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.

Can botanic gardens serve as refuges for taxonomic and functional diversity of Odonata? The case of the botanic garden of Castilla-La Mancha (Spain).

In a scenario with declining biodiversity and habitat loss, botanic gardens could serve as refuges for invertebrates, but the opportunities they offer for animal conservation are still poorly understood. Odonata is a good model group for conservation studies, because it includes threatened species and responses to habitat disturbance are well documented. In this study, we assessed the role of the botanic garden of Castilla-La Mancha in Spain as a refuge for members of Odonata by analysing their taxonomic and functional diversity. We explored if the small size of the botanic garden might constrain the taxonomic diversity of Odonata and if low habitat diversity might limit their functional diversity. We sampled adult Odonata from five water bodies along a gradient of human impact and characterized the Odonata communities based on 12 functional traits in Odonata. We used a species-area relationship to control for differences in the size of water bodies. Compared with natural lakes, the Odonata communities contained less species and their functional diversity was lower in the botanic garden ponds, where generalist species were basically hosted. Despite these limitations, the botanic garden ponds hosted the number of species expected for natural water bodies with the moderate surface area and functional diversity, thereby demonstrating that they are a valuable habitat for Odonata in an urban environment. Appropriate management involving the removal of exotic fish and habitat diversification, including creating lotic environments, would increase the taxonomic and functional diversity of Odonata in this urban system. Supplementary Information: The online version contains supplementary material available at 10.1007/s10201-022-00704-3.

Warming reverses directionality in the richness-abundance relationship in ephemeral Mediterranean plant communities.

Recent findings in forests worldwide have demonstrated how directionality in the richness-abundance causality shifts along global climate gradients: The so-called more-species hypothesis (richness determines abundance) prevails in Earth's most productive climates, whereas the opposite, the so-called more-individuals hypothesis (abundance determines richness), is more likely to prevail in climatically harsh conditions. Since temporal variability is the norm, a critical question is whether this directionality shift is also a function of temporal climatic fluctuations locally. Here, we analyze whether directionality in the richness-abundance relationship is contingent on temporal variability over 10 annual consecutive realizations in ephemeral plant assemblages. Our results support the idea that the more-species hypothesis prevailed in the most benign years, whereas the more-individuals hypothesis did so during less productive years, which were significantly linked to the warmest years. These results support the idea that rising temperatures can reverse directionality in the richness-abundance relationship in these annual plant communities, and therefore, climate warming can have a significant effect on the relationship between diversity and ecosystem functions, such as productivity, by altering the prevalence of primary mechanisms involved in species assembly.

Diverse phylogenetic neighborhoods enhance community resistance to drought in experimental assemblages.

Although the role played by phylogeny in the assembly of plant communities remains as a priority to complete the theory of species coexistence, experimental evidence is lacking. It is still unclear to what extent phylogenetic diversity is a driver or a consequence of species assembly processes. We experimentally explored how phylogenetic diversity can drive the community level responses to drought conditions in annual plant communities. We manipulated the initial phylogenetic diversity of the assemblages and the water availability in a common garden experiment with two irrigation treatments: average natural rainfall and drought, formed with annual plant species of gypsum ecosystems of Central Spain. We recorded plant survival and the numbers of flowering and fruiting plants per species in each assemblage. GLMMs were performed for the proportion of surviving, flowering, fruiting plants per species and for total proportion of surviving species and plants per pot. In water limited conditions, high phylogenetic diversity favored species coexistence over time with higher plant survival and more flowering and fruiting plants per species and more species and plants surviving per pot. Our results agree with the existence of niche complementarity and the convergence of water economy strategies as major mechanisms for promoting species coexistence in plant assemblages in semiarid Mediterranean habitats. Our findings point to high phylogenetic diversity among neighboring plants as a plausible feature underpinning the coexistence of species, because the success of each species in terms of surviving and producing offspring in drought conditions was greater when the initial phylogenetic diversity was higher. Our study is a step forward to understand how phylogenetic relatedness is connected to the mechanisms determining the maintenance of biodiversity.

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.

Effect of aridity on species assembly in gypsum drylands: a response mediated by the soil affinity of species.

Previous studies found that plant communities on infertile soils are relatively resistant to climatic variation due to stress tolerance adaptations. However, the species assemblies in gypsum soil habitats require further investigation. Thus, we considered the following questions. (1) Do harsher arid conditions determine the characteristics of the species that form plant assemblages? (2) Is the selection of the species that assemble in arid conditions mediated by their ability to grow on gypsum soils? (3) Is the selection of species that assemble in harsher conditions related to phylogenetically conserved functional traits? Perennial plant communities were analysed in 89 gypsum-soil sites along a 400 km climate gradient from the central to southeastern Iberian Peninsula. Each local assemblage was analysed in 30 × 30 m plots and described based on taxonomic, functional (soil plant affinity) and phylogenetic parameters. The mean maximum temperatures in the hottest month, mean annual precipitation and their interaction terms were used as surrogates for the aridity conditions in generalized linear models. In the hottest locations, the gypsophily range narrowed and the mean gypsophily increased at the community level, thereby suggesting the filtering of species and the dominance of soil specialists in the actual plant assemblies. Drier sites had higher taxonomic diversity. The species that formed the perennial communities were close in evolutionary terms at the two ends of the aridity gradient. The mean maximum temperatures in the hottest month had the main abiotic filtering effect on perennial plant communities, which was mediated by the ability of species to grow on gypsum soils, and thus gypsum specialists dominated the species assemblies in the hottest locations. In contrast, the perennial communities on gypsum soils were relatively resistant to changes in precipitation. Our findings suggest that the warmer environmental conditions predicted by global change models will favour gypsum specialists over generalists.

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.

Positive associations among rare species and their persistence in ecological assemblages.

According to the competitive exclusion principle, species with low competitive abilities should be excluded by more efficient competitors; yet, they generally remain as rare species. Here, we describe the positive and negative spatial association networks of 326 disparate assemblages, showing a general organization pattern that simultaneously supports the primacy of competition and the persistence of rare species. Abundant species monopolize negative associations in about 90% of the assemblages. On the other hand, rare species are mostly involved in positive associations, forming small network modules. Simulations suggest that positive interactions among rare species and microhabitat preferences are the most probable mechanisms underpinning this pattern and rare species persistence. The consistent results across taxa and geography suggest a general explanation for the maintenance of biodiversity in competitive environments.

Effects of lead shot ingestion on bone mineralization in a population of red-legged partridge (Alectoris rufa).

The effect of lead (Pb) toxicity on bone mineralization was investigated in a wild population of red-legged partridge (Alectoris rufa) inhabiting a farmland area contaminated with Pb-shot from recreational hunting activities in Albacete, a southeastern province of Spain. Femora from 40 specimens of red-legged partridge were analyzed for Pb by graphite furnace atomic absorption spectroscopy (GF-AAS), and for bone composition by Fourier transform infrared (FTIR) spectroscopy and X-ray diffraction (XRD). The FTIR and DRX data of bone were analyzed in detail to determine possible alterations in bone mineral chemistry and crystallinity due to Pb toxicity. Results showed a marked decrease in the degree of mineralization as Pb concentrations in bone tissue increased while XRD analyses showed that the crystallinity of apatite crystals increased with the Pb load in bone. These load-dependent effects are indicative that Pb contamination altered bone remodeling by reducing new bone mineral formation and demonstrate that bone quality is a sensitive indicator of adverse effects on wild bird populations exposed to Pb pollution.

Germination and soil seed bank traits of Podocarpus angustifolius (Podocarpaceae): an endemic tree species from Cuban rain forests.

Podocarpus angustifolius is an endangered recalcitrant-seeded small tree, endemic to mountain rain forests in the central and Pinar del Río regions in Cuba. In this study, the germination patterns of P. angustifolius seeds were evaluated and the nature of the soil seed bank was determined. Using a weighted two-factor design, we analyzed the combined germination response to seed source (i.e. freshly matured seeds directly collected from trees versus seeds extracted from soil samples) and pretreatment (i.e. seed water-immersion for 48h at room temperature). Germination was delayed for four weeks (= 30 days) in all cases, regardless of both factors analyzed. Moreover, nine additional days were necessary to achieve high germination values (in the case of fresh, pretreated seeds). These results overall may indicate the existence of a non-deep simple morphophysiological dormancy in P. angustifolius seeds. The water-immersion significantly enhanced seed germination, probably as a result of the hydration of recalcitrant seeds. Although germination of seeds extracted from soil samples was low, probably due to aging and pathogen effects throughout the time of burial, the study revealed the existence of a persistent soil seed bank (at least short-termed) of approximately 42 viable seeds per m2 in the upper 10cm of soil. Such a record is noteworthy since references to persistent soil seed banks in recalcitrant-seeded species are scarce in the literature. The population consequences derived from the formation of persistent soil seed banks in this endangered species are discussed.

Germination and soil seed bank traits of Podocarpus angustifolius (Podocarpaceae): an endemic tree species from Cuban rain forests

Podocarpus angustifolius is an endangered recalcitrant-seeded small tree, endemic to mountain rain forests in the central and Pinar del Río regions in Cuba. In this study, the germination patterns of P. angustifolius seeds were evaluated and the nature of the soil seed bank was determined. Using a weighted two-factor design, we analyzed the combined germination response to seed source (i.e. freshly matured seeds directly collected from trees versus seeds extracted from soil samples) and pretreatment (i.e. seed water-immersion for 48h at room temperature). Germination was delayed for four weeks (≈30 days) in all cases, regardless of both factors analyzed. Moreover, nine additional days were necessary to achieve high germination values (in the case of fresh, pretreated seeds). These results overall may indicate the existence of a non-deep simple morphophysiological dormancy in P. angustifolius seeds. The water-immersion significantly enhanced seed germination, probably as a result of the hydration of recalcitrant seeds. Although germination of seeds extracted from soil samples was low, probably due to aging and pathogen effects throughout the time of burial, the study revealed the existence of a persistent soil seed bank (at least short-termed) of ≈42 viable seeds per m² in the upper 10cm of soil. Such a record is noteworthy since references to persistent soil seed banks in recalcitrant-seeded species are scarce in the literature. The population consequences derived from the formation of persistent soil seed banks in this endangered species are discussed. Rev. Biol. Trop. 59 (3): 1061-1069. Epub 2011 September 01.

Podocarpus angustifolius es un árbol endémico de los bosques lluviosos de la región de Pinar del Río y la parte central de Cuba, que se encuentra en peligro de extinción. En este estudio se evaluó la germinación de sus semillas y la naturaleza del banco de semillas del suelo. Específicamente, se analizó la respuesta germinativa de las semillas a dos factores: su procedencia (recolectadas directamente de los árboles versus extraídas de muestras de suelo) y el pretratamiento (inmersión en agua durante 48h a temperatura ambiente). La germinación no comenzó hasta las cuatro semanas (≈30 días) en todos los casos. Además, fueron necesarios 9 días más para alcanzar un valor elevado de germinación (en el caso de semillas recolectadas del árbol con pretratamiento), lo que parece indicar la existencia de un letargo de tipo morfofisiológico simple no profundo. El pretratamiento incrementó significativamente la germinación, posiblemente por hidratación de las semillas recalcitrantes. El estudio demostró la existencia de un banco de semillas persistente en el suelo, al menos de corta duración de ≈42 semillas viables por m² en los 10 primeros cm del suelo. Dicho hallazgo es destacable, ya que los casos descritos en la literatura sobre bancos persistentes en especies con semillas recalcitrantes son escasos. Además, se discute la importancia que tiene para esta especie amenazada la existencia de un banco de semillas persistente.

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).

Lead-shot exposure in red-legged partridge (Alectoris rufa) on a driven shooting estate.

The goal of the study was to investigate the accumulation and spatial patterns of spent lead (Pb) shot pellets and the prevalence of shot ingestion in red-legged partridge in a driven shooting estate. Soil was collected using a regular sampling design perpendicular to three shooting lines. Factors involved in shot spatial distribution were investigated by a causal structural equation model (SEM). Shot ingestion prevalence and liver and bone Pb concentrations were studied in partridges hunted in 2004 and 2006. Shot soil-burden averaged 73,600 units/ha (i.e., 8.1 kg/ha). Shot density was significantly higher in front of than behind shooting lines, with greatest accumulation occurring at between 40-110 m and in certain ecotones (i.e., shrubland-dry cropland). Analyses revealed 7.8% of partridges with evidence of Pb shot ingestion. Particle size in diet, grit-size composition, and shot ingestion prevalence were significantly higher in 2004 than in 2006, indicating that supplying partridges with large seeds (i.e., corn) may increase the risk of Pb shot ingestion. Moving shooting lines into croplands and controlling seed size used for diet supplementation may reduce shot ingestion.