Biotechnological strains of Komagataella (Pichia) pastoris are Komagataellaphaffii as determined from multigene sequence analysis

Pichia pastoris was reassigned earlier to the genus Komagataella following phylogenetic analysis of gene sequences. Since that time, two additional species of Komagataella have been described, K. pseudopastoris and K. phaffii. Because these three species are unlikely to be resolved from the standard fermentation and growth tests used in yeast taxonomy, the identity of biotechnologically important strains of K. pastoris was determined from multigene sequence analyses. Results from this study show that the strain of ‘Pichia pastoris’ commonly used in gene expression studies is actually K. phaffii.     

The taxonomy of rhizobia: an overview

The taxonomy of rhizobia, bacteria capable of nodulating leguminous plants, has changed considerably over the last 20 years, with the original genus Rhizobium, a member of the alpha-Proteobacteria, now divided into several genera. The study of new geographically dispersed host plants, has been a source of many new species and is expected to yield many more. Here we provide an overview of the history of the rhizobia, but focus on the Rhizobium–Allorhizobium–Agrobacterium relationship. Finally, we review recent reports of nodulation and nitrogen fixation with legume hosts by bacteria that are outside the traditional rhizobial phylogenetic lineages. They include species of Methylobacterium and Devosia in the alpha- Proteobacteria and of Burkholderia and Ralstonia in the beta-Proteobacteria.     

Evolution and systematic relationships in theTriticeae (Poaceae)

The evolution and taxonomic relationships in theTriticeae are discussed with the view to highlight aspects of this agronomically important group of plants, which may be of interest to molecular biology. Some of these aspects are addressed in more detail in adjoining papers in which specific genomic loci have been examined at the DNA sequence or isozyme level. Aspects discussed include the systematics and geographic distribution of theTriticeae species, isozyme and chromosome pairing studies on some of the species as well as more recent developments in DNA analyses. A survey of the systematics of theTriticeae indicated that the genomic system ofLöve is probably the most useful starting point for interpreting molecular data even though the system has many problems from a taxonomic point of view. The geographical distribution ofTriticeae species, using both published and unpublished data, suggested that information of this type taken together with the theory of continental drift provides a broad time-span for considering data from DNA sequence studies. The significance, and modes of analyses, of isozyme studies were assessed because they often provide valuable characters in determining relationships between species. The main character underlyingLöve's andDewey's analyses of theTriticeae, namely chromosome pairing, is discussed with particular reference to isozyme studies to show that in some cases, such as species ofHordeum sensu lato, consistent relationships are obtained. Finally, new developments in understanding chromosome structure are considered in relation to the above variables in the taxonomy and evolution of theTriticeae.     

Poor genetic differentiation but clear cytoform divergence among cryptic species in Simulium damnosum complex (Diptera: Simuliidae)

A complex cryptic species taxonomy has been developed for the flies that vector Onchocerca volvulus L euckart in A frica. This taxonomy names cytoforms of Simulium damnosum T heobald s.l. based on chromosomal inversion polymorphisms. Researchers have linked variation in cytoforms to variation in geographical distributions and biological traits, but have failed to demonstrate that many of these cytoforms are differentiated genetically or morphologically. This may be explained by recent divergence and incomplete lineage sorting of ancestral polymorphisms. Alternatively, it may be the outcome of ongoing gene flow, with inversions maintaining sets of co‐adapted alleles and selection driving inversion frequency differences across habitats. Here, our goals were to test species delimitations in the S. damnosum complex, and determine if the poor differentiation between these species is better explained by the ancestral polymorphisms or gene‐flow hypotheses. Geometric morphometric analysis and more intensive genetic sampling of populations in N igeria only reinforced the lack of clear divergence among S. damnosum complex cytoforms. By contrast, the first statistical estimates of evolutionary relationships based on chromosomal inversion polymorphism data revealed clear signs of divergence, which lends support to the ancestral polymorphisms hypothesis. But the latter analyses may be biased by non‐neutral evolution of inversion polymorphisms; more rigorous tests of cytoform‐based species hypothesis will require deeper sampling across populations and neutral genetic markers along with explicit modelling of genetic isolation and migration.     

The Genera and Species of Tarsiidae

We revise the taxonomy of the primate family Tarsiidae. We classify extant tarsiers in 3 genera—Tarsius, Cephalopachus, and Carlito—each of which originated in the Miocene, or earlier, and each of which is allopatrically distributed within a distinct biogeographic region: Sulawesi, Sundaland, and Greater Mindanao, respectively. Within the genus Tarsius, formerly regarded as a single species, Tarsius spectrum, we recognize 8 allopatric and parapatric species, 9 if the inclusion of pumilus is warranted, and note that more are likely to be described in the near future. We restrict Tarsius tarsier, the senior taxon of the genus, to the island of Selayar, off the tip of the southwestern peninsula of Sulawesi. In doing so, it is required that we resurrect Tarsius fuscus Fischer 1804, for the population of tarsiers from the southwestern peninsula near the city of Makassar. We note that neither Cephalopachus nor Carlito has been the subject of anywhere near as much field research as has Tarsius; thus we question if the currently accepted α taxonomy for these genera is based on knowledge or ignorance.     

Assessing the Viability of Bacterial Species in Drinking Water by Combined Cellular and Molecular Analyses

The question which bacterial species are present in water and if they are viable is essential for drinking water safety but also of general relevance in aquatic ecology. To approach this question we combined propidium iodide/SYTO9 staining (“live/dead staining” indicating membrane integrity), fluorescence-activated cell sorting (FACS) and community fingerprinting for the analysis of a set of tap water samples. Live/dead staining revealed that about half of the bacteria in the tap water had intact membranes. Molecular analysis using 16S rRNA and 16S rRNA gene-based single-strand conformation polymorphism (SSCP) fingerprints and sequencing of drinking water bacteria before and after FACS sorting revealed: (1) the DNA- and RNA-based overall community structure differed substantially, (2) the community retrieved from RNA and DNA reflected different bacterial species, classified as 53 phylotypes (with only two common phylotypes), (3) the percentage of phylotpes with intact membranes or damaged cells were comparable for RNA- and DNA-based analyses, and (4) the retrieved species were primarily of aquatic origin. The pronounced difference between phylotypes obtained from DNA extracts (dominated by Betaproteobacteria, Bacteroidetes, and Actinobacteria) and from RNA extracts (dominated by Alpha-, Beta-, Gammaproteobacteria, Bacteroidetes, and Cyanobacteria) demonstrate the relevance of concomitant RNA and DNA analyses for drinking water studies. Unexpected was that a comparable fraction (about 21%) of phylotypes with membrane-injured cells was observed for DNA- and RNA-based analyses, contradicting the current understanding that RNA-based analyses represent the actively growing fraction of the bacterial community. Overall, we think that this combined approach provides an interesting tool for a concomitant phylogenetic and viability analysis of bacterial species of drinking water.     

A novel clade of sporocarp-forming species of glomeromycotan fungi in the <Emphasis Type="Italic">Diversisporales</Emphasis> lineage

In the early times of taxonomy of arbuscular mycorrhizal fungi (Glomeromycota), exclusively sporocarpic species were described. Since then the focus has mainly shifted to species forming spores singly. For many of the sporocarpic species, no molecular data have been made available, and their phylogenetic position has remained unclear. We obtained small subunit ribosomal rDNA and internal transcribed spacer data from specimens of glomeromycotan sporocarps from tropical areas that were assigned to three morphospecies. The complete sequence of the 18S small rDNA subunit sequence, internal transcribed spacers (ITS) 1 and 2 and 5.8S rDNA subunit, was determined from a sporocarp of Glomus fulvum. Partial sequences of the small subunit and the other regions were obtained from Glomus pulvinatum and the newly described species Glomus megalocarpum. Molecular phylogenetic analyses placed all species analyzed as a monophyletic sister group to the Diversispora spurca/Glomus versiforme clade group (“Glomus group C”) within the Diversisporales. The phylogenetic divergence from other known species suggests that this clade may constitute a new genus. These findings will have important consequences for taxon definition in the Diversisporales. They will facilitate identification of these fungi using rDNA sequences within colonized roots or the environment. Taxonomic novelties: Glomus megalocarpum D. Redecker     

Taxonomic revision of the dolphin genus Lagenorhynchus

Evolutionary relationships among cetaceans within the family Delphinidae have been difficult to resolve due to the high number of species and their relatively rapid radiation. This is the case for the dolphin species currently placed in the genus Lagenorhynchus, and their relations to Cephalorhynchus and Lissodelphis species. Phylogenetic relationships among these species have been investigated using multiple lines of evidence, and that evidence consistently suggests that the six species currently assigned to Lagenorhynchus do not form a monophyletic group. Here, we summarize findings from studies of morphology, genetics, historical biogeography, and acoustics that offer insight into the phylogenetics of these taxa. We present the taxonomic basis for revision of Lagenorhynchus, propose retention of Lagenorhynchus albirostris and reassignment of the remaining five species into other existing generic names, namely Leucopleurus acutus, Sagmatias australis, Sagmatias cruciger, Sagmatias obliquidens, and Sagmatias obscurus. Making these revisions now so that the taxonomy better reflects evolutionary relationships among these species will ensure that future studies include the most appropriate taxa for investigating the complex phylogenetic and systematic relationships among cetaceans. However, comprehensive analyses using multiple lines of evidence are still needed to clarify the phylogenetic relationships within and among the Lissodelphininae genera Cephalorhynchus, Lissodelphis, and Sagmatias.     

Taxonomic re-evaluation of the Taiwanese montane earthworm Amynthas wulinensis Tsai,Shen & Tsai, 2001 (Oligochaeta: Megascolecidae): Polytypic species or species complex?

Body size and colouration are two characters commonly used in the taxonomy of many animal taxa. However, they are seldom used by earthworm taxonomists because they are subject to environmental influences and tend to vary intraspecifically. In the present study, DNA sequences of the mitochondrial COI gene are used to evaluate whether specimens of the megascolecid earthworm Amynthas wulinensis Tsai, Shen & Tsai, 2001 that differ in body size and/or colouration belong to different genetic lineages. Phylogenetic analyses and morphological comparisons indicate that A. wulinensis in the previous broad sense is a species complex composed of three species differing in body size, colouration, and genital markings. Consequently, two new species, Amynthas lini and Amynthas meishanensis, are described. Taxonomic affinities of the A. wulinensis species complex are discussed, as is the feasibility of using body size and colouration in earthworm taxonomy.     

C4 photosynthetic modifications in the evolutionary transition from land to water in aquatic grasses

Cladistic analysis supports the conclusion that the Orcuttieae tribe of C₄ grasses reflect evolution from a terrestrial ancestry into seasonal pools. All nine species in the tribe exhibit adaptations to the aquatic environment, evident in the structural characteristics of the juvenile foliage, which persist submerged for 1–3 months prior to metamorphosis to the terrestrial foliage. Aquatic leaves of the least derived or basal genus Neostapfia have few morphological and anatomical characteristics specialized to the aquatic environment and have retained full expression of the C₄ pathway, including Kranz anatomy. Orcuttia species have many derived characteristics and are more specialized to the aquatic environment. These latter species germinate earlier in the season and persist in the submerged stage longer than Neostapfia and evidence from the literature indicates length of submergence is positively correlated with fitness components. Aquatic leaves of Orcuttia species lack Kranz or PCR bundle sheath anatomy, yet ¹⁴C-pulse chase studies indicate >95% malate + aspartate as the initial products of photosynthesis and these products turn over rapidly to phosphorylated sugars, indicating a tight coupling of the C₄ and C₃ cycles. Presence of the C₄ pathway is further supported by enzymological data. Contemporary dogma that Kranz anatomy is a sine qua non for operation of the C₄ pathway is contradicted by the patterns in Orcuttia; however, it is unknown whether the pathway acts as a CO₂ concentrating mechanism in these aquatic plants. Received: 12 June 1997 / Accepted: 10 February 1997     

Phylogenetic relationships among the black fly species (Diptera: Simuliidae) of Thailand based on multiple gene sequences

Simulium is a very speciose genus of the black fly family Simuliidae that includes many important pests of humans and animals. Cytotaxonomic and morphological studies have made substantial progress in Simulium systematics. 16S rRNA and ITS-1 DNA sequence studies have assisted this progress. Intensive multi-gene molecular systematic investigations will, however, be required for a comprehensive understanding of the genus’ taxonomy and evolution. Our research was conducted to investigate the relationships of Thai Simulium at the subgeneric, species group and species levels. We also examined the possibility of using mitochondrial DNA sequences to facilitate Simulium species identification. Data were collected from three mitochondrial genes (COI, ND4 and 16S rRNA) and two segments of the nuclear 28S ribosomal RNA (the D1 to D2 and the D4 expansion regions). The subgenera Simulium and Gomphostilbia were monophyletic in most analyses. Nevermannia included Montisimulium but was otherwise monophyletic in multigene analyses. In most analyses, Simulium and Nevermannia were more closely related to each other than to Gomphostilbia which was usually basal. Species groups were generally monophyletic. Within Gomphostilbia, however, the batoense species group was always paraphyletic to the other two species groups found in Thailand. Three species groups in Simulium were not monophyletic. The tendency to gill filament number reduction for some species groups in the subgenus Simulium was associated with a derived position in multigene analyses. Most species were monophyletic with two exceptions that probably represent species complexes and will present difficulties for rapid mitochondrial DNA identification.     

From shallow to deep divergences: mixed messages from Amazon Basin cichlids

Cichlids are a conspicuous component of Amazonian ichthyofauna, filling a wide range of niches. Yet taxonomy of many groups is still poorly known in the Amazon, and most of the yet-to-be discovered species are concentrated there. We analyzed 230 individuals sampled from six major Amazonian River Basins representing 56 morpho-species, 34 nominal and 22 undescribed species in 18 cichlid genera. We used four different single-locus species-discovery (SLSD) methods, delimiting between 53 (mPTP) and 57 (GMYC) species/lineages. When detected, species/lineages are hierarchically geographically structured. Many groups such as the Geophaginae and the Cichlinae have recently diversified, and species of genera such as Cichla and Symphysodon hybridize or have a history of hybridization; thus, these species will not be detected by SLSD methods. At the same time, for example, the genera Apistogramma and Biotodoma harbor cryptic species. For all these reasons, species/lineage diversity of Amazonian cichlids is significantly underestimated. The diversity of Amazonian cichlids is particularly remarkable given that the 570 species of Neotropical cichlids, many of which are from the Amazon Basin, are found in just 1.7% of the freshwater aquatic habitat in which the ~ 2,000 species of the East African rift lake cichlids evolved.

     

Environmental niche divergence between genetically distant lineages of an endangered water beetle

Historically, there has been considerable disagreement between researchers about the criteria used to discriminate among species. Decisions based on traditional morphological and genetic data alone can be potentially problematic, especially if the hypotheses are contradictory. Today, taxonomy is integrating new methods from different disciplines that study species' limits and evolution; this diverse range of evidence aids researchers in the recognition of species. Differences in niche characteristics could become a new and useful criterion in helping to decide the status of conflicting taxonomical entities. Ochthebius glaber (family Hydraenidae) is an endangered water beetle typical from southeast Iberian hypersaline streams that shows three clear discrete genetic units within its distribution range. However, there is no evidence to date that these lineages of O. glaber exhibit any adaptive morphological or ecological divergence. Using a modelling approach directed to generate niche representation from distributional data, we found a significant environmental niche divergence for allopatric lineages of O. glaber that followed an aridity gradient. Although we can not conclude firmly at present that the separate populations of O. glaber studied represent separate, reproductively isolated species, the present study complements and supports previous phylogeographic analyses through the inclusion of measures of another form of evolutionary change; in this case, ecological diversification. Despite the existence of some methodological limitations, also discussed in the present study, we emphasize the importance of recent conceptual advances that allow taxonomy to improve species delimitation practices through the integration of theory and methods from disciplines that study the origin and evolution of species. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 103 , 891–903.     

Putting incertae sedis taxa in their place: a proposal for ten new families and three new genera in Sphaeropleales (Chlorophyceae,Chlorophyta)

Best known for aquatic colonial algae such as Hydrodictyon, Pediastrum, or Scenedesmus, the order Sphaeropleales also contains numerous coccoid taxa from aquatic and terrestrial habitats. Recent findings indicate that coccoid lineages in this order are very diverse genetically and may be the prevalent form, although their diversity is often hidden morphologically. This study characterizes coccoid algae recently discovered from desert soil crusts that share morphological and ecological features with the genera Bracteacoccus, Pseudomuriella, and Chromochloris. Analyses of a multi‐gene data set that includes members from all sphaeroplealean families are used to examine the monophyly of these morphologically similar taxa, which are shown instead to be phylogenetically distinct and very divergent. We propose new generic names for these lineages: Bracteamorpha, Rotundella, and Tumidella. In addition, we propose an updated family‐level taxonomy within Sphaeropleales that includes ten new families of coccoid algae to accommodate the newly presented genera and many incertae sedis taxa in the order: Bracteamorphaceae, Chromochloridaceae, Dictyococcaceae, Dictyochloridaceae, Mychonastaceae, Pseudomuriellaceae, Rotundellaceae, Schizochlamydaceae, Schroederiaceae, and Tumidellaceae.     

Taxonomy as a hypothesis: testing the status of the Bermuda buckeye butterfly Junonia coenia bergi (Lepidoptera: Nymphalidae)

Species determination and definition in eukaryotes have traditionally been based on morphology, with little focus on genetic differentiation. Molecular methods allow for the independent assessment of morphology‐based taxonomic hypotheses. Three criteria used to define a full species for taxonomic purposes are morphological distinction, formation of a monophyletic lineage, and reproductive isolation. Junonia butterflies (Nymphalidae) are becoming an important experimental model system, but the taxonomy of many New World Junonia species is unclear. One of these species is J. coenia, which contains the subspecies J. coenia coenia, J. coenia grisea and J. coenia bergi. Previous studies suggest that J. coenia grisea may meet the criteria for full species status. Therefore, we evaluated the geographically isolated and rarely studied Bermuda buckeye butterfly J. coenia bergi to determine if it was similarly distinct. Physical examination of specimens and phylogenetic and population genetic analyses of mitochondrial cytochrome c oxidase subunit I, nuclear wingless, and complete mitochondrial genome sequences suggest that while J. coenia bergi is smaller in body size than many Junonia and has distinctive ventral hindwing colouration, it does not form a monophyletic lineage and shows indications of continued gene flow with North American mainland J. coenia coenia populations. Thus, J. coenia bergi does not meet the criteria for full species designation, but geographic isolation, morphological distinctiveness, and cultural importance suggest that it remain recognized as a subspecies of J. coenia. Similar analyses will be useful for addressing further taxonomic questions in Junonia and other taxa, especially where morphology‐based taxonomic determinations are ambiguous.     

Birding and DNA: species for the new millennium

Capsule Based on the 1999 Witherby Memorial Lecture – reviews how developments in molecular and population genetics have led to a reappraisal of species limits in birds.

The taxonomy of birds of the West Palearctic has moved from the comparative stability of the ‘Voous List’ into a period of serious activity, with new data emerging in almost every issue of every evolutionary and avian journal! This activity comes from two directions. Firstly, developments in population genetics, molecular biology, acoustics, behaviour and distributional studies have opened new avenues to measuring differentiation among groups of birds. This, in turn, has led to the recognition that earlier views of what constitutes a ‘species’ are in need of modification (‘improvement’), and the emergence of the ‘lineage concept’ of species. I review some of the species concepts most relevant to avian studies, and attempt to show how and why this change has happened, and its consequences for taxonomy and species limits. Examples are given in the form of ‘case studies’, and include Carrion/Hooded Crows Corvus corone/cornix, Green-winged/Eurasian Teals Anas carolinensis/crecca and Phylloscopus warblers.     

Cryptic diversity with wide salinity tolerance in the putative euryhaline Testudinella clypeata (Rotifera,Monogononta)

Aquatic faunas in fresh, brackish, and salt waters are usually well defined and differ amongst these three habitats. Nonetheless, some animals are known to be euryhaline, namely present across wide salinity ranges. The wide tolerance of putative euryhaline species has, however been refuted in some cases by DNA taxonomy, which has uncovered cryptic diversity with narrow ecological niches. We aim to improve knowledge on the putative euryhalinism of microinvertebrates and test whether it might actually be a real phenomenon or if euryhaline species are mostly a consequence of our previous inability to identify cryptic species with narrow salinity ranges, as discovered in Brachionus plicatilis. Using morphological analyses and DNA taxonomy, we investigated the species reality and distribution of a putative euryhaline rotifer species, Testudinella clypeata, and evaluated whether cryptic species are ecologically and/or geographically segregated. Different DNA taxonomy approaches concurred in revealing the presence of seven cryptic species within the T. clypeata morphospecies, which, in contrast to what has been previously detected, are actually euryhaline. Moreover, differences in analysed morphological traits were not significantly different amongst cryptic species. This suggests that DNA taxonomy improves our estimates of the actual diversity of microscopic species, in contrast to the morphological approach. © 2013 The Linnean Society of London     

Global diversity of butterflies (Lepidotera) in freshwater

In Lepidoptera, the subfamily Acentropinae and Pyraustinae of Crambidae (Pyraloidea) and the family Arctiidae (Noctuoidea) contain species with true aquatic larvae, which live submerged during larval development. In Pyraustinae and Arctiidae only a few species exhibit an aquatic life-history. From the latter, aquatic larvae are known from the Neotropical genus Paracles. The number of aquatic Paracles species is unknown. The Acentropinae are predominantly aquatic. They are distributed worldwide, and reach the highest diversity in tropical regions of South East Asia/Malesia and in the Neotropical Region. At present, the Acentropinae include a total of 50 genera and 737 described species. All genera, assigned to the subfamily, are listed in a table, and the numbers of included species are indicated. The taxonomy and phylogeney of the genera are inadequately known. The species have a minor economic importance, however, they are very sensitive to degradation of water quality and habitat destruction. Guest editors: E.V. Balian, C. Lévêque, H. Segers & K. Martens Freshwater Animal Diversity Assessment