In vitro cormlet production of saffron (Crocus sativus L. Kashmirianus) and their flowering response under greenhouse.

A complete protocol for the saffron cormlet production under in vitro conditions and subsequent flowering under greenhouse conditions is described. Highest number of cormlets (70.0 ± 0.30) per corm slice (explant) could be regenerated on Murashige and Skoog (MS) half strength medium supplemented with thidiazuron (TDZ) (20 µM), Indole acetic acid (IAA) (10 µM), and sucrose (40 g/l). Maximum germination (90%) of these cormlets could be achieved on MS medium containing 6-benzyl amino purine (BAP) (20 µM) and α-naphthalene acetic acid (NAA) (15 µM). In order to increase the size of the in vitro raised cormlets, these were cultured on MS medium containing TDZ (15 µM) and IAA in the range of 1.5-30 µM. Maximum increase in cormlet size could be attained on TDZ (15 µM) + IAA (12.5 µM) + sucrose (30 g/l), and the average size of cormlets was 2.5g. In another experiment, apical vegetative buds of actively growing corms were cultured for cormlet development, and corms of size 2.5g could be developed on MS medium with NAA (15 µM), BAP (20 µM), and sucrose (30 g/l). The in vitro developed cormlets were dried under shade at 25 ± 2°C for 7 d. These were then planted in small cups containing clay loam soil and kept in green house at 20 ± 2°C. In vitro developed cormlets with mean weight 2.5 g showed maximum flowering (25%) as well as vegetative growth (55%), while only 19% cormlets of 2.0 g flowered. To our knowledge this is the first report on successful flowering from in vitro raised cormlets under greenhouse.