Uzun Yapraklı Üçgülün Trifolium pannonicum ssp. elongatum Hipokotil ve Yaprak Sapı Eksplantından in vitro Çoğaltılması

Bu çalışmada, kinetin ve α- naftalin asetik asitin NAA ’in farklı konsantrasyonlarının kinetin ve 0.1+ 0.1, 0.2, 0.4 NAA mg/l , uzun yapraklı üçgül Trifolium pannonicum ssp. elongatum bitkisininhipokotil ve kotiledon yaprak sapı eksplantlarından gelişen sürgün yüzdesi, sürgün sayısı ve sürgün uzunluğu üzerine olan etkileri araştırılmıştır. Araştırmada, sürgün oluşturan eksplant yüzdesinin en yüksek oranda % 91.7 hipokotil eksplantından ve Murashige and Skoog MS içinde 1 mg/l Kinetin + 0.4 mg/l NAA içeren ortamdan; en düşük sürgün yüzdesinin % 25.0 hipokotil ve kotiledon yaprak sapı eksplantlarının her ikisinde de 1 mg/l Kinetin + 0.1 NAA içeren rejenerasyon ortamında geliştiği belirlenmiştir. Eksplant başına en fazla sürgün sayısı 11.2±1.3 adet ile 1 mg/l Kinetin+0.4 mg/l NAA içeren ortamdan, en düşük sürgün sayısı 2.5±0.6 adet ile 1 mg/l kinetin + 0.1 NAA uygulamasında gözlenmiştir. En uzun sürgün gelişimi 3.5±0.5 cm ile 1 mg/l kinetin+ 0,1 mg/L NAA içeren MS ortamında, en kısa sürgün gelişimi ise 1.5±0.2 cm ile 1 mg/l kinetin + 0.4 NAA MS ortamında belirlenmiştir. Trifolium pannonicum JACQ. ssp. elongatum bitkisinden maksimum kök gelişimi % 95±2.9 ile 1 mg/l IBA uygulamasından elde edilmiştir

Anahtar Kelimeler:

Trifolium pannonicum ssp. elongatum, hipokotil, kotiledon yaprak sapı, adventif sürgün rejenerasyonu, köklenme

In vitro Propagation of Long Leaved Clover Trifolium pannonicum ssp. elongatum Using Hypocotyle and Petiole Explants

The study reports effects of various concentrations of Kinetin and α- Naftalin asetik asit NAA kinetin and 0.1+ 0.1, 0.2, 0.4 NAA mg/l on shoot regeneration percentage, number and length of shoot of Trifolium pannonicum ssp. elongatum from hypocotyl and petiole of cotyledon leaf. The maximum 91.7% shoot regeneration frequency 25% was recorded on hypocotyl explant on Murashige and Skoog MS medium containing 1 mg/l Kinetin + 0.4 mg/l NAA; whereas, minimum shoot regeneration frequency 25% was in medium containing 1 mg/l Kinetin + 0.1 NAA respectively, in both hypcotyl and petiole of cotyledon leaf explant. The highest number of 11.2±1.3 shoots per explant was recorded on MS medium containing 1 mg/l Kinetin+0.4 mg/l NAA and the lowest number of 2.5±0.6 was in medium containing 1 mg/l Kinetin+0.1 mg/l NAA respectively. The longest shoots 3.5±0.5 cm were recorded on MS medium containing 1 mg/l kinetin+ 0,1 mg/L NAA, and the smallest shoots 1.5±0.2 cm were recorded on MS medium containing 1 mg/l kinetin + 0.4 NAA. Maximum frequency of 95% roots was recorded on MS medium containing 1 mg/l IBA

Keywords:

Trifolium pannonicum ssp.elongatum, hypocotyl, petiole of cotyledon, adventitious shoot regeneration, rooting,

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Açıkgöz, E. 2001. Yem Bitkileri. Uludağ Üniversitesi Ziraat Fakültesi Tarla Bitkileri Bölümü. 3. Baskı. 584, Bursa
Barik, D.P., S. K. Naik, U. Mohapatra and P. K. Chand. 2004. Hıgh frequency plant regeneration by in vitro shoot proliferation in cotyledonary node explants pf grasspea (Lathyrus sativus L.). In Vitro Cellular and Developmental Biology – Plant40: 467–470.
Beach, K.H. and R.R. Smith. 1979. Plant regeneration from callus of red and crimson clover. Plant Science Letters 16: 231-237.
Bisby, F.A, J. Buckingham and J.B. Harborne. 1994. Phytochemical Dictionary of the Leguminosae. Chapman and Hall, London. 673.
Fratini, R. and M.L. Ruiz. 2002. Comparative study of different cytokinins in the induction of morphogenesis in lentil (Lens culinaris Medik.). In Vitro Cellular and Developmental Biology – Plant 38: 46-51.
Komalavalli, N. and M.V. Rao. 1997. In vitro micropropagation of Gymnema elegans W& A, a rare medicinal plant. Indian Journal of Experimental Biology 35: 1088–1092.
Konieczny, R. 1995. Plant regeneration in callus culture of Trifolium nigrescens Viv. Acta Biologica Cracoviensia Series Botanica 37: 47-53.
McLean, N.L. and J. Nowak. 1989. Plant regeneration from hypocotyl and petiole callus of Trifolium pratense L. Plant Cell Reports 8: 395-398.
Mokhtarzadeh, A. and M.J. Constantin. 1978. Plant regeneration from hypocotyl-and. anther-derived callus of berseem clover.Crop Science 18:567-572.
Murashige, T. and F. Skoog. 1962. A revised medium for rapid growth and bioassay with tobacco tissue cultures. Plant Physiology 15: 473-497.
Pederson, G.A., K.H. Quesenberry, G.R. Smith and Y.K. Guteva. 1999. Collection of Trifolium sp. and other forage legumes in Bulgaria.Genetic Resources and Crop Evolution 46:325–33
Sevimay, C.S., K.M. Khawar, S. Çöçü and S. Özcan. 2005. Somatic embryogenesis in white clover (Trifolium repens L.). Periodicum Biologorum 107(1): 101-105..
Singha, S., B.S., Barker. and S.K. Bhatia.1988. Tissue culture propagation of runing buffalo clover (Trifolium stoloniferum Muhl. ex A. eatton). Plant Cell Tissue and Organ Culture 15:9-11.
Snedecor G.W. and W.G. Cochran. 1967. Statistical Methods. The Lowa State University Pess, Iowa, USA.
ahin Demirbağ N., H. Kendir and K.M. Khawar. 2008. InVitro regeneration of Turkish endemic Trifolium pannonicum JACQ. Subsp. Elongatum (WILLD).Biotechnology & Biotechnological Equipment 22(4):921-924.
Tubives 2008. Türkiye Bitkileri Veri Servisi
http://www.tubitak.gov.tr/tubives. (25.9.2008).
Uranbey, S., C.S. Sevimay and S. Özcan. 2005. Development of high frequency multiple shoot formation in persian clover (Trifolium resupinatum). Plant Cell Tissue and Organ Culture 80 (2) :229-232.
Wang, H. and F.B. Holl. 1988. In vitro culture and incidence of. somaclonal variation in regenerated plants of Trifolium pratense L.Plant Science 55: 159–167.
Zhang, D.Y., J. Li, X. Li, Q. Li, D. Y. Zhang, J.M. Li, W. Liıx. and Q. S. Li. 1999. Induction of cali and establishment of embryogenic cell suspension culture derived from red clover and white clover. Grassland of China 1: 15-18.
Zohary, M. 1972. Flora Palastina. 2: Trifolium, p. 157-193. Plates 231-276. The Israel Academy of Sciences and Humanities, Jerusalem.
Zohary, M. and D. Heller. 1984. The Genus Trifolium. The Israel Academy of Sciences and Humanities. 606.