Abstract

This study was performed in order to micropropagate the critically endangered Balkan endemic Dianthus ingoldbyi and to investigate the anatomical and morphological characteristic of shoots following the slowing down of the growth of in vitro shoots under cold and dark conditions. For this purpose, axillary buds isolated from aseptic seedlings were cultured on Murashige and Skoog (MS) medium including 1mg/L Benzylaminopurine (BAP) and 0.3mg/L Naphthaleneacetic acid (NAA) and a regeneration ratio of 84.8% was obtained. The number of shoots per explant was 5.9 after 30 days from culture initial and the mean shoot length was 3.6cm. These shoots were transferred to MS medium including 0.5mg/L NAA and stored at 4 oC in a refrigerator in total darkness for 1-6 months without subculturing. 58% of shoots survived after 6 months of cold storage conditions. Growth of shoots was significantly slowed down and there was no anatomical or morphological deformation at the end of the experimental period. Shoots transferred to normal conditions from cold storage developed better than the control group. Well rooted plants were acclimatized to outdoor conditions with a survival rate of 48%. In conclusion, the cold storage technique used in this study is suggested as an effective method for in vitro conservation of the critically endangered D. ingoldbyi.