Cengiz SANCAK, Sebahattin ÖZCAN, Serkan URANBEY, Khalid Mahmood KHAWAR

Effect of thidiazuron on shoot regeneration from different explants of lentil (Lens culunaris Medik.) via organogenesis

Thidiazuron (TDZ) en aktif sitokinin benzeri bileşiklerden olup, birçok bitki türünde sitokininlerden daha yüksek oranlarda sürgün rejenerasyonu sağlamaktadır. Yaygın olarak üretimi yapılan mercimek çeşitlerinden Ali Dayı ve Kayı 91 çeşitlerine ait yaprak, gövde, gövde bouğumu ve kotiledon boğum eksplantları farklı oranlarda TDZ içeren Murashige ve Skoog (MS) besin ortamlarında kültüre alınmıştır. Bu çalışma Mercimekte (lens Culinaris Medik.) gen aktarımına uygun hızlı ve etkili bir sürgün rejenerasyon sistemi geliştirmek için yürütülmüştür. Kotiledon ve gövde boğumlarından, başlangıç kallus gelişimini takiben organogenesis aracılığıyla prolifik adventif sürgün rejenerasyonu elde edilmiştir. Yaprak ve gövde eksplantlarında ise kallus ve sürgün gelişimi gözlenmemiştir. DMSO, TDZ çözücüsü olarak kullanıldığında bitki dokularına ölümcül etki yapmış ve çalışmalarda TDZ %50 ethanolda çözülmüştür. Kotiledon boğumları gövde boğumlarından daha yüksek sürgün rejenerasyon kabiliyetine sahip olmuştur. Her iki çeşitte de en yüksek sürgün rejenerasyonu 0.25 mg/l TDZ içeren MS besin ortamlarında kotiledon boğumlarından elde edilmiştir. Elde edilen sürgünler (10-20 mm uzunlukta) 0.25 mg/l indol-3-butrik asid (IBA) içeren MS ortamında köklendirilmiştir. Köklenen sürgünler son olarak kum içeren saksılara aktarılmıştır.

Organogenesis aracılığıyla farklı mercimek (Lens culunaris Medik.) eksplantlarından sürgün rejenerasyonuna TDZ' nin etkisi

Thidiazuron (TDZ) is among the most active cytokinin like substances and induces greater in vitro shoot proliferation than many other cytokinins in many plant species. Leaf, stem, stem node and cotyledonary node explarits of 2 extensively cultivated Turkish lentil cultivars, Ali Dayı and Kayi 91, were cultured on Murashige and Skoog (MS) media supplemented with various concentrations of TDZ. The present study was conducted to develop a rapid and efficient shoot regeneration system suitable for the transformation of lentil (Lens culinaris Medik.) using TDZ. Cotyledonary nodes and stem nodes after the initial callus stage regenerated prolific adventitious shoots via organogenesis. Shoot or callus formation was not achieved from leaf or stem explants. DMSO as a solvent for TDZ was necrotic on plant tissues and therefore TDZ was dissolved in 50% ethanol to carry out the studies. Cotyledonary nodes showed a higher shoot formation capacity than stem nodes. MS medium supplemented with 0.25 mg/l TDZ produced the highest frequency of shoot formation from cotyledonary nodes in both genotypes. Regenerated shoots (10-20 mm long) rooted in MS medium containing 0.25 mg/l indole-3-butyric acid (IBA). Rooted plantlets were finally transferred to sand in pots. Abbreviations: TDZ - thidiazuron [1 Phenyl 3-(l ,2,3-thiadiazol -5YL) urea], IBA - indole-3-butyric acid, MS - Murashige and Skoog, DMSO - dimethylsulphoxide

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