Cannabis Sativa L. (Kenevir)’de İn Vitro Mikroçoğaltım

Öz

Cannabis sativa (Kenevir) birçok endüstriyel alanda kullanılan bir bitkidir. Dioik (son yıllarda geliştirilen monoik çeşitler) ve yabancı döllenen bir bitki olması, ıslah çalışmalarında ve belirli bir amaca yönelik yetiştiricilikte birçok sorun yaşatmaktadır. Bu yüzden ana ve baba birey olarak kullanılacak genotiplerin klonal çoğaltılması büyük öneme sahiptir. Klonal çoğaltım için biyoteknolojik yöntem olan mikroçoğaltım uygulaması başarılı sonuçlar vermektedir. Daha kısa sürede aynı gen dizilimine sahip birçok bitki üretimi sunmaktadır. Bu çalışmada, kenevir üzerinde yapılmış mikroçoğaltım için eksplant kaynağı ve onun sterilizasyonu, kullanılan besin ortamları, büyüme düzenleyicileri ve köklendirme araştırmaları incelenmiş ve en uygun ortamlar tespit edilmiştir. Gelecekte, ıslah çalışmaları ve amaca uygun sera veya açık alandaki yetiştiricilik işlemleri için uygun klonların çoğaltılmasında yol gösterici olacaktır.

Anahtar Kelimeler

• Cannabis sativa
• in vitro
• mikroçoğaltım

In Vitro Micropropagation of Cannabis Sativa L. (Hemp)

Öz

Cannabis sativa (Hemp) is a plant used in many industrial areas. As hemp is a dioecious (and monoic varieties developed in recent years) and allogamy plant, it causes many problems in breeding and cultivation for a specific purpose. For this reason, the clonal reproduction of genotypes to be used as a parent is of great importance. Micropropagation application, which is a biotechnological method for clonal reproduction, gives successful results. Micropropagation offers the production of many plants with the same gene sequence in a shorter time. In this study, previous studies about explant source and its sterilization, nutrient media used, growth regulators and rooting for hemp micropropagation have been examined, and the most suitable micropropagation conditions of hemp were determined. In the future, this study will guide the reproduction of suitable clones for breeding and greenhouse or open field cultivation.

Anahtar Kelimeler

• Cannabis sativa
• in vitro
• micropropagation

Kaynakça

1. Berahmand, F., Beizaee, N., Dehghan Nayyeri, M., Sharafi, A., Kheiri Manjili, H., Danafar, H., et al. (2016). Cannabis sativa L. genetically transformed root based culture via Agrobacterium rhizogenes. Pharm Biomed Res., 2(3),13-18.
2. Bing, X., Ning, L., Jinfeng, T., & Nan, G. (2007). Rapid tissue culture method of Cannabis sativa for industrial uses. CN, 1887043, 9.
3. Chaohua, C., Gonggu, Z., Lining, Z., Chunsheng, G., Qing, T., Jianhua, C., et al. (2016). A rapid shoot regeneration protocol from the cotyledons of hemp (Cannabis sativa L.). Ind Crops Prod., 83, 61-65.
4. De Meijer, E. (1995). Fibre hemp cultivars: a survey of origin, ancestry, availabilityand brief agronomic characteristics. J. Int. Hemp Assoc., 2, 66-73.
5. Farag, S., & Kayser, O. (2015). Cannabinoids production by hairy root cultures of Cannabis sativa L. American Journal of Plant Sciences, 6,1874-1884.
6. Fitzcharles, M., Clauw, D.J., & Hauser, W. (2020). A cautious hope for cannabidiol (CBD) in rheumatology care. Arthritis Care & Research. Flores-Sancheza, I.J., Peˇc, J., Fei, J., Choia, Y.H., Duˇsek, J., & Verpoorte, R. (2009). Elicitation studies in cell suspension cultures of Cannabis sativa L. Journal of Biotechnology, 143, 157-168.
7. Galán-Ávila, A., García-Fortea, E., Prohens, J., & Herraiz, F.J. (2020). Development of a direct in vitro plant regeneration protocol from Cannabis sativa l. Seedling explants: developmental morphology of shoot regeneration and ploidy level of regenerated plants. Front. Plant Sci. https://doi.org/10.3389/fpls.2020.00645.
8. George, E.F., Hall, M.A., & Klerk, G.J.D. (2008). Plant growth regulators I: Introduction; auxins, their analogues and inhibitors. In: George EF, Hall MA, Klerk GJD, editors. Plant propagation by tissue culture. Dordrecht: Springer, p.175-204.

9. Hawley, D., Graham, T., Stasiak, M., & Dixon, M. (2018). Improving Cannabis bud quality and yield with subcanopy lighting. Hortscience, 53(11), 1593-9.
10. Jiang, Y., Xia, Z., Tang, Y., Han, Q., & Han, C. (2015). Preliminary studies on the tissue culture of Cannabis sativa L. (Industrial hemp). Agricultural science&Technology, 16(5), 923-925.
11. Lata, H., Chandra, S., Khan, I., & ElSohly, M.A. (2009a). Thidiazuron-induced high-frequency direct shoot organogenesis of Cannabis sativa L. In Vitro Cell Dev Biol-Plant, 45, 12-19.
12. Lata, H., Chandra, S., Khan, I., & ElSohly, M.A. (2010). High frequency plant regeneration from leaf derived callus of high Δ9-tetrahydrocannabinol yielding Cannabis sativa L. Planta Med., 76, 1629-1633.
13. Lata, H., Chandra, S., Khan, I.A., & Elsohly, M.A. (2009b). Propagation through alginate encapsulation of axillary buds of Cannabis sativa L. - an important medicinal plant. Physiol. Mol. Biol. Plants 15, 79-86.
14. Lata, H., Chandra, S., Techen, N., Khan, I.A., & ElSohly, M.A. (2016). In vitro mass propagation of Cannabis sativa L.: a protocol refinement using novel aromatic cytokinin meta-topolin and the assessment of eco-physiological, biochemical and genetic fidelity of micropropagated plants. J Appl Res Med Aromat Plants, 3, 18-26
15. Murashige, T., & Skoog, F. (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant, 15, 473-497. Papadopoulou, E., Bikiaris, D., Chrysafis, K., Wladyka-Przybylak, M., Wesolek, D., Mankowski, J., et al. (2015). Value added industrial products from bast fiber crops. Ind. Crops Prod., 68, 116-125.
16. Quimby, M. (1974). Botany of Cannabis sativa. In: Mateos-Gomez JL (ed) Archivos de investigacion medica. El Instituto Mexicano del Seguro Social.
17. Salentijn, E.M.J., Zhang, Q., Amaducci, S., Yang, M., & Trindade, L.M. (2015). New developments in fiber hemp (Cannabis sativa L.) breeding. Ind Crops Prod., 68, 32-41.
18. Schachtsiek, J., Warzecha, H., Kayser, O., & Stehle, F. (2018). Current perspectives on biotechnological cannabinoid production in plants. Planta Med., 84(04), 214-220.
19. Slusarkiewicz-Jarzina, A., Ponitka, A., & Kaczmarek, Z. (2005). Influence of cultivar, explant source and plant growth regulator on callus induction and plant regeneration of Cannabis sativa L. Acta Biol Cracov Ser Bot., 47, 145-51.
20. Wahby, I., Caba, J.M., & Ligero, F. (2013). Agrobacterium infection of hemp (Cannabis sativa L.): establishment of hairy root cultures. J Plant Interact., 8, 312-320.
21. Wang, R., He, L.S., Xia, B., Tong, J.F., Li, N., & Peng, F.A. (2009). A micropropagation system for cloning of hemp (Cannabis sativa L.) by shoot tip culture. Pak. J. Bot., 41, 603-608.
22. Wielgus, K., Luwanska, A., Lassocinski, W., & Kaczmarek, Z. (2008). Estimation of Cannabis sativa L. tissue culture conditions essential for callus induction and plant regeneration. J Nat Fibers., 5, 199-207.
23. Wróbel, T., Dreger, M., Wielgus, K, & Słomski, R. (2020). Modified Nodal Cuttings and Shoot Tips Protocol for Rapid Regeneration of Cannabis sativa L. Journal of Natural Fibers, DOI: 10.1080/15440478.2020.1748160.
24. Ye, X. (2015). Development and application of plant transformation techniques. J.Integr. Agric., 14, 411-413.