‘Kalender’ Yonca (Medicago sativa L.) Çeşidinin In vitro Çoğaltımı Üzerine Farklı Besin Ortamları, Sitokininler ve Eksplant Tiplerinin Etkisi

Abstract

Medicago sativa L. is one of the most important forage crops cultivated all over the world. In vitro propagation of some M. sativa cultivars has been studied earlier; however, it is highly cultivar dependent and necessary to develop different in vitro propagation procedures for each cultivar. So far, in vitro propagation of alfalfa ‘Kalender’ cultivar has not been reported. In this study, the effects of different basal media, cytokinins and explant types on in vitro propagation of alfalfa cultivar ‘Kalender’ were investigated. Initially, different explants types (meristem, shoot tip and node) excised from in vitro seedlings were culture on Gamborg medium (B5) containing different concentrations of kinetin (1.0 and 2.0 mg/L) to induce shoot regeneration. Besides high shoot regeneration rates (57.78% - 93.33%), high callus regeneration rates (82.22% – 93.33%) were also observed. Because of high callus formation, second experiment was conducted. Shoot tip explants excised from in vitro seedlings were transferred on B5, Murashige and Skoog (MS) and Woody Plant Medium (WPM) supplemented with different concentrations of 6-benzylaminopurine (BAP) (0, 0.125, 0.25, 0.5, and 1.0 mg/L). Although, the best shoot regeneration rate (100%) was obtained on WPM supplemented with 0.25 or 1.0 mg/L BAP and MS supplemented with 0.25 mg/L BAP, shoot tips cultured on BAP-containing media produced high callus regeneration. In contrast to BAP-containing media, on WPM medium, the development of shoots and roots occurred simultaneously and healthy and well-developed plantlets were obtained. In vitro shoots rooted best (71.11%) on WPM. All plantlets were successfully acclimatized. Thus, an efficient in vitro propagation protocol for M. sativa L. cultivar ‘Kalender’ was developed.