Callus Production in Geranium (Pelargonium quercetorum Agnew) Growing Naturally in Türkiye

Abstract

Pelargonium quercetorum Agnew grows naturally in the Hakkari province of Türkiye. Although P. quercetorum Agnew has potential use as a medicine and ornamental plant, it is especially used as a medicinal plant for the cure of various diseases by local people. In vitro tissue culture methods are favorable for the propagation, conservation, and breeding of medicinal plants. We aimed in this study to achieve regeneration of P. quercetorum Agnew from different explant types. Seeds of P. quercetorum Agnew were germinated in vitro conditions and explants were taken from these germinated sterile plantlets. Totally four different experiments, containing three of them embryogenic and one of them organogenic culture, were established to achieve regeneration in P. quercetorum Agnew. Leaf, petiole, cotyledon, cotyledon stalk, and root collar disc were used as explant. Different concentrations of 1-Naphthaleneacetic acid (NAA), 2,4-dichlorophenoxyacetic (2,4-D), 6-Benzylaminopurine (BA), 6-Furfurylaminopurine (Kinetin), 6-(γ,γ-Dimethylallylamino) purine (2iP), and Thidiazuron (TDZ) were used to induce embryogenic or organogenic regeneration. Explants were cultured in half-strength or full-strength Murashige and Skoog (MS) medium. In the embryogenic experiments, callus formation from different media ranged from 63.5% to 100%, and for explant types ranged from 39% to 100%. In the organogenic experiment, callus formation from different media ranged from 12.5% to 100%, and for explant types ranged from 71% to 93%. Also, embryo-like structures were obtained from embryogenic experiments. However, these structures could not grow more and transformed into plantlets.

Keywords

• Pelargonium quercetorum Agnew
• Somatic embryogenesis
• Organogenesis
• Callus

References

1. Agarwal, P. K., & Ranu, R. S. (2000). Regeneration of plantlets from leaf and petiole explants of Pelargonium × hortorum. In Vitro Cellular and Developmental Biology - Plant, 36(5). https://doi.org/10.1007/s11627-000-0070-y
2. Agnew, A. D. Q. (1967). Contributions to the Flora of Iraq: IV: Notes on the Geraniaceae of Iraq, with a New Species of Pelargonium. Kew Bulletin, 21(2). https://doi.org/10.2307/4108508
3. Altan, F., & Duru, M. (2017). Investigation of alkaloids in callus and multiple shoots obtained from Catharantus roseus L. YYU J AGR SCI, 27(3), 337-346.
4. Amer, A. M., Mohamed, G. M., Hussein, M. H., Sedik, M. Z., & Aly, U. I. (2017). Effect of some of the natural organic sources on rice tissue culture. Egyptian Pharmaceutical Journal, 16(3), 152. https://doi.org/10.4103/EPJ.EPJ_32_17
5. Bakhshaie, M., Babalar, M., Mirmasoumi, M., & Khalighi, A. (2010). Somatic embryogenesis and plant regeneration of Lilium ledebourii (Baker) Boiss., an endangered species. Plant Cell, Tissue and Organ Culture, 102(2), 229–235. https://doi.org/10.1007/S11240-010-9726-4
6. Benazir, J. F., Suganthi, R., Chandrika, P., & Mathithumilan, B. (2013). In vitro regeneration and transformation studies on Pelargonium graveolens (geranium) - an important medicinal and aromatic plant. Journal of Medicinal Plants Research, 7(38).
7. Benjamin, E. D., Ishaku, G. A., Peingurta, F. A., & Afolabi, A. S. (2019). Callus culture for the production of therapeutic compounds. American Journal of Plant Biology, 4(4), 76-84. https://doi.org/10.11648/j.ajpb.20190404.14
8. Brown, J. T., & Charlwood, B. V. (1986). The accumulation of essential oils by tissue cultures of Pelargonium fragrans (Willd.). FEBS Letters, 204(1). https://doi.org/10.1016/0014-5793(86)81397-7

9. Castillo, B., & Smith, M. A. L. (1997). Direct somatic embryogenesis from Begonia gracilis explants. Plant Cell Reports, 16(6). https://doi.org/10.1007/s002990050244
10. Choffe, K. L., Victor, J. M. R., Murch, S. J., & Saxena, P. K. (2000). In vitro regeneration of Echinacea purpurea L.: Direct somatic embryogenesis and indirect shoot organogenesis in petiole culture. In Vitro Cellular and Developmental Biology - Plant, 36(1). https://doi.org/10.1007/s11627-000-0008-4
11. Daneshvar, M. H., Havil, M., & Lotfi Jalal-Abadi, A. (2022). Micropropagation of Polianthes tuberosa L. through direct organogenesis. J. Plant Prod., 45(2).
12. Davis, P.H. (1967). Pelargonium L. In: Davis PH (Ed.) Flora of Turkey and the East Aegean Islands. Edinburgh University Press, Edinburgh, 451–474.
13. Ebrahimzadeh, A., Fathollahzadeh, M., Aazami, M. A., & Hassanpouraghdam, M. B. (2022). In vitro direct and indirect regeneration of plants from nodal and petiole explants in Pelargonium odoratissimum (L.) Herit. Acta Agriculturae Slovenica, 118(4). https://doi.org/10.14720/aas.2022.118.4.1723
14. Habiba, S. U., Ahasan, M. M., & Shimasaki, K. (2018). Effects of Ethrel on Organogenesis of Protocorm-like bodies in Dendrobium kingianum In vitro. Plant Tissue Culture and Biotechnology, 28(1). https://doi.org/10.3329/ptcb.v28i1.37205
15. Kaval, I., Behçet, L., & Cakilcioglu, U. (2014). Ethnobotanical study on medicinal plants in Geçitli and its surrounding (Hakkari-Turkey). Journal of Ethnopharmacology, 155(1). https://doi.org/10.1016/j.jep.2014.05.014
16. Kazeroonian, R., Mousavi, A., Kalate Jari, S., & Tohidfar, M. (2018). Factors Influencing in Vitro Organogenesis of Chrysanthemum morifolium cv. ‘Resomee Splendid.’ Iranian Journal of Biotechnology, 16(2). https://doi.org/10.21859/ijb.1454
17. Kocak, M., Izgu, T., Sevindik, B., Tutuncu, M., Curuk, P., Simsek, O., Aka Kacar, Y., Teixeira Da Silva, J. A., & Mendi, Y.Y., (2014). Somatic embryogenesis of Turkish Cyclamen persicum Mill. Scientia Horticulturae 172, 26–33.
18. Kumar, V., Moyo, M., & Van Staden, J. (2015). Somatic embryogenesis of Pelargonium sidoides DC. Plant Cell, Tissue and Organ Culture, 121(3), 571–577. https://doi.org/10.1007/S11240-015-0726-2/METRICS
19. Lü, J., Chen, R., Zhang, M., da Silva, J. A. T., & Ma, G. (2013). Plant regeneration via somatic embryogenesis and shoot organogenesis from immature cotyledons of Camellia nitidissima Chi. Journal of Plant Physiology, 170(13). https://doi.org/10.1016/j.jplph.2013.03.019
20. Madden, J. I., Jones, C. S., & Auer, C. A. (2005). Modes of regeneration in Pelargonium x hortorum (Geraniaceae) and three closely related species. In Vitro Cellular and Developmental Biology - Plant, 41(1). https://doi.org/10.1079/IVP2004605
21. Maślanka, M., & Bach, A. (2014). Induction of bulb organogenesis in in vitro cultures of tarda tulip (Tulipa tarda Stapf.) from seed-derived explants. In Vitro Cellular and Developmental Biology - Plant, 50(6). https://doi.org/10.1007/s11627-014-9641-1
22. Mendi, Y. Y., Curuk, P., Kocaman, E., Unek, C., Eldogan, S., Gencel, G., & Çetiner, S. (2009). Regeneration of begonia plantlets by direct organogenesis. African Journal of Biotechnology, 8(9).
23. Öktem, H., & Yücel, M. (2012). Bitki Biyoteknolojisi ve Genetik: İlkeler. Teknikler ve Uygulamalar. Ankara: Nobel Akademik Yayıncılık. p.113-134.
24. Pareek, A., & Kothari, S. L. (2003). Direct somatic embryogenesis and plant regeneration from leaf cultures of ornamental species of Dianthus. Scientia Horticulturae, 98(4). https://doi.org/10.1016/S0304-4238(03)00078-5
25. Pawar, B., Kale, P., Bahurupe, J., Jadhav, A., Kale, A., & Pawar, S. (2015). Proline and glutamine improve in vitro callus induction and subsequent shooting in rice. Rice Science, 22(6). https://doi.org/10.1016/j.rsci.2015.11.001
26. Qureshi, J. A., & Saxena, P. K. (1992). Adventitious shoot induction and somatic embryogenesis with intact seedlings of several hybrid seed geranium (Pelargonium x hortorum Bailey) varieties. Plant Cell Reports, 11(9). https://doi.org/10.1007/BF00232687
27. Robichon, M. P., Renou, J. P., & Jalouzot, R. (1997). Plant regeneration of ivy leaved geranium through shoot organogenesis. Plant Cell, Tissue and Organ Culture, 49(3). https://doi.org/10.1023/A:1005843901764
28. Röschenbleck, J., Albers, F., Müller, K., Weinl, S., & Kudla, J. (2014). Phylogenetics, character evolution and a subgeneric revision of the genus Pelargonium (Geraniaceae). Phytotaxa, 159(2). https://doi.org/10.11646/phytotaxa.159.2.1
29. Rout, G. R., Mohapatra, A., & Jain, S. M. (2006). Tissue culture of ornamental pot plant: A critical review on present scenario and future prospects. In Biotechnology Advances (Vol. 24, Issue 6). https://doi.org/10.1016/j.biotechadv.2006.05.001
30. Saxena, G., Banerjee, S., Rahman, L., Mallavarapu, G. R., Sharma, S., & Kumar, S. (2000). An efficient in vitro procedure for micropropagation and generation of somaclones of rose scented Pelargonium. Plant Science, 155(2). https://doi.org/10.1016/S0168-9452(00)00213-2
31. Sreedhar, D. (1999). Tissue culture of Pelargonium graveolans L.Her. Ex. A it (Geranium). A Role of Biotechnology in Medicinal and Aromatic Plants, 2, 177–184.
32. Sukhumpinij, P., Kakihara, F., & Kato, M. (2010). In vitro regeneration from mature leaf explants of Pelargonium rapaceum (L.) L’Hérit. Scientia Horticulturae, 126(3), 385–389. https://doi.org/10.1016/J.SCIENTA.2010.07.028
33. Taherpour, A., & Khojasteh, K. (2008). Comparison of the volatile constituents of Pelargonium quercetorum Agnew. of Iran with some of the other pelargonium species. Asian Journal of Chemistry, 20(8), 6335.
34. Vejsadová, H., & Kuchtová-Jadrná, P. (2008). Indukce organogeneze u diploidních kultivarů Pelargonium× hortorum LH Bailey. Acta Pruhoniciana, 89, 41–45.
35. Visser, C., Qureshi, J. A., Gill, R., & Saxena, P. K. (1992). Morphoregulatory role of thidiazuron: Substitution of auxin and cytokinin requirement for the induction of somatic embryogenesis in geranium hypocotyl cultures. Plant Physiology, 99(4). https://doi.org/10.1104/pp.99.4.1704
36. Wojtania, A., Gabryszewska, E., & Marasek, A. (2004). Regeneration of Pelargonium x hederaefolium “Bonete” from petiole explants. Acta Physiologiae Plantarum, 26(3), 255–262. https://doi.org/10.1007/S11738-004-0015-X/METRICS
37. Zhang, J., Gai, M. Z., Li, X. Y., Li, T. L., & Sun, H. M. (2016). Somatic embryogenesis and direct as well as indirect organogenesis in Lilium pumilum DC. Fisch., an endangered ornamental and medicinal plant. Bioscience, Biotechnology and Biochemistry, 80(10). https://doi.org/10.1080/09168451.2016.1194178
38. Zuraida, A., Shukri, M., Sabrina, M., & Nazreena, O. (2015). Improvement of Regeneration of Pelargonium radula via Somatic Embryogenesis. British Biotechnology Journal, 5(4). https://doi.org/10.9734/bbj/2015/15337