Rahma BEJAOUI, Görkem Eren ÖZDEMİR, Şeküre Şebnem ELLİALTIOĞLU

Doku Kültürü Tekniğiyle Çoğaltılan Kalanchoe blossfeldiana Poelln. Sürgünlerinin in vitro veya ex vitro Köklendirilmesi ve Dış Koşullara Alıştırılması

Bir süs bitkisi olan kalanşonun (Kalanchoe blossfeldiana Poelln.) doku kültürü yoluyla çoğaltılması incelenmiştir. Yaprak parçaları, kalanşo türünde in vitro çoğaltım için başlangıç eksplantı olarak kullanılmıştır. Çoğaltma ortamında (MS makro ve mikro elementleri + 0.3 mg/L GA3 + 1.0 mg/L BAP + 3 g/L sukroz + 7 g/L agar) elde edilen kalanşo sürgünleri, pişkinleştirme ortamına aktarılmıştır. Pişkinleştirme aşamasında hormonsuz MS + %0.2 aktif kömür ortamı kullanılmıştır. Bu aşamadan sonra köklenme amacıyla 1 mg/L IBA ilave edilmiş MS ortamı kullanılmıştır. Ayrıca hormonsuz MS ve ½ MS ortamları da köklendirme denemesinde yer almıştır. Tüm ortamlarda %100 köklenme sağlanmasına rağmen IBA katkısı kök yoğunluğunu arttırmış ve köklenme kapasitesini hızlandırmıştır. Vermikülit, kum, vermikülit + kum ortamında dış şartlara geçiş aşamasında; torf ve hidroponik sistem üzerine yerleştirilen viyollere yapılan tüm transfer uygulamalarında %100 başarı ve yaşama oranı sağlanmıştır. Doku kültüründe ex vitro köklendirme, üretim maliyetini azaltabilecek bir tekniktir. Kalanchoe bitkisinin in vitro klonal çoğaltımı, ex vitro köklendirme ve dış koşullara alıştırılması işlemleri başarıyla gerçekleştirilmiştir.

Anahtar Kelimeler:

Köklendirme, Kalanchoe, süs bitkisi, in vitro, ex vitro, iklimlendirme.

In vitro or ex vitro Rooting and Acclimatization of Kalanchoe blossfeldiana Poelln. Shoots Propagated by Tissue Culture Technique.

The propagation of kalanchoe (Kalanchoe blossfeldiana Poelln.), an ornamental plant, through tissue culture has been studied. Leaf pieces were used as initial explant for in vitro propagation of kalanchoe. Kalanchoe shoots obtained in proliferation medium included (MS macro and microelements + 0.3 mg/L GA3 + 1.0 mg/L BAP + 3 g/L sucrose + 7 g/L agar). The shoots were transferred to the hardening medium. For hardening of in vitro kalanchoe shoots, it was used hormon-free MS medium + %0.2 activated charcoal. After the hardening medium, 1 mg/L IBA was added to the MS-medium at the rooting stage were used. Hormone-free MS or hormone-free ½ MS medium were also used as rooting media. Although 100% rooting was achieved in all media, IBA additive increased root density and accelerated rooting capacity. In the media of vermiculite, sand, vermiculite + sand at the stage of transferring to external conditions; 100% success and survival rate was achieved in all applications of transfer to viols containing peat and to the float hydroponic system. Ex vitro rooting in tissue culture could be a technique that reduces the production cost. in vitro clonal propagation of kalanchoe plant, ex vitro rooting and acclimatization have been successfully carried out.

Keywords:

Rooting, Kalanchoe, ornamental plant, in vitro, ex vitro, acclimatization.,

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Altındağ Çelik, F. N. 2022. Investigations on in vitro regeneration and micropropagation from leaf explants of Kalanchoe blossfeldiana (Kalanchoe). Bartın University, Graduate School of Natural and Applied Sciences, Department of Horticulture, M.Sc. Thesis, 67 p., Bartın.
Anonim. 2004. T.C. Başbakanlık Devlet Meteoroloji İşleri Genel Müdürlüğü, Elazığ İl Md. Kayıtları.
Becker, H. C., Leon, J. 1988. Stability analysis in plant breeding. Plant Breeding 101: 1–23.
Bejaoui, R. 2022. In vitro Micropropagation of Kalanchoe (Kalanchoe blossfeldiana Poelln.). Ankara University, Graduate School of Natural and Applied Sciences, Department of Horticulture, Ph.D. Thesis, 103 p., Ankara.
Bejaoui, R., Gümüş, C., Sönmez, K., Kırbay, E. and Ellialtıoğlu, Ş. Ş. 2023.The effects of different PGR contents on in vitro organogenesis and shoot proliferation in kalanchoe (Kalanchoe blossfeldiana Poelln.). In: 11th International Conference on Agriculture, Animal Sciences and Rural Development, 03-05 March 2023, Muş Alparslan University, Muş – Türkiye. Conference Proceedings Book (Eds: Karadağ,Y., Seydoşoğlu, S.). pp: 995-1010. ISBN:978-625-7720-91-5.
Bhuiyan, M.S.U., Kim, T., In, J. G., Yang, D. C. and Choi, K. S. 2006. Plant regeneration from leaf explants of Kalanchoe daigremontiana Hamet & Perrier. Korean Journal of Medicinal Crop Science, 14(5), 293-298.
Bonga, J. and Von Aderkas, P. 1992. In vitro Culture of Trees. The Netherlands: Kluwer Academic Publishers,72-125, Dordrecht.
Chen, M. 2007. Study on in vitro rapid propagation of Kalanchoe blossfeldiana. Journal of Anhui Agricultural Sciences, 32. (Accessed on 23.04.2018): http://en.cnki.com.cn/Article_en/CJFDTOTAL-AHNY200732072.htm. Clapa, D., Fira, A. and Joshee, N. 2013. An efficient ex vitro rooting and acclimatization method for horticultural plants using float hydroculture. HortScience, 48(9), 1159-1167.
Clapa, D., Fira, A., Dumitraş, A. and Ciorchina, N. 2011. Înrădăcinarea şi aclimatizarea ex vitro în hidrocultură prin flotaţie a unor genotipuri de mur. Revista Botanică, 3(1), 133-139.
Cui, G. and Wei, B. 2003. Studies on propagation and rooting rate of Kalanchoe blossfeldiana in vitro. Special Wild Economic Animal and Plant Research, 4. (Accessed on 23.04.2018): http://en.cnki.com.cn/Article_en/CJFDTOTAL-TCYA200304004.htm.
De Klerk, G. J., Arnholdt-Schmitt, B., Lieberei, R. and Neumann, K. H. 1997. Regeneration of roots, shoots and embryos: physiological, biochemical and molecular aspects. Biologia Plantarum, 39(1), 53-66.
De Klerk, G. J., Van Der Krieken, W. and De Jong, J. C. 1999. Review the formation of adventitious roots: new concepts, new possibilities. In Vitro Cellular & Developmental Biology-Plant, 35(3), 189-199.
De Klerk, G.J. 2001. In: Plant Roots: The Hidden Half. (ed) Waisel, Y., Eshel, A. and Kafkafi, U. New York, Basel: Marcel Dekker Publishers, 349–357.
Deng, Q. Zhang, Y. and Wang, C. 2005. Study on in vitro propagation of Kalanchoe blossfeldiana. Journal of Sichuan Agricultural University, Vol.2.
Donnelly, D. J., and Tisdall, L. 1993. Acclimatization strategies for micropropagated plants. Micropropagation of Woody Plants, 153-166.
Duan, J. X., Duan, Q. X., Zhang, S. F., Cao, Y. M., Yang, C. D., and Cai, X. D. 2020. Morphological, physiological, anatomical and histochemical responses of micropropagated plants of Trichosanthes kirilowii to hydroponic and soil conditions during acclimatization. Plant Cell, Tissue and Organ Culture (PCTOC), 142(1), 177-186.
Eberhart, S. A. T., Russell, W. A. 1966. Stability parameters for comparing varieties. Crop Science 6: 36-40. Finlay, K. W., Wilkinson, G. N. 1963. Adaptation in a plant breeding programme. Australian Journal of Agricultural Research, 14: 742-754.
Fox, P., Skovmand, B., Thompson, B., Braun, H. I., Cormier, R. 1990. Yield and adaptation of hexaploid spring triticale. Euphytica, 47: 57-64.
Gajdošová, A., Ostrolucká, M.G., Libiaková, G. and Ondrušková, E. 2007. In: Protocols for Micropropagation of Woody Trees and Fruits. (eds) Jain, SM and Häggman, H. Springer, Dordrecht, 447–464.
Girish, M. H., Gasti, V. D., Mastiholi, A. B., Thammaiah, N., Shantappa, T., Mulge, R., Kerutagi, M. G. 2012. Correlation and path analysis for growth, pod yield, seed yield and quality characters in cluster bean (Cyamopsis tetragonoloba (L.) Taub.). Karnataka Journal of Agricultural Sciences, 25 (4), 498-502.
Gökkuş, A., Bakoğlu, A. ve Koç, A. 1996. Bazı Adı Fiğ (Vicia sativa L.) hat ve çeşitlerinin Erzurum sulu şartlarına adaptasyonu üzerine bir çalışma. Türkiye 3. Çayır-Mera ve Yembitkileri Kongresi, 17-19 Haziran, Erzurum, s. 674-678.
Haifan, H., Baoyin, L. and Ping, L. 2004. Several factors influencing in vitro culture and plantlet regeneration of Kalanchoe blossfediana. Chinese Agricultural Science Bulletin, Vol.02.
Hepaksoy, S. and Aksoy, U. 2006. Propagation of Ficus carica L. clones by in vitro culture. Biologia Plantarum, 50(3), 433-436.
Hocaoğlu, O., Akan, K., Akçura, M. 2020. Evaluating leaf rust reactions of pure bread wheat landrace lines using non-parametric statistics. Phytoparasitica, 48: 261-271.
Hong, S. Y., Kim, S. D., Lee, Y. S. and Choi, S. Y. 2002. Selection of transplanting soil for acclimatization of in vitro cultured grapevine plantlets. Horticultural Science & Technology, 20(1), 38-41.
Huhn, M. 1990. Non-parametric measures of phenotypic stability. Part 1: Theory. Euphytica, 47: 189-1990. Ibrahim, A. O. S. 2022. Researches on using of float hydroculture for ex vitro rooting of microcuttings and acclimatization of in vitro plantlets in some Pyrus species. Ankara University, Graduate School of Natural and Applied Sciences, Department of Horticulture, Ph.D. Thesis, 106 pp, Ankara.
Kang, M. S. 1988. A rank-sum method for selecting high-yielding, stable corn genotypes. Cereal Research Communication, 16: 113-115.
Kordi, M., Kaviani, B. and Hashemabadi, D. 2013. In vitro propagation of Kalanchoe blossfeldiana using BA and NAA. European Journal of Experimental Biology, 3(1), 285-288.
Lineberger, R. D. 1983. Shoot proliferation, rooting, and transplant survival of tissue-cultured ‘Hally Jolivette’cherry. HortScience, 18(2), 182-185.
Linjian, F., Dongpo, J. and Xinshuan, Z. 2006. Research of different culture conditions on plantlet regeneration from leaves of Kalanchoe blossfeldiana. Chinese Agricultural Science Bulletin, 5. (Accessed on 23.04.2018): http://en.cnki.com. cn/Article_en/CJFDTotal ZNTB200605021.htm.
Liu, H. 2010. High propagation and tissue culture system and outside-tube rooting technology of Kalanchoe blossfeldiana. Journal of Anhui Agricultural Sciences, 12. (Accessed on 23.04.2018): http://en.cnki. com.cn/Article_en/CJFDTOTAL-AHNY201012018.htm.
Matsubara, S. and Chen, D. 1989. In vitro production of garlic plants and field acclimatization. HortScience, 24(4), 677-679.
Németh, G. 1986. Induction of Rooting. In: Trees I. Biotechnology in Agriculture and Forestry, Bajaj, Y.P.S. (eds), vol 1. Springer, Berlin, Heidelberg.
Ostrolucká, M. G., Gajdošová, A. and Libiaková, G. 2007b. In: Protocols for Micropropagation of Woody Trees and Fruits. (eds) Jain, SM. and Häggman, H. Springer -Verlag, Berlin, Heidelberg, pp. 85–91.
Ostrolucká, M. G., Gajdošová, A., Libiaková, G., Hrubíková, K. and Bežo, M. 2007a. Protocol for micropropagation of selected Vaccinium spp, In: Protocols for Micropropagation of Woody Trees and Fruits Springer, Dordrecht, 445-455.
Pan, Mj. and Van Staden, J. 1998. The use of charcoal in in vitro culture-A review. Plant Growth Regulation, 26, 111-163.
Peng, C., Wang, L., Li, K., Xia, L. and Lu, F. 2008. Research on tissue culture and plantlet reproduction from leafstalk of Kalanchoe blossfeldiana with orange flower. Tropical Agricultural Science & Technology, 01. (Accessed on 23.04.2018): http://en.cnki.com.cn/Article_en/CJFDTOTAL-YNRJ200801012.htm.
Plaisted, R. L. 1960. A shorter method for evaluating the ability of selections to yield consistently over locations. American Potato Journal, 37: 166-172.
Preece, Y. JE. and Sutter, EG .1991. Acclimatization of micropropagated plants to the greenhouse and field. In: Micropropagation, Technology and Application, (eds) Deberg, PC. and Zimmerman, RH. Dordrecht: Kluwer Academic.
Rodrigues, P. H. V., Lima, A. M. L. P., Ambrosano, G. M. B. and Dutra, M. D. F. B. 2005. Acclimatization of micropropagated Heliconia bihai (Heliconiaceae) plants. Scientia Agricola, 62, 299-301.
Rohr, R., Iliev I., Scaltsoyiannes, A. and Tsoulpha, P. 2003. Acclimatization of micropropagated forest trees. Acta Horticulturae, 616, 59–69. doi:10.17660/ actahortic.2003.616.3.
Sanikhani, M., Frello, S. and Serek, M. 2006. TDZ induces shoot regeneration in various Kalanchoë blossfeldiana Poelln. cultivars in the absence of auxin. Plant Cell, Tissue and Organ Culture, 85(1), 75–82.
Shukla, G. K. 1972. Some statistical aspects of partitioning genotype-environmental components of variability. Heredity, 29: 237-245.
Skirvin, R. M., Chu, M. C. and Rukan, H. 1980. Rooting studies with Prunus spp. in vitro. HortScience 15 (3): 415.
Tang, J. 2007. Study on Tissue Culture and Rapid Propagation of Kalanchoe blossfeldiana Leaves. Northern Horticulture, 10, 188-189. (oriprobe.com)
Thennarasu, K. 1995. On certain non-parametric procedures for studying genotype-environment interactions and yield stability. PhD thesis, PJ School, IARI, New Delhi, India.
Tükel, T. ve Hatipoğlu, R. 1997. Mera Amenajmanı. Çukurova Üniversitesi Ziraat Fakültesi Genel Yayın No: 191. Ders Kitabı No: A59, Adana, 300 s.
Varga, A., Thoma, L. H. and Bruinsma, J. 1988. Effects of auxins and cytokinins on epigenetic instability of callus-propagated Kalanchoe blossfeldiana Poelln. Plant Cell, Tissue and Organ Culture, 15(3), 223-231.
Wafaa, A. F. and Wahdan, H. M. 2017. Influence of substrates on in vitro rooting and acclimatization of micropropagated strawberry (Fragaria × ananassa Duch). Middle East J Agric Research, 6(3), 682-691.
Wricke, G. 1962. Übereine Methode zur Erfassung der ökologischen Streubreite in Feldversuchen. Zeitschrift für Pflanzenzüchtung, 47: 92-96.
Xinzheng, S., Qingwei, L. and Mingquing, M. 2006. Research on the technology of Kalanchoe blossfeldiana with white flower tissue culture and rapid reproduction. Chinese Agricultural Science Bulletin, 3.
Yan, W. 2014. Crop Variety Trials: Data Management and Analysis John Wiley and Sons, 349.
Zhang, R. and Guo, X. 2005. Study on regeneration of vitro leaf of Kalanchoe blossfeldiana. Shanxi Forestry Science and Technology, 01. (Accessed on 23.04.2018): http://en.cnki.com.cn/Article_en/CJFDTotal-SXLK200501003. htm.
Zhang, Y. S., Gu, S. J., Chen, J. J., and Cai, X. D. 2019. Effects of different nutrient solutions on the acclimatization of in vitro Caladium plantlets using a simplified hydroponic system. Sains Malays, 48, 1627-1633.