Morphological differentiation supports the genetic pattern of the geographic structure of Juniperus thurifera (Cupressaceae)

Juniperus thurifera is an important component of woodland communities of dry sites within the West Mediterranean region and is characterised by a strongly disjunctive geographic range. Two subspecies were recognised, subsp. thurifera in Europe and subsp. africana in Africa. The aim of the study was the comparison of phenetic diversity to the pattern of AFLP geographic differentiation of the species described in the literature. The examination of phenetic diversity was based on the biometrical analysis of 17 populations using 12 morphological characters of cone and seed. The differences among populations were analysed using Student’s t test, analysis of discrimination, UPGMA agglomeration and hierarchical analysis of variance. The majority of morphological characters differentiated at a statistically significant level between populations and between J. thurifera subsp. thurifera and subsp. africana. Three groups of populations were detected using multivariate statistical analyses. The first, well separated, is subsp. africana, while the following two concern subsp. thurifera. The morphological differentiation of populations appeared similar to that described on the AFLP. The Gibraltar Straight appeared to be the most important barrier.     

Inter- and intraspecific morphometric variability in Juniperus L. seeds (Cupressaceae)

In this study, a statistical classifier for Mediterranean taxa of Juniperus, based on 98 seed morphometric parameters, was tested at interspecific, specific and intraspecific levels. Ripe cones of 10 taxa were collected in different regions of the Mediterranean Basin to compare and discuss their taxonomic treatments according to two different sources. High percentages of correct identification were reached for both taxonomic treatments at the specific and intraspecific level and from the comparison among taxa of the J. oxycedrus, J. communis and J. phoenicea complexes. Moreover, ripe cones of J. macrocarpa were collected from four Sardinian populations, in two seasons, and from plant and soil, in order to analyse inter-population, seasonal and source variability in seed morphology. This statistical classifier discriminated J. macrocarpa seeds collected in spring more accurately than those collected in autumn, but it failed to distinguish between the seeds collected from plants and soil, or between those collected from different populations of the same geographic region.     

Geographical Distribution of Diploid and Tetraploid Cytotypes of Thymus sect. Mastichina (Lamiaceae) in the Iberian Peninsula, Genome Size and Evolutionary Implications

The occurrence of diploid and tetraploid cytotypes in Thymus section Mastichina (Lamiaceae) was investigated by chromosome counting and flow cytometric measurements of DNA content. To cover the entire native distribution of the section across the Iberian Peninsula, plant material was sampled from 29 populations, representing all three taxa (Thymus albicans, T. mastichina subsp. mastichina and T. mastichina subsp. donyanae). We provide first estimates of genome size for taxa of this section. Analyses revealed the existence of new cytotypes in T. mastichina subsp. mastichina (2n?=?2x?=?28, 30) and confirmed the existence of previous ones (2n?=?4x?=?56, 58, 60). We also confirmed the presence of exclusively diploid cytotypes (2n?=?2x?=?30) in the southwestern Iberian endemics T. albicans and T. mastichina subsp. donyanae. We conclude that the southwestern Iberian Peninsula acted as a Pleistocene glacial refugium that may have permitted speciation processes within this section. Several hypotheses concerning these processes and the origin and distribution of the studied taxa are discussed.     

High level of genetic differentiation of Juniperus phoenicea (Cupressaceae) in the Mediterranean region: geographic implications

Fourteen natural populations of Juniperus phoenicea L. from the quite entire species range have been compared using isoenzyme polymorphism. Among 17 loci, 5 (Got1, 6Pgd3, Pgi2, Pgm2 and Shdh2) appeared to be differentiated sufficiently to provide useful information for discrimination between the subspecies phoenicea and turbinata (Guss.) Nyman. Two distinct groups of populations were detected using the Nei’s genetic distance unweighted pair group method with arithmetic mean (UPGMA) and discrimination analyses, one including the inland populations of the eastern Iberian Peninsula and southern France (subsp. phoenicea), and the second from the Mediterranean and Atlantic shores, and from the Atlas mountains in Africa (subsp. turbinata). The high level of differences confirms a long period of isolation, probably during the whole Pleistocene. The population from the Aegean Sea shore differed from the other Mediterranean shore plus Atlas mountain population. It also suggests spatial isolation between them, at least during the last Glaciation.     

Taxonomic and geographic differentiation of Juniperus phoenicea agg. based on cone,seed, and needle characteristics

The aim of this study was to examine the diversity of, and differentiation between, 41 natural populations representing Juniperus phoenicea complex within their geographic range using the morphological characteristics of cones, seeds, and sprouts with leaves. Seven populations of J. phoenicea s.s., 25 J. turbinata and nine J. canariensis, each represented by ～20–30 individuals, were studied. The characteristics describing the size of cones and seeds and their proportions were different between J. phoenicea s.s., J. turbinata, and J. canariensis, and were useful in distinguishing between these three taxa. The populations of J. canariensis had a significantly higher percentage of trimerous cones, a higher number of cone scales, but a lower number of seeds, which were also larger than the seeds from other taxa. Juniperus phoenicea s.s., J. turbinata, and J. canariensis revealed a high level of multivariate differentiation, but there were no single characteristics which allowed one to distinguish between these alone. We detected significant differences between populations of J. turbinata from Europe and Africa, which supports the role of the Gibraltar Straight in the structuring of conifer species. Significant differences were also detected between European and Asiatic populations in several characters examined, indicating the important role of the Aegean Sea in the structuring of this taxon.     

Reproductive isolation of two sympatric subspecies ofJuniperus phoenicea (Cupressaceae) in southern Spain

The phenology, reproductive cycle and cone crop of two sympatric subspecies ofJuniperus phoenicea (Cupressaceae) were studied during three consecutive years in southern Spain. The flowering pattern of each subspecies was constant during this period: inJ. phoenicea subsp.turbinata flowering was always in October–November, whileJ. phoenicea subsp.phoenicea flowered in February–March. There was no overlap between the flowering periods of the two taxa. The reproductive cycle of the two subspecies was similar, though there were some important differences because of their flowering times. Male cone development in both subspecies occurred in autumn, but the male cones ofJ. phoenicea subsp.phoenicea did not shed pollen until the end of February. InJ. phoenicea subsp.turbinata, the gap between pollination and fertilisation was seven months while inJ. phoenicea subsp.phoenicea it was only three months, fertilisation taking place at the same time in the two taxa. Post-fertilisation events were similar in the two subspecies, leading to a cycle of two years inJ. phoenicea subsp.turbinata and a shorter one of about 20 months inJ. phoenicea subsp.phoenicea. Alternating good and slight seed cone crops were found in both taxa during the three years studied, the good crops of one subspecies coinciding with the slight crops of the other one. Although the proximity of the two subspecies and method of pollination could make hybridisation between the taxa possible, the different flowering seasons prevent hybridisation isolating them reproductively.     

Morphological analysis,chorology and ecology of Juniperus navicularis Gand. in the Iberian Peninsula

Juniperus navicularis is an endemic taxon of Iberian Peninsula that develops on paleodunes of coastal ecosystems and whose distribution range is not well defined. In this paper, we present the results of the morphological analysis in populations attributed to J. navicularis of southern Spain. In order to assess the similarity between Portuguese and Spanish populations, the Mann–Whitney U test was performed, the same test was also applied between this taxon and Juniperus oxycedrus subsp. oxycedrus. In order to distinguish this species from other taxa of the Juniperus oxycedrus group (subsp. oxycedrus, subsp. badia, subsp. macrocarpa) in the southwestern Mediterranean region, a principal component analysis has been carried out. New data about the chorology and ecology of Spanish populations considered as a priority habitat in the Habitat Directive as well as number of individuals and conservation status are provided.     

Contribution to the Karyological Knowledge of Mediterranean Romulea Species (Iridaceae)

Eighteen chromosome counts are reported in nine Mediterranean Romulea species. Among them, four species have a wide distribution range (R. bulbocodium, R. columnae, R. ramiflora and R. rollii) and five are narrow central Mediterranean endemics (R. bocchierii, R. ligustica, R. linaresii, R. requienii and R. revelieri). Chromosome numbers range from 2n?=?3x?=?27 in one R. columnae accession to 2n?=?6x?=?54 in R. linaresii. Our results only partially agree with previous reports, and the chromosome number records of R. bocchierii, R. ligustica, R. linaresii subsp. linaresii and R. revelieri presented here are completely new. According to our data and contrary to previous statements, x?=?9 seems to be the basic chromosome number in this group of Mediterranean Romulea. Moreover, a highly significant negative correlation between the chromosome size and ploidy level was found.     

Diversity of Phytophthora Species from Declining Mediterranean Maquis Vegetation,including Two New Species,Phytophthora crassamura and P. ornamentata sp. nov.

The Mediterranean basin is recognized as a global biodiversity hotspot accounting for more than 25,000 plant species that represent almost 10% of the world’s vascular flora. In particular, the maquis vegetation on Mediterranean islands and archipelagos constitutes an important resource of the Mediterranean plant diversity due to its high rate of endemism. Since 2009, a severe and widespread dieback and mortality of Quercus ilex trees and several other plant species of the Mediterranean maquis has been observed in the National Park of La Maddalena archipelago (northeast Sardinia, Italy). Infected plants showed severe decline symptoms and a significant reduction of natural regeneration. First studies revealed the involvement of the highly invasive wide-host range pathogen Phytophthora cinnamomi and several fungal pathogens. Subsequent detailed research led to a better understanding of these epidemics showing that multiple Phytophthora spp. were involved, some of them unknown to science. In total, nine Phytophthora species were isolated from rhizosphere soil samples collected from around symptomatic trees and shrubs including Asparagus albus, Cistus sp., Juniperus phoenicea, J. oxycedrus, Pistacia lentiscus and Rhamnus alaternus. Based on morphological characters, growth-temperature relations and sequence analysis of the ITS and cox1 gene regions, the isolates were identified as Phytophthora asparagi, P. bilorbang, P. cinnamomi, P. cryptogea, P. gonapodyides, P. melonis, P. syringae and two new Clade 6 taxa which are here described as P. crassamura sp. nov. and P. ornamentata sp. nov. Pathogenicity tests supported their possible involvement in the severe decline that is currently threatening the Mediterranean maquis vegetation in the La Maddalena archipelago.     

Population genetic structure and adaptive differentiation of iron walnut Juglans regia subsp. sigillata in southwestern China

Southwestern (SW) China is an area of active tectonism and erosion, yielding a dynamic, deeply eroded landscape that influences the genetic structure of the resident populations of plants and animals. Iron walnut (Juglans regia subsp. sigillata) is a deciduous tree species endemic to this region of China and cultivated there for its edible nuts. We sampled 36 iron walnut populations from locations throughout the species' range in SW China and genotyped a total of 765 individuals at five chloroplast DNA regions and 22 nuclear microsatellite loci. Species distribution models were produced to predict the evolution and historical biogeography of iron walnut and to estimate the impacts of climate oscillations and orographic environments on the species' demography. Our results indicated that J. regia subsp. sigillata had relatively low genetic diversity, high interpopulation genetic differentiation, and asymmetric interpopulation gene flow. Based on DIYABC analysis, we identified two lineages of J. sigillata in southwestern China. The lineages (subpopulations) diverge during the last glacial period (~1.34 Ma). Southwestern China was a glacial refuge during the last glacial period, but increasingly colder and arid climates might have fostered the fragmentation of J. regia subsp. sigillata within this refugium. Finally, we found that recent habitat fragmentation has led to a reduction in population connectivity and increased genetic differentiation by genetic drift in isolated populations. Our results support a conclusion that geological and climatic factors since the Miocene triggered the differentiation, evolutionary origin, and range shifts of J. sigillata in the studied region.     

Ecological status of the Mediterranean Juniperus phoenicea L. Relicts in the desert mountains of North Sinai,Egypt

Juniperus phoenicea L. is listed as threatened tree by IUCN Red List. In Egypt, J. phoenicea L. is the only conifer tree that is restricted to the three mountains of northern Sinai: Gabal El-Halal, Gabal El-Maghara and Gabal Yelleq. As a Mediterranean relict it has been included in a national list as target for conservation and management. To provide baseline information for the development of a conservation strategy, the present study aims at comparing the isolated populations of J. phoenicea and their associated plant composition and diversity at the three mountains. The application of TWINSPAN and DCA analysis techniques has resulted in identifying of four vegetation types associated with juniper, and each could be related to a specific geomorphologic habitat on a topographic gradient. Chiliadenus montanus and Zygophyllum dumosum characterized the slopes of smooth-faced rock outcrops in Wadi Abu Seyal (at 350–470 m altitude of Gabal El-Halal), Deverra tortuosa, Ephedra aphylla and Gymnocarpos decander inhabited together with the target species the soil pockets of north-facing slope in Neqeb Abu Hamam (at 600–700 m altitude of Gabal El-Halal), Stachys aegyptiaca and Moricandia nitens characterized the juniper occurrences in the runnels of Wadi Arar (at 450–560 m altitude of Gabal El-Maghara), and Artemisia herba-alba, Atriplex halimus and Reaumuria hirtella represent the stands on slope runnels (at 900–960 m) of Gabal Yelleq. The two vegetation types recognized at Gabal El-Halal had, on average, the highest species diversity, juniper density and cover. Juniper shows generally poor conditions of vitality at higher elevation (600–960 m) with a higher proportions of old and recent dead trees, and with the predominance of male individuals, as compared with the populations of Gabal El-Maghara and Gabal Yelleq. In contrast, the juniper populations at lower elevation (350–470 m) of Gabal El-Halal proved to be in best condition with mostly living foliage and reproductive branches. The differences in rock types and elevation among the three mountains reflect serious limitation on recruitment of J. phoenicea due to moisture availability. The results of this study showed that J. phoenicea is an endangered species and its conservation in northern Sinai mountains is a priority. For a successful conservation of this community it is highly recommended to preserve in particular the suitable habitats at Gabal El-Halal, but also the other stands merit conservation measures.     

The biogeography and genetic relationships of Juniperus oxycedrus and related taxa from the Mediterranean and Macaronesian regions

Despite Juniperus spp. being an important component of Mediterranean arid and semi‐arid ecosystems, there is a lack of complex studies on their biogeographical patterns. Using 16 morphological cone and seed traits and three nuclear microsatellite markers, we investigated the morphological and genetic variability of seven Mediterranean and Macaronesian Juniperus taxa (J. oxycedrus ssp. oxycedrus, J. oxycedrus ssp. badia, J. brevifolia, J. cedrus, J. deltoides, J. macrocarpa and J. navicularis) to identify biogeographical trends and interspecific genetic relationships. The highest gene diversity was measured in J. oxycedrus ssp. oxycedrus (H_E = 0.716) and the lowest in J. brevifolia (H_E = 0.441). The west Mediterranean was characterized by a higher level of genetic diversity than the east Mediterranean. A lack of significant genetic differences between European and African populations of J. oxycedrus suggests that the Strait of Gibraltar was not a significant barrier to gene flow, but has promoted some morphological differentiation. The genetic and morphological results strongly support the recognition of J. macrocarpa, J. navicularis and J. deltoides at the species rank, whereas J. oxycedrus ssp. badia should be included in J. oxycedrus. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 174 , 637–653.     

A comparative analysis between SNPs and SSRs to investigate genetic variation in a juniper species (<Emphasis Type="Italic">Juniperus phoenicea</Emphasis> ssp. <Emphasis Type="Italic">turbinata</Emphasis>)

Genomic resources are a valuable research tool for understanding and forecasting the response of forest trees to global change and for developing science-based management strategies. Yet, many ecologically relevant tree species still lack such resources. The conifer genus Juniperus contains >?70 species that are widely distributed through the Northern Hemisphere, including several keystone species that form extensive forests in arid landscapes. To date, single-nucleotide polymorphism (SNP) markers have not been described for this ecologically important tree genus and the few described simple sequence repeat (SSR) markers result insufficient for performing reliable population demographic inference. Here, we report on the successful development of 19 new SSR and 147 SNP markers for Phoenician juniper (Juniperus phoenicea ssp. turbinata), a species widely distributed along the coasts of the Mediterranean Basin. We calculate a series of population genetic diversity estimates for each set of markers independently and for both sets combined. Our comparison shows that the higher per-locus information content of SSRs makes them the marker of choice for parentage and assignment studies, whereas SNPs provide more reliable demographic inferences (N_e and detection of a recent bottleneck). We also test and confirm the transferability of the new set of SNP markers to the closely related tetraploid species J. thurifera. Finally, we perform an orthology analysis with two gymnosperm model species to search for SNPs linked with functional genes.     

Allozyme Diversity and Geographic Variation among Populations of the Locally Endangered Taxon Carex magellanica subsp. irrigua (Cyperaceae)

Carex magellanica subsp. irrigua is a wet habitat taxon that is extinct or declining in the Baltic States and Central Europe, but still quite common in northern areas, in Fennoscandia and Alaska. We investigated the extent of genetic variation within and among populations and geographic regions of this subspecies. Isozyme electrophoresis in polyacrylamide gels was applied to characterize genetic diversity with allozymes as genetic markers. Of the nine putative isozyme loci assessed, five (56%) were found to be polymorphic. The genetic diversity in small and fragmented Estonian populations was lower (H _e?=?0.034) than in larger Fennoscandian and Alaskan populations (average H _e?=?0.082). All standard genetic parameters (A _e, H _o, H _e, P, F _is, t) showed the lowest values in Estonian populations. The heterozygosity level in Fennoscandian populations was low (H _o?=?0.01), whereas no heterozygotes were found in Estonian and Alaskan populations. High F _is values indicate that C. magellanica subsp. irrigua is predominantly inbreeding. The main reason for its decline in Estonia is the destruction of suitable habitats. More attention to the protection of Estonian habitats is needed to maintain genetic diversity and stop further decline of this taxon.     

Morphological Characters Useful for the Delimitation of Taxa Within Viola Subsect. Viola (Violaceae): A Morphometric Study from the West Carpathians

Forty-nine morphological characters were scored or measured on 44 populations (376 individuals) of Viola subsect. Viola from the West Carpathians and adjacent areas (Slovakia, Czech Republic, Austria and Hungary). The presence of six species, namely V. alba (represented by subsp. alba), V. ambigua, V. collina, V. hirta, V. odorata and V. suavis s.l. was revealed based on pollen fertility, cytological and morphometric analyses. The morphological characters traditionally used to delimit taxa within the subsection and those revealed by our study as most reliable are widely discussed. A key for identifying the taxa and most common hybrids of subsection Viola occurring in the West Carpathians is presented. Chromosome counting and flow cytometry were used to determine the ploidy levels of the populations studied. All individuals of V. alba subsp. alba, V. collina, V. hirta and V. odorata were tetraploid, while those of V. ambigua and V. suavis s.l. were octoploid.     

Genome size and chromosome number in Echinops (Asteraceae, Cardueae) in the Aegean and Balkan regions: technical aspects of nuclear DNA amount assessment and genome evolution in a phylogenetic frame

This work focuses on the representatives of genus Echinops (Asteraceae, Cardueae) in the Aegean and Balkan regions, from the perspective of their genome evolution. Chromosome numbers were determined by orcein staining in 14 populations of nine taxa, and DNA contents were assessed by flow cytometry in 24 populations of nine taxa. A molecular phylogeny based on the internal transcribed spacer (ITS) and trnL-trnF and including first sequences for two taxa (Echinops sphaerocephalus subsp. taygeteus and E.?spinosissimus subsp. neumayeri) provided a framework for discussing genome changes. From a methodological point of view, similar C-DNA value estimates were obtained when measuring, for a same population, fresh leaves from adult plants collected in the field and from cultivated seedlings. Conversely, despite giving the appearance of being correct (e.g., low coefficient of variation), genome size assessed using silica gel-preserved material differs significantly from values obtained for the same populations with fresh material. Nevertheless, silica gel-preserved material may still provide rough estimates of genome size for, e.g., inferring ploidy level. Suitable—non-silica gel-based—DNA amounts assessed for 23 populations range from 2C?=?6.52?pg (E.?spinosissimus subsp. neumayeri) to 2C?=?9.37?pg (E.?bannaticus). Chromosome counts were established for the first time for Echinops graecus (2n?=?32), E.?sphaerocephalus subsp. albidus (2n?=?32), E.?sphaerocephalus subsp. taygeteus (2n?=?ca.?30), and E.?spinosissimus subsp. neumayeri (2n?=?28). Genome size and chromosome number are confirmed as crucial parameters for deciphering lineage diversification within the genus Echinops.     

Frankia Populations in Soil and Root Nodules of Sympatrically Grown Alnus Taxa

The genetic diversity of Frankia populations in soil and in root nodules of sympatrically grown Alnus taxa was evaluated by rep-polymerase chain reaction (PCR) and nifH gene sequence analyses. Rep-PCR analyses of uncultured Frankia populations in root nodules of 12 Alnus taxa (n?=?10 nodules each) growing sympatrically in the Morton Arboretum near Chicago revealed identical patterns for nodules from each Alnus taxon, including replicate trees of the same host taxon, and low diversity overall with only three profiles retrieved. One profile was retrieved from all nodules of nine taxa (Alnus incana subsp. incana, Alnus japonica, Alnus glutinosa, Alnus incana subsp. tenuifolia, Alnus incana subsp. rugosa, Alnus rhombifolia, Alnus mandshurica, Alnus maritima, and Alnus serrulata), the second was found in all nodules of two plant taxa (A. incana subsp. hirsuta and A. glutinosa var. pyramidalis), and the third was unique for all Frankia populations in nodules of A. incana subsp. rugosa var. americana. Comparative sequence analyses of nifH gene fragments in nodules representing these three profiles assigned these frankiae to different subgroups within the Alnus host infection group. None of these sequences, however, represented frankiae detectable in soil as determined by sequence analysis of 73 clones from a Frankia-specific nifH gene clone library. Additional analyses of nodule populations from selected alders growing on different soils demonstrated the presence of different Frankia populations in nodules for each soil, with populations showing identical sequences in nodules from the same soil, but differences between plant taxa. These results suggest that soil environmental conditions and host plant genotype both have a role in the selection of Frankia strains by a host plant for root nodule formation, and that this selection is not merely a function of the abundance of a Frankia strain in soil.     

Intraspecific phylogenetic relationships in Ixeridium dentatum (Compositae) complex based on genetic and morphological evidence

Ixeridium dentatum (I. dentatum; family Compositae) forms a complex species through intraspecific differentiation. In this study, intraspecific taxa of I. dentatum were analyzed by counting chromosome numbers, sequencing internal transcribed spacer (ITS), and detecting amplified fragment length polymorphisms (AFLP) to explore phylogenetic relationships within I. dentatum. Analysis of ploidy levels showed that I. dentatum subsp. kitayamense and subsp. nipponicum were diploid (2n = 2x = 14), I. dentatum subsp. kimuranum was tetraploid (2n = 4x = 28), and other intraspecific taxa of I. dentatum were triploid (2n = 3x = 21). The ITS sequences contained only 13 variable sites (2.7 %) among intraspecific taxa, suggesting that the I. dentatum complex originated by autoploidy. Our AFLP-based analysis separated these taxa into five groups: (1) I. dentatum subsp. kimuranum and subsp. kitayamense; (2) subsp. nipponicum and var. albiflorum; (3) two types of subsp. dentatum and f. atropurpureum; (4) f. amplifolium; (5) subsp. ozense and new taxa, which were discovered in this study. The identified clusters significantly differed from morphological classifications. For example, morphologically similar variety and forms of I. dentatum subsp. nipponicum were separated into different groups. Taken together with the ploidy data, this study proposes a possible evolutionary history generating the current taxonomic combinations of the I. dentatum complex. The proposed model suggests that diverse taxa recently emerged during migration to lowlands.     

Morphometric analysis of Schizachyrium condensatum (Poaceae) and related species

Our aim was to assess the degree of morphological differentiation of a group of taxa of Schizachyrium Nees, which presents similar inflorescences and shares habitat and geographic areas: Schizachyrium bimucronatum, S. condensatum, S. lactiflorum, S. microstachyum subsp. microstachyum, S. microstachyum subsp. elongatum, and S. plumigerum. To accomplish this purpose, 22 exomorphological traits were analyzed using multivariate methods. The results obtained support the identity of Schizachyrium condensatum and related species as independent taxa. In addition, the analysis evidences the reliability of several inflorescence characters, which had not been previously considered in the identification of the different taxa. Based upon the information obtained, a new identification key for these taxa was constructed.     

Variation in seed packaging of a fleshy‐fruited conifer provides insights into the ecology and evolution of multi‐seeded fruits

• The study of intraspecific seed packaging (i.e. seed size/number strategy) variation across different populations may allow better understanding of the ecological forces that drive seed evolution in plants. Juniperus thurifera (Cupressaceae) provides a good model to study this due to the existence of two subspecies differentiated by phenotypic traits, such as seed size and cone seediness (number of seeds inside a cone), across its range.
• The aim of this study was to analyse seed packaging (seed mass and cone seediness) variation at different scales (subspecies, populations and individuals) and the relationship between cone and seed traits in European and African J. thurifera populations.
• After opening more than 5300 cones and measuring 3600 seeds, we found that seed packaging traits followed different patterns of variation. Large‐scale effects (region and population) significantly contributed to cone seediness variance, while most of the seed mass variance occurred within individuals. Seed packaging differed between the two sides of the Mediterranean Sea, with African cones bearing fewer but larger seeds than the European ones. However, no differences in seed mass were found between populations when taking into account cone seediness. Larger cones contained more pulp and seeds and displayed a larger variation in individual seed mass.
• We validated previous reports on the intraspecific differences in J. thurifera seed packaging, although both subspecies followed the same seed size/number trade‐off. The higher seediness and variation in seed mass found in larger cones reveals that the positive relationship between seed and cone sizes may not be straightforward.We hypothesise that the large variation of seed size found within cones and individuals in J. thurifera, but also in other fleshy‐fruited species, could represent a bet‐hedging strategy for dispersal.