Direct shoot regeneration from leaf explants of Digitalis lamarckii, an endemic medicinal species

Yücesan, Buhara; VERMA, Sandeep Kumar; Sahin, Gunce; Gürel, Ekrem; Gürel, Songül

Direct shoot regeneration from leaf explants of Digitalis lamarckii, an endemic medicinal species

Sandeep Kumar VERMA, (Abant İzzet Baysal Üniversitesi, Fen Edebiyat Fakültesi, Biyoloji Bölümü, Bolu, Türkiye)

Bahtiyar Buhara YÜCESAN, (Abant İzzet Baysal Üniversitesi, Fen Edebiyat Fakültesi, Biyoloji Bölümü, Bolu, Türkiye)

Günce ŞAHİN, (Abant İzzet Baysal Üniversitesi, Fen Edebiyat Fakültesi, Biyoloji Bölümü, Bolu, Türkiye)

Songül GÜREL, (Şeker Enstitüsü, Bitki Islahı Anabilim Dalı, 06790 Etimesgut, Ankara, Türkiye)

Ekrem GÜREL (Abant İzzet Baysal Üniversitesi, Fen Edebiyat Fakültesi, Biyoloji Bölümü, Bolu, Türkiye)

Turkish Journal of Botany

1 0

Yıl: 2011 Cilt: 35 Sayı: 6 Sayfa Aralığı: 689 - 695 Metin Dili: Türkçe İndeks Tarihi: 29-07-2022

Direct shoot regeneration from leaf explants of Digitalis lamarckii, an endemic medicinal species

Öz:

Digitalis lamarckii (bodur yüksükotu) için direkt sürgün organogenezisi yolu ile etkili bir in vitro rejenerasyon protokolu ilk defa bu çalışma ile rapor edilmektedir. İki farklı deney seti kurulmuştur; ilk sette, in vitro koşullarda çimlendirilmiş fi delerden alınan yaprak eksplantları kullanılarak, 6-benzilaminopürin (BAP), kinetin, thidiazuron (TDZ) ve zeatinin farklı konsantrasyonları karşılaştırılmış, ikinci sette ise sürgün çoğaltımı amacıyla, birinci sette kültüre alınan ve çok sayıda sürgün meydana getirmiş olan eksplantlar kullanılarak, indol-3-butirik asidin (IBA), BAP, kinetin, TDZ ve zeatin ile kombinasyonları test edilmiştir. Sürgün rejenerasyonu (birinci deney seti) sonuçlarına bakıldığında, TDZ’nin, BAP’den istatistiki olarak çok daha etkili olduğu bulunmuş, 1,0 mg L-1 dozunda kullanıldığında ise eksplant başına ortalama 10,3 sürgün üreterek, en başarılı sonucu vermiştir. Sürgün çoğaltımı (ikinci deney seti) sonuçlarına bakıldığında ise, 0,2 mg L-1 IBA’nın 0,2 mg L-1 TDZ ile kombinasyonu, eksplant başına ortalama 16,5 sürgün üreterek, BAP (11,0 sürgün), zeatin (5,5 sürgün) veya kinetin (4,0 sürgün) ile kombinasyonuna göre istatistiki olarak çok daha başarılı sonuçlar vermiştir. Rejenere olan sürgünler, 0,5 mg L-1 indol-3-asetik asit (IAA) içeren ortamda kolaylıkla köklendirilmiştir. Köklendirilmiş rejenerantlar, daha sonra saksılara aktarılmış ve normal bir gelişme seyri göstererek olgunlaşma sürecini tamamlamışlardır. Bu çalışmada tanımlanan protokol ile, bitki rejenerasyonunun direkt sürgün organogenezisi yoluyla başarıldığı ortaya konulmuş, ve bunun, tıbbi önemi olan bu endemik türde büyük ölçekli kardenolit üretimi, germplasm korunması ve genetik transformasyon çalışmalarına katkı sağlaması beklenmektedir.

Anahtar Kelime: Digitalis lamarckii filiz tıbbi bitkiler bitki özütü gençleştirme yapraklar endemik bitkiler bodur çeşitler

Konular: Biyoloji

Endemik tıbbi bir tür olan Digitalis lamarckii’nin yaprak eksplantlarından direkt sürgün rejenerasyonu

Öz:

This study reports, for the fi rst time, an effi cient in vitro plant regeneration protocol for Digitalis lamarckii Ivan. (dwarf foxglove) via direct shoot organogenesis. Two sets of experiments were carried out; the fi rst compared diff erent concentrations of 6-benzylaminopurine (BAP), kinetin, thidiazuron (TDZ), and zeatin alone using leaf explants excised from in vitro germinated seedlings, while the second set tested the combinations of indole-3-butyric acid (IBA) with BAP, kinetin, TDZ, and zeatin for shoot multiplication from the leaf explants, which were already cultured and developed numerous shoots during the fi rst set of experiments. For shoot regeneration (the fi rst set of experiments), TDZ was the most eff ective at 1.0 mg L-1 concentration, producing a mean of 10.3 shoots per explant and was signifi cantly more eff ective than BAP. For shoot multiplication (the second set of experiments), a combination of 0.2 mg L-1 IBA with 0.2 mg L-1 TDZ produced signifi cantly more shoots per explant (16.5 shoots) than with BAP (11.0 shoots), zeatin (5.5 shoots), or kinetin (4.0 shoots). Th e regenerated shoots were readily rooted on medium containing 0.5 mg L-1 indole- 3-acetic acid (IAA). Rooted regenerants were then transferred to the pots, where they grew well and attained maturity. Th e described protocol provides a simple way to regenerate plants through direct shoot organogenesis, which would be useful for a large-scale production of cardenolides, germplasm conservation, and genetic transformation studies in this medicinally important endemic species.

Anahtar Kelime: regeneration leaves endemic plants dwarf cultivars Digitalis lamarckii shoots medicinal plants plant extracts

Konular: Biyoloji

Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık

Ahmadabadi M & Bock R (2010). Development of a highly responsive leaf-based regeneration system for Peperomia species. Turk J Bot 33: 329-334.
Arrillaga I, Brisa MC & Segura J (1986). Somatic embryogenesis and plant regeneration from hypocotyl cultures of Digitalis obscura L. J Plant Physiol 124: 425-430.
Bacchetta L, Remotti PC, Bernardini C & Saccardo F (2003). Adventitious shoot regeneration from leaf explants and stem nodes of Lilium. Plant Cell Tiss Org 74: 37-44.
Baytop T (1999). Türkiye’de Bitkilerle Tedavi (2nd ed, in Turkish). İstanbul: Nobel Tıp Kitabevi.
Bogaert I, Van Cauter S, Werbrouck SPO & Dolezal K (2006). New aromatic cytokinins can make the diff erence. Acta Hortic 725: 265-270.
Brisa MC & Segura J (1987). Isolation, culture and plant regeneration from mesophyll protoplasts of Digitalis obscura. Physiol Plantarum 69: 680-686.
Blakesey D (1991). Uptake and metabolism of 6-benzyladenine in shoot proliferation of Musa and Rhododendron. Plant Cell Tiss Org 25: 69-74.
Cacho M, Moran M, Herrera MT, Tarrago JF & Corchete MP (1991). Morphogenesis in leaf, hypocotyl and root explants of Digitalis thapsi L. cultured in vitro. Plant Cell Tiss Org 25: 117-123.
Chen J, Chang C & Chang WC (1999). Direct somatic embryogenesis from leaf explants of Oncidium ‘Gower Ramsey’ and subsequent plant regeneration. Plant Cell Rep 19: 143-149.
Chen JT & Chang WC (2001). Eff ect of auxin and cytokinins on direct somatic embryogenesis from leaf explants of Oncidium ‘Gower Ramsey’. Plant Growth Regul 34: 229-232.
Chen JT & Chang WC (2006). Direct somatic embryogenesis and plant regeneration from leaf explants of Phalaenopsis amabilis. Biol Plantarum 50: 169-173.
Chang C & Chang WC (1998). Plant regeneration from callus culture of Cymbidium ensifolium var. misericors. Plant Cell Rep 17: 251-255.
Chen Y & Piluek C (1995). Eff ects of thiaiazuron and N6– benzylaminopurine on shoot regeneration of Phalaenopsis. Plant Growth Regul 16: 99-101.
Davis PH (1978). Digitalis L. In: Davis PH (ed.), Flora of Turkey and the East Aegean Islands. Vol. 11. Edinburgh: Edinburgh Univ Press.
Ernst R (1994). Eff ect of thidiazuron on in vitro propagation of Phalaenopsis and Doritaenopsis (Orchidaceae). Plant Cell Tiss Org 39: 273-275.
Fatima Z, Mujib A, Fatima S, Arshi A & Umar S (2009). Callus induction, biomass growth, and plant regeneration in Digitalis lanata Ehrh.: infl uence of plant growth regulators and carbohydrates. Turk J Bot 33: 393-405.
Gurel E, Yucesan B, Aglic E, Gurel S, Verma SK, Sokmen M & Sokmen A (2011). Regeneration and cardiotonic glycoside production in Digitalis davisiana Heywood (Alanya foxglove). Plant Cell Tiss Org 104: 217-225.
Herrera MT, Cacho M, Corchete MP & Fernandez-Tarrago J (1990). One step shoot tip multiplication and rooting of Digitalis thapsi L. Plant Cell Tiss Org 22: 179-182.
Huetteman CA & Preece JE (1993). Th idiazuron: a potent cytokinin for plant tissue culture. Plant Cell Tiss Org 33: 105-119.
IUCN (2001). IUCN Red List Categories and Criteria: Version 3.1. IUCN Species Survival Commission. IUCN, Gland, Switzerland and Cambridge, UK.
Kaminek M (1992). Progress in cytokinin research. Trends Biotechnol 10: 159-164.
Khurana-Kaul V, Kanchhwaha S & Kothari SL (2010). Direct shoot regeneration from leaf explants of Jatropha curcas in response to thidiazuran and high copper contents in the medium. Biol Plantarum 54: 369-372.
Lin HS, De Jeu MJ & Jacobsen E (1997). Direct shoot regeneration from excised leaf explants of in vitro grown seedlings of Alstroemeria L. Plant Cell Rep 16: 770-774.
Lincy A & Sasikumar B (2010). Enhanced adventitious shoot regeneration from aerial stem explants of ginger using TDZ and its histological studies. Turk J Bot 34: 21-29.
Ling Fei X, Feng Wang M & Dong L (2009). Plant regeneration from in vitro cultured leaves of Lanzhou lily (Lilium davidii var. unicolor). Sci Hortic 119: 458-461.
Lopez-Lazaro M (2007). Digitoxin as an anticancer agent with selectivity for cancer cells: Possible mechanisms involved. Expert Opin Th er Tar 11: 1043-1053.
Malabadi RB, Mulgund GS & Nataraja K (2004). Effi cient regeneration of Vanda coarulea, an endangered orchid using thidiazuron. Plant Cell Tiss Org 76: 289-293.
Mirici S, Parmaksiz I, Ozcan S, Sancak C, Uranbey S, Sarihan EO, Gumuscu A, Gurbuz B & Arslan N (2005). Effi cient in vitro bulblet regeneration from immature embryos of endangered Sternbergia fi scheriana. Plant Cell Tiss Org 80: 239-246.
Mok MC & Mok DWS (1985). Th e metabolism of [14C] thidiazuron in callus tissues of Phaseolus lunatus. Physiol Plantarum 65: 425-432.
Murashige T & Skoog FA (1962). Revised for rapid growth and bioassays with tobacco tissue cultures. Physiol Plantarum 15: 473-497.
Newman RA, Yang P, Pawlus AD & Block KI (2008). Cardiac glycosides as novel cancer therapeutic agents. Mol Interv 8: 36- 49.
Perez-Bermudez P, Cornejo MJ & Seguara J (1983). In vitro propagation of Digitalis obscura L. Plant Sci Lett 30: 77-82.
Perez-Bermudez P, Falco JM & Segura J (1987). Morphogenesis in root tip meristem cultures of Digitalis obscura L. J Plant Physiol 130: 87-91.
Perez-Bermudez P, Cornejo MJ & Segura J (1990). Digitalis spp.: In vitro production of haploids. In: Bajaj YPS (ed.) Biotechnology in Agriculture and Forestry, Vol. 12, Haploids in Crop Improvement I, pp. 277-288. Berlin: Springer-Verlag.
Pradel H, Dumke-Lehmann U, Diettrich B & Luckner M (1997). Hairy root cultures of Digitalis lanata. Secondary metabolism and plant regeneration. J Plant Physiol 151: 209-215.
Rout GR, Saxena C, Samantary S & Das P (1999). Rapid plant regeneration from callus cultures of Plumbago zeylanica. Plant Cell Tiss Org 56: 47-51.
Sahai A, Shahzad A & Anis M (2010). High frequency plant production via shoot organogenesis and somatic embryogenesis from callus in Tylophora indica, an endangered plant species. Turk J Bot 34: 11-20.
Sanikhani M, Frello S & Serek M (2006). TDZ induces shoot regeneration in various Kalanchoe blossfeldiana Poelln. cultivars in the absence of auxin. Plant Cell Tiss Org 85: 75-82.
Saxena PK, Malik KA & Gill LG (1992). Induction by thidiazuron of somatic embryogenesis in intact seedling of peanut. Planta 187: 421-424.
Turker A, Yucesan B & Gurel E (2009a). In vitro regeneration of Achillea millefolium L. from shoot-tips and root segments of seedlings. J Plant Biochem Biot 18: 61-69.
Turker A, Yucesan B & Gurel E (2009b). Plant regeneration via adventitious shoot development from cultured leaf explants of purple loosestrife (Lythrum salicaria L.): A medicinal wetland plant. Biol Plantarum 54: 750-754.
Tran Th anh Van K & Trinh TH (1990). Organogenic diff erentiation. In: Bhojwani SS (ed.) Plant Tissue Culture: Application and Limitations, pp. 34-53, Amsterdam: Elsevier.
Yeh JY, Huang WJ, Kan SF & Wang PS (2001). Inhibitory eff ects of Digitalis on the proliferation of androgen dependent and independent prostate cancer cells. J Urol 166: 1937-1942.
Yucesan B, Turker A & Gurel E (2007). TDZ-induced high frequency plant regeneration through multiple shoot formation in witloof chicory (Cichorium intybus L.). Plant Cell Tiss Org 91: 243-250.

APA	VERMA S, Yücesan B, Sahin G, Gürel S, Gürel E (2011). Direct shoot regeneration from leaf explants of Digitalis lamarckii, an endemic medicinal species. , 689 - 695.
Chicago	VERMA Sandeep Kumar,Yücesan Buhara,Sahin Gunce,Gürel Songül,Gürel Ekrem Direct shoot regeneration from leaf explants of Digitalis lamarckii, an endemic medicinal species. (2011): 689 - 695.
MLA	VERMA Sandeep Kumar,Yücesan Buhara,Sahin Gunce,Gürel Songül,Gürel Ekrem Direct shoot regeneration from leaf explants of Digitalis lamarckii, an endemic medicinal species. , 2011, ss.689 - 695.
AMA	VERMA S,Yücesan B,Sahin G,Gürel S,Gürel E Direct shoot regeneration from leaf explants of Digitalis lamarckii, an endemic medicinal species. . 2011; 689 - 695.
Vancouver	VERMA S,Yücesan B,Sahin G,Gürel S,Gürel E Direct shoot regeneration from leaf explants of Digitalis lamarckii, an endemic medicinal species. . 2011; 689 - 695.
IEEE	VERMA S,Yücesan B,Sahin G,Gürel S,Gürel E "Direct shoot regeneration from leaf explants of Digitalis lamarckii, an endemic medicinal species." , ss.689 - 695, 2011.
ISNAD	VERMA, Sandeep Kumar vd. "Direct shoot regeneration from leaf explants of Digitalis lamarckii, an endemic medicinal species". (2011), 689-695.

APA	VERMA S, Yücesan B, Sahin G, Gürel S, Gürel E (2011). Direct shoot regeneration from leaf explants of Digitalis lamarckii, an endemic medicinal species. Turkish Journal of Botany, 35(6), 689 - 695.
Chicago	VERMA Sandeep Kumar,Yücesan Buhara,Sahin Gunce,Gürel Songül,Gürel Ekrem Direct shoot regeneration from leaf explants of Digitalis lamarckii, an endemic medicinal species. Turkish Journal of Botany 35, no.6 (2011): 689 - 695.
MLA	VERMA Sandeep Kumar,Yücesan Buhara,Sahin Gunce,Gürel Songül,Gürel Ekrem Direct shoot regeneration from leaf explants of Digitalis lamarckii, an endemic medicinal species. Turkish Journal of Botany, vol.35, no.6, 2011, ss.689 - 695.
AMA	VERMA S,Yücesan B,Sahin G,Gürel S,Gürel E Direct shoot regeneration from leaf explants of Digitalis lamarckii, an endemic medicinal species. Turkish Journal of Botany. 2011; 35(6): 689 - 695.
Vancouver	VERMA S,Yücesan B,Sahin G,Gürel S,Gürel E Direct shoot regeneration from leaf explants of Digitalis lamarckii, an endemic medicinal species. Turkish Journal of Botany. 2011; 35(6): 689 - 695.
IEEE	VERMA S,Yücesan B,Sahin G,Gürel S,Gürel E "Direct shoot regeneration from leaf explants of Digitalis lamarckii, an endemic medicinal species." Turkish Journal of Botany, 35, ss.689 - 695, 2011.
ISNAD	VERMA, Sandeep Kumar vd. "Direct shoot regeneration from leaf explants of Digitalis lamarckii, an endemic medicinal species". Turkish Journal of Botany 35/6 (2011), 689-695.