The genus Narcissus has several endemic, rare and/or threatened species in the Iberian Peninsula and North Africa. In vitro propagation is a useful tool for threatened plants conservation used in ex situ strategies. Thus, the aim of this work was to study the propagation in vitro of bulb scale explants of five endemic, rare and/or endangered Narcissus species from the Iberian Peninsula, treated with different PGR combinations. Initiation was achieved in half-strength Murashige and Skoog (MS) basal salts and vitamins, 10 g/L sucrose, 500 mg/L casein hydrolysate, 2 mg/L adenine, 10 mg/L glutathione and 5.5 g/L plant agar. In the multiplication phase, the highest bulblet proliferation was obtained in MS medium supplemented with 30 g/L sucrose and the combination of 10 μM 6-Benzylaminopurine (BAP) + 5 μM α-Naphthaleneacetic acid (NAA) in N. alcaracensis, N. eugeniae and N. hedraeanthus; 20 μM BAP + 5 μM NAA in N. jonquilla and N. yepesii. The highest rooting was obtained with 5 μM NAA + 1 μM Indole-3-butyric acid (IBA) for all species (>75%) and more than 80% of the produced bulblets were successfully acclimatized.

The achene morphology and pericarp anatomy of 12 taxa representing three genera ( Anemone, Hepatica, and Pulsatilla) of the subtribe Anemoninae were investigated using microtome and light microscopy to evaluate the taxonomic implications of achene characters. The achenes of Anemone were elliptical or obovoid and beaked, whereas the achene of Hepatica and Pulsatilla were obovoid and elliptical, respectively. Noticeable variations in both quantitative and qualitative features of achenes were observed among the species of the three genera. One-way analysis of variance indicated that the quantitative achene variables among the species were highly significant (P<0.001). Pearson’s correlation coefficient also showed a significant correlation between different achene variables. The pericarp structure, particularly the number of cell layers and cell forms in the exocarp and endocarp, seems to be very useful for species delimitation in Anemone and Hepatica. The nature of the endotesta could provide substantial proof for sub-generic classification in Anemone. Unweighted paired group analysis showed the utility of achene features for taxonomic groupings of the species within the studied genera. Although the specimen samples represented a limited range of taxa, the achene features and pericarp anatomy provided a reasonable source for the taxonomic treatment of the studied genera within the subtribe.

In this study, female gametophytes of Silene muradica, which is a gynodioecious species, were examined histologically. Buds and blossoms of S. muradica were used as the research material. They were collected in the Sivas province (Turkey) in July 2019, and fixed with ethanol:acetic acid solution (3:1, v/v). Flower parts were dissected under a stereo microscope. They were dehydrated in rising alcohol series and then embedded in Historesin. The sections were taken by a rotary microtome and stained with 0.5% Toluidine blue O. The ovary of S. muradica has three carpels and a single chamber, the ovules are arranged on a central column. The mature ovule is of the campylotropous type, crassinucellate and bitegmig. The megaspore mother cell undergoes regular meiotic division and forms a linear megaspore tetrad after meiosis. The development of the embryo sac is monosporic. The chalazal megaspore is functional and the others degenerate. The mature embryo sac is eight-nucleated and of the Polygonum type. The synergid cells and the egg cell are completely surrounded by the cell wall. Antipodal cells are temporary cells, which degenerate immediately after fertilization. Before fertilization, polar nuclei are fused in the central cell and form the secondary nucleus. The endosperm development is of the nuclear type. Nucellar tissue is permanent and forms perisperm in mature seeds. The embryo development is of the Caryophyllad type. In this study, the development of the female gametophyte of S. muradica, which was determined to be a gynodioecious species, was reported for the first time.