Sea daffodil is a perennial medicinal plant with several beneficial health properties that is enlisted as an endangered and protected species. For the exploitation and conservation of such valuable species, sustainable propagation is a key factor. For this purpose, the establishment of in vitro cultures using botanical seeds is an effective way to produce a large number of microplants which can be used as propagating material and/or bulblets with further uses for the isolation–production of biocactive substances. In this work, we performed two separate experiments where we studied (a) the effect of temperature (10 °C, 15 °C, 20 °C, 25 °C, and 30 °C) on seed germination in vitro and (b) the effect of temperature (10 °C, 20 °C, and 30 °C) and sucrose concentration (40 and 80 g/L) in growth medium on microplant development and the production of bulblets. Seed germination rates were high (over 70%) at a wide range of temperatures (15–30 °C), although the fastest and the most uniform germination was observed at 20–25 °C. Regarding the effect of temperature on microplant development and bulblet production, low temperatures (10 °C) had a negative effect on the number of leaves, bulb diameter, and fresh weight (F.W.) and dry weight (D.W.) of leaves, roots, and bulblets. The best growth of microplants (number of leaves, F.W. and D.W. of leaves, roots) and bulblet production (diameter, F.W., and D.W.) were observed in culture media with sucrose concentration 40 g/L and temperature of 30 °C. In culture medium with a sucrose concentration of 80 g/L the production of bulbs (diameter, F.W., and D.W.) was not inferior to culture media with a sucrose concentration of 40 g/L, although microplants had less root growth. It is concluded that the best treatment for the production of well-rooted microplants with large bulblets that could be used as propagating material or for the production of bulblets as a source for twin scales explants or pharmaceutical substances was the culture at 30 °C in culture medium with sucrose concentration 40 g/L. On the other hand, culture medium with a sucrose concentration of 80 g/L at 30 °C or 20 °C could be also used for the production of bulblets.

The aim of the study was to develop an efficient micropropagation protocol of Origanum scabrum, which will allow its commercial exploitation in the pharmaceutical and horticultural industries. First, the effect of the date of the explant collection (20 April, 20 May, 20 June, 20 July, 20 August) and the position of the explant on the plant stem (shoot apex, 1st node, 3rd node, 5th node) on the establishment of in vitro cultures was studied (1st experiment: Stage I). Next, the effect of temperature (15 °C, 25 °C) and the node position (microshoot apex, 1st node, 5th node) on the microplant production and ex vitro survival of plantlets was studied (2nd experiment: Stage II). The optimum season to collect explants from wild plants was shown to be during the vegetative growth of the plants (April to May), while the shoot apex and the 1st node were the most suitable explants. For the proliferation and production of rooted microplants, the best results were obtained from single-node explants excised from microshoots produced from 1st node-explants collected on 20th of May. Temperature did not affect microshoot number, leaf number and the percentage of rooted microplants, while microshoot length was higher at 25 °C. Moreover, microshoot length and the percentage of rooted microplants were higher in those derived from apex explants, while the survival of plantlets was not affected by treatments and ranged between 67% and 100%.