lower concentrations (1 μg ml−1) and inhibited it at higher concentrations, while CYN had only inhibitory effects. Low CYN concentrations (0.01 μg ml−1) stimulated lateral root formation, whereas low MCY-LR concentrations increased only the number of lateral root primordia. Both inhibited lateral root development at higher concentrations. They induced lignifications, abnormal cell swelling and inhibited xylem differentiation in roots and shoots. MCY-LR and CYN induced the disruption of metaphase and anaphase spindles, causing altered cell divisions. Similar alterations could be related to decreased protein phosphatase (PP1 and PP2A) activities in shoots and roots. However, in vitro phosphatase assay with purified PP1 catalytic subunit proved that CYN in contrast to MCY-LR, decreased phosphatase activities of mustard in a non-specific way. This study intends to contribute to the understanding of the mechanisms of toxic effects of a protein phosphatase (MCY-LR) and a protein synthesis (CYN) inhibitory cyanotoxin in vascular plants.



Blue-Green Sinapis Test




4’,6’-diamidino-2 phenylindole


Glutathione-peroxidase enzymes


Glutathione-S-transferase enzymes


fifty percent inhibitory concentration


microtubule associated proteins






programmed cell death


serine/threonine protein phosphatases of type 1


catalytic subunit of protein phosphatase 1


serine/threonine protein phosphatases of type 2A


reactive oxygen species


  1. 1.

    Abe, T., Lawson, T., Weyers, J. D. B., Codd, G. A. (1996) Microcystin-LR inhibits photosynthesis of Phaseolus vulgaris primary leaves: Implications for current spray irrigation practice. New Phytol. 133, 651–658.

    CAS  Google Scholar 

  2. 2.

    Allen, M. M. (1968) Simple conditions for the growth of unicellular blue-green algae on plates. J. Phycol. 4, 1–4.

    CAS  PubMed  PubMed Central  Google Scholar 

  3. 3.

    Armstrong, J., Armstrong, W. (2001) An overview of the effects of phytotoxins on Phragmites aus-tralis in relation to die-back. Aquatic Bot. 69, 251–268.

    CAS  Google Scholar 

  4. 4.

    Banker, R., Carmeli, S., Hadas, O., Teltsch, B., Porat, R., Sukenik, A. (1997) Identification of cylin-drospermopsin in Aphanizomenon ovalisporum (Cyanophyceae) isolated from lake Kinneret, Israel. J. Phycol. 33, 613–616.

    CAS  Google Scholar 

  5. 5.

    Beyer, D., Surányi, G., Vasas, G., Roszik, J., Erdődi, F., M-Hamvas, M., Bácsi, I., Bátori, R., Serfőző, Z., Szigeti, Z. M., Vereb, G., Demeter, Z., Gonda, S., Mathe, C. (2009) Cylindrospermopsin induces alterations of root histology and microtubule organization in common reed (Phragmites australis) plantlets cultured in vitro. Toxicon 54, 440–449.

    CAS  Google Scholar 

  6. 6.

    Campos, A., Vasconcelos, V. (2010) Molecular mechanisms of microcystin toxicity in animal cells. Review. Int. J. Mol. Sci. 11, 268–287.

    CAS  PubMed  PubMed Central  Google Scholar 

  7. 7.

    Carmichael, W. W., Azevedo, S. M. F. O., An, J. S., Molica, R. J. R., Jochimsen, E. M., Lau, S., Rinehart, K. L., Shaw, G. R., Eaglesham, G. K. (2001) Human fatalities from cyanobacteria: chemical and biological evidence for cyanotoxins. Environ. Health Persp. 109, 663–668.

    CAS  Google Scholar 

  8. 8.

    Chaffey, N., Cholewa, E., Regan, S., Sundberg, B. (2002) Secondary xylem development in Arabidopsis: a model for wood formation. Physiol. Plan. 114, 594–600.

    CAS  Google Scholar 

  9. 9.

    Chen, J., Han, F. X., Wang, F., Zhang, H., Shi, Z. (2012) Accumulation and phytotoxicity of micro-cystin-LR in rice (Oryza sativa). Ecotoxicol. Environ. Saf. 76, 193–199.

    CAS  PubMed  Google Scholar 

  10. 10.

    Chen, J., Song, L., Dai, J., Gan, N., Liu, Z. (2004) Effects of microcystins on the growth and the activity of superoxide dismutase and peroxidase of rape (Brassica napus L.) and rice (Oryza sativa L.). Toxico. 43, 393–400.

    CAS  Google Scholar 

  11. 11.

    Chong, M. W. K., Wong, B. S. E., Lam, P. K. S., Shaw, G. R., Seawright, A. A. (2002) Toxicity and uptake mechanism of cylindrospermopsin and lophyrotomin in primary rat hepatocytes. Toxico. 40, 205–211.

    CAS  Google Scholar 

  12. 12.

    Conrath, U., Pieterse, C. M. J., Mauch-Mani, B. (2002) Priming in plant-pathogen interactions. Trends Plant Sci. 7, 210–216.

    CAS  PubMed  Google Scholar 

  13. 13.

    Demeter, A., Vámosi, J., Peregovits, L., Topál, G. (1995) An image capture and data collection system for morphometric studies. In: Marcus, L. F., Corti, M., Loy, A., Naylor, G., Slice, D. E. (eds) Advances in Morphometries. Plenum Press, New York, pp. 91–101.

    Google Scholar 

  14. 14.

    Erdődi, F., Tóth, B., Hirano, K., Hirano, M., Hartshorne, D. J., Gergely, P. (1995) Endothall thioan-hydride inhibits protein phosphatases-1 and -2A in vivo. Am. J. Physiol. 269, C1176–C1184.

    PubMed  Google Scholar 

  15. 15.

    Froscio, S. M., Humpage, A. R., Wickramasinghe, W., Shaw, G., Falconer, I. R. (2008) Interaction of the cyanobacterial toxin cylindrospermopsin with the eukaryotic protein synthesis system. Toxico. 51, 191–198.

    CAS  Google Scholar 

  16. 16.

    Fukuda, H. (2004) Signals that control plant vascular cell differentiation. Nat. Rev. Mol. Cell Bio. 5, 379–391.

    CAS  Google Scholar 

  17. 17.

    Gácsi, M., Antal, O., Vasas, G., Máthé, C., Borbély, G., Saker, M. L., Győri, J., Farkas, A., Vehovszky, Á., Bánfalvi, G. (2009) Comparative study of cyanotoxins affecting cytoskeletal and chromatin structures in CHO-K1 cells. Toxicol, in Vitr. 23, 710–718.

    Google Scholar 

  18. 18.

    Gehringer, M. M. (2004) Microcystin-LR and okadaic acid-induced cellular effects: a dualistic response. FEES Lett. 557, 1–8.

    CAS  Google Scholar 

  19. 19.

    Gergely, P., Erdődi, F., Bot, G. (1984) Heparin inhibits the activity of protein phosphatase-1. FEES Lett. 169, 45–48.

    CAS  Google Scholar 

  20. 20.

    Huang, W., Xing, W., Li, D., Liu, Y. (2008) Microcystin-RR induced apoptosis in tobacco BY-2 suspension cells is mediated by reactive oxygen species and mitochondrial permeability transition pore status. Toxicol, in Vitr. 22, 328–337.

    CAS  Google Scholar 

  21. 21.

    Kinnear, S. H. W., Fabbro, L. D., Duivenvoorden, L. J. (2008) Variable growth responses of water thyme (Hydrilla verticillata) to whole-cell extracts of Cylindrospermopsis raciborskii. Arc. Environ Contain. Toxicol. 54, 187–194.

    CAS  Google Scholar 

  22. 22.

    Kós, R., Gorzó, G., Suranyi, G., Borbély, G. (1995) Simple and efficient method for isolation and measurement of cyanobacterial hepatotoxins by plant tests (Sinapis alba). Anal. Biochem. 225, 49–53.

    Google Scholar 

  23. 23.

    Kováts, N., Ács, A., Paulovits, G., Vasas, G. (2011) Response of Lemna minor clones to microcystis toxicity. Appl. Ecol. Env. Res. 9, 17–26.

    Google Scholar 

  24. 24.

    Kurki-Helasmo, K., Meriluoto, J. (1998) Microcystin uptake inhibits growth and protein phosphatase activity in mustard (Sinapis alba L.) seedlings. Toxico. 36, 1921–1926.

    CAS  Google Scholar 

  25. 25.

    Luan, S. (2003) Protein phosphatases in plants. Annu. Rev. Plant Biol. 42, 63–92.

    Google Scholar 

  26. 26.

    MacKintosh, C., Beattie, K. A., Klumpp, S., Cohen, P., Codd, G. A. (1990) Cyanobacterial microcys-tin-LR is a potent and specific inhibitor of protein phosphatases 1 and 2A from both mammals and higher plants. FEES Lett. 264, 187–192.

    CAS  Google Scholar 

  27. 27.

    Máthé, C., M-Hamvas, M., Vasas, G., Surányi, G., Bácsi, I., Beyer, D., Tóth, S., Tímár, M., Borbely, G. (2007) Microcystin-LR, a cyanobacterial toxin, induces growth inhibition and histological alterations in common reed (Phragmites australis) plants regenerated from embryonic calli. New Phyto. 176, 824–835.

    Google Scholar 

  28. 28.

    Máthé, C., Beyer, D., Erdődi, F., Serfőző Z., Székvölgyi, L., Vasas, G., M-Hamvas, M., Jámbrik, K., Gonda, S., Kiss, A., Szigeti Z. M., Suranyi, G. (2009) Microcystin-LR induces abnormal root development by altering its microtubule organization in tissue-cultured common reed (Phragmites australis) plantlets. Aquat. Toxicol. 92, 122–130.

    PubMed  Google Scholar 

  29. 29.

    Metcalf J. S., Barakate, A., Codd, G. A. (2004) Inhibition of plant protein synthesis by the cyanobacterial hepatotoxin, cylindrospermopsin. FEMS Microbiol. Lett. 235, 125–129.

    CAS  Google Scholar 

  30. 30.

    Metcalf, J. S., Lindsay, J., Beattie, K. A., Birmingham, S., Saker, M. L., Törökné, A. K., Codd, G. A. (2002) Toxicity of cylindrospermopsin to the brine shrimp Artemia salina: comparisons with protein synthesis inhibitors and microcystins. Toxico. 40, 1115–1120.

    CAS  Google Scholar 

  31. 31.

    M-Hamvas, M., Máthé, Cs., Molnár, E., Vasas, G., Grigorszky, I., Borbely, G. (2003) Microcystin-LR alters the growth, anthocyanin content and single-stranded DNase enzyme activities in Sinapis alba L. seedlings. Aquat. Toxicol. 62, 1–9.

    CAS  PubMed  Google Scholar 

  32. 32.

    M-Hamvas, M., Máthé, C., Vasas, G., Jámbrik, K., Papp, M., Beyer, D., Mészaros, I., Borbély, G. (2010) Cylindrospermopsin and microcystin-LR alter the growth, development and peroxidase enzyme activity of white mustard (Sinapis alba L.) seedlings, a comparative analysis. Acta Biol Hung. 61 (Suppl), 35–48.

    CAS  PubMed  Google Scholar 

  33. 33.

    Padisak, J. (1997) Cylindrospermopsis raciborkii (Woloszynska) Seenaya et Subba Raju, an expanding, highly adaptive cyanobacterium: worldwide distribution and review of its ecology. Arch. Hidrobiol. Suppl. 4, 563–593.

    Google Scholar 

  34. 34.

    Peuthert, A., Chakrabarti, S., Pflugmacher, S. (2007) Uptake of microcystin-LR and -LF (cyanobacterial toxins) in seedlings of several important agricultural plant species and the correlation with cellular damage (lipid peroxidation). Environ. Toxicol. 22, 436–442.

    CAS  Google Scholar 

  35. 35.

    Pflugmacher, S. (2004) Promotion of oxidative stress in aquatic macrophyte Ceratophyllum demer-sum during biotransformation of the cyanobacterial toxin microcystin-LR. Aquat. Toxicol. 70, 169–178.

    CAS  Google Scholar 

  36. 36.

    Prieto, A., Campos, A., Camean, A., Vasconcelos, V. (2011) Effects on growth and oxidative stress status of rice plants (Oryza sativa) exposed to two extracts of toxin-producing cyanobacteria (Aphanizomenon ovalisporum and Microcystis aeruginosa). Ecotox. Environ. Safe. 74, 1973–1980.

    CAS  Google Scholar 

  37. 37.

    Puerto, M., Campos, A., Prieto, A., Cameán, A., de Almeida, A. M., Coelho, A. V., Vasconcelos, V. (2011) Differential protein expression in two bivalve species: Mytilus galloprovincialis and Corbicula fluminea; exposed to Cylindrospermopsis raciborskii cells. Aquat. Toxicol. 101, 109–116.

    CAS  PubMed  Google Scholar 

  38. 38.

    Ros Barceló, A. (1998) The generation of H2O2 in the xylem of Zinnia elegans is mediated by an NADPH-oxidase-like enzyme. Plant. 207, 207–216.

    Google Scholar 

  39. 39.

    Runnegar, M. T., Kong, S. M., Zhong, Y. Z., Lu, S. C. (1995) Inhibition of reduced glutathion synthesis by cyanobacterial alkaloid cylindrospermopsin in cultured rat hepatocytes. Biochem. Pharmacol. 49, 219–225.

    CAS  PubMed  Google Scholar 

  40. 40.

    Smertenko, A. P., Chang, H. Y., Sonobe, S., Fenyk, S. I., Weingartner, M., Bogre, L., Hussey, P. J. (2006) Control of the AtMAP65-l interaction with microtubules through the cell cycle. J. Cell Sci. 119, 3227–3237.

    CAS  PubMed  Google Scholar 

  41. 41.

    Törökné, A. K., László, E., Chorus, I., Fastner, I., Heinze, R., Padisák, J., Barbosa, F. A. R. (2000) Water quality monitoring by Thamnotoxkit F(™) including cyanobacterial blooms. Water Sci. Techno. 42, 381–385.

    Google Scholar 

  42. 42.

    Vasas, G., Cáspár, A., Páger, C., Surányi, G., Máthé, C., M-Hamvas, M., Borbely, G. (2004) Analysis of cyanobacterials toxins (anatoxin-a, cylindrospermopsin, microcystin-LR) by capillary electrophoresis. Electrophoresi. 25, 108–115.

    CAS  Google Scholar 

  43. 43.

    Vasas, G., Gáspár, A., Surányi, G., Batta, G., Gyémánt, G., M-Hamvas, M., Máthé, C., Grigorszky, I., Molnár, E., Borbély, G. (2002) Capillary electrophoretic assay and purification of cylindrospermopsin, a cyanobacterial toxin from Aphanizomenon ovalisporum by plant test (Blue-Green Sinapis Test). Anal. Biochem. 302, 95–103.

    CAS  Google Scholar 

  44. 44.

    Žegura, B., Štraser, A., Filipič, M. (2011) Genotoxicity and potential carcinogenicity of cyanobacterial toxins - a review. Mutat. Res.-Rev Mutat. 727, 16–41.

    Google Scholar 

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We acknowledge Dr. Ola Hadas for kindly helping with Aphanizomenon ovalisporum ILC164. We thank Dr. István Grigorszky for his help in taking microscopical photos. The work was supported by the Hungarian National Research Foundation Grants OTKA F46493, K68416, K81370, GVOP-3.2.1.-2004-04-0110/3.0. M. M-Hamvas, C. Máthé, G. Surányi and G. Vasas greatly acknowledge the personal support from Bolyai Janos Scholarship of the Hungarian Academy of Sciences. Mrs. Ágnes Németh, Éva Barabás and Katalin Havelant are acknowledged for their excellent technical assistance.

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Correspondence to Márta M-Hamvas.

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Máthé, C., Vasas, G., Borbély, G. et al. Histological, Cytological and Biochemical Alterations Induced by Microcystin-LR and Cylindrospermopsin in White Mustard (Sinapis alba L.) Seedlings. BIOLOGIA FUTURA 64, 71–85 (2013). https://doi.org/10.1556/ABiol.64.2013.1.7

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  • Microcystin-LR
  • cylindrospermopsin
  • Sinapis alba
  • protein phosphatase
  • root histology