Öz

Colchicum calcedonicum Azn, pek çok endemik bitki türünün görüldüğü Türkiye'de yayılış gösteren endemik türlerden biridir. Toprak altında uzun-oval şekilli soğanı ile genellikle 3-4 yapraklıdır. Kallus kültürü kullanarak endemik türlerin in vitro üretimi, bu türlerin korunmasında umut verici bir çalışma haline gelmiştir. Bu çalışmanın amacı, in vitro C. chalcedonicum üretimi için verimli kallus protokolünün oluşturulabilmesidir. Explantların sterilizasyonunda, 20 dk %0.25 (w/v) cıva klorür (HgCl2) kullanılmıştır. Cıva klorüre ilaveten, yüzey sterilizasyonunda 30 dk boyunca Tween 80, %6,5 NaCl ile birlikte kullanılmıştır. Bu çalışmada, 19 farklı besiyeri kullanılmış olup primer kallus oluşumu 2,4-D (2 mg L-1), 2IP (0,5 mg L-1), %3 sükroz ve %0,05 aktif karbon içeren Murashige & Skoog bazal besiyerinde elde edilmiştir. Çalışmamız, aktif karbon kullanımının primer kallus oluşumunda etkili olduğunu göstermiştir. Bu çalışma, C. chalcedonicum’un primer kallus oluşumu için ilk rapordur. Bununla birlikte, çalışmamız endemik tür olan C. chalcedonicum'un in vitro korunması ve kallus oluşum protokolünün geliştirilmesinde öncü bir çalışmadır. 

PIONEERING in vitro STUDIES FOR CALLUS FORMATION OF Colchicum chalcedonicum Azn.

Abstract

Colchicum calcedonicum Azn is one of the endemic species distributed in Turkey, where many endemic plant species occur. It has long-oval shaped corm under the soil, and usually 3-4 leaves on it. In vitro production of endemic species using callus culture has become promising study for conservation. The aim of this study is to generate an efficient callus protocol for in vitro production of C. chalcedonicum. To sterilize the explants, 0.25% (w/v) mercuric chloride (HgCl2) was used for 20 min. In addition to mercuric chloride, surface sterilization was conducted by using 6.5% NaCl with Tween 80 for 30 min. We used 19 different mediums and the primary callus formation was obtained in Murashige & Skoog’s basal medium (MS) supplemented with 2,4-D (2 mg L-1), 2IP (0.5 mg L-1), 3% sucrose and 0.05% active carbon. Our study demonstrated the active carbon usage was effective for the primary callus formation. This study is the first report for primary callus formation of C. chalcedonicum. However, our work is a pioneering study to improve callus formation protocol system for in vitro conservation of endemic species C. chalcedonicum.

Keywords

• Colchicum chalcedonicum
• Tissue Culture
• Callus Formation
• Conservation

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