EFFICIENT IN VITRO BULBLET PRODUCTION OF ENDANGERED HYACINTHELLA Micrantha (BOISS.) CHOUARD
B. Bulut1, S. Uranbey2*, H. A. A. Ahmed3, and M. Çalişkan4
1Cankiri Karatekin University. Institute of Science Forestry Engineering Çankırı/Turkey
2Ankara University, Agriculture Faculty, Department of Field Crops, 06100 Ankara/Turkey
3Uşak University, Faculty of Agriculture and Natural Sciences Department of Field Crops, 64200 Uşak/Turkey
4Central Research Institute for Field Cronps, Ministry of Food, Agriculture and Livestock, Ankara, Turkey
*Corresponding author’s e-mail: uranbey@ankara.edu.tr
ABSTRACT
Hyacinthella micrantha (BOISS.) Chouard with their attractive flowers is one of the important endemic and endangered species of Turkey. We aimed to improve a standard in vitro multiplication protocol using basal scales for bulblet production and conservation of the species. Bulb scales of the species collected from natural habitat were cultured in the different basal Murashige and Skoog (MS), Orchimax and Lindeman Orchid Medium embryonic callus induction medium. Then, they were transferred to MS, Orchimax and Lindeman Orchid Medium basal bulblet induction medium supplemented with cytokinins (1.0, 2.0, 4.0 and 8.0 mg/l N6-benzylamino-purine (BAP), Zeatin and 2-isopentyladenine (2-iP) at 16 °C. After ten months of culture initiation, high frequency bulblet regeneration was developed using bulb scales on different basal media containing different concentrations of cytokinintypes. The highest ratio of bulb scales forming shoot/bulblets per explant (87%) and bulblets per explants (5.60) were induced on a Orchimax media supplemented with 8 mg/L Zeatin. MS and Orchimax medium gave the best result in terms of embryonic calli formation and in vitro bulblet production compared to Lindeman Orchid medium. 2,4-D, BAP and Zeatin were more effective than2-iPfor in vitro bulblet production. Also, regenerated bulblets subjected to pre-cold treatments for 3 months at 4-6 ºC resulted in increasing of new mini bulblets (approximately 15%). Regenerated bulblets were acclimatized with high survival ratio (82 %).
Keywords: bulblets, Hyacinthella micrantha, in vitro, micropropagation. |
