Plant Production Protocols from Seeds of Threatened Atropa baetica and Widespread A. belladonna, Both Rich in Alkaloids.

Members of the genus Atropa contain various tropane alkaloids, including atropine ((±)-hyoscyamine) and scopolamine, which possess medicinal properties. Preserving the diverse genetic background of wild populations via optimal plant production from seeds could be essential for avoiding the loss of potential uses. We analyzed the germination ecology of two Atropa species comprising the threatened A. baetica and widespread A. belladonna to determine the: (1) influence of temperature, light, and seed age on germination patterns; (2) effects of cold stratification and gibberellic acid (GA3); (3) phenology of seedling emergence in outdoor conditions; (4) phenology of dormancy break and loss of viability in buried seeds; and (5) ability to form persistent soil seed banks. Freshly matured seeds exhibited conditional physiological dormancy, with germination at high temperatures (32/18 °C) but not at low and cold ones (5, 15/4, 20/7 °C). The germination ability increased with time of dry storage and with GA3, thereby suggesting nondeep physiological dormancy. Under outdoor conditions, no seedlings emerged during the first post-sown autumn, but emergence peaks occurred in late winter-early spring. Both species could form small persistent soil seed banks with short durations (3-5 years). A plant production protocol from seeds was established for both taxa.

Optimization of Plant Production by Seed Treatment in Two Wild Subspecies of Narcissus pseudonarcissus Rich in Alkaloids.

The daffodil Narcissus pseudonarcissus L. contains alkaloids of pharmaceutical interest. Wild daffodil populations have diverse genetic backgrounds and various genetic traits of possible importance. Developing protocols for plant production from seeds may ensure the availability of a large reservoir of individuals as well as being important for species with bulbs that are difficult to acquire. The closely related Narcissus pseudonarcissus subsp. munozii-garmendiae and subsp. nevadensis were investigated in this study because the alkaloids isolated from both are of high pharmacological interest. At the dispersal time, the seeds of both were dormant with underdeveloped embryos, i.e., morphophysiological dormancy (MPD). Experiments were conducted outdoors and under controlled laboratory conditions. Embryo growth and the percentages of radicle and seedling emergence were calculated under different temperature-light stratifications. In N. munozii-garmendiae, embryo growth occurred during warm stratification (28/14 °C or 25/10 °C) and the radicle then emerged when the temperature decreased, but the shoot was dormant. In N. nevadensis, the seeds germinated when cold stratified (5 °C) and then incubated at cool temperatures. Thus, N. munozii-garmendiae and N. nevadensis exhibit different levels of MPD, i.e., deep simple epicotyl and intermediate complex, respectively. Plant production protocols from seeds were established for both taxa in this study.

Seed germination in Narcissus yepesii (Amaryllidaceae): clinal variation in the morphophysiological dormancy levels.

Seed dormancy classes determine both population and species-level processes which can be crucial in the life cycle of many plants. However, there are no studies of a dormancy cline between levels of morphophysiological dormancy (MPD). We aimed to determine the class of seed dormancy of Narcissus yepesii exhibits in order to explore links between different dormancy levels, previously characterized in two closely related phylogenetic congeners, N. alcaracensis and N. longispathus. Experiments were carried out under both near-natural temperature and controlled laboratory conditions. The parameters calculated were mean embryo length, radicle and shoot emergence percentages. The effects of different periods of storage; and different periods with or without GA3 of warm, cold or warm plus cold were analysed. The Narcissus populations from the Baetic System of mountain ranges in south-eastern Spain show clinal variation in a northeast-southwest gradient from intermediate to non-deep complex MPD, through the coexistence of intermediate and non-deep complex MPD in N. yepesii (21 % and 74 %, respectively). In addition, 54 % of stored seeds were able to show both levels of MPD. Narcissus yepesii occupies an intermediate position between N. alcaracensis and N. longispathus in the geographical distribution and in the clinal germination ranges. It strongly suggests an evolutionary gradient, which connects the intermediate complex MPD with the non-deep complex MPD in southern Iberian daffodils. This is the first study showing a gradient in the evolution between levels of MPD. Our results demonstrate a cline in these levels in response to both an environmental gradient and genetic differences.

Physiology, morphology and phenology of seed dormancy break and germination in the endemic Iberian species Narcissus hispanicus (Amaryllidaceae).

BACKGROUND AND AIMS: Only very few studies have been carried out on seed dormancy/germination in the large monocot genus Narcissus. A primary aim of this study was to determine the kind of seed dormancy in Narcissus hispanicus and relate the dormancy breaking and germination requirements to the field situation. METHODS: Embryo growth, radicle emergence and shoot growth were studied by subjecting seeds with and without an emerged radicle to different periods of warm, cold or warm plus cold in natural temperatures outdoors and under controlled laboratory conditions. KEY RESULTS: Mean embryo length in fresh seeds was approx. 1·31 mm, and embryos had to grow to 2·21 mm before radicle emergence. Embryos grew to full size and seeds germinated (radicles emerged) when they were warm stratified for 90 d and then incubated at cool temperatures for 30 d. However, the embryos grew only a little and no seeds germinated when they were incubated at 9/5, 10 or 15/4 °C for 30 d following a moist cold pre-treatment at 5, 9/5 or 10 °C. In the natural habitat of N. hispanicus, seeds are dispersed in late May, the embryo elongates in autumn and radicles emerge (seeds germinate) in early November; however, if the seeds are exposed to low temperatures before embryo growth is completed, they re-enter dormancy (secondary dormancy). The shoot does not emerge until March, after germinated seeds are cold stratified in winter. CONCLUSION: Seeds of N. hispanicus have deep simple epicotyl morphophysiological dormancy (MPD), with the dormancy formula C(1b)B(root) - C(3)(epicotyl). This is the first study on seeds with simple MPD to show that embryos in advanced stages of growth can re-enter dormancy (secondary dormancy).