Research Article
BibTex RIS Cite

PIONEERING in vitro STUDIES FOR CALLUS FORMATION OF Colchicum chalcedonicum Azn.

Year 2020, Volume: 21 Issue: 2, 131 - 137, 15.10.2020
https://doi.org/10.23902/trkjnat.776974

Abstract

Colchicum calcedonicum Azn is one of the endemic species distributed in Turkey, where many endemic plant species occur. It has long-oval shaped corm under the soil, and usually 3-4 leaves on it. In vitro production of endemic species using callus culture has become promising study for conservation. The aim of this study is to generate an efficient callus protocol for in vitro production of C. chalcedonicum. To sterilize the explants, 0.25% (w/v) mercuric chloride (HgCl2) was used for 20 min. In addition to mercuric chloride, surface sterilization was conducted by using 6.5% NaCl with Tween 80 for 30 min. We used 19 different mediums and the primary callus formation was obtained in Murashige & Skoog’s basal medium (MS) supplemented with 2,4-D (2 mg L-1), 2IP (0.5 mg L-1), 3% sucrose and 0.05% active carbon. Our study demonstrated the active carbon usage was effective for the primary callus formation. This study is the first report for primary callus formation of C. chalcedonicum. However, our work is a pioneering study to improve callus formation protocol system for in vitro conservation of endemic species C. chalcedonicum.

Supporting Institution

Kadıköy Belediyesi

Thanks

This study was supported by Kadıköy Municipality. Also, the authors thank Merve Albayrak and Prof. Dr. NESLIHAN YESIM YALCIN MENDI for sharing their results and studies on Colchicum species. Also, the authors thank Dr. Stuart James Lucas for his kind revision.

References

  • 1. Akan, H. & Eker, İ. 2005. Check-list of the genus Colchicum in the flora of Turkey. Turkish Journal of Botany, 29: 327-331.
  • 2. Aznavour, G.Y. 1897. Note sur la flore des environs dc Constantinople. Bulletin de la Société Botanique de France, 44: 164-177.
  • 3. Bisi, A., Gobbi, S., Merolle, L., Farruggia, G., Belluti, F., Rampa, A., Molnar, J., Malucelli, E. & Cappadone, C. 2015. Design, synthesis and biological profile of new inhibitors of multidrug resistance associated proteins carrying a polycyclic scaffold. European Journal of Medicinal Chemistry, 92: 471-80. https://doi.org/10.1016/j.ejmech.2015.01.004.
  • 4. Brickell, C.D. 1984. Colchicum L, In: Davis PH (ed) Flora of Turkey and the East Aegean Islands. Vol. 8, University Press, Edinburgh.
  • 5. Chang, C., Chen, C.T., Tsai, Y.C. & Chang, W.C. 2000. A tissue culture protocol for propagation of a rare plant, Lilium speciosum Thunb. var. gloriosoides Baker. Botanical bulletin of Academia Sinica, 41: 139-142. https://doi.org/10.7016/BBAS.200004.0139.
  • 6. Cocco, G., David, C.C. & Pandolfi, S. 2010. Colchicine in Clinical Medicine. A Guide for Internists. European Journal of Internal Medicine, 21: 503-508. https://doi.org/10.1016/j.ejim.2010.09.010.
  • 7. Combeau, C., Provost, J., Lancelin, F., Tournoux, Y., Prod’homme, F., Herman, F., Lavelle, F., Leboul, J. & Vuilhorgne, M. 2000. RPR112378 and RPR115781: two representatives of a new family of microtubule assembly inhibitors. Molecular Pharmacology, 57(3): 553-63. https://doi.org/10.1124/mol.57.3.553.
  • 8. Dahlgren, R.M.T., Clifford, H.T. & Yeo, P.F. 1985. The Families of the Monocotyledons -Structure, Evolution and Taxonomy. Springer-Verlag Berlin Heidelberg, New York.
  • 9. Daradkeh, N.Q., Shibli, R.A., Makhadmeh, I.M., Alali, F. & Al-Qudah, T.S. 2012. Cell suspension and in vitro production of colchicine in wild Colchicum hierosolymitanum Feib. The Open Conference Proceedings Journal, 3: 52-59. https://doi.org/10.2174/1876326X01203020052.
  • 10. Dumontet, C. & Sikic, B.I. 1999. Mechanisms of action of and resistance to antitubulin agents: microtubule dynamics, drug transport, and cell death. Journal of Clinical Oncology, 17(3): 1061-1070. https://doi.org/10.1200/JCO.1999.17.3.1061.
  • 11. Engelmann, F. 1997. In vitro conservation methods. In: Ford-Lloyd BV, Newburry HJ, Callow JA, (Eds) Biotechnology and plant genetic resources conservation and use. Wallingford: CABI, pp. 119-162.
  • 12. Engelmann F. 1998. In vitro conservation of horticultural genetic resources: Review of the state of the art. World Conference on Horticultural Research; 1998. Rome. 17-20.
  • 13. Evans, W.C. 2002. Trease and Evans’ Pharmacognosy, WB Saunders, Edinburgh, London.
  • 14. Filova, A. 2014. Production of Secondary Metabolites in Plant Tissue Cultures. Research Journal of Agricultural Science, 46(1): 236-45.
  • 15. Gulsoy-Toplan, G., Göger, F., Yildiz-Peko, A., Gibbons, S., Sariyar, G. & Mat A. 2018. Chemical Constituents of the Different Parts of Colchicum micranthum and C. chalcedonicum and their Cytotoxic Activities. Natural Product Communications, 13:535-538. https://doi.org/10.1177/1934578X1801300506.
  • 16. Hayashi, T., Yoshida, K. & Sano, K. 1988. Formation of alkaloids in suspension-cultured Colchicum autumnale. Phytochemistry, 27(5): 1371-1374. https://doi.org/10.1016/0031-9422(88)80196-1.
  • 17. Kapadia, V.H., Dev, S., Rao, R.S. & Ansari, M.Y. 1972. New source of colchicine in Iphigenia. Phytochemistry, 11: 1193
  • 18. Khan, H., Tariq S.A. & Khan, M.A. 2011. Biological and phytochemical studies on corms of Colchicum luteum Baker. Journal of Medicinal Plants Research, 5(32):7031-7035. https://doi.org/10.5897/JMPR11.1159.
  • 19. Kováč, J. 1995. Explantátové kultury rostlin. Skript, UP, Olomouc.
  • 20. Küçüker, O. 1984. Liliaceae (Colchicum) in IOPB chromosome number reports. Taxon, 33: 536.
  • 21. Levy, M., Spino, M. & Read, S.E. 1991. Colchicine: A State of the Art Review. Pharmacotherapy: The Journal of Human Pharmacology and Drug Therapy, 11: 196-211. https://doi.org/10.1002/j.1875-9114.1991.tb02629.x.
  • 22. Masseti, M. 2009. Protected Areas and Endemic Species. In: Gherardi, F., Corti, C., & Gualtieri, M. (Eds), Biodiversity Conservation and Habitat Management. Vol. 1, EOLSS Publications, pp. 118-127.
  • 23. Murashige, T. & Skoog, F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, 15: 473-497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x.
  • 24. Nagaraju, V., Bhowmik, G. & Parthasarathy, V. 2002. Effect of paclobutrazol and sucrose on in vitro cormel formation in gladiolus. Acta Botanica Croatica, 61: 27-33.
  • 25. Persson, K. 1988. New Species of Colchicum (Colchicaceae) from the Greek mountains. Wildenowia, 18: 29-46.
  • 26. Persson, K. 1998. Comments on Some Tesselated Colchicum Species in the East Mediterranean area. Candollea, 53(2): 399-417.
  • 27. Persson, K. 1999a. The Genus Colchicum in Turkey I. New Species. Edinburgh Journal of Botany, 56(1): 85-102. https://doi.org/10.1017/S0960428600002390.
  • 28. Persson, K. 1999b. The Genus Colchicum in Turkey II. Revision of the Large-Leaved Autumnal Species. Edinburgh Journal of Botany, 56(1): 103-142. https://doi.org/10.1017/S0960428600002407.
  • 29. Persson, K. 1999c. New and Revised Species Colchicum L. (Colchicaceae) from the Balkan Peninsula. Plant Systematics and Evolution, 217: 55-80. https://doi.org/10.1007/BF00984922.
  • 30. Persson, K. 2000. Colchicum L. In: Güner, A. (Ed) Flora of Turkey and the East Aegean Islands. Vol. 11. University Press, Edinburg.
  • 31. Persson, K. 2001. A New Soboliferous Species of Colchicum in Turkey. Botanical Journal of the Linnean Society, 135: 85-88. https://doi.org/10.1006/bojl.2000.0384.
  • 32. Persson, K. 2007. Nomenclatural synopsis of the genus Colchicum (Colchicaceae), with some new species and combinations. Botanische Jahrbücher für Systematik, 127: 165-242. https://doi.org/10.1127/0006-8152/2007/0127-016.
  • 33. Pirildar, S., Sutlupinar, N., Atasever, B., Erdem Kuruca, S., Papouskova, B. & Śimánek, V. 2010. Chemical constituents of the different parts of Colchicum baytopiorum and their cytotoxic activities on K562 and HL60 cell lines. Pharmaceutical Biology, 48(1): 32-39. https://doi.org/10.3109/13880200903029373.
  • 34. Shibli, R.A. & Ajlouni, M.M. 2000. Somatic embryogenesis in the endemic black iris. Plant Cell, Tissue and Organ Culture, 61: 15-21. https://doi.org/10.1023/A:1006468122819.
  • 35. The Wealth of India. 1962. A dictionary of Indian raw materials, Vol. VI., C.S.I.R New Delhi, India, 1962.
  • 36. Thomas, T.D. 2008. The role of activated charcoal in plant tissue culture. Biotechnology Advances, 26: 618-631. https://doi.org/10.1016/j.biotechadv.2008.08.003.
  • 37. Verpoorte, R., Contin, A. & Memelink, J. 2002. Biotechnology for the Production of Plant Secondary Metabolites. Phytochemistry Reviews, 1: 13-25. https://doi.org/10.1023/A:1015871916833.
  • 38. Wagh, P.J., Patil, V.W. & Kale, M.C. 2015. Effect of Soluble Chitosan Application on in vitro Germination and Growth of Dormant Corms of Colchicum Luteum Baker. International Journal of Researches in Biosciences, Agriculture and Technology, 3:179-182.
  • 39. Yalçin Mendi, N.Y., Özkaya, A., Sari, M., Özdemir, M.Ş., Sevindik, B., Şimşek, O. & et al, 2017. Somatic embryogenesis of some Colchicum species. Paper presented at International Molecular Biology and Biotechnology Congress, 22-25 December 2017, Afyon, Turkey.
  • 40. Yoshida, K., Hayashi, T. & Sano, K. 1988. Colchicine precursors and the formation of alkaloids in suspension-cultured Colchicum autumnale. Phytochemistry, 27: 1375-1378. https://doi.org/10.1016/0031-9422(88)80197-3.
Year 2020, Volume: 21 Issue: 2, 131 - 137, 15.10.2020
https://doi.org/10.23902/trkjnat.776974

Abstract

Colchicum calcedonicum Azn, pek çok endemik bitki türünün görüldüğü Türkiye'de yayılış gösteren endemik türlerden biridir. Toprak altında uzun-oval şekilli soğanı ile genellikle 3-4 yapraklıdır. Kallus kültürü kullanarak endemik türlerin in vitro üretimi, bu türlerin korunmasında umut verici bir çalışma haline gelmiştir. Bu çalışmanın amacı, in vitro C. chalcedonicum üretimi için verimli kallus protokolünün oluşturulabilmesidir. Explantların sterilizasyonunda, 20 dk %0.25 (w/v) cıva klorür (HgCl2) kullanılmıştır. Cıva klorüre ilaveten, yüzey sterilizasyonunda 30 dk boyunca Tween 80, %6,5 NaCl ile birlikte kullanılmıştır. Bu çalışmada, 19 farklı besiyeri kullanılmış olup primer kallus oluşumu 2,4-D (2 mg L-1), 2IP (0,5 mg L-1), %3 sükroz ve %0,05 aktif karbon içeren Murashige & Skoog bazal besiyerinde elde edilmiştir. Çalışmamız, aktif karbon kullanımının primer kallus oluşumunda etkili olduğunu göstermiştir. Bu çalışma, C. chalcedonicum’un primer kallus oluşumu için ilk rapordur. Bununla birlikte, çalışmamız endemik tür olan C. chalcedonicum'un in vitro korunması ve kallus oluşum protokolünün geliştirilmesinde öncü bir çalışmadır. 

References

  • 1. Akan, H. & Eker, İ. 2005. Check-list of the genus Colchicum in the flora of Turkey. Turkish Journal of Botany, 29: 327-331.
  • 2. Aznavour, G.Y. 1897. Note sur la flore des environs dc Constantinople. Bulletin de la Société Botanique de France, 44: 164-177.
  • 3. Bisi, A., Gobbi, S., Merolle, L., Farruggia, G., Belluti, F., Rampa, A., Molnar, J., Malucelli, E. & Cappadone, C. 2015. Design, synthesis and biological profile of new inhibitors of multidrug resistance associated proteins carrying a polycyclic scaffold. European Journal of Medicinal Chemistry, 92: 471-80. https://doi.org/10.1016/j.ejmech.2015.01.004.
  • 4. Brickell, C.D. 1984. Colchicum L, In: Davis PH (ed) Flora of Turkey and the East Aegean Islands. Vol. 8, University Press, Edinburgh.
  • 5. Chang, C., Chen, C.T., Tsai, Y.C. & Chang, W.C. 2000. A tissue culture protocol for propagation of a rare plant, Lilium speciosum Thunb. var. gloriosoides Baker. Botanical bulletin of Academia Sinica, 41: 139-142. https://doi.org/10.7016/BBAS.200004.0139.
  • 6. Cocco, G., David, C.C. & Pandolfi, S. 2010. Colchicine in Clinical Medicine. A Guide for Internists. European Journal of Internal Medicine, 21: 503-508. https://doi.org/10.1016/j.ejim.2010.09.010.
  • 7. Combeau, C., Provost, J., Lancelin, F., Tournoux, Y., Prod’homme, F., Herman, F., Lavelle, F., Leboul, J. & Vuilhorgne, M. 2000. RPR112378 and RPR115781: two representatives of a new family of microtubule assembly inhibitors. Molecular Pharmacology, 57(3): 553-63. https://doi.org/10.1124/mol.57.3.553.
  • 8. Dahlgren, R.M.T., Clifford, H.T. & Yeo, P.F. 1985. The Families of the Monocotyledons -Structure, Evolution and Taxonomy. Springer-Verlag Berlin Heidelberg, New York.
  • 9. Daradkeh, N.Q., Shibli, R.A., Makhadmeh, I.M., Alali, F. & Al-Qudah, T.S. 2012. Cell suspension and in vitro production of colchicine in wild Colchicum hierosolymitanum Feib. The Open Conference Proceedings Journal, 3: 52-59. https://doi.org/10.2174/1876326X01203020052.
  • 10. Dumontet, C. & Sikic, B.I. 1999. Mechanisms of action of and resistance to antitubulin agents: microtubule dynamics, drug transport, and cell death. Journal of Clinical Oncology, 17(3): 1061-1070. https://doi.org/10.1200/JCO.1999.17.3.1061.
  • 11. Engelmann, F. 1997. In vitro conservation methods. In: Ford-Lloyd BV, Newburry HJ, Callow JA, (Eds) Biotechnology and plant genetic resources conservation and use. Wallingford: CABI, pp. 119-162.
  • 12. Engelmann F. 1998. In vitro conservation of horticultural genetic resources: Review of the state of the art. World Conference on Horticultural Research; 1998. Rome. 17-20.
  • 13. Evans, W.C. 2002. Trease and Evans’ Pharmacognosy, WB Saunders, Edinburgh, London.
  • 14. Filova, A. 2014. Production of Secondary Metabolites in Plant Tissue Cultures. Research Journal of Agricultural Science, 46(1): 236-45.
  • 15. Gulsoy-Toplan, G., Göger, F., Yildiz-Peko, A., Gibbons, S., Sariyar, G. & Mat A. 2018. Chemical Constituents of the Different Parts of Colchicum micranthum and C. chalcedonicum and their Cytotoxic Activities. Natural Product Communications, 13:535-538. https://doi.org/10.1177/1934578X1801300506.
  • 16. Hayashi, T., Yoshida, K. & Sano, K. 1988. Formation of alkaloids in suspension-cultured Colchicum autumnale. Phytochemistry, 27(5): 1371-1374. https://doi.org/10.1016/0031-9422(88)80196-1.
  • 17. Kapadia, V.H., Dev, S., Rao, R.S. & Ansari, M.Y. 1972. New source of colchicine in Iphigenia. Phytochemistry, 11: 1193
  • 18. Khan, H., Tariq S.A. & Khan, M.A. 2011. Biological and phytochemical studies on corms of Colchicum luteum Baker. Journal of Medicinal Plants Research, 5(32):7031-7035. https://doi.org/10.5897/JMPR11.1159.
  • 19. Kováč, J. 1995. Explantátové kultury rostlin. Skript, UP, Olomouc.
  • 20. Küçüker, O. 1984. Liliaceae (Colchicum) in IOPB chromosome number reports. Taxon, 33: 536.
  • 21. Levy, M., Spino, M. & Read, S.E. 1991. Colchicine: A State of the Art Review. Pharmacotherapy: The Journal of Human Pharmacology and Drug Therapy, 11: 196-211. https://doi.org/10.1002/j.1875-9114.1991.tb02629.x.
  • 22. Masseti, M. 2009. Protected Areas and Endemic Species. In: Gherardi, F., Corti, C., & Gualtieri, M. (Eds), Biodiversity Conservation and Habitat Management. Vol. 1, EOLSS Publications, pp. 118-127.
  • 23. Murashige, T. & Skoog, F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, 15: 473-497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x.
  • 24. Nagaraju, V., Bhowmik, G. & Parthasarathy, V. 2002. Effect of paclobutrazol and sucrose on in vitro cormel formation in gladiolus. Acta Botanica Croatica, 61: 27-33.
  • 25. Persson, K. 1988. New Species of Colchicum (Colchicaceae) from the Greek mountains. Wildenowia, 18: 29-46.
  • 26. Persson, K. 1998. Comments on Some Tesselated Colchicum Species in the East Mediterranean area. Candollea, 53(2): 399-417.
  • 27. Persson, K. 1999a. The Genus Colchicum in Turkey I. New Species. Edinburgh Journal of Botany, 56(1): 85-102. https://doi.org/10.1017/S0960428600002390.
  • 28. Persson, K. 1999b. The Genus Colchicum in Turkey II. Revision of the Large-Leaved Autumnal Species. Edinburgh Journal of Botany, 56(1): 103-142. https://doi.org/10.1017/S0960428600002407.
  • 29. Persson, K. 1999c. New and Revised Species Colchicum L. (Colchicaceae) from the Balkan Peninsula. Plant Systematics and Evolution, 217: 55-80. https://doi.org/10.1007/BF00984922.
  • 30. Persson, K. 2000. Colchicum L. In: Güner, A. (Ed) Flora of Turkey and the East Aegean Islands. Vol. 11. University Press, Edinburg.
  • 31. Persson, K. 2001. A New Soboliferous Species of Colchicum in Turkey. Botanical Journal of the Linnean Society, 135: 85-88. https://doi.org/10.1006/bojl.2000.0384.
  • 32. Persson, K. 2007. Nomenclatural synopsis of the genus Colchicum (Colchicaceae), with some new species and combinations. Botanische Jahrbücher für Systematik, 127: 165-242. https://doi.org/10.1127/0006-8152/2007/0127-016.
  • 33. Pirildar, S., Sutlupinar, N., Atasever, B., Erdem Kuruca, S., Papouskova, B. & Śimánek, V. 2010. Chemical constituents of the different parts of Colchicum baytopiorum and their cytotoxic activities on K562 and HL60 cell lines. Pharmaceutical Biology, 48(1): 32-39. https://doi.org/10.3109/13880200903029373.
  • 34. Shibli, R.A. & Ajlouni, M.M. 2000. Somatic embryogenesis in the endemic black iris. Plant Cell, Tissue and Organ Culture, 61: 15-21. https://doi.org/10.1023/A:1006468122819.
  • 35. The Wealth of India. 1962. A dictionary of Indian raw materials, Vol. VI., C.S.I.R New Delhi, India, 1962.
  • 36. Thomas, T.D. 2008. The role of activated charcoal in plant tissue culture. Biotechnology Advances, 26: 618-631. https://doi.org/10.1016/j.biotechadv.2008.08.003.
  • 37. Verpoorte, R., Contin, A. & Memelink, J. 2002. Biotechnology for the Production of Plant Secondary Metabolites. Phytochemistry Reviews, 1: 13-25. https://doi.org/10.1023/A:1015871916833.
  • 38. Wagh, P.J., Patil, V.W. & Kale, M.C. 2015. Effect of Soluble Chitosan Application on in vitro Germination and Growth of Dormant Corms of Colchicum Luteum Baker. International Journal of Researches in Biosciences, Agriculture and Technology, 3:179-182.
  • 39. Yalçin Mendi, N.Y., Özkaya, A., Sari, M., Özdemir, M.Ş., Sevindik, B., Şimşek, O. & et al, 2017. Somatic embryogenesis of some Colchicum species. Paper presented at International Molecular Biology and Biotechnology Congress, 22-25 December 2017, Afyon, Turkey.
  • 40. Yoshida, K., Hayashi, T. & Sano, K. 1988. Colchicine precursors and the formation of alkaloids in suspension-cultured Colchicum autumnale. Phytochemistry, 27: 1375-1378. https://doi.org/10.1016/0031-9422(88)80197-3.
There are 40 citations in total.

Details

Primary Language English
Subjects Genetics
Journal Section Research Article/Araştırma Makalesi
Authors

Elif Karlık 0000-0003-0669-2725

Meltem Değer This is me 0000-0002-8860-2756

Erdal Üzen 0000-0002-7654-6881

Nermin Gozukirmizi 0000-0002-7129-3045

Publication Date October 15, 2020
Submission Date August 4, 2020
Acceptance Date September 4, 2020
Published in Issue Year 2020 Volume: 21 Issue: 2

Cite

APA Karlık, E., Değer, M., Üzen, E., Gozukirmizi, N. (2020). PIONEERING in vitro STUDIES FOR CALLUS FORMATION OF Colchicum chalcedonicum Azn. Trakya University Journal of Natural Sciences, 21(2), 131-137. https://doi.org/10.23902/trkjnat.776974
AMA Karlık E, Değer M, Üzen E, Gozukirmizi N. PIONEERING in vitro STUDIES FOR CALLUS FORMATION OF Colchicum chalcedonicum Azn. Trakya Univ J Nat Sci. October 2020;21(2):131-137. doi:10.23902/trkjnat.776974
Chicago Karlık, Elif, Meltem Değer, Erdal Üzen, and Nermin Gozukirmizi. “PIONEERING in Vitro STUDIES FOR CALLUS FORMATION OF Colchicum Chalcedonicum Azn”. Trakya University Journal of Natural Sciences 21, no. 2 (October 2020): 131-37. https://doi.org/10.23902/trkjnat.776974.
EndNote Karlık E, Değer M, Üzen E, Gozukirmizi N (October 1, 2020) PIONEERING in vitro STUDIES FOR CALLUS FORMATION OF Colchicum chalcedonicum Azn. Trakya University Journal of Natural Sciences 21 2 131–137.
IEEE E. Karlık, M. Değer, E. Üzen, and N. Gozukirmizi, “PIONEERING in vitro STUDIES FOR CALLUS FORMATION OF Colchicum chalcedonicum Azn”., Trakya Univ J Nat Sci, vol. 21, no. 2, pp. 131–137, 2020, doi: 10.23902/trkjnat.776974.
ISNAD Karlık, Elif et al. “PIONEERING in Vitro STUDIES FOR CALLUS FORMATION OF Colchicum Chalcedonicum Azn”. Trakya University Journal of Natural Sciences 21/2 (October 2020), 131-137. https://doi.org/10.23902/trkjnat.776974.
JAMA Karlık E, Değer M, Üzen E, Gozukirmizi N. PIONEERING in vitro STUDIES FOR CALLUS FORMATION OF Colchicum chalcedonicum Azn. Trakya Univ J Nat Sci. 2020;21:131–137.
MLA Karlık, Elif et al. “PIONEERING in Vitro STUDIES FOR CALLUS FORMATION OF Colchicum Chalcedonicum Azn”. Trakya University Journal of Natural Sciences, vol. 21, no. 2, 2020, pp. 131-7, doi:10.23902/trkjnat.776974.
Vancouver Karlık E, Değer M, Üzen E, Gozukirmizi N. PIONEERING in vitro STUDIES FOR CALLUS FORMATION OF Colchicum chalcedonicum Azn. Trakya Univ J Nat Sci. 2020;21(2):131-7.

You can reach the journal's archive between the years of 2000-2011 via https://dergipark.org.tr/en/pub/trakyafbd/archive (Trakya University Journal of Natural Sciences (=Trakya University Journal of Science)


Creative Commons Lisansı

Trakya University Journal of Natural Sciences is licensed under Creative Commons Attribution 4.0 International License.