In vitro production of Phalaenopsis orchids

Yıl 2024, Cilt: 7 Sayı: 1, 41 – 53, 30.06.2024

Mir Abdullatif Yahya Dilek Killi Haworth Emre Özden Fatma Tunalı Atalay Sökmen

https://doi.org/10.38059/biodiversity.1452374

Öz

Phalaenopsis – known as moth orchids – are the most popular orchids cultivated indoors as decorative house plants. This makes propagation and cultivation of Phalaenopsis important for commercial growers. Enhancements to the micropropagation of Phalaenopsis would have pronounced economic benefits through reduced losses and wastage. We examined the effects of several nutrient media and specific plant growth regulators (PGRs) belonging to the gibberellic acid and cytokinin groups on the in vitro germination of Phalaenopsis seeds, utilizing a single group pretest-posttest model. The effects of several nutrient media such as: Knudson C (KCM), Lindemann (LM), Orchimax (-OM), Orchimax + activated charcoal (+OM), Murashige & Skoog (MS), as well as various PGRs such as 6-Benzylaminopurine (6BA), 6-Furfurylaminopurine (KIN), Adenin hemisulfate (AHS), Thidiazuron (TDZ), 2-Isopentenyl adenine (2iP), and Gibberellic acid (GA3), on the process of germination were also investigated. The explants obtained from the germinating seedlings were subjected to direct organogenesis, and the optimal PGR and tissue fragments were determined. The +OM medium facilitated the shortest germination period (in days). An inverse relationship between the concentration of TDZ and the percentage of germination in the context of the employed PGRs was observed. Apart from TDZ, the remaining PGRs exhibited a positive correlation with concentration. However, no significant difference in germination was observed in comparison to the control. The findings of direct organogenesis investigations revealed that the medium that exhibited the highest productivity was enriched with 5.0 ppm of 6BA. The media containing TDZ exhibited a reduced level of efficiency. Particularly, the group treated with 1.0 ppm of TDZ exhibited reduced efficacy compared to the control group. All concentrations of cytokinin in root elongation stage exhibited a favorable impact in comparison to the control. The variance between these PGRs was not statistically significant.

Anahtar Kelimeler

Cytokinin, direct organogenesis, gibberellic acid (GA3), nutrient media, Plant growth regulators (PGRs)

Kaynakça

• Akça Ş (2021). İç mekân süs bitkileri tüketici tercihlerinin belirlenmesi; Zonguldak kenti örneği. Türk Tarım ve Doğa Bilimleri Dergisi 427-435.
• Arditti J (1967). Factors affecting orchid seeds, Botanical Review 33(1): 1-97.
• Arditti J (1980). Aspects of the Physiology of Orchids. Advances in Botanical Research 7:421-655.
• Arditti J, Ghani A (2000). Numerical and physical properties of orchid seeds and their biological implications. Tansley Review, New Phytologist 145(3): 367-421.
• Aytar E, Kömpe Y (2021). Orkidelerin Geleneksel Kullanımları, Fitokimyasal İçerikleri ve Biyolojik Aktiviteleri. Black Sea Journal of Engineering and Science 4 (3):141-152.
• Bektaş E (2014). Orchis sancta l. ve Serapias vomeracea (BURM. F.) briq. türlerinin bitki doku kültürü yöntemiyle üretimi. Doktora, Karadeniz Teknik Üniversitesi, Trabzon, Türkiye.
• Bektaş E, Cüce M, Sökmen A (2013). İn vitro germination, protocorm formation and plantlet development of Orchis coriophora (Orchidaceae), a naturally growing orchid species in Turkey. Turkish Journal of Botany 37:336-342.
• Bhojwani S S, & Razdan M K (1986). Plant tissue culture: theory and practice. Elsevier.
• Chang C, Chang W (1998). Plant regeneration from callus culture of Cymbidium ensifolium var. misericors. Plant Cell Reports 17: 251-255.
• Chen JT, Chang WC, (2000). Efficient plant regeneration through somatic embryogenesis from callus cultures of Oncidium (Orchidaceae). Plant Science 160(1): 87-93. doi: https://doi.org/10.1016/S0168-9452(00)00367-8.
• Chen TY, Chen JT, Chang WC (2004). Plant regeneration through direct shoot bud formation from leaf cultures of Paphiopedilum orchids. Plant Cell, Tissue and Organ Culture 76(1): 11-15.
• Chen Y, Piluek C (2004). Effects of thidiazuron and N6-benzylaminopurine on shoot regeneration of Phalaenopsis. Plant Growth Regulation 16: 99-101.
• Crafts C, Miller C (1974). Detection and identification of cytokinins produced by mycorrhizal fungi. Plant Physiology 54(4): 586-588.
• Díaz M, Álvarez C (2009). Plant regeneration through direct shoot formation from leaf cultures and from protocorm-like bodies derived from callus of Encyclia mariae (Orchidaceae), a threatened Mexican orchid. İn vitro Cellular and Developmental Biology-Plant 45(2): 162-170.
• Ernst R (1994). Effect of thidiazuron on in vitro propagation of Phalaenopsis and Doritaenopsis (Orchidaceae). Plant Cell Tissue and Organ Culture 39: 273–275.
• Eymar E, Alegre J, Toribio M, Lo’ pez-Vela D (2000). Effect of activated charcoal and 6-benzyladenine on in vitro nitrogen uptake by Lagerstroemia indica. Plant Cell Tissue and Organ Culture 63(1): 57-65.
• Ferreira W, Kerbauy G, Costa A (2006). Micropropagation and genetic stability of a Dendrobium hybrid (Orchidaceae). İn vitro Cellular & Developmental Biology-Plant 42(6): 568-571.
• Gaspar T, Kevers C, Penei C, Greppin H, Reid D, Thorpe T (1996). Plant Hormones and Plant Growth Regulators in Plant Tissue Culture. İn vitro Cellular and Developmental Biology-Plant 32: 272-289.
• Haberer G, Kieber J (2002). Cytokinins: New Insights into a Classic Phytohormone. Plant Physiology 128: 354-362. Hinsley A, de Boer H, Fay M, Gale S, Gardiner L, Gunasekara R, Phelps J (2017). A review of the trade in orchids and its implications for conservation. Botanical Journal of the Linnean Society 186(4): 435-455.
• Lauzer D, St-Arnaud M, Barabe D (1994). Tetrazolium staining and in vitro germination of mature seeds of Cypripedium acaule (Orchidaceae). Lindleyana 9(3): 197-204.
• Mansuroğlu S, Gürel E (2001). Mikroçoğaltım. In: Babaoğlu, M., Gürel, E. ve Özcan, S. (eds.), Bitki Biyoteknolojisi I – Doku Kültürü ve Uygulamaları. pp: 262-281, Selçuk Üniversitesi Vakfı Yayınları, Konya.
• Mead J, Bulard C (1975). Effects of vitamins and nitrogen sources on asymbiotic germination and development of Orchis laxiflora and Ophrys sphegodes. New Phytologist 74(1):33-40.
• Nayak NR, Rath SP, Patnaik S (1997). İn vitro propagation of three epiphytic orchids, Cymbidium aloifolium (L.) Sw., Dendrobium aphyllum (Roxb.) Fisch. and Dendrobium moschatum (Buch-Ham) Sw. through thidiazuron-induced high frequency shoot proliferation. Scientia Horticulturae 71(3-4): 243-250.
• Paek K, Murthy H (1977). Temperate Oriental Cymbidium species. In: Kull, T., Arditti, J. (eds.), Orchid Biology: Reviews and Perspectives. VIII. Edition, 287, Kluwer Academic Publishers, Dordrecht.
• Park SY, Huh YS, Paek KY, (2018). Common protocols in orchid micropropagation. Orchid Propagation: From Laboratories to Greenhouses-Methods and Protocols pp: 179-193. DOI: 10.1007/978-1-4939-7771-0_8.
• Raghavan V, Torrey JG (1964). Inorganic nitrogen nutrition of the seedlings of the orchid, Cattleya. American Journal of Botany 51(3): 264-274.
• Silva A, Fernandes MM, Fereira AG (2000). Ontogenetic events in androgenesis of Brazilian barley genotypes. Revista Brasileira de Biologia 60(2): 315- 319.
• Stewart L, Kane M (2006). Asymbiotic seed germination and in vitro seedling development of Habenaria macroceratitis (Orchidaceae), a rare Florida terrestrial orchid. Plant Cell Tissue and Organ Culture 86(2): 147-158. DOI: 10.1007/s11240-006-9098-y.
• Valletta A, Attorre F, Bruno F, Pasqua G (2008). İn vitro asymbiotic germination of Orchis mascula L. Plant Biosystems – An International Journal Dealing with all Aspects of Plant Biology 142(3): 653-655. DOI: 10.1080/11263500802411205.
• Van Waes J, Debergh P (1986). İn vitro germination of some Western European Orchids. Physiologia Plantarum 67(2): 253-261.
• Waes J (1987). Effect of Activated Charcoal on İn vitro Propagation of Western European Orchids. Symposium on İn vitro Problems Related to Mass Propagation of Horticultural Plants. Acta Horticulturae 212:131-138. Yuan S, Bolaños-Villegas P, Tsao C, Chen F (2021). The breeding of Phalaenopsis hybrids. The Orchid Genome pp: 29-40, Springer.

Phalaenopsis orkidelerinin in vitro üretimi

Yıl 2024, Cilt: 7 Sayı: 1, 41 – 53, 30.06.2024

Mir Abdullatif Yahya Dilek Killi Haworth Emre Özden Fatma Tunalı Atalay Sökmen

https://doi.org/10.38059/biodiversity.1452374

Öz

Phalaenopsis, ev dekorasyonu için iç mekanlarda yetiştirilen en popüler orkidelerdir. Bu nedenle, Phalaenopsis’in çoğaltılması ve yetiştirilmesi ticari yetiştiriciler için büyük önem taşımaktadır. Phalaenopsis’in mikro çoğaltılmasındaki iyileştirmeler, kayıpların ve israfın azaltılmasını sağlayarak ekonomik faydalar sunabilir. Bu çalışmanın amacı, bir grup ön test-son test modeli kullanarak Phalaenopsis tohumlarının in vitro çimlenmesi üzerinde çeşitli besin ortamlarının ve bazı bitki büyüme düzenleyicilerinin (PGR’ler) etkilerini incelemektir. Araştırmada kullanılan besin ortamları arasında Knudson C (KCM), Lin-demann (LM), Orchimax (-OM), Orchimax + aktif kömür (+OM) ve Murashige & Skoog (MS) yer alırken, PGR’ler olarak 6-Benzilaminopurin (6BA), 6-Furfurylaminopurin (KIN), Adenin hemisülfat (AHS), Thidiazuron (TDZ), 2-İsopentenil adenin (2iP) ve Gibberellik asit (GA3) incelenmiştir. Çimlenen fidelerden elde edilen eksplantlar, doğrudan organogenez işlemine tabi tutularak optimal PGR ve doku fragmanları belirlenmiştir. Sonuçlar, +OM ortamının en kısa çimlenme süresini (gün cinsinden) sağladığını göstermiştir. TDZ konsantrasyonu ile çimlenme yüzdesi arasında ters bir ilişki gözlenmiş, diğer PGR’ler ise konsantrasyon ile pozitif bir korelasyon göstermiştir. Ancak, kontrol ile karşılaştırıldığında çimlenme açısından önemli bir fark bulunmamıştır. Doğrudan organogenez araştırmalarının bulguları, en yüksek üretkenliğe sahip olan ortamın 5.0 ppm 6BA ile zenginleştirilmiş olduğunu ortaya koymuştur. TDZ içeren ortamlar düşük düzeyde bir verimlilik sergilemiş, özellikle 1.0 ppm TDZ grubu, kontrol grubuna kıyasla azalmış etkinlik göstermiştir. Kök uzatma aşamasındaki tüm sitokinin konsantrasyonları, kontrole kıyasla olumlu bir etki göstermiş, ancak bu PGR’ler arasındaki fark istatistiksel olarak anlamlı bulunmamıştır.

Anahtar Kelimeler

besin ortamı, Bitki büyüme düzenleyicileri (PGR, doğrudan organogenez, gibberellik asit (GA3), sitokinin

Kaynakça

• Akça Ş (2021). İç mekân süs bitkileri tüketici tercihlerinin belirlenmesi; Zonguldak kenti örneği. Türk Tarım ve Doğa Bilimleri Dergisi 427-435.
• Arditti J (1967). Factors affecting orchid seeds, Botanical Review 33(1): 1-97.
• Arditti J (1980). Aspects of the Physiology of Orchids. Advances in Botanical Research 7:421-655.
• Arditti J, Ghani A (2000). Numerical and physical properties of orchid seeds and their biological implications. Tansley Review, New Phytologist 145(3): 367-421.
• Aytar E, Kömpe Y (2021). Orkidelerin Geleneksel Kullanımları, Fitokimyasal İçerikleri ve Biyolojik Aktiviteleri. Black Sea Journal of Engineering and Science 4 (3):141-152.
• Bektaş E (2014). Orchis sancta l. ve Serapias vomeracea (BURM. F.) briq. türlerinin bitki doku kültürü yöntemiyle üretimi. Doktora, Karadeniz Teknik Üniversitesi, Trabzon, Türkiye.
• Bektaş E, Cüce M, Sökmen A (2013). İn vitro germination, protocorm formation and plantlet development of Orchis coriophora (Orchidaceae), a naturally growing orchid species in Turkey. Turkish Journal of Botany 37:336-342.
• Bhojwani S S, & Razdan M K (1986). Plant tissue culture: theory and practice. Elsevier.
• Chang C, Chang W (1998). Plant regeneration from callus culture of Cymbidium ensifolium var. misericors. Plant Cell Reports 17: 251-255.
• Chen JT, Chang WC, (2000). Efficient plant regeneration through somatic embryogenesis from callus cultures of Oncidium (Orchidaceae). Plant Science 160(1): 87-93. doi: https://doi.org/10.1016/S0168-9452(00)00367-8.
• Chen TY, Chen JT, Chang WC (2004). Plant regeneration through direct shoot bud formation from leaf cultures of Paphiopedilum orchids. Plant Cell, Tissue and Organ Culture 76(1): 11-15.
• Chen Y, Piluek C (2004). Effects of thidiazuron and N6-benzylaminopurine on shoot regeneration of Phalaenopsis. Plant Growth Regulation 16: 99-101.
• Crafts C, Miller C (1974). Detection and identification of cytokinins produced by mycorrhizal fungi. Plant Physiology 54(4): 586-588.
• Díaz M, Álvarez C (2009). Plant regeneration through direct shoot formation from leaf cultures and from protocorm-like bodies derived from callus of Encyclia mariae (Orchidaceae), a threatened Mexican orchid. İn vitro Cellular and Developmental Biology-Plant 45(2): 162-170.
• Ernst R (1994). Effect of thidiazuron on in vitro propagation of Phalaenopsis and Doritaenopsis (Orchidaceae). Plant Cell Tissue and Organ Culture 39: 273–275.
• Eymar E, Alegre J, Toribio M, Lo’ pez-Vela D (2000). Effect of activated charcoal and 6-benzyladenine on in vitro nitrogen uptake by Lagerstroemia indica. Plant Cell Tissue and Organ Culture 63(1): 57-65.
• Ferreira W, Kerbauy G, Costa A (2006). Micropropagation and genetic stability of a Dendrobium hybrid (Orchidaceae). İn vitro Cellular & Developmental Biology-Plant 42(6): 568-571.
• Gaspar T, Kevers C, Penei C, Greppin H, Reid D, Thorpe T (1996). Plant Hormones and Plant Growth Regulators in Plant Tissue Culture. İn vitro Cellular and Developmental Biology-Plant 32: 272-289.
• Haberer G, Kieber J (2002). Cytokinins: New Insights into a Classic Phytohormone. Plant Physiology 128: 354-362. Hinsley A, de Boer H, Fay M, Gale S, Gardiner L, Gunasekara R, Phelps J (2017). A review of the trade in orchids and its implications for conservation. Botanical Journal of the Linnean Society 186(4): 435-455.
• Lauzer D, St-Arnaud M, Barabe D (1994). Tetrazolium staining and in vitro germination of mature seeds of Cypripedium acaule (Orchidaceae). Lindleyana 9(3): 197-204.
• Mansuroğlu S, Gürel E (2001). Mikroçoğaltım. In: Babaoğlu, M., Gürel, E. ve Özcan, S. (eds.), Bitki Biyoteknolojisi I – Doku Kültürü ve Uygulamaları. pp: 262-281, Selçuk Üniversitesi Vakfı Yayınları, Konya.
• Mead J, Bulard C (1975). Effects of vitamins and nitrogen sources on asymbiotic germination and development of Orchis laxiflora and Ophrys sphegodes. New Phytologist 74(1):33-40.
• Nayak NR, Rath SP, Patnaik S (1997). İn vitro propagation of three epiphytic orchids, Cymbidium aloifolium (L.) Sw., Dendrobium aphyllum (Roxb.) Fisch. and Dendrobium moschatum (Buch-Ham) Sw. through thidiazuron-induced high frequency shoot proliferation. Scientia Horticulturae 71(3-4): 243-250.
• Paek K, Murthy H (1977). Temperate Oriental Cymbidium species. In: Kull, T., Arditti, J. (eds.), Orchid Biology: Reviews and Perspectives. VIII. Edition, 287, Kluwer Academic Publishers, Dordrecht.
• Park SY, Huh YS, Paek KY, (2018). Common protocols in orchid micropropagation. Orchid Propagation: From Laboratories to Greenhouses-Methods and Protocols pp: 179-193. DOI: 10.1007/978-1-4939-7771-0_8.
• Raghavan V, Torrey JG (1964). Inorganic nitrogen nutrition of the seedlings of the orchid, Cattleya. American Journal of Botany 51(3): 264-274.
• Silva A, Fernandes MM, Fereira AG (2000). Ontogenetic events in androgenesis of Brazilian barley genotypes. Revista Brasileira de Biologia 60(2): 315- 319.
• Stewart L, Kane M (2006). Asymbiotic seed germination and in vitro seedling development of Habenaria macroceratitis (Orchidaceae), a rare Florida terrestrial orchid. Plant Cell Tissue and Organ Culture 86(2): 147-158. DOI: 10.1007/s11240-006-9098-y.
• Valletta A, Attorre F, Bruno F, Pasqua G (2008). İn vitro asymbiotic germination of Orchis mascula L. Plant Biosystems – An International Journal Dealing with all Aspects of Plant Biology 142(3): 653-655. DOI: 10.1080/11263500802411205.
• Van Waes J, Debergh P (1986). İn vitro germination of some Western European Orchids. Physiologia Plantarum 67(2): 253-261.
• Waes J (1987). Effect of Activated Charcoal on İn vitro Propagation of Western European Orchids. Symposium on İn vitro Problems Related to Mass Propagation of Horticultural Plants. Acta Horticulturae 212:131-138. Yuan S, Bolaños-Villegas P, Tsao C, Chen F (2021). The breeding of Phalaenopsis hybrids. The Orchid Genome pp: 29-40, Springer.

Toplam 31 adet kaynakça vardır.

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