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.     

Cryptic speciation between Juniperus deltoides and Juniperus oxycedrus (Cupressaceae) in the Mediterranean

Analyses of individuals classically treated as Juniperus oxycedrus L. var. oxycedrus from Morocco, Portugal, Spain, France, Italy, Greece and Turkey, using DNA sequencing of nrDNA (ITS 1, 5.8S, ITS 2) plus RAPDs, leaf terpenoids and morphology revealed that two cryptic, genetically distinct but morphologically almost identical species are present. These species, J. oxycedrus L. var. oxycedrus and Juniperus deltoides R.P. Adams, are about as different from each other as Juniperus navicularis and Juniperus macrocarpa are from J. oxycedrus var. oxycedrus. Examination of herbarium specimens revealed that the two species are largely allopatric with J. deltoides occurring from Italy eastward through Turkey into the Caucasus Mts. and Iran. J. oxycedrus var. oxycedrus appears to be largely concentrated west of Italy (France, Spain, Portugal, Morocco). Cryptic speciation is discussed.     

Past,present, and future geographic range of the relict Mediterranean and Macaronesian Juniperus phoenicea complex

AimThe aim of this study is to model the past, current, and future distribution of J. phoenicea s.s., J. turbinata, and J. canariensis, based on bioclimatic variables using a maximum entropy model (Maxent) in the Mediterranean and Macaronesian regions.LocationMediterranean and Macaronesian.TaxonCupressaceae, Juniperus.MethodsData on the occurrence of the J. phoenicea complex were obtained from the Global Biodiversity Information Facility (GBIF.org), the literature, herbaria, and the authors’ field notes. Bioclimatic variables were obtained from the WorldClim database and Paleoclim. The climate data related to species localities were used for predictions of niches by implementation of Maxent, and the model was evaluated with ENMeval.ResultsThe potential niches of Juniperus phoenicea during the Last Interglacial period (LIG), Last Glacial Maximum climate (LGM), and Mid‐Holocene (MH) covered 30%, 10%, and almost 100%, respectively, of the current potential niche. Climate warming may reduce potential niches by 30% in RCP2.6 and by 90% in RCP8.5. The potential niches of Juniperus turbinata had a broad circum‐Mediterranean and Canarian distribution during the LIG and the MH; its distribution extended during the LGM when it was found in more areas than at present. The predicted warming in scenarios RCP2.6 and RCP8.5 could reduce the current potential niche by 30% and 50%, respectively. The model did not find suitable niches for J. canariensis during the LIG and the LGM, but during the MH its potential niche was 30% larger than at present. The climate warming scenario RCP2.6 indicates a reduction in the potential niche by 30%, while RCP8.5 so indicates a reduction of almost 60%.Main conclusionsThis research can provide information for increasing the protection of the juniper forest and for counteracting the phenomenon of local extinctions caused by anthropic pressure and climate changes.     

Phylogeography and genetic structure of the threatened Canarian Juniperus cedrus (Cupressaceae)

We used plastid sequences (trnL, trnL‐trnF, petN‐psbM and trnT‐trnL) to infer the phylogenetic relationships and inter‐island connections of the Canarian Juniperus cedrus, and AFLP fingerprints to assess its genetic diversity patterns. Maximum Likelihood, Maximum Parsimony and Bayesian methods suggest independent colonization events for the three Macaronesian junipers and support the monophyly of J. cedrus. Plastid sequences reveal a low genetic diversity (three haplotypes) and do not provide sufficient information to resolve its temporal and geographical origin. AFLPs indicate a greater isolation in J. cedrus than in other Macaronesian trees with similar distributions and dispersal syndromes. Gran Canaria harbours the least genetically diverse population, which justifies immediate conservation actions. This island and Tenerife also show independent genetic structure, meaning that genetic exchange from other islands should be avoided in eventual reinforcements. Populations from La Palma and La Gomera show the highest genetic diversity levels and number of polymorphic AFLPs, probably because a lower incidence of felling has allowed a less dramatic influence of genetic bottlenecks. We suggest that management efforts should prioritize populations from these islands to preserve the evolutionary potential of the species, but we also stress the importance of knowledge of the evolutionary history, genetic structure and ecological interactions in conservation strategies. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 175 , 376–394.     

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.     

Using genetic evaluation to guide conservation of remnant Juniperus communis (Cupressaceae) populations

• Many critically endangered plant species exist in small, genetically depauperate or inbred populations, making assisted gene flow interventions necessary for long‐term population viability. However, before such interventions are implemented, conservation practitioners must consider the genetic and demographic status of extant populations, which are strongly affected by species’ life‐history traits. In northwestern Europe, Juniperus communis, a dioecious, wind‐pollinated and bird‐dispersed gymnosperm, has been declining for the past century and largely exists in small, isolated and senescent populations.
• To provide useful recommendations for a recovery plan involving translocation of plants, we investigated genetic diversity and structure of populations in Belgium using four microsatellite and five plastid single‐nucleotide polymorphism (SNP) markers.
• We detected no clonality in the populations, suggesting predominantly sexual reproduction. Populations exhibited high genetic diversity (H_e = 0.367–0.563) and low to moderate genetic differentiation (F_ST ≤ 0.133), with no clear geographic structure. Highly positive inbreeding coefficients (F_IS = 0.221–0.507) were explained by null alleles, population substructuring and biparental inbreeding. No isolation by distance was observed among distant populations, but isolation at close geographic proximity was found. Patterns were consistent with high historical gene flow through pollen and seed dispersal at both short and long distances. We also tested four pre‐germination treatments among populations to improve germination rates; however, germination rates remained low and only cold‐stratification treatments induced germination in some populations.
• To bolster population regeneration, introductions of cuttings from several source populations are recommended, in combination with in situ management practices that improve seedling survival and with ex situ propagation.

     

Genetic variation in five Mediterranean populations of Juniperus phoenicea as revealed by inter-simple sequence repeat (ISSR) markers

BACKGROUND AND AIMS: The assessment of the genetic variability and the identification of isolated populations within a given species represent important information to plan conservation strategies on a genetic basis. In this work, the genetic variability in five natural populations of Juniperus phoenicea, three from Sardinia, one from Cyprus and the last one in the Maritime Alps was analysed by means of ISSRs, on the hypothesis that the latter could have been a refugial one during the last glaciation. METHODS: ISSRs were chosen because of their ability to detect variation without any prior sequence information. The use of three primers yielded 45 reproducible, polymorphic bands, which were utilized to estimate the basic parameters of genetic variability and diversity. KEY RESULTS: All of the populations analysed harboured an adequate amount of genetic variability, with H(S) = 0.1299. The proportion of genetic diversity between populations has been estimated by G(ST) = 0.12. The three Sardinian populations are separated, as tested by AMOVA, from the Cyprus and the continental ones. CONCLUSIONS: The results indicate that geographical isolation has represented a major barrier to gene flow in Juniperus phoenicea. This work represents a first step towards a full genetic characterization of a conifer from the Mediterranean, a world biodiversity hotspot confronted with climate change, and thus contributes towards the planning of genetics-informed conservation strategies.     

Effect of the Aegean Sea barrier between Europe and Asia on differentiation in Juniperus drupacea (Cupressaceae)

Juniperus drupacea is an eastern Mediterranean mountain tree with a disjunct geographical range. We hypothesized that this disjunct occurrence (the Peloponnese in Europe and the Taurus and Lebanon Mountains in Asia) should be reflected in the patterns of genetic and morphological diversity and differentiation. Nuclear microsatellite markers (nSSR) and biometric variables of the cones and seeds were examined on material sampled from four populations in Europe and eight in Asia. The Asian populations were characterized by a higher level of genetic diversity than the European populations. The genetic differentiation among populations was moderate but significant (F_ST = 0.101, P < 0.001). According to the clustering performed with BAPS, six genetically and geographically groups of populations were found: I and II from the Peloponnese; III from the Taurus Mountains; IV and V from the Anti‐Taurus Mountains; and VI from the Lebanon Mountains. The level of genetic differentiation among these six groups (4.30%, P = 0.012) probably reflects long‐lasting genetic isolation during the Pleistocene, as limited genetic admixture was found. In accordance with genetic analysis, the biometric investigations indicated a high level of morphological divergence between the European and Asian populations of the species, with further differentiation between the populations from the Taurus and Lebanon Mountains.     

Morphological and ecological comparison of populations of Juniperus turbinata Guss. and J. phoenicea L. from the Mediterranean region

The aim of the present study was the biometrical comparison and ecological analysis of six populations of Juniperus phoenicea complex from the western part of the species' geographic range, each represented by about 30 individuals. This is the first study to include the Sicilian population of J. turbinata sensu stricto. Seven features of the cones and seeds, two of the shoots and leaves and eight ratios were used to describe the individuals and populations which were then compared using statistical methods. Four of the analysed populations (representatives of J. turbinata s.s.) are closely related to each other. The two other populations representing J. phoenicea s.s. appear to be much more variable but on the whole the two taxa treated at a species level seem to be quite distinct from each other. It was also noted that there are two quite different ecologies of these species: the first is typical of the thermomediterranean belt which is only found at high altitudes in the mountains of north Africa, and the second circumscribed to the meso- or supramediterranean belt of western Spain, France and northwestern Italy. In many parts of its coastal distribution, Juniperus turbinata is endangered due to the construction of summer resorts and therefore actions to protect this species are needed.     

Genetic Diversity and Population Structure of Juniperus rigida (Cupressaceae) and Juniperus coreana

Enzyme electrophoresis was used to estimate genetic diversity and population structure of Juniperus rigida (Cupressaceae) and Juniperus coreana in Korea. In J. rigida, 16 of the 22 loci (72.7%) showed detectable polymorphism. Genetic diversity (0.224) was higher than average values for species with similar life history traits. The endemic species (J. coreana) was found to have fewer alleles per locus (1.39 vs. 1.61), fewer alleles per polymorphic locus (2.42 vs. 2.63), lower percent polymorphic locus (54.6 vs. 72.7%), and lower diversity (0.199 vs. 0.224) than J. rigida. These genetic diversity parameters indicated that J. coreana was genetically depauperate relative to its presumptive progenitor, J. rigida. Analysis of fixation indices showed a substantial deficiency of heterozygotes relative to Hardy–Weinberg expectations suggesting inbreeding in J. coreana. The G _ST values of J. rigida and J. coreana were 0.173 and 0.118 respectively. The indirect estimate of gene flow based on mean G _ST was moderate (Nm = 1.19 for J. rigida and 1.86 for J. coreana). This revised version was published online in July 2006 with corrections to the Cover Date.     

The colonization history of Juniperus brevifolia (Cupressaceae) in the Azores Islands

Background

A central aim of island biogeography is to understand the colonization history of insular species using current distributions, fossil records and genetic diversity. Here, we analyze five plastid DNA regions of the endangered Juniperus brevifolia, which is endemic to the Azores archipelago.

Methodology/Principal Findings

The phylogeny of the section Juniperus and the phylogeographic analyses of J. brevifolia based on the coalescence theory of allele (plastid) diversity suggest that: (1) a single introduction event likely occurred from Europe; (2) genetic diversification and inter-island dispersal postdated the emergence of the oldest island (Santa Maria, 8.12 Ma); (3) the genetic differentiation found in populations on the islands with higher age and smaller distance to the continent is significantly higher than that on the younger, more remote ones; (4) the high number of haplotypes observed (16), and the widespread distribution of the most frequent and ancestral ones across the archipelago, are indicating early diversification, demographic expansion, and recurrent dispersal. In contrast, restriction of six of the seven derived haplotypes to single islands is construed as reflecting significant isolation time prior to colonization.

Conclusions/Significance

Our phylogeographic reconstruction points to the sequence of island emergence as the key factor to explain the distribution of plastid DNA variation. The reproductive traits of this juniper species (anemophily, ornithochory, multi-seeded cones), together with its broad ecological range, appear to be largely responsible for recurrent inter-island colonization of ancestral haplotypes. In contrast, certain delay in colonization of new haplotypes may reflect intraspecific habitat competition on islands where this juniper was already present.     

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.     

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.     

Isolation and characterization of microsatellite DNA primers in Juniperus przewalskii Kom (Cupressaceae)

Juniperus przewalskii (Cupressaceae) is a dominant tree species endemic to the northeast Qinghai-Tibetan Plateau. This species plays an important role in maintaining the arid ecosystem in this region. However, natural distributions of this species have been declined. In order to develop effective conservation methods, it is important to know the distribution of the genetic diversity within and among populations. In this study, we developed nine new microsatellite loci for this species. We used the combining biotin capture method to enrich AG/CT/AC/GGT microsatellites. The polymorphisms of each locus were further assessed in 12 individuals from four geographically distant populations. The number of alleles per locus varied from three to six and expected heterozygosity ranged from 0.58 to 0.70. These loci together provide a useful tool to investigate the genetic diversity of this species. In addition, all markers have been crossly checked in the other four congeneric species.     

The coastal plant communities of Juniperus macrocarpa in the Mediterranean region

Juniperus macrocarpa is a threatened taxon growing on coastal areas of the Mediterranean region. A remarkable part of its communities are included in Juniperion turbinatae, considered as priority habitats for conservation in the Habitats Directive 92/43 EEC. We have summarized the current status of this type of vegetation, including the described associations of this alliance in order to analyse the variability of the syntaxa, based on its floristic composition and geographic distribution. The available relevés were arranged in a synthetic table and subjected to a detendred correspondence analysis. Two new suballiances (Juniperenion turbinatae and Asparago orientalis-Juniperenion macrocarpae) have been proposed. A typological classification of these communities with ecological and biogeographical diagnoses is given.     

基于叶绿体DNA trnT-trnF序列研究祁连圆柏的谱系地理学

由于青藏高原的地貌效应,第四纪冰期气候的反复变化应对现今该地区生物的地理分布及其居群遗传结构产生重大影响。本文对这一地区特有分布物种祁连圆柏Juniperus przewalskii Kom.整个分布区内20居群392个个体的trnT-trnF序列变化进行了研究;共发现3种单倍型(haplotype),构成两种地理区域:高原台面上的居群主要固定Hap A,而Hap A、Hap B和Hap C在高原边缘居群均有分布。所有居群总的遗传多样性HT = 0.511,GST= 0.847。在低海拔的高原边缘,Hap A、Hap B和Hap C高频率固定在不同的居群中,表明可能存在多个不同的避难所,居群反复缩小和扩张的瓶颈效应造成了遗传多样性的丢失。而边缘的一个居群含有两种相似单倍型频率则可能是冰期后迁移融合而成或者该居群在冰期经受的瓶颈作用更弱。高原台面东部间断分布的居群只固定Hap A,表明它们可能经历了冰期后共同的回迁过程和由此产生的奠基者效应。我们的研究结果表明祁连圆柏在冰期可能存在多个避难所,瓶颈效应和奠基者效应造成了这些居群现在的遗传多样性分布式样。     

柏科4个属(Fokienia、Cupressus、Chamaecyparis和Juniperus)植物雌球果的发育(英文)

通过扫描电镜和常规石蜡切片技术,观察了柏科4个属(Fokienia、Cupressus、Chamaecyparis和Juniperus)植物雌球果中胚珠的发育及苞片的结构变化。在所有研究的种类中,可育苞片腋部最先观察到的结构是一扁平的突起,并在其上分化出发育为胚珠的胚珠原基。在雌球果的发育过程中,未观察到独立的珠鳞发育。不同的种中,胚珠的数量和发育顺序有所不同,但苞片的发育是相似的。传粉前,苞片的结构与叶相似。传粉后,由于剧烈的居间生长,苞片发育为盾形,形成球形的球果。另外,在发育后期,苞片内维管系统变得复杂,并且在近轴面有反向的维管束发育。我们还对柏科植物雌球果的形态学特性及其可能的演化趋势进行了讨论。     

柏科4个属(Fokienia、Cupressus、Chamaeyparis 和Juniperus)植物雌球果的发育

通过扫描电镜和常规石蜡切片技术,观察了柏科4个属(Fokienia、Cupressus、Chamaeyparis 和Juniperus)植物雌球果中胚珠的发育及苞片的结构变化.在所有研究的种类中,可育苞片腋部最先观察到的结构是一扁平的突起,并在其上分化出发育为胚珠的胚珠原基.在雌球果的发育过程中,未观察到独立的珠鳞发育.不同的种中,胚珠怕数量和发育顺序有所不同,但苞片的发育是相似的.传粉前,苞片的结构与叶相似.传粉后,由于剧烈的居间维管束发育.我们还对柏科植物雌球果的形态学特性及其可能的演化趋势进行了讨论.     

Morphological variation of Juniperus oxycedrus subsp. oxycedrus (Cupressaceae) in the Mediterranean region

The intra- and inter-population variation of Juniperus oxycedrus subsp. oxycedrus were examined biometrically on the basis of morphological characters of needles, seeds and cones. Cluster analysis of Euclidean distances and discriminant analysis were performed to verify the hypothesis of geographic differentiation and origin from the West- and East-Mediterranean Pleistocene refugial regions. The West-Mediterranean populations of J. oxycedrus subsp. oxycedrus differed from the eastern ones. The intra-populational differentiation of three of the West-Mediterranean populations was significantly higher than within the other West- and all the East-Mediterranean ones.Results confirm the hypothesis of probable different origins of the East- and West-Mediterranean populations of the taxon.