Genotoxicity of metaphase-arresting methods in barley

Amiprophos-methyl (APM) and colchicine (COL) are therapeutic agents and are also used in plant science. Cell cycle synchronization (CCS) is mediated by hydroxyurea (HU) and APM. The genotoxic effects of APM are unclear. In order to assess the genotoxicity of APM separately and with HU and then compare it with COL, we treated 2-day-old barley (Hordeum vulgare 'Bornova-92') seedlings with 8 µM APM for 2 h, 1.25 mM COL for 18 h, or 1.25 mM HU for 18 h and 4 µM APM for 2 h, with a final ice-cold water incubation. Following treatment, seedlings were recovered without any substance for 2 days. Genomic DNA samples were analyzed by random amplified polymorphic DNA, inter-simple sequence repeat, and inter-retrotransposon amplified polymorphisms. Polypeptide profiles were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Mean genomic stability (GTS) values at the end of APM, COL, and CCS treatments were 83.6%, 83%, and 82.3%, respectively. Mean GTS values after recovery from APM, COL, and CCS treatments were 78.5%, 83.9%, and 78.7%, respectively. APM treatment resulted in altered polypeptide profiles during both treatment and recovery. The genotoxicity of APM and COL was similar, while the genotoxic effects of APM continued after removal. Only COL treatment caused retrotransposon activation. APM treatment caused appearance of new polypeptides, which continued after the removal of APM. Although the microtubule inhibitory effect of APM has been reported to be reversible, APM may cause genotoxic effects on plants, and the molecular effects of APM may not be reversible.

Genotoxicity of metaphase-arresting methods in barley

Amiprophos-methyl (APM) and colchicine (COL) are therapeutic agents and are also used in plant science. Cell cycle synchronization (CCS) is mediated by hydroxyurea (HU) and APM. The genotoxic effects of APM are unclear. In order to assess the genotoxicity of APM separately and with HU and then compare it with COL, we treated 2-day-old barley (Hordeum vulgare 'Bornova-92') seedlings with 8 µM APM for 2 h, 1.25 mM COL for 18 h, or 1.25 mM HU for 18 h and 4 µM APM for 2 h, with a final ice-cold water incubation. Following treatment, seedlings were recovered without any substance for 2 days. Genomic DNA samples were analyzed by random amplified polymorphic DNA, inter-simple sequence repeat, and inter-retrotransposon amplified polymorphisms. Polypeptide profiles were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Mean genomic stability (GTS) values at the end of APM, COL, and CCS treatments were 83.6%, 83%, and 82.3%, respectively. Mean GTS values after recovery from APM, COL, and CCS treatments were 78.5%, 83.9%, and 78.7%, respectively. APM treatment resulted in altered polypeptide profiles during both treatment and recovery. The genotoxicity of APM and COL was similar, while the genotoxic effects of APM continued after removal. Only COL treatment caused retrotransposon activation. APM treatment caused appearance of new polypeptides, which continued after the removal of APM. Although the microtubule inhibitory effect of APM has been reported to be reversible, APM may cause genotoxic effects on plants, and the molecular effects of APM may not be reversible.

___

  • Atienzar FA, Cheung VV, Jha AN, Depledge MH (2001). Fitness parameters and DNA effects are sensitive indicators of copper- induced toxicity in Daphnia magna. Toxicol Sci 59: 241–250.
  • Atienzar FA, Jha AN (2006). The random amplified polymorphic DNA (RAPD) assay and related techniques applied to genotoxicity and carcinogenesis studies: a critical review. Mutat Res 613: 76–102.
  • Attia SM, Kliesch U, Schriever-Schwemmer G, Badary OA, Hamada FM, Adler ID (2003). Etoposide and merbarone are clastogenic and aneugenic in the mouse bone marrow micronucleus test complemented by fluorescence in situ hybridization with the mouse minor satellite DNA probe. Environ Mol Mutagen 41: 99–103.
  • Bell A (1998). Microtubule inhibitors as potential antimalarial agents. Parasitol Today 14: 234–240.
  • Binarova P, Dolezel J (1993). Effect of anti-microtubular drug amiprophos-methyl on somatic embryogenesis and DNA ploidy levels in alfalfa and carrot cell suspension cultures. Biol Plantarum 35: 329–339.
  • Bradford MM (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254.
  • de Wolf H, Blust R, Backeljau T (2004). The use of RAPD in ecotoxicology. Mutat Res 566: 249–262.
  • Dhooghe E, van Laere K, Eeckhaut T, Leus L, van Huylenbroeck J (2011). Mitotic chromosome doubling of plant tissues in vitro. Plant Cell Tissue Org 104: 359–373.
  • Dolezel J, Cihalikova J, Weiserova J, Lucretti S (1999). Cell cycle synchronization in plant root meristems. Methods Cell Sci 21: 95–107.
  • Erturk FA, Ay H, Nardemir G, Agar G (2012). Molecular determination of genotoxic effects of cobalt and nickel on maize (Zea mays L.) by RAPD and protein analyses. Toxicol Ind Health 29: 662–671.
  • Falconer MM, Seagull RW (1987). Amiprophos-methyl (APM): a rapid, reversible, anti-microtuble agent for plant cell cultures. Protoplasma 136: 118–124.
  • Fennell BJ, Carolan S, Pettit GR, Bell A (2003). Effects of the antimitotic natural product dolastatin 10, and related peptides, on the human malarial parasite Plasmodium falciparum. J Antimicrob Chemoth 51: 833–841.
  • Fennell BJ, Naughton JA, Dempsey E, Bell A (2006). Cellular and molecular actions of dinitroaniline and phosphorothioamidate herbicides on Plasmodium falciparum: tubulin as a specific antimalarial target. Mol Biochem Parasit 145: 226–238.
  • Fernandez ME, Figueiras AM, Benito C (2002). The use of ISSR and RAPD markers for detecting DNA polymorphism, genotype identification and genetic diversity among barley cultivars with known origin. Theor Appl Genet 104: 845–851.
  • Francis A, Jones RN (1989). Heritable nature of colchicine induced variation in diploid Lolium perenne. Heredity 62: 407–410.
  • Gilbert SK, Patterson FL (1965). Colchicine-induced mutants of Decatur barley. Crop Sci 5: 44–47.
  • Grzebelus E, Adamus A (2004). Effect of anti-mitotic agents on development and genome doubling of gynogenic onion (Allium cepa L.) embryos. Plant Sci 167: 569–574.
  • Habibi L, Shokrgozar MA, Tabrizi M, Modarressi MH, Akrami SM (2013). Mercury specifically induces LINE-1 activity in a human neuroblastoma cell line. Mutat Res 759: 9–20.
  • Hague LM, Jones RN (1987). Cytogenetics of Lolium perenne. Theor Appl Genet 74: 233–241.
  • Hamat-Mecbur H, Yilmaz S, Temel A, Sahin K, Gozukirmizi N (2014). Effects of Epirubicin on barley seedlings. Toxicol Ind Health 30: 52–59.
  • Hansen AL, Gertz A, Joersbo M, Andersen SB (1998). Antimicrotubule herbicides for in vitro chromosome doubling in Beta vulgaris L. ovule culture. Euphytica 101: 231–237.
  • Hantzschel KR, Weber G (2010). Blockage of mitosis in maize root tips using colchicine-alternatives. Protoplasma 241: 99–104.
  • Harpstead DD, Ross JG, Franzke CJ (1954). The nature of chromatin changes of colchicine-induced variants in Sorghum. J Hered 45: 255–258.
  • Henny RJ, Holm JR, Chen JJ, Scheiber M (2009). In vitro induction of tetraploids in Dieffenbachia × ‘Star Bright M-1’ by colchicine. HortScience 44: 646–650.
  • Izzatullayeva V, Akparov Z, Babayeva S, Ojaghi J, Abbasov M (2014). Efficiency of using RAPD and ISSR markers in evaluation of genetic diversity in sugar beet. Turk J Biol 38: 429–438.
  • Khayat AS, Antunes LM, Guimaraes AC, Bahia MO, Lemos JAR, Cabral IR, Lima PDL, Amorim MIM, Cardoso PCS, Smith MAC et al. (2006). Cytotoxic and genotoxic monitoring of sickle cell anaemia patients treated with hydroxyurea. Clin Exp Med 6: 33–37.
  • Khosravi AR, Kadir MA, Kadzemin SB (2009). RAPD analysis of colchicine induced variation of the Dendrobium Serdang beauty. Afr J Biotechnol 8: 1455–1465.
  • Laemmli UK (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680– 685.
  • Lama R, Sandhu R, Zhong B, Li B, Su B (2012). Identification of selective tubulin inhibitors as potential anti-trypanosomal agents. Bioorg Med Chem Lett 22: 5508–5516.
  • Liu W, Li PJ, Qi XM, Zhou QX, Zheng L, Sun TH, Yang YS (2005). DNA changes in barley (Hordeum vulgare) seedlings induced by cadmium pollution using RAPD analysis. Chemosphere 61: 158–167.
  • Liu XZ, Lin H, Mo XY, Long T, Zhang HY (2009). Genetic variation in colchicine-treated regenerated plants of Eucalyptus globulus Labill. J Genet 88: 345–348.
  • Marakli S, Yilmaz S, Gozukirmizi N (2012). BARE1 and BAGY2 retrotransposon movements and expression analyses in developing barley seedlings. Biotechnol Biotec Eq 26: 3451– 3456.
  • Morejohn L, Fosket D (1991). The biochemistry of compounds with anti-microtubule activity in plant cells. Pharmacol Therapeut 51: 217–230.
  • Morejohn LC, Bureau TE, Tocchi LP, Fosket DE (1984). Tubulins from different higher-plant species are immunologically nonidentical and bind colchicine differentially. P Natl Acad Sci USA 81: 1440–1444.
  • Pan WH, Houben A, Schlegel R (1993). Highly effective cell synchronization in plant roots by hydroxyurea and amiprophos-methyl or colchicine. Genome 36: 387–390.
  • Portis E, Acquadro A, Comino C, Lanteri S (2004). Analysis of DNA methylation during germination of pepper (Capsicum annuum L.) seeds using methylation-sensitive amplification polymorphism (MSAP). Plant Sci 166: 169–178.
  • Puigderrajols P, Jofre A, Mir G, Pla M, Verdaguer D, Huguet G, Molinas M (2002). Developmentally and stress-induced small heat shock proteins in cork oak somatic embryos. J Exp Bot 53: 1445–1452.
  • Rauf S, Khan IA, Khan FA (2006). Colchicine-induced tetraploidy and changes in allele frequencies in colchicine-treated populations of diploids assessed with RAPD markers in Gossypium arboreum L. Turk J Biol 30: 93–100.
  • Reddy MP, Sarla N, Siddiq EA (2002). Inter simple sequence repeat (ISSR) polymorphism and its application in plant breeding. Euphytica 128: 9–17.
  • Rubuluza T, Nikolova RV, Smith MT, Hannweg K (2007). In vitro induction of tetraploids in Colophospermum mopane by colchicine. S Afr J Bot 73: 259–261.
  • Saeidi H, Rahiminejad MR, Heslop-Harrison JS (2008). Retroelement insertional polymorphisms, diversity and phylogeography within diploid, D-genome Aegilops tauschii (Triticeae, Poaceae) sub-taxa in Iran. Ann Bot 101: 855–861.
  • Santos JL, Bosquesi PL, Almeida AE, Chin CM, Varanda EA (2011). Mutagenic and genotoxic effect of hydroxyurea. Int J Biomed Sci 7: 263–267.
  • Sapra S, Bhalla Y, Nandani, Sharma S, Singh G, Nepali K (2012). Colchicine and its various physicochemical and biological aspects. Med Chem Res 22: 531–547.
  • Schulman AH, Flavell AJ, Paux E, Ellis THN (2012). The application of LTR retrotransposons as molecular markers in plants. Methods Mol Biol 859: 115–153.
  • Sukumaran S, Grant A (2013). Effects of genotoxicity and its consequences at the population level in sexual and asexual Artemia assessed by analysis of inter-simple sequence repeats (ISSR). Mutat Res 757: 8–14.
  • Suzuki K, Takatsu Y, Gonai T, Kasumi M (2005). Plant regeneration and chromosome doubling of wild Gladiolus species. Acta Hortic 673: 175–181.
  • Tchorbadjieva MI, Pantchev IY (2004). DNA methylation and somatic embryogenesis of orchardgrass (Dactylis glomerata L.). Bulg J Plant Physiol 30: 3–13.
  • Verhoeven HA, Sree Ramulu K, Dijkhuis PA (1990). Comparison of the effects of various spindle toxins on metaphase arrest and formation of micronuclei in cell-suspension cultures of Nicotiana plumbaginifolia. Planta 182: 408–414.
  • Wan Y, Duncan DR, Rayburn AL, Petolino JF, Widholm JM (1991). The use of antimicrotubule herbicides for the production of doubled haploid plants from anther-derived maize callus. Theor Appl Genet 81: 205–211.
  • Werbovetz KA (2002). Tubulin as an antiprotozoal drug target. Mini Rev Med Chem 2: 519–529.
  • Yunus MF, Aziz MA, Kadir MA, Daud SK, Rashid AA (2013). In vitro mutagenesis of Etlingera elatior (Jack) and early detection of mutation using RAPD markers. Turk J Biol 37: 716–725.
Turkish Journal of Biology-Cover
  • ISSN: 1300-0152
  • Yayın Aralığı: Yılda 6 Sayı
  • Yayıncı: TÜBİTAK
Sayıdaki Diğer Makaleler

In vitro antiproliferative/cytotoxic activity of 2,3'-biindole against various cancer cell lines

Tuğba KÖPRÜLÜ KUL, Salih ÖKTEN, Ramazan ERENLER, Şaban TEKİN

Evaluation of the genotoxic or mutagenic effects of thermal stress on cultured human lymphocytes

Hasan Basri İLA, Mehmet TOPAKTAŞ, Mehmet ARSLAN, Mehmet BÜYÜKLEYLA, Erman Salih İSTİFLİ

Genotoxicity of metaphase-arresting methods in barley

Aslıhan TEMEL, Nermin GÖZÜKIRMIZI

Cloning and expression of chitinase A, B, and C (chiA, chiB, chiC) genes from Serratia marcescens originating from Helicoverpa armigera and determining their activities

Mehtap DANIŞMAZOĞLU, İsmail DEMİR, Kazım SEZEN, Hacer MURATOĞLU, Remziye NALÇACIOĞLU

Identification and pathogenicity of bacteria in the Mediterranean corn borer Sesamia nonagrioides Lefebvre (Lepidoptera: Noctuidae)

Ardahan ESKİ, Filiz Özkan ÇAKICI, Mustafa GÜLLÜ, Hacer MURATOĞLU, Zihni DEMİRBAĞ, İsmail DEMİR

Biosurfactant production by Pleurotus ostreatus in submerged and solid-state fermentation systems

Zülfiye VELİOĞLU, Raziye Öztürk ÜREK

Improvement of atropine production by different biotic and abiotic elicitors in hairy root cultures of Datura metel

Zahra SHAKERAN, Mehrnaz KEYHANFAR, Gholamreza ASGHARI, Mustafa GHANADIAN

In vitro plant regeneration of 4 Capsicum spp. genotypes using different explant types

Marta ORLINSKA, Pawel NOWACZYK

Inhibitory effects of salicylic acid on A549 human lung adenocarcinoma cell viability

Djanan VEJSELOVA, Hatice Mehtap KUTLU

Genome-wide distribution of superoxide dismutase (SOD) gene families in Sorghum bicolor

Ertuğrul FİLİZ, Hüseyin TOMBULOĞLU