Abstract
Immortelle grass (Helichrysum italicum (Roth) G. Don), which spreads in the Southern Marmara and Aegean regions, can be grown in arid and semi-arid regions. In addition, due to its rich essential oil and secondary metabolite content, it has an important place in modern medicine and cosmetics, including traditional treatment methods. Although the propagation of plants by shoot regeneration in vitro has been achieved in many plant species, studies on tissue culture in immortelle grass are limited. This study aims to optimize the tissue culture study in immortelle grass and provide a basis for the next in vitro, molecular, and secondary metabolite studies. In addition, it promotes the plant by optimizing the healthy and disease-free seedling production method for cultural agriculture in the region. Three different (15%, 25%, and 35%) NaOCl concentrations were tested for 10 and 20 minutes during the sterilization phase of the explants. The most successful result was obtained in the medium containing 35% NaOCl for 10 minutes. Sterilized explants were transferred to MS and Gamborg B5 nutrient media containing BAP, GA, and NAA plant growth regulators for shoot regeneration. The best regeneration in explants was obtained in MS medium containing 0.5 mg L-1 BAP, 1 mg L-1 GA, and 0.2 mg L-1 NAA. No growth was observed in trials containing Gamborg B5, and vitrification and darkening occurred in the explants. After four weeks, the shoots reaching a length of 3 cm were taken into MS and ½MS medium containing 0 MS, 0.5 mg L-1 IBA, 1 mg L-1 IBA, 1.5 mg L-1, and 2 mg L-1 IBA as a rooting medium. 100% rooting was observed in all prepared media within four weeks. As a result of micropropagation studies, the rooted plants were transferred to the acclimatization stage within three months and then moved to the pots in the greenhouse and to the field one month later.
References
- Albayrak, S., Aksoy, A., Sagdic, O. and Hamzaoglu, E. (2010). Compositions, antioxidant and antimicrobial activities of Helichrysum (Asteraceae) species collected from Turkey. Food Chemistry, 119(1): 114-122.
- Andrade, L. B., Echeverrigaray, S., Fracaro, F., Pauletti, G. F. and Rota, L. (1999). The effect of growth regulators on shoot propagation and rooting of common lavender (Lavandula vera DC). Plant Cell, Tissue and Organ Culture 56: 79-83.
- Clasquin, S. and Henry, M. (2002). Micropropagation Helichrysum arenarium (L.) Moench. Acta Botanica Gallica. 149(2). 189–195.
- Daneshvar Royandazagh, S. and Pehlivan, E. C. (2016). In vitro micropropagation of Lilium candidum L. and alkaloids. Journal of Tekirdag Agricultural Faculty, 13(3): 100-110 (In Turkish).
- Dimitrova, N. G. and Nacheva, L. (2018). Micropropagation of Helichrysum italicum (ROTH) G. Don–a medicinal plant with ornamental value. Journal of BioScience and Biotechnology, 7(2-3): 97-101.
- El-Banna, H. Y. (2017). Micropropagation of thyme plant (Thymus vulgaris). Journal of Plant Production, 8(11): 1221-1227.
- Figas, A., Tomaszewska-Sowa, M., Sawilska, A. and Keutgen, A. J. (2016). Improvement of In vitro propagation and acclimation of Helichrysum arenarium L. Moench. Acta Scientiarum Polonorum Hortorum Cultus, 15(4): 17-26.
- Gamborg, O. L., Murashige, T., Thorpe, T. A. and Vasil, I. K. (1976). Plant tissue culture media. In vitro, 12(7): 473-478.
- Giovannini, A., Amoretti, M., Savona, M., di Guardo, A. and Ruffoni, B. (2003). Tissue culture in Helichrysum spp. Acta Horticulturae, 616: 339–342.
- Gupta, N., Jain, V., Joseph, M. R. and Devi, S. (2020). A review on micropropagation culture method. Asian Journal of Pharmaceutical Research and Development, 8(1): 86-93.
- Hirakawa, T. and Tanno, S. (2022). In vitro Propagation of Humulus lupulus through the induction of axillary bud Development. Plants, 11(8): 1066.
- Khan, S., Shende, S. M. and Bonde, D. R. (2021). In vitro micropropagation of mint (Mentha). World Journal of Pharmacy and Pharmaceutical Sciences, 10: 1688-1692.
- Morone Fortunato, I., Montemurro, C., Ruta, C., Perrinia, R., Sabetta, W., Blanco, A., Lorusso, E., ve Avato, P. (2010). Essential oils, genetic relationships and In vitro establishment of Helichrysum italicum (Roth) G. Don ssp. italicum from wild Mediterranean germplasm. Industrial Crops and Products, 32(3): 639–649.
- Murashige, G. and Skoog, F. (1962). A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiologia Plantarum,15: 473-497.
- Ninčević, T., Grdiša, M., Šatović, Z. and Jug-Dujaković, M. (2019). Helichrysum italicum (Roth) G. Don: Taxonomy, Biological Activity, Biochemical and Genetic Diversity. Industrial Crops and Products, 138, 111487.
- Perrinia, R., Ruta, C. and Morone Fortunato, I. (2009). Regeneration through organogenesis from leaves of Helichrysum italicum (Roth) G. Don. Acta Horticulturae, 812: 217–222.
- Petrova, M., Nikolova, M., Dimitrova, M. and Dimitrova, L. (2021). Assessment of the effect of plant growth regulators on In vitro micro propagation and metabolic profiles of Melissa officinalis (lemon balm). Journal of Microbiology, Biotechnology and Food Sciences, 11(3): 1–5.
- Ramírez-Mosqueda, M. A., Iglesias-Andreu, L. G. and Bautista-Aguilar, J. R. (2017). The effect of light quality on growth and development of in vitro plantlet of Stevia rebaudiana Bertoni. Sugar Tech, 19: 331-336.
- Silva, M. M., de Oliveira, A. L., Oliveira-Filho, R. A., Gouveia-Neto, A. S., Camara, T. J. and Willadino, L. G. (2014). Effect of blue/red LED Light Combination on Growth and Morphogenesis of Saccharum officinarum Plantlets in vitro. In Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XII, 894, 373-380.
- Snedecor, G. W. and Cochran, W. G. (1967). Statistical Methods, 593 pp. Iowa State Univ., Ames. Staver, M. M., Gobin, I., Ratkaj, I., Petrovic, M., Vulinovic, A., Dinarina-Sablic, M. and Broznic, D. (2018). In vitro antiproliferative and antimicrobial activity of the essential oil from the flowers and leaves of Helichrysum italicum (Roth) G. Don growing in Central Dalmatia (Croatia). Journal of Essential Oil-Bearing Plants, 21(1): 77–91.
- Tastekin, D. (2020). Comparison of the phytochemical content of St. John's wort (Hypericum perforatum L.) grown in externel and In vitro conditions. (Master thesis). Tekirdag Namik Kemal University, The Institute of Natural Sciences, Tekirdag, Turkey.
- Umarusman, M. A., Aysan, Y. and Özgüven, M. (2019). Investigation of The Antibacterial Effects of Different Plant Extracts Against Pea Bacterial Leaf Blight Disease Caused by Pseudomonas syringae pv. pisi. Journal of Tekirdag Agricultural Faculty, 16(3): 297-314 (In Turkish).
- Umaz, A. and Umaz, K. (2020). Determination and Comparison of Volatile Compounds of Gold Grass (Helichrysum arenarium) from Two Different Locations. Gümüşhane University Journal of Science, 10(3): 592-600 (In Turkish).
- Yesmin, S. (2019). In Vitro Micropropagation of Stevia rebaudiana Bertoni. Plant Tissue Culture and Biotechnology, 29(2): 277–284.
Abstract
Güney Marmara ve Ege bölgesinde yayılış gösteren ölmez otu bitkisi (Helichrysum italicum (Roth) G. Don) kurak ve yarı kurak bölgelerde yetiştirilebilmektedir. Ayrıca zengin uçucu yağ ve sekonder metabolit içeriğinden dolayı geleneksel tedavi yöntemleri dahil modern tıp ve kozmetik alanında önemli bir yere sahiptir. Bitkilerin in vitro koşullarda sürgün rejenerasyonu yoluyla çoğaltılması birçok bitki türünde elde edilmiş olmasına rağmen, ölmez otunda doku kültürü üzerine yapılan çalışmalar oldukça sınırlıdır. Bu çalışmanın amacı, ölmez otu bitkisinde doku kültürü çalışmasının optimizasyonunun yapılması ve bir sonraki in vitro, moleküler ve sekonder metabolit çalışmalarına zemin sağlamaktır. Bunun yanı sıra bölgede kültür tarımı için sağlıklı ve hastalıksız fide üretimi yönteminin optimize edilerek bitkiyi tanıtmaktır. Eksplantların sterilizasyon aşamasında üç farklı (%15, %25 ve %35) NaOCl konsantrasyonu 10 ve 20 dakika süre ile denenmiştir. En başarılı sonuç 10 dakika %35 NaOCl içeren ortamdan elde edilmiştir. Steril edilen eksplantlar sürgün rejenerasyonu için BAP, GA ve NAA bitki büyüme düzenleyicileri içeren MS ve Gamborg B5 besin ortamlarına aktarılmıştır. Eksplantlarda en iyi rejenerasyon 0.5 mg L-1 BAP, 1 mg L-1 GA ve 0.2 mg L-1 NAA içeren MS ortamında elde edilmiştir. Gamborg B5 içeren denemelerde herhangi gelişim gözlenememiş olup eksplantlarda vitrifikasyon ve kararma oluşmuştur. Dört hafta sonunda 3 cm uzunluğa ulaşan sürgünler köklendirme ortamı olarak 0 MS, 0.5 mg L-1 IBA, 1 mg IBA, 1.5 mg L-1 ve 2 mg L-1 IBA içeren MS ve ½MS ortamına alınmıştır. Hazırlanan tüm ortamlarda dört hafta içerisinde %100 köklenme görülmüştür. Mikroçoğaltım çalışmaları sonucu üç ay içerisinde köklenmiş bitkiler aklimatizasyon aşamasına alındıktan sonra seradaki saksılara ve bir ay sonra tarlaya aktarılmıştır.
Thanks
Doç. Dr. Sefer DEMİRBAŞ ve bu çalışmanın yapıldığı Tekirdağ Namık Kemal Üniversitesi Ziraat Fakültesi Tarımsal Biyoteknoloji Araştırma ve Üretim Birimi'ne (Ziraatbiyotek) teşekkür ederiz.
References
- Albayrak, S., Aksoy, A., Sagdic, O. and Hamzaoglu, E. (2010). Compositions, antioxidant and antimicrobial activities of Helichrysum (Asteraceae) species collected from Turkey. Food Chemistry, 119(1): 114-122.
- Andrade, L. B., Echeverrigaray, S., Fracaro, F., Pauletti, G. F. and Rota, L. (1999). The effect of growth regulators on shoot propagation and rooting of common lavender (Lavandula vera DC). Plant Cell, Tissue and Organ Culture 56: 79-83.
- Clasquin, S. and Henry, M. (2002). Micropropagation Helichrysum arenarium (L.) Moench. Acta Botanica Gallica. 149(2). 189–195.
- Daneshvar Royandazagh, S. and Pehlivan, E. C. (2016). In vitro micropropagation of Lilium candidum L. and alkaloids. Journal of Tekirdag Agricultural Faculty, 13(3): 100-110 (In Turkish).
- Dimitrova, N. G. and Nacheva, L. (2018). Micropropagation of Helichrysum italicum (ROTH) G. Don–a medicinal plant with ornamental value. Journal of BioScience and Biotechnology, 7(2-3): 97-101.
- El-Banna, H. Y. (2017). Micropropagation of thyme plant (Thymus vulgaris). Journal of Plant Production, 8(11): 1221-1227.
- Figas, A., Tomaszewska-Sowa, M., Sawilska, A. and Keutgen, A. J. (2016). Improvement of In vitro propagation and acclimation of Helichrysum arenarium L. Moench. Acta Scientiarum Polonorum Hortorum Cultus, 15(4): 17-26.
- Gamborg, O. L., Murashige, T., Thorpe, T. A. and Vasil, I. K. (1976). Plant tissue culture media. In vitro, 12(7): 473-478.
- Giovannini, A., Amoretti, M., Savona, M., di Guardo, A. and Ruffoni, B. (2003). Tissue culture in Helichrysum spp. Acta Horticulturae, 616: 339–342.
- Gupta, N., Jain, V., Joseph, M. R. and Devi, S. (2020). A review on micropropagation culture method. Asian Journal of Pharmaceutical Research and Development, 8(1): 86-93.
- Hirakawa, T. and Tanno, S. (2022). In vitro Propagation of Humulus lupulus through the induction of axillary bud Development. Plants, 11(8): 1066.
- Khan, S., Shende, S. M. and Bonde, D. R. (2021). In vitro micropropagation of mint (Mentha). World Journal of Pharmacy and Pharmaceutical Sciences, 10: 1688-1692.
- Morone Fortunato, I., Montemurro, C., Ruta, C., Perrinia, R., Sabetta, W., Blanco, A., Lorusso, E., ve Avato, P. (2010). Essential oils, genetic relationships and In vitro establishment of Helichrysum italicum (Roth) G. Don ssp. italicum from wild Mediterranean germplasm. Industrial Crops and Products, 32(3): 639–649.
- Murashige, G. and Skoog, F. (1962). A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiologia Plantarum,15: 473-497.
- Ninčević, T., Grdiša, M., Šatović, Z. and Jug-Dujaković, M. (2019). Helichrysum italicum (Roth) G. Don: Taxonomy, Biological Activity, Biochemical and Genetic Diversity. Industrial Crops and Products, 138, 111487.
- Perrinia, R., Ruta, C. and Morone Fortunato, I. (2009). Regeneration through organogenesis from leaves of Helichrysum italicum (Roth) G. Don. Acta Horticulturae, 812: 217–222.
- Petrova, M., Nikolova, M., Dimitrova, M. and Dimitrova, L. (2021). Assessment of the effect of plant growth regulators on In vitro micro propagation and metabolic profiles of Melissa officinalis (lemon balm). Journal of Microbiology, Biotechnology and Food Sciences, 11(3): 1–5.
- Ramírez-Mosqueda, M. A., Iglesias-Andreu, L. G. and Bautista-Aguilar, J. R. (2017). The effect of light quality on growth and development of in vitro plantlet of Stevia rebaudiana Bertoni. Sugar Tech, 19: 331-336.
- Silva, M. M., de Oliveira, A. L., Oliveira-Filho, R. A., Gouveia-Neto, A. S., Camara, T. J. and Willadino, L. G. (2014). Effect of blue/red LED Light Combination on Growth and Morphogenesis of Saccharum officinarum Plantlets in vitro. In Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XII, 894, 373-380.
- Snedecor, G. W. and Cochran, W. G. (1967). Statistical Methods, 593 pp. Iowa State Univ., Ames. Staver, M. M., Gobin, I., Ratkaj, I., Petrovic, M., Vulinovic, A., Dinarina-Sablic, M. and Broznic, D. (2018). In vitro antiproliferative and antimicrobial activity of the essential oil from the flowers and leaves of Helichrysum italicum (Roth) G. Don growing in Central Dalmatia (Croatia). Journal of Essential Oil-Bearing Plants, 21(1): 77–91.
- Tastekin, D. (2020). Comparison of the phytochemical content of St. John's wort (Hypericum perforatum L.) grown in externel and In vitro conditions. (Master thesis). Tekirdag Namik Kemal University, The Institute of Natural Sciences, Tekirdag, Turkey.
- Umarusman, M. A., Aysan, Y. and Özgüven, M. (2019). Investigation of The Antibacterial Effects of Different Plant Extracts Against Pea Bacterial Leaf Blight Disease Caused by Pseudomonas syringae pv. pisi. Journal of Tekirdag Agricultural Faculty, 16(3): 297-314 (In Turkish).
- Umaz, A. and Umaz, K. (2020). Determination and Comparison of Volatile Compounds of Gold Grass (Helichrysum arenarium) from Two Different Locations. Gümüşhane University Journal of Science, 10(3): 592-600 (In Turkish).
- Yesmin, S. (2019). In Vitro Micropropagation of Stevia rebaudiana Bertoni. Plant Tissue Culture and Biotechnology, 29(2): 277–284.
Details
| Primary Language | English |
|---|---|
| Subjects | Agricultural Biotechnology (Other) |
| Journal Section | Articles |
| Authors | |
| Early Pub Date | January 24, 2024 |
| Publication Date | January 30, 2024 |
| Submission Date | April 27, 2023 |
| Acceptance Date | September 13, 2023 |
| Published in Issue | Year 2024 Volume: 21 Issue: 1 |
