Species Of Araucaria Research Articles

Gastroprotective and anti-Helicobacter pylori potentials of essential oils from the oleoresins of Araucaria bidwillii and Araucaria heterophylla

Plant resins or oleoresins comprise a chemically complex mixture of different classes of compounds. Oleoresin of the genus Araucaria combines essential oil (EO) and resin. It possesses gastroprotective, cytotoxic, and timicrobial, antipyretic, and anti-inflammatory activities. The study aimed to investigate the EOs from the oleoresins of two Araucaria species, A. bidwillii and A. heterophylla, chemically and biologically for their gastroprotective, anti-inflammatory, antioxidant, and anti-Helicobacter pylori potentials. The chemical composition of both species cultivated in Egypt was analyzed with GC-MS and compared with those cultivated abroad using principal component analysis (PCA). There were 37 and 17 secondary metabolites identified in A. heterophylla and A. bidwillii, respectively. The EOs of both species showed a pronounced inhibitory effect on Helicobacter pylori activity in vitro. The gastroprotective effect was assessed in vivo using ethanol-induced gastric ulcer model in rats. Inflammatory cytokines, oxidative stress, and the nuclear factor-kappa B (NF-κB) biomarkers were assessed in the stomach tissues. The ulcer index and percentage of ulcer protection were determined. Stomach sections were examined histopathologically by staining with (H/E) and periodic acid Schiff (PAS). Moreover, the proliferative index was determined using the Ki-67 immunostaining. The treatment of rats with EOs (50, 100, and 200 mg/kg, orally) 1 hour prior to ethanol administration showed promising gastroprotective, anti-inflammatory, and antioxidant potentials. These findings declared the gastroprotective role played by both EOs with the superiority of A. bidwillii over A. heterophylla via modulation of oxidative stress/NF-κB/inflammatory cytokines. Their use can be recommended to protect against the recurrence of peptic ulcers.

Potential biomedical applications of Araucaria araucana as an antispasmodic, bronchodilator, vasodilator, and antiemetic: Involvement of calcium channels

Ethnopharmacological relevanceSince pre-Columbian era, the resin of Araucaria araucana tree has been used traditionally for the treatment of ulcers and wounds. Araucaria species have also been used to treat inflammation, respiratory problems, viral infections, ulcers, and rheumatoid, cardiovascular, and neurological disorders. Aims and objectiveDue to its popular use, the authors aimed to scrutinize the potential of this plant as an antispasmodic and an antiemetic agent. Furthermore broncho- and vasodilatory effects of this plant was explored to rationalize its folkloric uses. Materials and methodsAraucaria araucana crude extract (Aa.Cr) was evaluated in isolated preparations of rabbit jejunum, trachea, aorta, and atria to investigate the antispasmodic, bronchodilator, and vasodilator effects. The potential mechanistic approaches were compared with the standard drug ‘verapamil’. The antiemetic activity was determined and compared with the standard drug ‘domperidone’ via chick emesis model. ResultsAa.Cr dose-dependently relaxed both spontaneous and K+-induced contractions in the isolated jejunum preparations of rabbits. In concentration–response curves of calcium (Ca++), Aa.Cr also triggered the rightward shift like verapamil. Applying carbachol and phenylephrine (1 μM) and K+ (80 mM) to the isolated tracheal and aortic tissue preparation, respectively, resulted in broncho- and vasodilatory activities, respectively which may be due to the inhibition of Ca++ channels. Aa.Cr inhibited atrial force and spontaneous contractions in the rabbit's right atria. Aa.Cr exhibited significant antiemetic activity (P < 0.001 vs. saline) in dose-dependent (50–150 mg/kg) manner like domperidone. In silico molecular docking was performed to investigate the biological targets of purified components of Aa.Cr which revealed that cadinol dominantly targets β2 receptors to cause bronchodilation, however, eudesmin binds non-specifically to all the selected targets, while secoisolariciresinol mediated high hydrogen bonding with muscarinic receptors (M1 and M3) and Ca++ channels, thus shows the suggested mechanistic pathways of targeted activities. ConclusionsThe results of this study indicates that Aa.Cr may exhibit antispasmodic activity, bronchodilation, and vasodilation by inhibiting voltage-dependent Ca++ channels and release of subcellular calcium. This explains its folkloric use in hypertension, bronchospasms, gastrointestinal spasms, and emesis.

Standardization and quantitative analysis of Araucaria Heterophylla extract via an UPLC-MS/MS method and its formulation as an antibacterial phytonanoemulsion gel

Skin infections are among the bacterial infections that present significant therapeutic challenges due to antibiotic resistance. Recently, herbal products clutched a significant attention as safe replacements for other medications but their low aqueous solubility and poor bioavailability are considered major challenges which could be circumvented via formulation. As a species of genera Araucaria, Araucaria Heterophylla possesses pharmacological activities such as antioxidant and antibacterial actions, and this study aimed to standardize the extract of the plant against 4ʹʹʹmethoxyamentoflavone (as a main component of the extract) through a validated UPLC-MS/MS method and evaluate its antibacterial activity, which was followed by loading the standardized extract into a nanoemulsion to form a phytonanoemulsion (PNE), where the design analysis and optimization were performed through a simplex lattice design. The optimized PNE (PNE 3) was then loaded into HPMC/Pluronic F-127 gel (in ratio 1:4) to sustain the release of the active constituent. The heightened penetrability of PNE 3 gel was visualized via confocal laser scanning microscopy, and its prolonged effect was proved thru an in vivo study conducted on male Wistar rats. A histopathological study revealed the safety of the formulation when applied topically. Thus, PNE gel could be a potentially broad-spectrum antibacterial drug delivery system.

Metabolomic profiling of three Araucaria species, and their possible potential role against COVID-19

The COVID-19 pandemic in Egypt is a part of the worldwide global crisis of coronavirus 2 (SARS-CoV-2). The contagious life-threatening condition causes acute respiratory syndrome. The present study aimed to assess the compounds identified by LC-MS of the methanolic leaves extracts from three conifers trees cultivated in Egypt (Araucaria bidwillii, Araucaria. cunninghamii and Araucaria heterophylla) via docking technique as potential inhibitor of COVID-19 virus on multiple targets; viral main protease (Mpro, 6LU7), non-structural protein-16 which is a methyl transferase (nsp16, 6W4H) and RNA dependent RNA polymerase (nsp12, 7BV2). Among the three targets, nsp16 was the best target recognized by the tested compounds as can be deduced from docking studies. Moreover, the methanolic extract of A. cunninghamii showed the highest radical-scavenging activity using (DPPH test) with 53.7 µg/mL comparable to ascorbic acid with IC50 = 46 µg/mL The anti-inflammatory potential carried using enzyme linked immunoassay showed the highest activity for A. cunninghamii and A. bidwillii followed by A. heterophylla with IC50 = 23.20 ± 1.17 µg/mL, 82.83 ± 3.21 µg/mL and 221.13 ± 6.7 µg/mL, respectively (Celecoxib was used as a standard drug with IC50 = 141.92 ± 4.52 µg/mL). Moreover, a molecular docking study was carried for the LC-MS annotated metabolites to validate their anti-inflammatory inhibitory effect using Celecoxib as a reference compound and showed a high docking score (−7.7 kcal/mol) for Octadecyl (E) P-coumarate and (−7.3 kcal/mol) for secoisolariciresinol rhamnoside. Communicated by Ramaswamy H. Sarma

A new specimen of Araucaria fricii from the early Coniacian of the Bohemian Massif, Central Europe

A specimen of Araucaria fricii is described from the upper part of the Teplice Formation in the Bohemian Cretaceous Basin. It extends the first occurrence of A. fricii from the mid-Coniacian back to the early Coniacian. Found in the Radovesice locality near Kučlín in the northern part of the Czech Republic, it is characterised by a deltoid cone scale complex with a centrally placed seed. It is compared to the type material of A. fricii from the mid-Coniacian Březno Formation and other European Cretaceous species of Araucaria. The taphonomy and palaeoecology of A. fricii is briefly discussed.

Phylogenetic biogeography of Araucaria (Araucariaceae)

The analysis of molecular data of all 19 recent species of the genus Araucaria Juss. (Araucariaceae) wascarried out and original cladogramm was build. Based on the cladogramm, the map of correlations of the phylogeneticrelationships and the distribution of all recent Araucaria species is proposed.

Diversification of crown group Araucaria: the role of Araucaria famii sp. nov. in the mid-Cretaceous (Campanian) radiation of Araucariaceae in the Northern Hemisphere.

Exceptional anatomical preservation of a fossil araucarian seed cone from a marine carbonate concretion from Vancouver Island, British Columbia, Canada provides unusually complete evidence for cone structure including seeds, megagametophytes, microgametophytes, and embryos of an Upper Cretaceous (Campanian) species of Araucaria, providing important new insights into the structure and relationships of Cretaceous Northern Hemisphere Araucariaceae. The cone was studied from serial thin sections prepared by the coal ball peel technique. Phylogenetic analysis using a modified morphological matrix with both discrete and continuous characters was performed using TNT version 1.5. The nearly spherical cone, 6 × 6 cm in diameter, has helically arranged cone-scale complexes, consisting of a large bract with an upturned tip and a small, fleshy ovuliferous scale. Vascularization of the cone-scale complex is single at its origin. Widely winged bracts, with a bulging base, contain numerous vascular bundles, interspersed with transfusion tissue, and a large number of resin canals. Seeds are ovoid, 1.2 cm long, 1.2 cm in diameter. Nucellus is free from the integument, except at its base, with a convoluted apex, containing possible pollen tubes. Megagametophytes and mature cellular embryos occur in several seeds. This small cone with attached, imbricate leaves, wide bracts, and unusually large seeds, most closely resembles those of Araucaria Section Eutacta. Width and continuity of secondary xylem in the cone axis, and intact cone-scale complexes indicate that this cone probably did not disarticulate readily at maturity. When added to a modified, previously published phylogenetic analysis, Araucaria famii sp. nov. enhances our understanding of the Cretaceous radiation of Northern Hemisphere Araucaria Section Eutacta.

Eocene Araucaria Sect. Eutacta from Patagonia and floristic turnover during the initial isolation of South America.

Eocene floras of Patagonia document biotic response to the final separation of Gondwana. The conifer genus Araucaria, distributed worldwide during the Mesozoic, has a disjunct extant distribution between South America and Australasia. Fossils assigned to Australasian Araucaria Sect. Eutacta usually are represented by isolated organs, making diagnosis difficult. Araucaria pichileufensis E.W. Berry, from the middle Eocene Río Pichileufú (RP) site in Argentine Patagonia, was originally placed in Sect. Eutacta and later reported from the early Eocene Laguna del Hunco (LH) locality. However, the relationship of A. pichileufensis to Sect. Eutacta and the conspecificity of the Araucaria material among these Patagonian floras have not been tested using modern methods. We review the type material of A. pichileufensis alongside large (n = 192) new fossil collections of Araucaria from LH and RP, including multi-organ preservation of leafy branches, ovuliferous complexes, and pollen cones. We use a total evidence phylogenetic analysis to analyze relationships of the fossils to Sect. Eutacta. We describe Araucaria huncoensis sp. nov. from LH and improve the whole-plant concept for Araucaria pichileufensis from RP. The two species respectively resolve in the crown and stem of Sect. Eutacta. Our results confirm the presence and indicate the survival of Sect. Eutacta in South America during early Antarctic separation. The exceptionally complete fossils significantly predate several molecular age estimates for crown Eutacta. The differentiation of two Araucaria species demonstrates conifer turnover during climate change and initial South American isolation from the early to middle Eocene.

Cuticle ultrastructure in Brachyphyllum garciarum sp. nov (Lower Cretaceous, Argentina) reveals its araucarian affinity

A detailed and extensive study of a new species, Brachyphyllum garciarum sp. nov., was carried out through the analysis of the gross morphology and the cuticle fine details, structure and ultrastructure characters of its leaves using light microscope and scanning and transmission electron microscope. The fossils consist of compressions of incomplete twigs with well-preserved cuticle, collected from pelitic levels of the Springhill Formation (lower Hauterivian/lower Barremian) at the Río Correntoso locality in the Santa Cruz province, Argentina. The twigs have adpressed scale-like leaves spirally disposed. Leaves have a rhomboidal to pyramidal shape, a width and length always in a 1:1 ratio, margin entire and apex mostly rounded. Leaves are amphistomatic with stomatal apparatuses occurring in groups of narrow-wedge shape along the leaf axis. Stomatal apparatuses are close to each other with subsidiary cells in contact; the guard cells are sunken, with marked polar extensions and thickened mouth. Remnants of hypodermis cells are present in both foliar epidermis. Ultrastructurally, four types of cuticles were observed and evaluated statistically in detail: the ordinary epidermal cell upper and lower cuticles and the subsidiary cell cuticle are compounded of A2 granular layer and a spongy B1 layer somewhat mixed with cell wall remnants, while the guard cell cuticle presents only a spongy B1 layer. An Energy Dispersive Spectroscopy study was also made revealing the presence of 10 characteristic ratios with taxonomic importance, at least at the species level: N/Na, S/Cl, S/Na, Cl/K, K/Na as the most important; followed by N/Ca, S/K, Cl/Ca, Cl/Na; and S/Ca being the least reliable. The combination of morphological and cuticle ultrastructure features of Brachyphyllum garciarum sp. nov. are unique and clearly different from other contemporary species of the genus from Western Gondwana. Moreover, the ultrastructure of the foliar cuticle suggests a highly probable affinity with the Araucariaceae Family, more precisely with the genus Araucaria. Of the four recognized sections among the living species of Araucaria, the leaf morphology of B. garciarum sp. nov. is mostly similar with some species of the Eutacta section.

Overlooked Parrot Seed Dispersal in Australia and South America: Insights on the Evolution of Dispersal Syndromes and Seed Size in Araucaria Trees

While Psittaciformes (parrots and allies) are well recognized as highly-mobile seed predators, their role as seed dispersers has been overlooked until very recently. It remains to be determined whether this role is anecdotic or is a key mutualism for some plant species. We recently found that the large nut-like seeds of the two South American Araucaria tree species (A. araucana in Andean forests and A. angustifolia in Atlantic forests, weighing c. 3.5 and 7g, respectively) are frequently dispersed, and to long distances, by parrots. Moreover, both observational and experimental work demonstrated that dispersed seeds can germinate faster after partial predation by parrots. Here, we hypothesized that a third, even larger-seeded (17.5g) congeneric Australian species (A. bidwillii) is also dispersed by parrots. We surveyed 52 A. bidwillii and 42 A. cunninghamii (a sympatric species with small winged seeds, c. 0.2g) during the seeding period. We found that sulphur-crested cockatoos (Cacatua galerita) consumed large amounts of seeds from all of the A. bidwillii trees surveyed. Cockatoos dispersed ca. 30% of the seeds they removed from the mother tree, carrying the seeds to distant perches for handling or dropped them while flying. Dispersal distances ranged between 10 and 153 m (mean = 61m). Most seeds handled for consumption (93%) were fully eaten but others were dropped intact (3%) or only partially eaten (4%), and germination was confirmed for both intact and partially-eaten dispersed seeds. Moreover, seeds dropped by cockatoos facilitated secondary seed dispersal by conspecifics and another three bird species. We found no evidence of other primary dispersal species for A. bidwillii, while the small, winged seeds of A. cunninghamii were just dispersed through barochory and anemochory. The seed weight of the three Araucaria species dispersed by zoochory is strongly related to the body mass of their main seed-dispersing parrot species. These results support a role for parrots as key dispersers of the three large-seeded Araucaria species around the world, and suggest that large seeds may have evolved–at least partially–as an adaptation that allows trees to attract parrots, satiate them, and benefit from their long-distance seed dispersal services.

Evaluación del estado sanitario de individuos de Araucaria araucana a través de imágenes hiperespectrales

&lt;p&gt;The &lt;em&gt;Araucaria araucana&lt;/em&gt; is an endemic species from Chile and Argentina, which has a high biological, scientific and cultural value and since 2016 has shown a severe affection of leaf damage in some individuals, causing in some cases their death. The purpose of this research was to detect, from hyperspectral images, the individuals of the Araucaria species (&lt;em&gt;Araucaria araucana&lt;/em&gt; (Molina and K. Koch)) and its degree of disease, by isolating its spectral signature and evaluating its physiological state through indices of vegetation and positioning techniques of the inflection point of the red edge, in a sector of the Ralco National Reserve, Biobío Region, Chile. Seven images were captured with the HYSPEX VNIR-1600 hyperspectral sensor, with 160 bands and a random sampling was carried out in the study area, where 90 samples of Araucarias were collected. In addition, from the remote sensing techniques applied, spatial data mining was used, in which Araucarias were classified without symptoms of disease and with symptoms of disease. A 55.11% overall accuracy was obtained in the classification of the image, 53.4% in the identification of healthy Araucaria and 55.96% in the identification of affected Araucaria. In relation to the evaluation of their sanitary status, the index with the best percentage of accuracy is the MSR (70.73%) and the one with the lowest value is the SAVI (35.47%). The positioning technique of the inflection point of the red edge delivered an accuracy percentage of 52.18% and an acceptable Kappa index.&lt;/p&gt;

Araucaria bract-scale complex and associated foliage from the early-middle Eocene of Antarctica and their implications for Gondwanan biogeography

ABSTRACTThe early-middle Eocene Fossil Hill flora from King George Island in the Antarctic Peninsula regions is one of the most diverse Cenozoic plant assemblages in Antarctica. It represents a rich Nothofagus and conifer dominated vegetation. These plant fossils, especially conifers are of crucial importance for understanding the biogeographic history of the Gondwanan plants. Here we describe the first Araucaria bract-scale complex, A. fildesensis sp. nov., and associated foliage from the Fossil Hill flora and tentatively assigned them to the Section Eutacta, which is today restricted to Australasia, based on comparison with extant material. This study confirms that Araucaria species with small bract-scale complexes, small scale-like mature leaves and awl-shaped juvenile leaves like those of extant Section Eutacta, lived in Antarctica in the Eocene. These fossils provide potential evidence for the trans-Antarctic floristic changes of Araucaria species in the Section Eutacta between southern Australia and southern South America during the Eocene, when Antarctica was ice-free and forested.

Chemical profiling and antiproliferative effect of essential oils of two Araucaria species cultivated in Egypt

Araucaria is a small genus of evergreen coniferous trees, widely used for ornamental and timber purposes. Araucaria species are rich in essential oils. In this study, the essential oils obtained from the foliage of Araucaria heterophylla (Salisb.) and Araucaria bidwillii Hook were analyzed using GC and GC/MS. Cytotoxicity of both essential oils was assessed on three human cancer cell lines using MTT assay. Twenty nine compounds were annotated in A. heterophylla essential oil, whereas forty three were identified in A. bidwillii accounting for 95.16% and 81.06% of the total volatile oil composition, respectively. α-Pinene (70.85%), d-limonene (4.26%) and germacrene D (2.99%) represented the major compounds in A. heterophylla. Beyerene (35.65%), trans-nerolidol (13.66%) and ɣ-elemene (6.09%) dominated in A. bidwillii. Both oils showed potent cytotoxicity against all tested cancer cell lines (Hep-G2, MCF-7 and Caco-2) exhibiting IC50 values of 0.70, 3.20 and 1.10 μg/ml for A. heterophylla; and 1.67, 1.10 and 1.32 μg/ml for A. bidwillii, respectively. Caspase-3 activation indicated that the cytotoxicity of A. bidwillii essential oil is mediated via caspase-dependent apoptosis. These findings suggest that the essential oils of A. heterophylla and A. bidwilli offer promising anticancer drug candidates pertaining to the synergistic effects of their individual components.

Changes of wood anatomical characters of selected species of Araucaria- during artificial charring - implications for palaeontology

ABSTRACT Charcoal is widely accepted as evidence of the occurrence of palaeo-wildfire. Although fossil charcoal remains have been used in many studies, investigation into the anatomical changes occurring during charring are few. The present study analyses changes in selected anatomical characters during artificial charring of modern wood of three species of the genus Araucaria (i.e. Araucaria angustifolia, Araucaria bidwillii and Araucaria columnaris). Wood samples of the studied species was charred under controlled conditions at varying temperatures. Measurements of anatomical features of uncharred wood and artificial charcoal were statistically analysed. The anatomical changes were statistically correlated with charring temperatures and most of the parameters showed marked decreases with increasing charring temperature. Compared to the intrinsic variability in anatomical features, both within and between growth rings of an individual plant, the changes induced by temperature account only for a comparatively small percentage of the observed variability. Regarding Araucaria charcoal, it seems possible that at least general taxonomic and palaeoenvironmental implications can be drawn from such material. However, it is not clear so far whether these results and interpretations based on only three taxa, can be generalized for the entire family and anatomically similar fossil taxa or not.

ARAUCARIA GOROENSIS (ARAUCARIACEAE), A NEW MONKEY PUZZLE FROM NEW CALEDONIA, AND NOMENCLATURAL NOTES ON ARAUCARIA MUELLERI

Araucaria goroensis R.R.Mill &amp; Ruhsam sp. nov., a new monkey puzzle species from New Caledonia, is described and illustrated with photographs from the field and from herbarium specimens. Previously confused with Araucaria muelleri, it is more similar to A. rulei. It is distinguished from the latter species by its larger leaves, microsporophylls without a shouldered base, and shorter female cone bracts. It occurs in a very limited area of south-east New Caledonia, where its existence is threatened by nickel mining. Using the guidelines of the International Union for Conservation of Nature, we propose an assessment of Endangered for the new species and reassess Araucaria muelleri also as Endangered. A key to the seven species in the ‘large-leaved clade’ of New Caledonian species of Araucaria is given. The name Eutassa latifolia de Laub. is synonymised with Araucaria muelleri, and the recent typification of the latter name by Vieillard 1276 is rejected. Detailed reasoning is given for these nomenclatural acts.

On the Evolutionary History of Uleiella chilensis, a Smut Fungus Parasite of Araucaria araucana in South America: Uleiellales ord. nov. in Ustilaginomycetes.

The evolutionary history, divergence times and phylogenetic relationships of Uleiella chilensis (Ustilaginomycotina, smut fungi) associated with Araucaria araucana were analysed. DNA sequences from multiple gene regions and morphology were analysed and compared to other members of the Basidiomycota to determine the phylogenetic placement of smut fungi on gymnosperms. Divergence time estimates indicate that the majority of smut fungal orders diversified during the Triassic–Jurassic period. However, the origin and relationships of several orders remain uncertain. The most recent common ancestor between Uleiella chilensis and Violaceomyces palustris has been dated to the Lower Cretaceous. Comparisons of divergence time estimates between smut fungi and host plants lead to the hypothesis that the early Ustilaginomycotina had a saprobic lifestyle. As there are only two extant species of Araucaria in South America, each hosting a unique Uleiella species, we suggest that either coevolution or a host shift followed by allopatric speciation are the most likely explanations for the current geographic restriction of Uleiella and its low diversity. Phylogenetic and age estimation analyses, ecology, the unusual life-cycle and the peculiar combination of septal and haustorial characteristics support Uleiella chilensis as a distinct lineage among the Ustilaginomycotina. Here, we describe a new ustilaginomycetous order, the Uleiellales to accommodate Uleiella. Within the Ustilaginomycetes, Uleiellales are sister taxon to the Violaceomycetales.

Species-level determination of closely related araucarian resins using FTIR spectroscopy and its implications for the provenance of New Zealand amber.

Some higher plants, both angiosperms and gymnosperms, can produce resins and some of these resins can polymerize and fossilize to form ambers. Various physical and chemical techniques have been used to identify and profile different plant resins and have then been applied to fossilized resins (ambers), to try to detect their parent plant affinities and understand the process of polymerization, with varying levels of success. Here we focus on resins produced from today’s most resinous conifer family, the Araucariaceae, which are thought to be the parent plants of some of the Southern Hemisphere’s fossil resin deposits. Fourier transform infrared (FTIR) spectra of the resins of closely related Araucariaceae species were examined to test whether they could be distinguished at genus and species level and whether the results could then be used to infer the parent plant of a New Zealand amber. The resin FTIR spectra are distinguishable from each other, and the three Araucaria species sampled produced similar FTIR spectra, to which Wollemia resin is most similar. Interspecific variability of the FTIR spectra is greatest in the three Agathis species tested. The New Zealand amber sample is similar in key shared features with the resin samples, but it does differ from the extant resin samples in key distinguishing features, nonetheless it is most similar to the resin of Agathis australis in this dataset. However on comparison with previously published FTIR spectra of similar aged amber and older (Eocene) resinites both found in coals from New Zealand and fresh Agathis australis resin, our amber has some features that imply a relatively immature resin, which was not expected from an amber of the Miocene age.

Does complete plastid genome sequencing improve species discrimination and phylogenetic resolution in Araucaria?

Obtaining accurate phylogenies and effective species discrimination using a small standardized set of plastid genes is challenging in evolutionarily young lineages. Complete plastid genome sequencing offers an increasingly easy-to-access source of characters that helps address this. The usefulness of this approach, however, depends on the extent to which plastid haplotypes track morphological species boundaries. We have tested the power of complete plastid genomes to discriminate among multiple accessions of 11 of 13 New Caledonian Araucaria species, an evolutionarily young lineage where the standard DNA barcoding approach has so far failed and phylogenetic relationships have remained elusive. Additionally, 11 nuclear gene regions were Sanger sequenced for all accessions to ascertain the success of species discrimination using a moderate number of nuclear genes. Overall, fewer than half of the New Caledonian Araucaria species with multiple accessions were monophyletic in the plastid or nuclear trees. However, the plastid data retrieved a phylogeny with a higher resolution compared to any previously published tree of this clade and supported the monophyly of about twice as many species and nodes compared to the nuclear data set. Modest gains in discrimination thus are possible, but using complete plastid genomes or a small number of nuclear genes in DNA barcoding may not substantially raise species discriminatory power in many evolutionarily young lineages. The big challenge therefore remains to develop techniques that allow routine access to large numbers of nuclear markers scaleable to thousands of individuals from phylogenetically disparate sample sets.

Evolutionary diversification of new Caledonian Araucaria.

New Caledonia is a global biodiversity hotspot. Hypotheses for its biotic richness suggest either that the island is a ‘museum’ for an old Gondwana biota or alternatively it has developed following relatively recent long distance dispersal and in situ radiation. The conifer genus Araucaria (Araucariaceae) comprises 19 species globally with 13 endemic to this island. With a typically Gondwanan distribution, Araucaria is particularly well suited to testing alternative biogeographic hypotheses concerning the origins of New Caledonian biota. We derived phylogenetic estimates using 11 plastid and rDNA ITS2 sequence data for a complete sampling of Araucaria (including multiple accessions of each of the 13 New Caledonian Araucaria species). In addition, we developed a dataset comprising 4 plastid regions for a wider taxon sample to facilitate fossil based molecular dating. Following statistical analyses to identify a credible and internally consistent set of fossil constraints, divergence times estimated using a Bayesian relaxed clock approach were contrasted with geological scenarios to explore the biogeographic history of Araucaria. The phylogenetic data resolve relationships within Araucariaceae and among the main lineages in Araucaria, but provide limited resolution within the monophyletic New Caledonian species group. Divergence time estimates suggest a Late Cretaceous-Cenozoic radiation of extant Araucaria and a Neogene radiation of the New Caledonian lineage. A molecular timescale for the evolution of Araucariaceae supports a relatively recent radiation, and suggests that earlier (pre-Cenozoic) fossil types assigned to Araucaria may have affinities elsewhere in Araucariaceae. While additional data will be required to adequately resolve relationships among the New Caledonian species, their recent origin is consistent with overwater dispersal following Eocene emersion of New Caledonia but is too old to support a single dispersal from Australia to Norfolk Island for the radiation of the Pacific Araucaria sect. Eutacta clade.

Evolutionary dynamics of emblematic Araucaria species (Araucariaceae) in New Caledonia: nuclear and chloroplast markers suggest recent diversification, introgression, and a tight link between genetics and geography within species.

BackgroundNew Caledonia harbours a highly diverse and endemic flora, and 13 (out of the 19 worldwide) species of Araucaria are endemic to this territory. Their phylogenetic relationships remain largely unresolved. Using nuclear microsatellites and chloroplast DNA sequencing, we focused on five closely related Araucaria species to investigate among-species relationships and the distribution of within-species genetic diversity across New Caledonia.ResultsThe species could be clearly distinguished here, except A. montana and A. laubenfelsii that were not differentiated and, at most, form a genetic cline. Given their apparent morphological and ecological similarity, we suggested that these two species may be considered as a single evolutionary unit. We observed cases of nuclear admixture and incongruence between nuclear and chloroplast data, probably explained by introgression and shared ancestral polymorphism. Ancient hybridization was evidenced between A. biramulata and A. laubenfelsii in Mt Do, and is strongly suspected between A. biramulata and A. rulei in Mt Tonta. In both cases, extensive asymmetrical backcrossing eliminated the influence of one parent in the nuclear DNA composition. Shared ancestral polymorphism was also observed for cpDNA, suggesting that species diverged recently, have large effective sizes and/or that cpDNA experienced slow rates of molecular evolution. Within-species genetic structure was pronounced, probably because of low gene flow and significant inbreeding, and appeared clearly influenced by geography. This may be due to survival in distinct refugia during Quaternary climatic oscillations.ConclusionsThe study species probably diverged recently and/or are characterized by a slow rate of cpDNA sequence evolution, and introgression is strongly suspected. Within-species genetic structure is tightly linked with geography. We underline the conservation implications of our results, and highlight several perspectives.Electronic supplementary materialThe online version of this article (doi:10.1186/s12862-014-0171-6) contains supplementary material, which is available to authorized users.