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Ferro glukonatın sitotoksik etkisinin Allium cepa kök ucunda değerlendirilmesi

Year 2021, Volume: 11 Issue: 3, 744 - 750, 15.07.2021
https://doi.org/10.17714/gumusfenbil.843648

Abstract

Araştırmada, Allium cepa L ve ferro glukonat (E579) kullanılmıştır. A. cepa kökleri, farklı konsantrasyonlarda ferro glukonat ile muamele edilmiştir. A. cepa kök uzunlukları ölçülmüştür. Böylece EC50 değeri 0,068 g / l olarak belirlenmiştir. Daha sonra A. cepa kökleri 24, 48, 72 saat EC50 / 2 (0.034 g / l), EC50 (0.068 g / l), 2XEC50 (0.136 g / l) ile muamele edilmiştir. Bu sürelerin sonunda kök uçları kesilerek ışık mikroskobunda gözlem için hazırlanmıştır. Her muamele periyodu ve konsantrasyon için en az 5000 hücre sayılmıştır. Sitotoksisitenin belirlenmesinde mitotik indeks kullanılmıştır. Mitotik indeks (MI) = bölünen hücre sayısı / toplam hücre sayısı X 100 formülü ile hesaplanmıştır. TUKEY çoklu karşılaştırma testi ve tekrarlanan ölçümlü ANOVA kullanılmıştır. Ferro glukonat konsantrasyonları arasındaki farkın muamele süresine göre değiştiği belirlenmiştir. Mitotik indeks açısından, tüm uygulama periyodu sonunda kontrol dozlarının ortalamasının EC50/2, EC50, EC50X2 dozlarına göre yüksek olduğu bulunmuştur ve artan dozla azaldığı ortaya konmuştur. EC50X2 konsantrasyonunun 24, 48 ve 72 saat süresince muamelesinin ardından, muamele süresi uzadıkça mitotik indeksin azalmış olduğu saptanmıştır. Kontrol grubuna göre ferro glukonatın kök uçlarında hücre bölünmesini azalttığı bulunmuştur. Böylece, ferro glukonatın A. cepa kök uçlarında sitotoksik olduğu ortaya konmuştur. Ayrıca, muamele süresi ve dozunun artması nedeniyle ferro glukonatın sitotoksik etkisinin arttığı ve bunun nedeninin de mitotik indeksin azaldığından dolayı olduğu saptanmıştır. Araştırma sonuçları, literatürdeki gıda katkı maddelerinin sitotoksik etkisinin olabileceğinin saptanması sonuçlarıyla paralellik göstermektedir.

Supporting Institution

Çanakkale Onsekiz Mart Üniversitesi Bilimsel Araştırmalar Projeleri Birimi

References

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  • Akı, C. ve Çördük N. (2011). Genetik laboratuvar kılavuzu, Kriter Yayınları, İstanbul.
  • Altuğ, T., Boyacıoğlu, D., Kurtcan, Ü. ve Demirağ, K. (2000) Gıda katkı maddeleri analiz yöntemleri, Ege Üniversitesi Mühendislik Fakültesi Yayınları, Ege Üniversitesi Basımevi, Bornova, İzmir.
  • Amer, S.M. and Ali, E. M. (1974). Cytological effects of pesticides V. Effects of some herbicides on Vicia faba. Cytologia, 39, 633-643.
  • Amer S.M. and Aly, F.A. (1992). Cytogenetic Effects of Pesticides. IV. Cytogenetic effects of the insecticides gardona and dursban. Mutation Research, 279(3), 165-170. https://doi.org/10.1016/0165-1218(92)90063-6
  • Anderson, D., Jenkinson, P.C., Dewdeney, R.S., Franis, A.J., Godbert, P. and Butterworth, K.R. (1988). Chromosome aberrations, mitogen-induced blastogenesis and proliferative rate index in peripheral lymphocytes from 106 control individuals of U.K. population. Mutation Research/Genetic Toxicology 204(3), 407-20. https://doi.org/10.1016/0165-1218(88)90150-4
  • Anonymous. (1995). Code of Federal Regulations. Office of the federal register national archives and records administration, Washington, DC.
  • Arslan, M. ve Rencüzoğulları, E. (2004). Borik asit’in insan periferal lenfositlerinde in vitro kromozom aberasyonu ve kardeş kromatid değişimi üzerindeki etkileri. Science Institute, Cukurova University, Adana, 2004.
  • Cruess, W.V. (1962). Experiments on color fixation with iron salts, in 41 Annual Technical Report of California Olive Association, Sacramento, CA.
  • Dwivedi, H. and Kumar, G. (2017). Genomic distortion induced by food dyes on meristematic cells of Trachyspermum ammi (L.) Sprague (Ajwain). Chromosome Botany, 12(3), 46-51. https://doi.org/10.3199/iscb.12.46
  • Dwivedi, K. and Kumar, G. (2015). Genetic damage induced by a food coloring dye (sunset yellow) on meristematic cells of Brassica campestris L. Journal of Environmental and Public Health, 2015, 1-5. https://doi.org/10.1155/2015/319727
  • Fiskesjö, G. (1993). The Allium cepa test in wastewater monitoring. Environmental Toxicology & Water Quality, 8(3), 291-298. https://doi.org/10.1002/tox.2530080306
  • Fiskesjö, G. (1985). The Allium-test as a standard in environmental monitoring, Hereditas, 102, 99-112. https://doi.org/10.1111/j.1601-5223.1985.tb00471.x
  • Fiskesjö, G. (1997) Allium test for screening chemicals; evaluation of cytological parameters. Ln: Wang W, Gorsuch JW and Hughes JS (Eds) Plants for Environmental Studies. New York, NY: Lewis Publishers, pp. 308-333. https://doi.org/10.1201/9781420048711.ch11
  • Garcia, P., Brenes, M. and Garrido, A. (1986). Use of ferrous lactate in the elaboration of ripe olives. Grasas Aceites, 37, 33-38.
  • Gomes, K.M.S. and Oliveira, M.V.G.A. (2013). Citotoxicity of food dyes sunset yellow (E-110), bordeaux red (E-123), and tatrazine yellow (E-102) on Allium cepa L. root meristematic cells. Food Science and Technology, Campinas, 33(1), 218-223. https://doi.org/10.1590/s0101-20612013005000012
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  • Hurrell, R.F (1997). Preventing iron deficiency through food fortification. Nutrition Reviews, 55(6), 210–222. https://doi.org/10.1111/j.1753-4887.1997.tb01608.x
  • Iganci, J.R.V, Bobrowski, V.L., Heiden, G., Stein, V.C. and Rocha, B.H.G. (2006). Efeito do extrato aquoso de diferentes espécies de boldo sobre a germinação e índice mitótico de Allium cepa L. Arquivos do Instituto Biológico 73, 79-82.
  • Kayraldız, A. and Topaktaş, M. (2007). The in vivo genotoxic effects of sodium metabisulfite in bone marrow cells of rats. Russian journal of genetics, 43(8), 905- 909. https://doi.org/10.1134/s1022795407080121
  • Kumar G. and Pandey A. (2015). Genotoxic and mito-depressive effects of food preservatives on root meristems of barley (Hordeum vulgare L.). Chromosome Botany, 10: 51-60. https://doi.org/10.3199/iscb.10.51
  • Kumar, G. and Singh, S. (2017). Genotoxic effects of food dyes on mitotic chromosomal entity in root meristems of Cluster bean (Cyamopsis tetragonoloba (L.) Taub.). International Journal of ChemTech Research, 10(13), 81-89.
  • Kuş, E. and Eroğlu, H.E (2015). Genotoxic and cytotoxic effects of sunset yellow and brilliant blue, colorant food additives, on human blood lymphocytes. Pakistan Journal of Pharmaceutical Sciences 28(1), 227-30.
  • Lerda, D. (2017). The effects of tartrazine in Allium cepa L. Journal of Food Nutrition, 3(101), 1-5. https://doi.org/10.17303/jfn.2017.3.101
  • Linnainmaa, K., Meretoja, T., Sorsa, M. and Vainto, H. (1978). Cytogenetic effects of styrene and styrene oxide. Mutation Research/Genetic Toxicology, 58, 277–286. https://doi.org/10.1016/0165-1218(78)90020-4
  • Liman, R., Akyıl, D., Eren, Y. and Konuk, M. (2010) Testing of mutagenicity and genotoxicity of metolcarb by using both Ames/Salmonella and Allium test. Chemosphere, 80:1056-1061. https://doi.org/10.1016/j.chemosphere.2010.05.011
  • Ma, T.H., Xu, C., Liao, S., McConnell H., Jeong, B.S. and Won C.D. (1996). In Situ monitoring with the tradescantia bioassays on the genotoxicity of gaseous Emissions from a closed landfill site and an incinerator. Mutation Research/Environmental Mutagenesis and Related Subjects, 359, 39–52. https://doi.org/10.1016/s0165-1161(96)90008-4
  • Marques, G.S. and Silva S.O., (2015b). Cytotoxic and genotoxic potential of liquid synthetic food flavorings evaluated alone and in combination. Food Science Technology, Campinas, 35(1), 183-188. https://doi.org/10.1590/1678-457x.6596
  • Marques, G.S., Sousa, J.J.A. and Peron, A.P. (2015a). Action of ponceau 4R (E-124) food dye on root meristematic cells of Allium cepa L. Acta Scientiarum. Biological Science, 37(1), 101-106. https://doi.org/10.4025/actascibiolsci.v37i1.23119
  • Misik, M., Solenska, K., Micieta, K., Misikova, K. and Knasmüler S. (2006). In situ monitoring of mutagenic effects of contamj­nated air in Bratislava (Slovakia) with Tradescantia micronucleus (Trad-MCN) and abortive microspore assays. Mutation Research, 605: 1-6.
  • Moura, A.G., Santana, G.M., Ferreirab, P.M.P, Sousaa, J.M.C and Peron, A.P. (2016). Cytotoxicity of cheese and cheddar cheese food flavorings on Allim cepa L root meristems. Brazilian Journal of Biology, 76(2), 439-443. https://doi.org/10.1590/1519-6984.20514
  • Nunes, R. D. M., Sales, I. M. S., Silva, S. I. O., Sousa, J. M. C. and Peron, A. P. (2017). Antiproliferative and genotoxic effects of nature identical and artificial synthetic food additives of aroma and flavor. Brazilian Journal of Biology, 77(1), 150-154. https://doi.org/10.1590/1519-6984.12115
  • Oliveira, V.A., Oliveira, V.M., Oliveira, T.W.N., Damasceno, A.N.C., Silva, C.E.O., Medeiros, S.R.A., Soares, B.M., Silva, F.C.C., Aguiar, R.P.S., Islam, M.T., MeloCavalcante, A.A.C., Peron, A.P. and Sousa, J.M.C. (2017). Evaluation of cytotoxic and mutagenic effects of two artificial sweeteners by using eukaryotic test systems. African Journal of Biotechnology, 16(11), 547-551. https://doi.org/10.5897/ajb2016.15695
  • Özkara, A., Akyil, D., Eren, Y., Erdogmus, S. F., Konuk, M. and Saglam, E. (2015). Assessment of cytotoxic and genotoxic potential of pyracarbolid by Allium test and micronucleus assay. Drug and Chemical Toxicology, 38(3), 337-341. https://doi.org/10.3109/01480545.2014.966831
  • Pandey, H., Vikas Kumar, V. and Roy, B.K. (2014). Assessment of genotoxicity of some common food preservatives using Allium cepa L. as a test plant. Toxicology Reports, 1, 300-308. https://doi.org/10.1016/j.toxrep.2014.06.002
  • Rencuzogulları, E., Kayraldız, A., Ila, H.B., Çakmak, T. and Topaktaş, M. (2001). The cytogenetic effects of sodium metabisulfite, a food preservative in root tip cells of Allium cepa L. Turkish Journal of Biology, 25, 361-370.
  • Rojas, E, Herrera, LA, Sordo, M, Gonsebatt, ME, Montero, R, Rodriguez, R. and Ostrosky-Wegman, P. (1993). Mitotic index and cell proliferation kinetics for the identification of antineoplastic activity. Anti-Cancer Drugs. 4(6), 637-40. https://doi.org/10.1097/00001813-199312000-00005
  • Sales, I.M.S., Oliveira, J.D., Santos, F.K.S., Feitoza, L.L., Sousa, J.M.C. and Peron, A.P. (2016). Toxicity at the cellular level of artificial synthetic flavorings. Acta Scientiarum. Biological Sciences, 38(3), 297-303. https://doi.org/10.4025/actascibiolsci.v38i3.30751
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Evaluation of cytotoxic effect of ferrous gluconate on Allium cepa root tip

Year 2021, Volume: 11 Issue: 3, 744 - 750, 15.07.2021
https://doi.org/10.17714/gumusfenbil.843648

Abstract

It was used Allium cepa L and ferrous gluconate (E579). A. cepa roots were treated with ferrous gluconate of different concentrations. A. cepa root lengths were measured. So, the EC50 value was determined as 0.068 g/l. Then, A. cepa L. roots were treated with EC50 / 2 (0.034 g/l), EC50 (0.068 g/l), 2XEC50 (0.136 g/l) for 24, 48, 72 hours. The root tips were cut and prepared for observation in the light microscope. For each group, at least 5000 cell counts were made. Mitotic index was used in the determination of cytotoxicity. TUKEY multiple comparison test and Repeated measurement ANOVA was used. It was determined that the difference between ferrous gluconate concentrations varied as far as the treatment duration. In the way of mitotic index, it was stated that mean of control doses at all treatment duration were significantly higher than the EC50/2, EC50, EC50X2 doses and decreased with the increasing dose. After 24, 48 and 72 hours treatment of EC50X2 dose, it was found that mitotic index decreased due to increasing in application duration. It was detected that ferrous gluconate reduced mitotic division at the root of A. cepa in comparison to the control group. In this way, it was established that ferrous gluconate can be cytotoxic. In addition to this, it was determined that the cytotoxic effect of ferrous gluconate increased and mitotic index decreased due to increasing of treatment duration and dose. Results of this research are parallel with the literatüre that food additives might cause cytotoxicity.

References

  • Adeyemo, O.A. and Farinmade, A. E. (2013). Genotoxic and cytotoxic effects of food flavor enhancer, monosodium glutamate (MSG) using Allium cepa assay. African Journal of Biotechnology, 12(13), 1459-1466. https://doi.org/ 10.5897/AJB12.2927
  • Akı, C. ve Çördük N. (2011). Genetik laboratuvar kılavuzu, Kriter Yayınları, İstanbul.
  • Altuğ, T., Boyacıoğlu, D., Kurtcan, Ü. ve Demirağ, K. (2000) Gıda katkı maddeleri analiz yöntemleri, Ege Üniversitesi Mühendislik Fakültesi Yayınları, Ege Üniversitesi Basımevi, Bornova, İzmir.
  • Amer, S.M. and Ali, E. M. (1974). Cytological effects of pesticides V. Effects of some herbicides on Vicia faba. Cytologia, 39, 633-643.
  • Amer S.M. and Aly, F.A. (1992). Cytogenetic Effects of Pesticides. IV. Cytogenetic effects of the insecticides gardona and dursban. Mutation Research, 279(3), 165-170. https://doi.org/10.1016/0165-1218(92)90063-6
  • Anderson, D., Jenkinson, P.C., Dewdeney, R.S., Franis, A.J., Godbert, P. and Butterworth, K.R. (1988). Chromosome aberrations, mitogen-induced blastogenesis and proliferative rate index in peripheral lymphocytes from 106 control individuals of U.K. population. Mutation Research/Genetic Toxicology 204(3), 407-20. https://doi.org/10.1016/0165-1218(88)90150-4
  • Anonymous. (1995). Code of Federal Regulations. Office of the federal register national archives and records administration, Washington, DC.
  • Arslan, M. ve Rencüzoğulları, E. (2004). Borik asit’in insan periferal lenfositlerinde in vitro kromozom aberasyonu ve kardeş kromatid değişimi üzerindeki etkileri. Science Institute, Cukurova University, Adana, 2004.
  • Cruess, W.V. (1962). Experiments on color fixation with iron salts, in 41 Annual Technical Report of California Olive Association, Sacramento, CA.
  • Dwivedi, H. and Kumar, G. (2017). Genomic distortion induced by food dyes on meristematic cells of Trachyspermum ammi (L.) Sprague (Ajwain). Chromosome Botany, 12(3), 46-51. https://doi.org/10.3199/iscb.12.46
  • Dwivedi, K. and Kumar, G. (2015). Genetic damage induced by a food coloring dye (sunset yellow) on meristematic cells of Brassica campestris L. Journal of Environmental and Public Health, 2015, 1-5. https://doi.org/10.1155/2015/319727
  • Fiskesjö, G. (1993). The Allium cepa test in wastewater monitoring. Environmental Toxicology & Water Quality, 8(3), 291-298. https://doi.org/10.1002/tox.2530080306
  • Fiskesjö, G. (1985). The Allium-test as a standard in environmental monitoring, Hereditas, 102, 99-112. https://doi.org/10.1111/j.1601-5223.1985.tb00471.x
  • Fiskesjö, G. (1997) Allium test for screening chemicals; evaluation of cytological parameters. Ln: Wang W, Gorsuch JW and Hughes JS (Eds) Plants for Environmental Studies. New York, NY: Lewis Publishers, pp. 308-333. https://doi.org/10.1201/9781420048711.ch11
  • Garcia, P., Brenes, M. and Garrido, A. (1986). Use of ferrous lactate in the elaboration of ripe olives. Grasas Aceites, 37, 33-38.
  • Gomes, K.M.S. and Oliveira, M.V.G.A. (2013). Citotoxicity of food dyes sunset yellow (E-110), bordeaux red (E-123), and tatrazine yellow (E-102) on Allium cepa L. root meristematic cells. Food Science and Technology, Campinas, 33(1), 218-223. https://doi.org/10.1590/s0101-20612013005000012
  • Gömürgen, A.N. (2005). Cytological effect of the potassium metabisulphite and potassium nitrate food preservative on root tips of Allium cepa L. Cytologia, 70 (2), 119–128. https://doi.org/10.1508/cytologia.70.119
  • Grant, W.F. (1978). Chromosome aberrations in plants as a monitoring system. Environmental Health Perspectives, 27, 37-43. https://doi.org/10.1289/ehp.782737
  • Gürsoy, S. ve Aktaç, T. (2001). Besinlerde katkı maddelerinin kullanımı ve sitrik asit toksisitesi, Yüksek Lisans Tezi, Trakya Üniversitesi Fen Bilimleri Enstitüsü, Tekirdağ.
  • Hurrell, R.F (1997). Preventing iron deficiency through food fortification. Nutrition Reviews, 55(6), 210–222. https://doi.org/10.1111/j.1753-4887.1997.tb01608.x
  • Iganci, J.R.V, Bobrowski, V.L., Heiden, G., Stein, V.C. and Rocha, B.H.G. (2006). Efeito do extrato aquoso de diferentes espécies de boldo sobre a germinação e índice mitótico de Allium cepa L. Arquivos do Instituto Biológico 73, 79-82.
  • Kayraldız, A. and Topaktaş, M. (2007). The in vivo genotoxic effects of sodium metabisulfite in bone marrow cells of rats. Russian journal of genetics, 43(8), 905- 909. https://doi.org/10.1134/s1022795407080121
  • Kumar G. and Pandey A. (2015). Genotoxic and mito-depressive effects of food preservatives on root meristems of barley (Hordeum vulgare L.). Chromosome Botany, 10: 51-60. https://doi.org/10.3199/iscb.10.51
  • Kumar, G. and Singh, S. (2017). Genotoxic effects of food dyes on mitotic chromosomal entity in root meristems of Cluster bean (Cyamopsis tetragonoloba (L.) Taub.). International Journal of ChemTech Research, 10(13), 81-89.
  • Kuş, E. and Eroğlu, H.E (2015). Genotoxic and cytotoxic effects of sunset yellow and brilliant blue, colorant food additives, on human blood lymphocytes. Pakistan Journal of Pharmaceutical Sciences 28(1), 227-30.
  • Lerda, D. (2017). The effects of tartrazine in Allium cepa L. Journal of Food Nutrition, 3(101), 1-5. https://doi.org/10.17303/jfn.2017.3.101
  • Linnainmaa, K., Meretoja, T., Sorsa, M. and Vainto, H. (1978). Cytogenetic effects of styrene and styrene oxide. Mutation Research/Genetic Toxicology, 58, 277–286. https://doi.org/10.1016/0165-1218(78)90020-4
  • Liman, R., Akyıl, D., Eren, Y. and Konuk, M. (2010) Testing of mutagenicity and genotoxicity of metolcarb by using both Ames/Salmonella and Allium test. Chemosphere, 80:1056-1061. https://doi.org/10.1016/j.chemosphere.2010.05.011
  • Ma, T.H., Xu, C., Liao, S., McConnell H., Jeong, B.S. and Won C.D. (1996). In Situ monitoring with the tradescantia bioassays on the genotoxicity of gaseous Emissions from a closed landfill site and an incinerator. Mutation Research/Environmental Mutagenesis and Related Subjects, 359, 39–52. https://doi.org/10.1016/s0165-1161(96)90008-4
  • Marques, G.S. and Silva S.O., (2015b). Cytotoxic and genotoxic potential of liquid synthetic food flavorings evaluated alone and in combination. Food Science Technology, Campinas, 35(1), 183-188. https://doi.org/10.1590/1678-457x.6596
  • Marques, G.S., Sousa, J.J.A. and Peron, A.P. (2015a). Action of ponceau 4R (E-124) food dye on root meristematic cells of Allium cepa L. Acta Scientiarum. Biological Science, 37(1), 101-106. https://doi.org/10.4025/actascibiolsci.v37i1.23119
  • Misik, M., Solenska, K., Micieta, K., Misikova, K. and Knasmüler S. (2006). In situ monitoring of mutagenic effects of contamj­nated air in Bratislava (Slovakia) with Tradescantia micronucleus (Trad-MCN) and abortive microspore assays. Mutation Research, 605: 1-6.
  • Moura, A.G., Santana, G.M., Ferreirab, P.M.P, Sousaa, J.M.C and Peron, A.P. (2016). Cytotoxicity of cheese and cheddar cheese food flavorings on Allim cepa L root meristems. Brazilian Journal of Biology, 76(2), 439-443. https://doi.org/10.1590/1519-6984.20514
  • Nunes, R. D. M., Sales, I. M. S., Silva, S. I. O., Sousa, J. M. C. and Peron, A. P. (2017). Antiproliferative and genotoxic effects of nature identical and artificial synthetic food additives of aroma and flavor. Brazilian Journal of Biology, 77(1), 150-154. https://doi.org/10.1590/1519-6984.12115
  • Oliveira, V.A., Oliveira, V.M., Oliveira, T.W.N., Damasceno, A.N.C., Silva, C.E.O., Medeiros, S.R.A., Soares, B.M., Silva, F.C.C., Aguiar, R.P.S., Islam, M.T., MeloCavalcante, A.A.C., Peron, A.P. and Sousa, J.M.C. (2017). Evaluation of cytotoxic and mutagenic effects of two artificial sweeteners by using eukaryotic test systems. African Journal of Biotechnology, 16(11), 547-551. https://doi.org/10.5897/ajb2016.15695
  • Özkara, A., Akyil, D., Eren, Y., Erdogmus, S. F., Konuk, M. and Saglam, E. (2015). Assessment of cytotoxic and genotoxic potential of pyracarbolid by Allium test and micronucleus assay. Drug and Chemical Toxicology, 38(3), 337-341. https://doi.org/10.3109/01480545.2014.966831
  • Pandey, H., Vikas Kumar, V. and Roy, B.K. (2014). Assessment of genotoxicity of some common food preservatives using Allium cepa L. as a test plant. Toxicology Reports, 1, 300-308. https://doi.org/10.1016/j.toxrep.2014.06.002
  • Rencuzogulları, E., Kayraldız, A., Ila, H.B., Çakmak, T. and Topaktaş, M. (2001). The cytogenetic effects of sodium metabisulfite, a food preservative in root tip cells of Allium cepa L. Turkish Journal of Biology, 25, 361-370.
  • Rojas, E, Herrera, LA, Sordo, M, Gonsebatt, ME, Montero, R, Rodriguez, R. and Ostrosky-Wegman, P. (1993). Mitotic index and cell proliferation kinetics for the identification of antineoplastic activity. Anti-Cancer Drugs. 4(6), 637-40. https://doi.org/10.1097/00001813-199312000-00005
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There are 46 citations in total.

Details

Primary Language English
Journal Section Articles
Authors

Nergis Kaya 0000-0002-4206-1149

Publication Date July 15, 2021
Submission Date December 20, 2020
Acceptance Date April 21, 2021
Published in Issue Year 2021 Volume: 11 Issue: 3

Cite

APA Kaya, N. (2021). Evaluation of cytotoxic effect of ferrous gluconate on Allium cepa root tip. Gümüşhane Üniversitesi Fen Bilimleri Dergisi, 11(3), 744-750. https://doi.org/10.17714/gumusfenbil.843648