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Background: Obtaining chloroplast genome sequences is important to increase the knowledge about the fundamental biology of plastids, to understand evolutionary and ecological processes in the evolution of plants, to develop biotechnological applications (e.g. plastid engineering) and to improve the efficiency of breeding schemes. Extraction of pure chloroplast DNA is required for efficient sequencing of chloroplast genomes. Unfortunately, most protocols for extracting chloroplast DNA were developed for eudicots and do not produce sufficiently pure yields for a shotgun sequencing approach of whole plastid genomes from the monocot grasses., Methodology/principal Findings: We have developed a simple and inexpensive method to obtain chloroplast DNA from grass species by modifying and extending protocols optimized for the use in eudicots. Many protocols for extracting chloroplast DNA require an ultracentrifugation step to efficiently separate chloroplast DNA from nuclear DNA. The developed method uses two more centrifugation steps than previously reported protocols and does not require an ultracentrifuge., Conclusions/significance: The described method delivered chloroplast DNA of very high quality from two grass species belonging to highly different taxonomic subfamilies within the grass family (Lolium perenne, Pooideae; Miscanthus x giganteus, Panicoideae). The DNA from Lolium perenne was used for whole chloroplast genome sequencing and detection of SNPs. The sequence is publicly available on EMBL/GenBank.

Background and Aims: Representatives from Papaver, Roemeria, Stylomecon and Meconopsis were studied to elucidate phylogenetic relationships between Papaver and these closely allied genera., Methods: Two molecular data sets were used individually and combined and included sequences from the internally transcribed spacer region (ITS) of 18S-26S nuclear ribosomal DNA and the trnL intron and the trnL-trnF intergenic spacer region of plastid DNA., Key Results: Parsimony analysis demonstrated that the genus is not monophyletic unless the closely related Roemeria, Stylomecon and Meconopsis cambrica are included in a revised circumscription of Papaver. Three distinct clades are resolved in a combined ITS and trnL-F analysis. Clade 1 consists of Papaver sect. Meconella and Asian Meconopsis. Clade 2 contains a group here identified as Papaver s.s., comprising sections Carinatae, Meconidium, Oxytona, Papaver, Pilosa, Pseudopilosa and Rhoeadium. Clade 3 consists of Papaver sect. Argemonidium and Roemeria refracta. A number of diagnostic indels support these groupings. Within clade 2, sects. Papaver and Rhoeadium are either not monophyletic or lack evidence supporting their monophyly., Conclusions: The results of this molecular analysis indicate that a number of morphological characters such as valvate capsule dehiscence, dark or light filaments and sessile stigmatic discs have arisen in parallel. The phylogenetic trees are incongruent with the existing taxonomy of Papaver, and a revised classification is suggested.

Taxus is a genus of trees and shrubs with high value in horticulture and medicine as a source of the anticancer drug paclitaxel. The taxonomy of the group is complex due to the lack of diagnostic morphological characters and the high degree of similarity among species. Taxus has a wide global geographic distribution and some taxonomists recognize only a single species with geographically defined subgroups, whereas others have described several species. To address these differences in taxonomic circumscription, phylogenetic analyses were conducted on DNA sequences using Maximum Likelihood, Bayesian Inference and TCS haplotype networks on single and combined gene regions obtained for the nuclear ribosomal ITS region and the plastid trnL intron and trnL-F intergenic spacer. Evidence is presented for the sister group status of Pseudotaxus to Taxus and the inclusion of Amentotaxus, Austrotaxus, Cephalotaxus and Torreya within Taxaceae. Results are consistent with the taxonomic recognition of nine species: T. baccata, T. brevifolia, T. canadensis, T. cuspidata, T. floridana, T. fuana, T. globosa, T. sumatrana and T. wallichiana, but evidence is found for less species distinction and considerable reticulation within the T. baccata, T. canadensis and T. cuspidata group. We compare the results to known taxonomy, biogeography, present new leaf anatomical data and discuss the origins of the hybrids T. Xmedia and T. Xhunnewelliana. [ABSTRACT FROM AUTHOR]

New and comprehensive collections of the perennial rhizomatous reed canary grass ( Phalaris arundinacea) were made in NW Europe along north-to-south and east-to-west clines from Denmark, Germany, Ireland, Poland, Sweden and the United Kingdom. Rhizome, seed and leaf samples were taken for analysis and genetic resource conservation. A subsample covering the geographical range was characterized using plastid genome sequencing and SNP discovery generated using a long-read PCR and MiSeq sequencing approach. Samples were also subject to flow cytometry and all found to be tetraploid. New sequences were assembled against a Lolium perenne (perennial ryegrass) reference genome, and an average of approximately 60% of each genome was aligned (81 064 bp). Genetic variation was high among the 48 sequenced genotypes with a total of 1793 SNPs, equating to 23 SNPs per kbp. SNPs were subject to principal coordinate and Structure analyses to detect population genetic groupings and to examine phylogeographical pattern. Results indicate substantial genetic variation and population genetic structuring of this allogamous species at a broad geographical scale in NW Europe with plastid genetic diversity organized more across an east-to-west than a north-to-south cline. [ABSTRACT FROM AUTHOR]

Aim We examined phylogenetic relationships and spatio-temporal diversification in Indian Ocean Primulaceae, assessing correlations between speciation rates, geographical expansion and ecomorphological specialization. Location Madagascar and Indian Ocean Islands. Methods We conducted phylogenetic analyses with plastid and nuclear DNA sequences of Primulaceae using maximum likelihood and Bayesian algorithms, and estimated divergence times using a Bayesian relaxed molecular clock. Temporal changes in diversification rate and possible correlations with the biogeographical history of the group were examined. We performed parametric ancestral area reconstruction incorporating a stratified palaeogeographical model that reflects changes in terrestrial configuration and the presence of phytogeographical connections through time in the western Indian Ocean Basin. Shifts in diversification rate were compared with ancestral area assignments and divergence age estimates. Results Indian Ocean Primulaceae were recovered as monophyletic with a sister relationship to Asian Ardisia. Oncostemum, a genus confined to Madagascar and the Comoros, was resolved as paraphyletic by the inclusion of a monophyletic Mascarene Badula group consisting of single-island endemics. We found evidence for diversification bursts early in the history of Indian Ocean Primulaceae that correspond closely to the sequence of dispersal and the appearance of newly formed Mascarene Islands. Age estimates suggest a dispersal to Rodrigues that is older than the estimated geological age of the island. Main conclusions Results suggest a Madagascan origin of Indian Ocean Primulaceae with subsequent dispersal to the Mascarenes in the middle to late Miocene, with initial establishment on either Mauritius or Rodrigues and subsequent stepping-stone dispersal to the other two Mascarene islands within the last 2 Myr. Analyses suggest that diversification has slowed over time, with significant rate changes following dispersal to new geographical areas. Onset of diversification in species-rich Oncostemum appears to have been recent, with major cladogenesis commencing in the early Pliocene. [ABSTRACT FROM AUTHOR]

With 14 species, Badula (Primulaceae) is the most species-rich endemic angiosperm genus of the Mascarene Archipelago. The relationship between Badula and its ally Oncostemum ( c. 100 spp; Madagascar and the Comoros Islands) is uncertain, with implications for the circumscription of Badula as a Mascarene endemic. Within Badula, species rarity (several being critically endangered) and a paucity of herbarium specimens hamper proper species delimitations. Here, we estimate the phylogenetic relationships of Badula based on DNA sequence data from the nuclear ribosomal internal transcribed spacer (ITS) and plastid trnS-trnG-trnG regions with complete taxon sampling of the genus and three samples or more of each taxon. The results strongly supported the monophyly of Badula. Paraphyly of Oncostemum was inferred with weak support; explicit hypothesis testing did not favour this hypothesis over one that forced the monophyly of Oncostemum. Monophyly of several Badula spp. was supported, particularly for taxa from the older islands of Mauritius and Rodrigues. Badula is inferred to have reached the Mascarene Archipelago through a single colonization event. The majority of species segregated into island clades, implying that few, rather than multiple, colonization events have occurred in Badula among the islands of the archipelago. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 169, 284-296. [ABSTRACT FROM AUTHOR]

Background: Arundinarieae are a large tribe of temperate woody bamboos for which phylogenetics are poorly understood because of limited taxon sampling and lack of informative characters. Aims: This study assessed phylogenetic relationships, origins and classification of Arundinarieae. Methods: DNA sequences (plastid trnL-F; nuclear ITS) were used for parsimony and Bayesian inference including 41 woody bamboo taxa. Divergence dates were estimated using a relaxed Bayesian clock. Results: Arundinarieae were monophyletic but their molecular divergence was low compared to the tropical Bambuseae. Ancestors of the Arundinarieae lineage were estimated to have diverged from the other bamboos 23 (15-30) million years ago (Mya). However, the Arundinarieae radiation occurred 10 (6-16) Mya compared to 18 (11-25) Mya for the tropical Bambuseae. Some groups could be defined within Arundinarieae, but they do not correspond to recognised subtribes such as Arundinariinae or Shibataeinae. Conclusions: Arundinarieae are a relatively ancient bambusoid lineage that underwent a rapid radiation in the late Miocene. The radiation coincides with the continental collision of the Indo-Australian and Eurasian Plates. Arundinarieae are distributed primarily in East Asia and the Himalayas to northern Southeast Asia. It is unknown whether they were present in Asia long before their radiation, but we believe recent dispersal is a more likely scenario. [ABSTRACT FROM AUTHOR]

Cyperaceae are the third largest monocotyledon family, with considerable economic and conservation importance. In subfamily Mapanioideae there is particular specialization of the inflorescence into units termed spicoids. The structural homology of the spicoid is difficult to interpret, making determination of intrafamilial relationships problematic. To address this, pollen from eight species in Mapanioideae was investigated using light microscopy and scanning and transmission electron microscopy. Pollen development was also examined to identify the type of pollen present in these species. We also analyzed DNA sequence data using the trnL-F and rps16 regions from 25 genera and 35 species of Cyperaceae, Juncaceae, and Thurniaceae. Two types of pollen, Mapania-type and pseudomonad, were identified. Analysis of combined DNA and pollen data resolved a clade sister to the rest of Cyperaceae, corresponding to Mapanioideae. Within this, two further clades were resolved. One comprised taxa assigned to tribe Hypolytreae, which had Mapania-type pollen. The other comprised taxa mainly assigned to tribe Chrysitricheae, but included two taxa from Hypolytreae, Capitularina and Exocarya. All taxa in this clade had pseudomonad pollen. Thus new groupings within the subfamily have been discovered based on the specialization of some taxa in terms of their pollination biology. [ABSTRACT FROM AUTHOR]