Population ecology of the endangered aquatic carnivorous macrophyte Aldrovanda vesiculosa at a naturalised site in North America

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Dormancy,germination and seed bank storage: a study in support of ex situ conservation of macrophytes of southwest Australian temporary pools

1. Vernal pools and rock pools (gnammas) in the Southwest Australian Floristic Region are forms of temporary wetlands that are under threat. Some of their aquatic macrophytes are rare and/or endemic, and there is a need to develop off‐site seed banks to assure their conservation. Here, we report results of the first comprehensive study of the seed germination, dormancy and seed storage behaviour of nine indigenous macrophyte species. 2. Seeds of Glossostigma drummondii, Myriophyllum balladoniense, M. lapidicola, M. petreaum and Triglochin linearis were non‐dormant, whereas those of Damasonium minus, Glossostigma sp. (currently undescribed), G. trichodes and Myriophyllum crispatum were dormant. Non‐dormant seeds germinated over a range of temperatures (5–20 °C) but temperatures at which highest germination occurred were species specific. All species demonstrated a germination preference for the light. Warm stratification substantially increased germination of dormant Glossostigma trichodes seeds and all dormancy‐breaking treatments partially overcame dormancy in Glossostigma spp. 3. Seeds possessed orthodox storage behaviour (tolerating drying to 5% moisture content and storage at ?18 °C) and are thus amenable to seed banking as a means of ex situ conservation. 4. It appears that seeds of most species are able to germinate upon inundation as long as they are situated at the soil surface. Thus, species are opportunistic and respond to the first rains of the season providing prompt ecological cuing in an environment vulnerable to rapid drying events. Maintaining the integrity of the soil crust may be an important first step for on‐site conservation if seeds are in the superficial layers.     

Seed banking of terrestrial orchids: evaluation of seed quality in Anacamptis following 4‐year dry storage

• Good‐quality dry seeds of some orchids have the potential to survive for decades under conventional seed bank conditions, but further research is needed to fill existing gaps in knowledge regarding seed behaviour under long‐term dry storage. The objectives of this study were to evaluate germination ability on two asymbiotic culture media with different nitrogen source; to assess seed desiccation tolerance needed for the storage at sub‐zero temperatures; and to study the effects of dry storage at low temperature.
• Asymbiotic seed germination tests of four Anacamptis species were carried out to evaluate the effects of different culture media, dehydration and dry storage on germination ability. Viability of 4‐year‐stored seeds was assessed by means of the tetrazolium test.
• Generalised linear model (GLM) analysis detected significant effects (P < 0.01) of the species, medium and storage time on total germination, while dehydration did not significantly affect it. Except for A. palustris, germination percentage was minimum after 1‐month storage and increased with longer storage periods. Tetrazolium viability tests detected high percentages of viable seed (>90%) following 4‐year storage in three out of four species.
• Seeds of the four Anacamptis species proved to be desiccation tolerant and have orthodox storage behaviour. The consequence of these findings is of interest to practical conservation approaches for orchids in seed‐banking. The results highlight the importance of multiple assessments of seed quality, both viability and germination, to understand seed storage behaviour.

     

Different plant provenance same seed tolerance to abiotic stress: implications for <Emphasis Type="Italic">ex situ</Emphasis> germplasm conservation of a widely distributed coastal dune grass (<Emphasis Type="Italic">Uniola paniculata</Emphasis> L.)

Preservation of genetic diversity within germplasm repositories represents an important tool for plant conservation. However, seeds must tolerate extreme levels of post-harvest desiccation and cold to realize benefits of ex situ storage. Factors including local climate and habitat impact expression of desiccation and freezing tolerance especially for widely distributed species. Our aim here was to understand the influence of a latitudinal gradient on seed desiccation and cryo-freezing tolerance. We sampled mature U. paniculata seeds from two geographically and genetically distinct populations then examined seed-water relations and germination following desiccation via equilibrium drying assays (0.5 to 91% RH; ?797 to ?12.9 MPa). Germination ability after drying and subsequent cryo-freezing treatments (?196?°C, 1 to 1440 min) was also evaluated. Seeds of both populations displayed similar reverse sigmoid moisture sorption isotherms characteristic of desiccation tolerant tissues. Furthermore, initial seed water potential (?63 and ?90 MPa) was considerably lower than the lethal limit (?20 MPa) identified for desiccation sensitive tissues. Final germination (range 58–93%) and temporal patterns differed significantly between populations following desiccation and cryo-freezing stress, but these germination responses were similar to initial germination. A higher proportion of non-germinated, yet viable seeds remained for the northern compared to southern population. Location does influence germination response, but differential germination is related to seed dormancy rather than desiccation or cryo-freezing sensitivity. Ex situ conservation of U. paniculata is therefore feasible across the latitudinal gradient studied here.     

Desiccation and storage studies on Dipterocarpm seeds

The relationship of seed moisture content (fresh weight basis) to germination, and the effect on viability of various storage conditions were examined for five species of the tropical forest tree genus Dipterocarpus. It was shown that seeds fall into two groups with regard to desiccation tolerance. Firstly, D. obtusifolius and D. turbinatus cannot be dried below about 45% moisture content without damage; a sigmoid curve was found to fit the relationship between germination and moisture content for the latter species. Secondly, D. intricatus, D. tuberculatus and D. alatus can be safely dried to 10%, 12% and 17% moisture contents respectively, but desiccation to near 7% moisture content reduced viability by at least a half. Storage studies showed that seed of D. intricatus and D. tuberculatus possessed increased longevity as moisture contents were reduced within the range 6–20%. It was concluded that seeds in the first group are ‘recalcitrant’ and that those in the second group are ‘orthodox’ in their storage physiology, according to the categories described by Roberts (1973). Wide differences between species in seed desiccation rates were observed. In 15% relative humidity D. intricatus dried to 7% moisture content within a week, whilst D. obtusifolius retained 30% moisture content even after 5 wk; other species had intermediate desiccation rates. Seed size and structure may partly account for the differences observed. Correlations were observed between seed storage physiology and other factors which were investigated. ‘Orthodox’ seeds had quicker desiccation rates, were derived from drier habitats, and had smaller embryos than those of ‘recalcitrant’ seeds. ‘Orthodox’ seeds, with the possible exception of D. alatus, should be kept at 0–3°C with about 12% moisture content in the short term and, provided less than 10% germination is lost on freezing, at-18°C with about 8% moisture content in the long term. ‘Recalcitrant’ seeds should be stored in ventilated containers at 21°C and with moisture contents above 45–50%.     

A comparative study of seed morphology in relation to desiccation tolerance and other physiological responses in 71 Eastern Australian rainforest species

Seed characteristics were measured in 71 Eastern Australian rainforest species representing 30 families. Sensitivity to desiccation to low moisture contents (< 10%) occurred in 42% of species. We estimate, based on findings from 100 species from this present study and previously published reports, that 49% of Eastern Australian rainforest species have non‐orthodox seeds. Germination level and time to 50% germination were not significantly different between desiccation sensitive (DS) and desiccation tolerant (DT) seeds. The estimation of seed desiccation sensitivity based on predictors is an important tool underpinning ex situ conservation efforts. Seed characteristics differed significantly between DS and DT seeds; that is, DS seeds had: (i) larger fruits (19 949 mg vs 8322 mg); (ii) larger seeds (1663 mg vs 202 mg); (iii) higher seed moisture contents (49.7% vs 35.5% fresh weight); (iv) lower oil content (7.3% vs 24.8% yield); and (v) less investment in seed coats (0.19 vs 0.48 seed coat ratio). Only 25% of DS seeded species had oily seeds compared with 87% of DT seeded species. Most green embryos were DS. Seed coat ratio was the best predictor of seed DS (80% correctly predicted). Seed moisture content at maturity was also related to germination time. Mean seed size was correlated (?0.657, P = 0.01) with mean seed oil content in 46 species. Further research on seed storage physiology of possible oily and/or DS seeded species is crucial to ensure future long‐term security of this biodiversity, particularly for species currently threatened in situ and/or of socioeconomic importance in Eastern Australian rainforests.     

Ecophysiology of seed dormancy and the control of germination in early spring‐flowering Galanthus nivalis and Narcissus pseudonarcissus (Amaryllidaceae)

Seed dormancy induction and alleviation in the winter‐flowering, moist temperate woodland species Galanthus nivalis and Narcissus pseudonarcissus are complex and poorly understood. Temperature, light and desiccation were investigated to elucidate their role in the germination ecophysiology of these species. The effect of different seasonal temperatures, seasonal durations, temperature fluctuations, the presence of light during different seasons and intermittent drying (during the summer period) over several ‘years’ on seed germination was investigated with outdoor and laboratory experiments. Warm summer‐like temperatures (20 °C) were necessary for germination at subsequent cooler autumn‐like temperatures (greatest at 15 °C in G. nivalis and 10 °C in N. pseudonarcissus). As the warm temperature duration increased, so did germination at subsequent cooler temperatures; further germination occurred in subsequent ‘years’ at cooler temperatures following a second, and also third, warm period. Germination was significantly greater in darkness, particularly in G. nivalis. Dormancy increased with seed maturation period in G. nivalis, because seeds extracted from green capsules germinated more readily than those from yellow capsules. Desiccation increased dormancy in an increasing proportion of N. pseudonarcissus seeds the later they were dried in ‘summer’. Seed viability was only slightly reduced by desiccation in N. pseudonarcissus, but was poor and variable in G. nivalis. Shoot formation occurred both at the temperature at which germination was greatest and also if 5 °C cooler. In summary, continuous hydration of seeds of both species during warm summer‐like temperatures results in the gradual release of seed dormancy; thereafter, darkness and cooler temperatures promote germination. Cold temperatures, increased seed maturity (G. nivalis) and desiccation (N. pseudonarcissus) increase dormancy, and light inhibits germination. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 177 , 246–262.     

Effect of storage time,site and floral morph on seed germination of the threatened distylous primrose Primula cusickiana var. maguirei

Ex situ conservation of rare plant species requires an understanding of germination requirements. We report the first successful germination trials for Maguire primrose (Primula cusickiana var. maguirei[L.O. Williams] N.H. Holmgren & S. Kelso), a threatened perennial herb narrowly restricted to a 20‐km canyon in northern Utah, USA. Seeds collected from different sites (upper or lower canyon populations) and flower morphs (pin or thrum) that had been stored for either 1 or 2 years were cold, moist stratified in darkness at 1–2°C for 270 days. Independent of treatment, time to first germination was approximately 75 days, time to 50% germination (T₅₀) was approximately 165 days, and total germination was 55.8%. Higher mean germination was observed in seeds stored for 2 years compared with seeds stored for 1 year, as well as in seeds from pin flowers compared with thrum flowers. There was not a significant effect of site on mean germination, but there was a significant storage time × site interaction. Seeds stored for 2 years also had a lower mean T₅₀ versus seeds stored for 1 year. In addition, seeds from upper canyon populations had a higher mean T₅₀ than those from lower canyon populations. Our results suggest that short‐term storage should not significantly decrease seed viability, and that both flower morph and canyon location may influence germination. Evidence suggests that germination patterns are adaptive. Our results contribute to our understanding of the germination biology of P. cusickiana var. maguirei and inform future conservation efforts for this threatened species.     

Seed germination and seedling emergence of Euryodendron excelsum H. T. Chang: implications for species conservation and restoration

Euryodendron excelsum H. T. Chang is a critically endangered plant from the family Theaceae that is endemic to China. It is now present in only one remnant population in a very narrow range and with a highly isolated and fragmented distribution pattern. The species is distributed close to the local villages and faces extinction because of the high level of anthropogenic disturbance. Thus, conservation and restoration of this species is urgent. The species reproduces by seed, but its germination requirements and seedling emergence are rarely understood. In this study, the germination requirements, desiccation tolerance and seedling emergence of the species were studied under controlled laboratory conditions. The results indicated that seeds of E. excelsum were non‐dormant. Optimal temperatures for germination were 15°C to 25°C; the germination percentage decreased and the mean germination time increased at high temperature. Seed germination was inhibited in the dark, suggesting that the seeds were positive photoblastic. Water stress also significantly inhibited germination percentage; no seeds germinated at 15% polyethylene glycol 6000. The fresh seeds had moderate moisture content of 28.6% and showed strong tolerance of dehydration. Thus, the seeds of E. excelsum tended to be orthodox. Seeds on the soil surface had the highest emergence percentage, which declined with depth of burial. Soil types significantly affected seedling emergence; seeds had higher emergence percentage in sandy soil than in mixed soil and clayed soil. Our study demonstrated that ex situ conservation and reintroduction of E. excelsum using seedling propagation from seeds is feasible.     

Seed germination of the carnivorous plant Byblis gigantea (Byblidaceae) is cued by warm stratification and karrikinolide

Byblidaceae is one of the most poorly studied carnivorous plant families, with seed dormancy and germination biology remaining unresolved. This knowledge deficit has significant conservation and management implications, particularly as the most southerly distributed species, the south‐west Western Australian endemic Byblis gigantea, is listed as critically endangered. This study examined the ecophysiology of seed dormancy and germination in B. gigantea in concert with a study of the population dynamics of a single plant community. Mass seedling emergence and plant establishment were observed after a wildfire in 2007 in the natural population, followed by a rapid decline in mature individuals (91%) over a 4‐year monitoring period, with almost no inter‐fire recruitment (1.4% of all emergence) observed. Seeds possessed a fully developed embryo, and the germination characteristics of fresh seeds classified them as showing physiological dormancy. Seed dormancy was partially alleviated by warm stratification (30 °C) for 8 weeks prior to incubation at 15 °C, with c. 40% germination observed. With the additional exposure of seeds to the germination‐active chemical in smoke, karrikinolide, the germination of warm‐stratified seeds increased to 89%. Seeds also displayed orthodox storage behaviour and appeared to be amenable to long‐term seed banking for conservation. These results present the first observation of the stimulation of the germination of a carnivorous plant by a smoke‐derived compound. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 172 , 143–152.     

Effect of water content and temperature on seed longevity of seven Brassicaceae species after 5 years of storage

Maximising seed longevity is crucial for genetic resource preservation and longevity of orthodox seeds is determined by environmental conditions (water content and temperature). The effect of water content (down to 0.01 g·H₂O·g^?1) on seed viability was studied at different temperatures for a 5‐year storage period in taxonomically related species. Seeds of seven Brassicaceae species (Brassica repanda, Eruca vesicaria, Malcolmia littorea, Moricandia arvensis, Rorippa nasturtium‐aquaticum, Sinapis alba, Sisymbrium runcinatum) were stored at 48 environments comprising a combination of eight water contents, from 0.21 to 0.01 g·H₂O·g^?1 DW and six temperatures (45, 35, 20, 5, ?25, ?170 °C). Survival curves were modelled and P50 calculated for those conditions where germination was reduced over the 5‐year assay period. Critical water content for storage of seeds of six species at 45 °C ranged from 0.02 to 0.03 g·H₂O·g^?1. The effect of extreme desiccation at 45 °C showed variability among species: three species showed damaging effects of drying below the critical water content, while for three species it was neither detrimental nor beneficial to seed longevity. Lipid content could be related to longevity, depending on the storage conditions. A variable seed longevity response to water content among taxonomically related species was found. The relative position of some of the species as long‐ or short‐lived at 45 °C varied depending on the humidity at which storage behaviour was evaluated. Therefore, predictions of survival under desiccated conditions based on results obtained at high humidity might be problematic for some species.     

Improved assessment of viability and germination of Cattleya (Orchidaceae) seeds following storage

Effective seed storage methods for orchid species provide a unique opportunity to preserve large amounts of germplasm. Early evaluation of the physiological status of seeds is useful for selecting seed lots for conservation. To score seed quality, seeds from nine Cattleya species (Cattleya granulosa, Cattleya hegeriana, Cattleya intermedia, Cattleya mossiae, Cattleya purpurata, Cattleya sanguiloba, Cattleya tenuis, Cattleya tigrina and Cattleya walkeriana) were germinated on Knudson C medium and viability was measured using a tetrazolium (TZ) test following pre-conditioning in a 10% sucrose solution, both before and after storage at −18°C for 3 mo. Tetrazolium test results were quantified using digital imaging of germinating seeds on a flatbed scanner. Seed viability (with or without pre-conditioning in sucrose), germination, Germination Velocity Index (GVI) and the correlation between viability measurements and germination were also evaluated. Both the tetrazolium test and the germination results showed that there was no loss of viability after storage, although in five seed lots, the GVI decreased. Pre-conditioning improved the correlation between viability exhibited in the tetrazolium test and seed germination. Image analysis proved to be a powerful tool for analysing these small seeds.     

Comparative longevity of Australian orchid (Orchidaceae) seeds under experimental and low temperature storage conditions

Seeds from ten terrestrial orchid species, nine from the south‐west Australian biodiversity hotspot (Caladenia arenicola, Caladenia flava, Caladenia huegelii, Diuris laxiflora, Microtis media ssp. media, Pterostylis recurva, Pterostylis sanguinea, Thelymitra crinita and Thelymitra macrophylla) and one from south‐east Australia (Diuris fragrantissima), were placed into experimental storage to assess their relative longevity and likely optimal conditions for long‐term conservation seed banking. Seeds from all species were desiccation tolerant, germinating after drying at 23% relative humidity (C. arenicola, C. huegelii, P. sanguinea and T. macrophylla) or 5% relative humidity (C. flava, D. laxiflora, M. media ssp. media, P. recurva and T. crinita) at 23 °C. From automatedly determined moisture adsorption and desorption isotherms at 23 °C, these equate to tolerance of drying to 0.03–0.06 g water g^?1 dry weight or 0.013–0.028 g water g^?1 dry weight, respectively. Results of storage experiments at a range of moisture contents and temperatures suggest conventional seed bank storage at ?18 °C after equilibration at c. 23% relative humidity (at 23 °C) may be suitable for most of the species, although there was higher germination of P. recurva seeds stored at ?80 °C and of M. media ssp. media seeds equilibrated at 75% relative humidity. However, there was considerable variation in germination of seeds sampled after different storage periods, making it difficult to identify optimal storage conditions definitively. Results of comparative longevity storage experiments at 60% relative humidity and 40 °C suggest seeds from these orchid species are short‐lived compared with non‐orchid species, and with Australian species in particular. © 2010 The Linnean Society of London, Botanical Journal of the Linnean Society, 2010, 164 , 26–41.     

Physiological characteristics underpinning successful cryopreservation of endemic and endangered species of Bromeliaceae from the Brazilian Atlantic Forest

The germination requirements and the basis of the optimal water content before and after cryopreservation were studied for ten endangered Brazilian species of Bromeliaceae. Constant and alternating temperature regimes were used to determine the best conditions for seed germination. The relationship between seed water content and relative humidity was evaluated using water sorption isotherms at 15 °C. Seeds were cryostored at four water contents (3, 5, 7 and 9%) and three storage periods (0, 180 and 365 days), and loss in viability and vigour were estimated. Fresh seeds of all species showed maximum germination in < 30 days at temperatures between 20 and 30 °C, indicating the absence of a physical/morphological dormancy. A sigmoidal relationship between seed water content and relative humidity was observed with no apparent differences in sorption characteristics among the species. The optimum water content for cryopreservation of most of these species was c. 7%. Ultra‐drying (3% seed water content) had a detrimental effect on seed viability and vigour. Our experiments suggested orthodox storage behaviour for all species of Bromeliaceae examined as they are able to survive desiccation and freezing. This study has shown the feasibility of ex situ conservation in seed cryobanks of endangered bromeliads from the Brazilian Atlantic Forest to support future reintroduction of these species in nature. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 176 , 567–576.     

Embryo growth and seed germination of four taxa of Narcissus (section Pseudonarcissi) conserved under non-recalcitrant seed conditions in germplasm banks

The aim of this study was to determine the germinative ability of the seeds of four Narcissus taxa belonging to Section Pseudonarcissi after they had been conserved under the conditions of non-recalcitrant seed storage protocols. For each taxon (N. alcaracensis, N. longispathus, N. radinganorum and N. pseudonarcissus subsp. munozii-garmendiae), one seed lot was desiccated to 4% moisture content (MC) and stored under laboratory conditions (22°C, 40–50% relative humidity (RH), whereas another was dehydrated to 3% MC and stored at −10°C. The latter treatment simulated standard conservation conditions for non-recalcitrant seeds. After 26 months, embryo growth and germination were evaluated. Seed responses were correlated with their MC upon dispersal. Seeds of N. alcaracensis, N. longispathus and N. radinganorum left to dry on the mother plant during maturation had 8–10% MC when dispersed, tolerated non-recalcitrant seed conservation and germinated to >90% under the most favorable incubation conditions. Narcissus pseudonarcissus subsp. munozii-garmendiae seeds did not undergo maturation drying and had 46.7% MC upon dispersal. They reached 100% germination after being desiccated to 4% and stored at 22°C, were not recalcitrant, but failed to germinate when stored at −10°C under non-recalcitrant seed conservation conditions. Therefore, N. alcaracensis, N. longispathus and N. radinganorum seeds can be conserved under non-recalcitrant seed conditions in germplasm banks, whereas those of N. pseudonarcissus subsp. munozii-garmendiae are moderately recalcitrant. Seed storage behavior is influenced primarily by the extent of maturation drying of the seeds on the mother plant.     

Dormancy and viability of Ferocactus peninsulae (Cactaceae) seeds

Knowledge on seed dormancy is crucial for the understanding of plant population dynamics, as it controls seed germination and seed bank formation. Dormant seeds have high potential to establish in soil seed banks, but such information within Cactaceae is scarce, although it is essential for conservation programs. The aim of this study was to determine if seeds of Ferocactus peninsulae showed any kind of dormancy and to test their germination capacity after storage. This was assessed with 15 seed sowing experiments done over 4 years with seeds stored under room conditions (20 ± 2°C). We demonstrated the existence of physiological dormancy in F. peninsulae seeds that is broken with an after-ripening period. Germination was low during the first 3 months of storage (d = 0.206) but increased after 10 months of storage (d = 0.654), and seeds maintained their viability at 48 months (d = 0.707). Also, their speed of germination increased with storage time. Ferocactus peninsulae seeds are positively photoblastic, and the requirement for light for germination persisted over all experiments. The results provide crucial information for propagation and conservation research and may allow us to infer that F. peninsulae seeds are able to form a persistent soil seed bank, as they maintained their viability after dormancy is released.     

Does perianth colour affect the seed germination of two desert shrubs under different storage periods and conditions?

Haloxylon salicornicum and Salsola vermiculata (Amaranthaceae) are two perennial shrubs producing fruits with differently coloured perianths (yellow vs pink) on separate individuals. To test the impact of different storage periods (four, eight and twelve months) and temperature conditions (freezing, room temperature and natural field temperatures) on the seed germination of the two seed categories, germination tests were carried out on both species. For both species, collected from wild plants in the Arabian desert (Qatar), seed germination was tested under two photoperiods (light 12/12; dark 24 hours) at 20/30°C for winged (intact) and de‐winged seeds. For each species, all the considered factors (perianth colour, wing presence, photoperiod, storage period and storage conditions) had significant effects on seed germination (p < 0.001), while the interactions among factors varied in their level of significance depending on the species. In both species, yellow seeds germinated considerably better than the pink ones, the presence of wings slowed the germination, and both storage period and temperature conditions highly influenced their germination ability. Our results indicate that seeds from different perianth colours have different germination strategies to delay their germination and distribute the species through time by escaping germination time by imposing some kind of after‐ripening or seed dormancy. The variation in storage time and storage conditions might help in extending their germination period, where one set of seeds germinate immediately, and the other set of seeds becomes dormant and thus contributes to the formation of a soil seed bank for the long‐term recruitment of seedlings. This study highlights an ecological advantageous strategy for these two species growing in the unpredictable desert habitat.     

Consequences of habitat disturbance on seed fate of a Brazilian tropical dry forest tree Cavanillesia arborea (Malvaceae)

We compared seed fate (survival, mortality by rodent predators, desiccation and other causes) of the tree Cavanillesia arborea (Malvaceae) in preserved old‐growth tropical dry forests and in nearby abandoned pasturelands in Brazil. For this purpose, we performed an experiment where 15 seeds were placed in the surroundings of 15 parental individuals in each habitat. These 450 seeds were monitored over four months. At the end of the experiment, seed predation by rodents was higher in forest (56.5%) than in pasture (8.9%) areas, but seed desiccation showed the opposite pattern (8.9% vs. 80.4%). Mortality by desiccation was also faster in pasture than in forest areas, probably reducing their attractiveness to predators in these areas. None of the seeds placed in the pasture survived, whereas 26.2% of seeds became seedlings in forest areas. The absence of seedling recruitment of Cavanillesia arborea in pastures is likely a consequence of their incapacity to tolerate the harsh abiotic conditions in this habitat. Thus, forest conversion imposes a strong limitation to the long‐term population viability of this species. However, restoration and natural regeneration of abandoned pastures can recreate the forest structure and microclimatic conditions favourable to seed germination and seedling establishment. Remnant individuals of Cavanillesia arborea in agricultural landscapes may have a negligible contribution to current recruitment, but they can attract potential dispersers of pioneer species, with positive feedbacks to future recruitment during secondary succession.     

Analysis of diaspore morphology and seed germination in Bubon macedonicum L., a rare species in Italy

Bubon macedonicum L. is a chasmophytic species of south-eastern Europe. In Italy, it has been detected only in Rocca Monforte (Campobasso, central Italy). This rare species is included in the IUCN Red Lists of Critically Endangered Italian Flora, and there are no studies relating to B. macedonicum biology. The seed germination dynamics of this species was studied with the aim of building up an appropriate germination protocol to be used in ex situ conservation. On the basis of an ISTA protocol, about 3,000 seeds were collected from Rocca Monforte in August 2013. Fifty seeds were measured. The considered parameters were seed length, width, thickness, seed surface, volume, density, surface/mass ratio and eccentricity index. The morphometric parameters examined showed morphological dormancy, where a short warm period is necessary for embryo growth and seed germination. The results showed high germination percentages under the different conditions of temperature, pH, GA₃ and photoperiod. Only at 5 °C was there no germination. Finally, the seeds maintain high germination percentages from the seed storage process after 130 and 390 days. This factor can be considered of great importance for the conservation of B. macedonicum over the medium and long term.