Premise The Amazonian hyperdominant genus Eperua (Fabaceae) currently holds 20 described species and has two strongly different inflorescence and flower types, with corresponding different pollination syndrome. The evolution of these vastly different inflorescence types within this genus was unknown and the main topic in this study. Methods We constructed a molecular phylogeny, based on the full nuclear ribosomal DNA and partial plastome, using Bayesian inference and maximum likelihood methods, to test whether the genus is monophyletic, whether all species are monophyletic and if the shift from bat to bee pollination (or vice versa) occurred once in this genus. Results All but two species are well supported by the nuclear ribosomal phylogeny. The plastome phylogeny, however, shows a strong geographic signal suggesting strong local hybridization or chloroplast capture, rendering chloroplast barcodes meaningless in this genus. Conclusions With our data, we cannot fully resolve the backbone of the tree to clarify sister genera relationships and confirm monophyly of the genus Eperua . Within the genus, the shift from bat to bee and bee to bat pollination has occurred several times but, with the bee to bat not always leading to a pendant inflorescence.

During the environmental impact study for a proposed nickel mine near Weda Bay on Halmahera in North Moluccas (Maluku Utara Province), Indonesia, two unknown Euphorbiaceae were discovered. Morphological comparisons and molecular phylogenetic analyses using four markers (plastid trnL-F and rbcL, and nuclear ribosomal internal transcribed spacer and external transcribed spacer) indicated that they should be recognized as constituting a new, distinct genus of two species, which are described and illustrated here as Weda fragarioides and Weda lutea. The new taxa are members of the Australasian tribe Ricinocarpeae in subfamily Crotonoideae, and they are most closely related to Alphandia. In contrast with the otherwise mostly sclerophyllous Ricinocarpeae, Weda possesses stellate to dendritic hairs, large, long-petiolate, glandular leaves, and inflorescences with a pair of large, leafy, subopposite bracts. The two narrowly distributed species are distinguished from each other by vegetative and floral features, molecular data, and elevational preferences. Leaf elemental analysis of Weda indicated manganese, but not nickel, accumulation. Newly resolved generic relationships and potential morphological synapomorphies within Crotonoideae are discussed, and the circumscription of Ricinocarpeae is expanded from 7 to 11 genera.

In 2001 and 2002 an excavation conducted in the town of Alphen aan den Rijn in the Netherlands revealed the vestiges of a Roman fort, Castellum Albaniana, situated along the historical Rhine delta and used for centuries during the Roman occupation (41 AD 275 CE). Among the animal bones retrieved from the surrounding defensive ditches, remains of Lynx lynx bones were found. Lynx is currently not native to the Netherlands but might have been in historical times and it could have been transported to the Limes region by the Romans or caught in the direct surroundings of the castellum. In the present study, we describe the retrieved lynx bones initially identified based on morphology. We performed ancient DNA amplification, sequencing and alignment to confirm species identification and to determine the haplotype. Previous haplotyping of lynx from various sites reported by other studies has shown that lynx distribution in Europe during Roman times was very different from its current distribution. DNA analysis of cytochrome oxidase I and cytochrome B confirmed the identification of the animal species as Lynx lynx. Sequencing of the mitochondrial control region revealed that the animal carried a DNA haplotype, different from those from North Sea fossil lynx remains, but comparable to a haplotype found in southern France. Analysis of stable isotope of the bone materials, to determine the region where the animal lived, suggests the provenance of the animal from a region which comprises southern and central Europe including a part of the Netherlands.

'Here for you a smiling garden of everlasting flowers' is the inscription in Latin of a 16th century Italian herbarium kept in Leiden, the Netherlands, since 1690. The origin and botanical content of this herbarium, one of the oldest in existence today, have remained largely unknown. Here we present the plants included in this so-called 'En Tibi' herbarium, which comprises 473 specimens (455 taxa, 97 families), and discuss the geographical provenance of the book based on certain plant traits. The En Tibi is of great historical value as it contains some of the earliest herbarium records of numerous species, among which are useful plants such as oregano, thyme, tomato and hot pepper. Although prepared as a present, the En Tibi is a fine example of new botanical trends that arose in 16th century Italy. It is an attempt to reconstruct the herbals of classical authors such as Dioscorides, Theophrastus and Pliny, not with illustrations but with actual plant individuals. More than just a collection of medicinal plants, the En Tibi shows an emerging interest in the study of taxonomy and the discovery of new plants, unknown to classical authors. Analysis of the intrinsic and extrinsic traits of the plants reveals a temperate-Mediterranean origin, suggesting that the book was made in central or north-central Italy. Our botanical identification is the first necessary step to further elucidate the origin of the En Tibi and trace the mysterious compiler of this magnificent collection.

Middens (nests and caches) of Late Pleistocene arctic ground squirrels (Urocitellus parryii) that are preserved in the permafrost of Beringia archive valuable paleoecological data. Arctic ground squirrels selectively include the plant material placed in middens. To account for this selectivity bias, we used a multi-proxy approach that includes ancient DNA (aDNA) and macro- and microfossil analyses. Here, we provide insight into Pleistocene vegetation conditions using macrofossils, pollen, phytoliths and non-pollen palynomorphs, and aDNA collected from one such midden from the Yukon Territory (Canada), which was formed between 30,740 and 30,380 cal yr BP. aDNA confirmed the midden was constructed by U. parryii. We recovered 39 vascular plant and bryophyte genera and 68 fungal genera from the midden samples. Grass and other herbaceous families dominated vegetation assemblages according to all proxies. aDNA data yielded several records of vascular plants that are outside their current biogeographic range, while some of the recovered fungi yielded additional evidence for local occurrence of Picea trees during glacial conditions. We propose that future work on fossil middens should combine the study of macro- and microfossils with aDNA analysis to get the most out of these environmental archives.

The phylogenetic relationships within Specklinia (Pleurothallidinae; Orchidaceae) and related genera are re-evaluated using Bayesian analyses of nrITS and chloroplast matK sequence data of a wide sampling of species. Specklinia is found paraphyletic in the DNA based trees, with species alternatively assigned to Muscarella proven distinct, monophyletic and easily recognizable. Specklinia as such includes about 100 morphologically highly diverse species. Their phenotypic differences have prompted the creation of up to eleven generic names within this relatively small group. Here we show not only that these morphologically divergent species are closely related, but also that they can still be recognized by certain conserved morphological traits. The genera Acostaea, Areldia, Empusella, Cucumeria, Gerardoa, Pseudoctomeria, Sarcinula, Sylphia, Tribulago and Tridelta are found embedded within Specklinia, and therefore reduced under the synonymy of the latter. Specklinia is confirmed as sister to a clade that includes Platystele, Scaphosepalum and Teagueia. Five well-supported subgenera are proposed for Specklinia and are characterized both geographically and morphologically. The species belonging to each subgenus are listed. Incaea is synonymized with Dryadella, while Rubellia is reduced under Platystele. New combinations for several species are proposed. The criteria for the generic delimitation of Specklinia and other genera in the Pleurothallidinae are discussed.

Slimming cactus (Hoodia), found only in southwestern Africa, is a well-known herbal product for losing weight. Consequently, Hoodia extracts are sought-after worldwide despite a CITES Appendix II status. The failure to eradicate illegal trade is due to problems with detecting and identifying Hoodia using morphological and chemical characters. Our aim was to evaluate the potential of molecular identification of Hoodia based on DNA barcoding. Screening of nrITS1 and psbA-trnH DNA sequences from 26 accessions of Ceropegieae resulted in successful identification, while conventional chemical profiling using DLI-MS led to inaccurate detection and identification of Hoodia. The presence of Hoodia in herbal products was also successfully established using DNA sequences. A validation procedure of our DNA barcoding protocol demonstrated its robustness to changes in PCR conditions. We conclude that DNA barcoding is an effective tool for Hoodia detection and identification which can contribute to preventing illegal trade.

I ndian s nakeroot ( R auvolfia serpentina ) is a valuable forest product, root extracts of which are used as an antihypertensive drug. Increasing demand led to overharvesting in the wild. Control of international trade is hampered by the inability to identify root samples to the species level. We therefore evaluated the potential of molecular identification by searching for species‐specific DNA polymorphisms. We found two species‐specific indels in the rps 16 intron region for R . serpentina . Our DNA barcoding method was tested for its specificity, reproducibility, sensitivity and stability. We included samples of various tissues and ages, which had been treated differently for preservation. DNA extractions were tested in a range of amplification settings and dilutions. Species‐specific rps 16 intron sequences were obtained from 79 herbarium accessions and one confiscated root, encompassing 39 different species. Our results demonstrate that molecular analysis provides new perspectives for forensic identification of I ndian s nakeroot.

Of four species of Sargassaceae, representing the genera Carpophyllum, Cystoseira, Landsburgia, and Sargassum, the intergenic spacer of the ribosomal cistron was amplified using a forward primer annealing at the 3′-end of the large subunit (LSU) of the ribosomal cistron and a reverse primer annealing at the 5S rRNA gene. The PCR products were cloned and the DNA sequences of multiple clones were determined. Almost each clone showed a unique DNA sequence. Intra-individual variation of this LSU-5S intergenic spacer was extremely high and was characterized by great length variation and a high number of short tandem repeats. Sequences were unalignable and therefore it was concluded that the LSU-5S intergenic spacer is unsuitable for phylogenetic and phylogeographic studies of sargassacean taxa.

The National Herbarium of the Netherlands houses a 17th century, bound herbarium containing 51 dried specimens from Suriname, which was composed by the well-known botanist Paul Hermann (1646-1695). This is considered as the oldest documented herbarium collection not only for Suriname but for the Guianas region. Most specimens are accompanied by (pre-Linnaean) Latin or vernacular names and sometimes by Latin descriptions of the plants and their uses. To assess the importance of this collection for the present-day flora and ethnobotany of Suriname, we identified all specimens (one by using ancient DNA analysis), translated the Latin texts, traced back the origin of the herbarium in national archives, 17th century and modern literature and compared plant names and uses with present-day ethnobotanical data. We digitized the entire herbarium and made it available online (http://www.hermann-herbarium.nl). The specimens were probably collected around 1687 by a certain Hendrik Meyer, who had a keen interest in botany and indigenous plant use. The 48 species in the herbarium are almost all useful plants: cultivated crops, wild edible fruits, medicinal plants, timber trees, fish poison, colorants and roof thatch material. Most species are used similarly today, and more than half of the vernacular names still exist in the region. The presence of Abelmoschus esculentus and Sesamum indicum in the herbarium prove the early establishment of African food plants in the emerging plantation economy of Suriname. Unlike Hermann's collections from Ceylon and the Cape, this herbarium was never seen by Linnaeus and therefore does not contain any type specimens.

Tropical agarwood (Aquilaria) is in danger of extinction in the wild due to illegal logging. Its resin (Gaharu) is used for the production of highly valued incense throughout Asia. We have isolated and characterized microsatellite loci of Aquilaria crassna to detect the geographic origin of agarwood for forensic applications using a modified enrichment procedure based on the capture of repetitive sequences from restricted genomic DNA. We assessed the polymorphisms of five microsatellites amplified from fresh leaves of 22 trees from seven plantations in Vietnam and Thailand and dried leaves of a herbarium specimen of one wild tree. Cross specificity of these markers was confirmed on two related Aquilaria species occurring in China and Vietnam and one microsatellite locus was successfully amplified from wood and incense samples. Four of the loci were polymorphic and the number of alleles ranged from 3 to 15. The loci characterized here can provide a starting point for forensic identification of traded material and certification of sustainably produced agarwood.

Omdat we wilden weten welke boomsoort hier als waardplant gebruikt werd, vergeleken we de schors van de geinfecteerde stammetjes met de schors van bomen uit de omgeving. Het bleek echter voor divere professionele plantentaxonomen onmogelijk om op deze manier de identiteit van het hout vast te stellen. Met behulp van DNA sequenties van het chloroplast rbcL gen en trnL intron kon de schors echter geidentificeerd worden als ruwe iep (Ulmus glabra). Determineren met DNA lijkt een veelbelovende nieuwe techniek om waardplanten mee op naam te brengen als bloemen en/of vruchten ontbreken en traditionele determinatiesleutels te kort schieten.

Sequences of three different genes in 69 taxa of Amorphophallus were combined to reconstruct the molecular phylogeny of this species-rich Aroid genus. The data set was analyzed by three different methods, Maximum Parsimony, Maximum Likelihood and Bayesian analysis, producing slightly different tree topologies. Three major clades identified in all analyses reflect the biogeographical distribution of Amorphophallus. Some clades were supported by morphological characters such as sessile/nonsessile stigma, pollen opening mechanism, shape of the main segments of the lamina, growth cycle, and berry colour. When optimised, a nonsessile stigma may have evolved from a sessile one with several reversals. Pollen opening by connective rupturing evolved from pollen opening by pores. Unequally shaped segments of the lamina evolved from equally shaped segments. Simultaneously existing leaf and inflorescences evolved from alternating leaves and inflorescences. Blue, purple, green, and yellow berries evolved from red/orange/white ones.

The tribe Thomsonieae (Araceae) comprise two genera of palaeotropical herbs: Amorphophallus with approximately 200 species and Pseudodracontium with seven species. A phylogenetic study of the Thomsonieae is presented. Nucleotide sequences of the chloroplast gene matK and the trnL intron were analyzed for 46 Amorphophallus species, two Pseudodracontium species and six outgroups using maximum parsimony. The two data sets were analyzed separately and in combination. Amorphophallus can be divided into five major well-supported clades, which provide new hypotheses on infrageneric relationships. The branches connecting these clades, however, are poorly supported. Results are compared with previous morphology-based divisions of Amorphophallus. Remarkable results from this study are the monophyly of the African Amorphophallus species and the nesting of Pseudodracontium within Amorphophallus, making Amorphophallus in the traditional sense paraphyletic to Pseudodracontium. Communicating Editor: Gregory M. ...