Population genetics of wood-feeding cockroaches in the genus Cryptocercus

Members of the genus Cryptocercus are xylophagous, wingless, subsocial cockroaches that inhabit decaying logs in temperate forests. Given their winglessness, subsocial living, and the patchy distribution of food resources (decomposing logs), it is likely that Cryptocercus populations are substructured. Allozyme variation at eight polymorphic loci was assayed for 10 subpopulations of Cryptocercus darwini and 13 subpopulations of Cryptocercus wrighti, both of which are distributed in the Appalachian Mountains. The mean F(IS) was 0.13 and F(ST) was about 0.25 for both C. darwini and C. wrighti. The relatedness among individuals of a subpopulation of both species was not significantly different from that expected among full sibs. In terms of how genetic variation is partitioned, C. darwini and C. wrighti differed from each other substantially. Most of the genetic variation occurred among subpopulations of C. wrighti in the same region and among subpopulations of C. darwini in different regions. We discuss the factors that may have contributed to the observed similarities and differences in the breeding structure of the two species.     

Cockroaches that lack Blattabacterium endosymbionts: the phylogenetically divergent genus Nocticola

Phylogenetic relationships among termites, mantids and the five traditionally recognized cockroach families have been the subject of several studies during the last half-century. One cockroach lineage that has remained notably absent from such studies is the Nocticolidae. This group of small, elusive surface- and cave-dwelling species from the Old World Tropics has been proposed to represent an additional family. Using molecular sequences, we performed an initial phylogenetic examination of Nocticola spp. The hypothesis that they are phylogenetically divergent was confirmed from the analyses of three genes and a combined dataset. To supplement our phylogenetic analyses, we attempted to amplify 16S rRNA from the obligate mutualistic endosymbiont Blattabacterium cuenoti, present in all cockroaches studied to date. Unexpectedly, amplification was unsuccessful in all Nocticola spp. examined. This result was confirmed by microscopic examinations of fat body tissue. These Nocticola spp. are the first cockroaches found to be uninfected by B. cuenoti, which raise questions about when the bacterium first infected cockroaches.     

Phylogenetic relationship among cockroach families inferred from mitochondrial12S rRNA gene sequence

A number of phylogenies exist for cockroaches that differ in the postulated relationships among families and genera. The relationship of the wood-feeding genus, Cryptocercus, to other cockroach families and to termites, has generated considerable debate. Grandcolas (1994), based on morphological analysis, synonymized the family Cryptocercidae with Polyphagidae and placed the genus Cryptocercus in the subfamily Polyphaginae. To determine if an independent set of characters supports the placement of Cryptocercus in Polyphaginae, a phylogenetic analysis of relationships among representative genera of the five cockroach families was undertaken. DNA sequence of a -430 base pair portion of the mitochondrial small ribosomal subunit gene from representatives of Blattidae, Blattellidae, Blaberidae and Cryptocercus, previously published by Kambhampati (1995) and Kambhampati et al. (1996), and the homologous sequence from representatives of Polyphagidae were used in the analysis. A total of twenty cockroach taxa and three termite genera were included in the study. Because a recent study showed that Cryptocercus punctulatus consists of a species complex, DNA sequence from four individuals collected in different parts of the U.S.A. was included in the study. The trees estimated from parsimony and neighbour-joining analyses indicated that Cryptocercus is a monophyletic clade which is most closely related to members of Blattidae. Polyphagidae is indicated as a sister group to the Blattidae + Cryptocercus complex, suggesting that Polyphagidae may belong to the superfamily Blattoidea rather than to Blaberoidea as proposed by McKittrick (1964). Blaberidae and Blattellidae were sister groups as previously proposed. Based on the present analysis, I propose that the genus Cryptocercus be retained in the family Cryptocercidae. Cockroaches     

Evidence for cocladogenesis between diverse dictyopteran lineages and their intracellular endosymbionts

Bacteria of the genus Blattabacterium are intracellular symbionts that reside in specialized cells of cockroaches and the termite Mastotermes darwiniensis. They appear to be obligate mutualists, and are transmitted vertically in the eggs. Such characteristics are expected to lead to equivalent phylogenies for host and symbiont, and we tested this hypothesis using recently accumulated data on relationships among termites and cockroaches and their Blattabacterium spp. Host and symbiont topologies were found to be highly similar, and various tests indicated that they were not statistically different. A close relationship between endosymbionts from termites and members of the wood-feeding cockroach genus Cryptocercus was found, supporting the hypothesis that the former evolved from subsocial, wood-dwelling cockroaches. The majority of the Blattabacterium spp. sequences appear to have undergone similar rates of evolution since their divergence from a common ancestor, and an estimate of this rate was determined based on early Cretaceous host fossils. The results support the idea that the stem group of modern cockroaches radiated sometime between the late Jurassic and early Cretaceous-not the Carboniferous, as has been suggested on the basis of roach-like fossils from this epoch.     

Coevolution between a cockroach and its bacterial endosymbiont: a biogeographical perspective

Cryptocercus are subsocial, xylophagous cockroaches that live in temperate forests. Like other cockroaches, Cryptocercus harbour endosymbiotic bacteria in their fat bodies. Two species of Cryptocercus occur in the palaearctic, one each in eastern Russia and south-central China. In the USA, there are five species: one in the north-west and four in the south-east. Little is known about the relationship between the Eurasian and North American Cryptocercus or the causes of the disjunct distribution. Here, a molecular phylogeny for six out of the seven Cryptocercus species and their endosymbionts is inferred in an attempt to understand the evolution and biogeography of the genus. Our analysis showed that the North American Cryptocercus are monophyletic, suggesting that a single colonization event was followed by vicariance. There was complete concordance between the host and endosymbiont phylogenetic trees. Divergence estimates based on endosymbiont DNA sequences suggested that the palaearctic and nearctic Cryptocercus diverged 70-115 million years (Myr) ago and the eastern- and western-USA species diverged 53-88 Myr ago. These divergence estimates were correlated with biogeographical events, and a hypothesis is presented to explain the current distribution of Cryptocercus. Our findings suggest that Cryptocercus has had a long evolutionary history, dating back to the Jurassic.     

Canaleparolina darwiniensis, gen. nov., sp. nov., and other pillotinaceous spirochetes from insects.

We describe two new pillotinaceous spirochetes (Canaleparolina darwiniensis, Diplocalyx cryptotermitidis) and identify for the first time Hollandina pterotermitidis from both the subterranean termite Cryptotermes cavifrons and the wood-eating cockroach Cryptocercus punctulatus based on morphometric analysis of transmission electron micrographic thin sections. C. darwiniensis, gen. nov., sp. nov., limited to near Darwin, Australia, invariably is present on the surface of the treponeme-studded trichomonad Mixotricha paradoxa, a consistent inhabitant of the hindgut of healthy termite Mastotermes darwiniensis. The spirochete both attached to the surface of protists and free-swimming in the paunch (hindgut) lumen of the insect has 16 periplasmic flagella (16:32:16) and imbricated wall structures that resemble flattened crenulations of Pillotina. The flagella surround half the protoplasmic cylinder. C. darwiniensis is the largest (0.5 microm diameter x 25 microm length) of the three epibiotic bacteria (two spirochetes, one rod) that comprise the complex cortex of its host Mixotricha paradoxa. Several criteria distinguish Diplocalyx cryptotermitidis sp. nov. isolated from Cryptotermes cavifrons intestine: smaller diameter, fewer flagella, absence of inner and outer coats of the outer membrane, wider angle subtended by its flagella and, most notably, cytoplasmic tubule-associated centers, which are periodic electron dense spheres within the protoplasmic cylinder from which emanate cytoplasmic tubules up to 24 nm in diameter. This is also the first report of abundant populations of Hollandina in Cryptotermes cavifrons (those populations belong to the species H. pterotermitidis). Morphometric analysis of the first thin sections of any spirochetes (published nearly 40 years ago by A.V. Grimstone) permits us to identify the large (0.9 microm diameter) free-swimming intestinal symbiont of Cryptocercus punctulatus also as Hollandina pterotermitidis.     

Ecological niche conservation and differentiation in the wood-feeding cockroaches, Cryptocercus, in the United States

The distribution of a species is determined by a set of ecological correlates (biotic and abiotic factors) and historical phenomena that define and constrain its ecological niche. By examining the geographical projection of the ecological niche, it is possible to identify areas that are suitable for the species ecologically but that are not actually inhabited. Previous studies have used such discords to anticipate the existence of undescribed species; the absence of species may also result from historical barriers that have prevented colonization of the area. Thus, discords between potential and realized niches of a species may result from aspects of the species’s biology (e.g. lack of dispersal ability, specific habitat requirements) or from barriers to dispersal. In this study, we developed ecological niche models of the five Nearctic species of wood‐feeding cockroaches in the genus Cryptocercus. One species, C. clevelandi, occurs in the western USA and four (C. darwini, C. garciai, C. punctulatus, C. wrighti) are distributed in the Appalachian Mountains in the eastern USA. In general, modelled niches for all species showed ecological restriction relative to the total range of available conditions. Niche models for the eastern USA species predicted the geographical distributions of the other eastern USA species, with the exception of C. garciai, suggesting general overall conservatism in ecological characteristics. Some interspecific differences in modelled niches were also apparent. In general, C. clevelandi was predicted to occupy cooler and drier areas relative to the eastern USA species; among the eastern USA species, C. darwini and C. garciai were predicted to inhabit warmer areas relative to C. punctulatus and C. wrighti. Interspecific differences and similarities were consistent with the phylogenetic relationships among Cryptocercus species previously reported. © 2007 The Linnean Society of London, Biological Journal of the Linnean Society, 2007, 90 , 457–466.     

Evolution of host- and habitat association in the wood-feeding cockroach, Cryptocercus

Members of the cockroach genus Cryptocercus are subsocial insects that live in temperate forests and feed on decomposing logs. At present, seven species are recognized worldwide: four in the eastern USA, one in the western USA, and one each in Russia and China. Genetic variation within and among the Nearctic species has been characterized extensively in previous studies. However, whether there has been a corresponding divergence in the host and habitat association of Cryptocercus species is not known. Here, we report on differences in host and habitat association among six of the seven Cryptocercus species, estimated from field observations, elevation data, and land cover data. Our results indicated that the eastern and western USA species differ from one another in their distribution patterns, abundance, and habitat association. The eastern USA species are associated largely with deciduous forests, whereas the western USA and the Russian species are associated with evergreen forests. Thus, the eastern USA species, which are evolutionarily the most recent ones, have adapted to a different set of tree species relative to the basal species. There were also differences in the habitat association of the various species. Specifically, in the eastern USA, Cryptocercus darwini , evolutionarily the most recent species, occupied a habitat that is predominantly at low elevation [<400 m above sea level (ASL)] while all the other Nearctic species and the Russian species occupied a habitat that is at relatively higher elevations (>400 m ASL). Mapping of the above traits on a phylogenetic tree revealed that the evolutionary trend in Cryptocercus with regard to host and habitat association has been toward the utilization of low elevation habitats dominated by deciduous forests. © 2002 The Linnean Society of London, Biological Journal of the Linnean Society , 2002, 75 , 163–172.     

Inheritance and diversification of symbiotic trichonymphid flagellates from a common ancestor of termites and the cockroach Cryptocercus

Cryptocercus cockroaches and lower termites harbour obligate, diverse and unique symbiotic cellulolytic flagellates in their hindgut that are considered critical in the development of social behaviour in their hosts. However, there has been controversy concerning the origin of these symbiotic flagellates. Here, molecular sequences encoding small subunit rRNA and glyceraldehyde-3-phosphate dehydrogenase were identified in the symbiotic flagellates of the order Trichonymphida (phylum Parabasalia) in the gut of Cryptocercus punctulatus and compared phylogenetically to the corresponding species in termites. In each of the monophyletic lineages that represent family-level groups in Trichonymphida, the symbionts of Cryptocercus were robustly sister to those of termites. Together with the recent evidence for the sister-group relationship of the host insects, this first comprehensive study comparing symbiont molecular phylogeny strongly suggests that a set of symbiotic flagellates representative of extant diversity was already established in an ancestor common to Cryptocercus and termites, was vertically transmitted to their offspring, and subsequently became diversified to distinct levels, depending on both the host and the symbiont lineages.     

Molecular phylogeny of Cryptocercus wood-roaches based on mitochondrial COII and 16S sequences, and chromosome numbers in Palearctic representatives

Woodroaches of the genus Cryptocercus are subsocial and xylophagous cockroaches, distributed in North America and Asia. Studies on male chromosome number in Nearctic species have shown that diploid numbers vary from 2n=37 to 2n=47; numbers from Palearctic species were heretofore unknown. Two hypotheses have been proposed to explain the varying number of chromosomes among Nearctic species: the serial reduction hypothesis, and the parallel scenario. We performed phylogenetic analyses of the COII gene in these species and found evidence for the topology (47(45(43(39,37), which is congruent with the serial reduction hypothesis. We also determined chromosome numbers for the first time in Palearctic species, and found Cryptocercus primarius and Cryptocercus relictus to have relatively low chromosome numbers (2n=17-21) compared to their Nearctic relatives. Finally, our study determined the phylogenetic position of Cryptocercus primarius among other Asian taxa.     

Genome shrinkage and loss of nutrient-providing potential in the obligate symbiont of the primitive termite Mastotermes darwiniensis

Beneficial microbial associations with insects are common and are classified as either one or a few intracellular species that are vertically transmitted and reside intracellularly within specialized organs or as microbial assemblages in the gut. Cockroaches and termites maintain at least one if not both beneficial associations. Blattabacterium is a flavobacterial endosymbiont of nearly all cockroaches and the termite Mastotermes darwiniensis and can use nitrogenous wastes in essential amino acid and vitamin biosynthesis. Key changes during the evolutionary divergence of termites from cockroaches are loss of Blattabacterium, diet shift to wood, acquisition of a specialized hindgut microbiota, and establishment of advanced social behavior. Termite gut microbes collaborate to fix nitrogen, degrade lignocellulose, and produce nutrients, and the absence of Blattabacterium in nearly all termites suggests that its nutrient-provisioning role has been replaced by gut microbes. M. darwiniensis is a basal, extant termite that solely retains Blattabacterium, which would show evidence of relaxed selection if it is being supplanted by the gut microbiome. This termite-associated Blattabacterium genome is ～8% smaller than cockroach-associated Blattabacterium genomes and lacks genes underlying vitamin and essential amino acid biosynthesis. Furthermore, the M. darwiniensis gut microbiome membership is more consistent between individuals and includes specialized termite gut-associated bacteria, unlike the more variable membership of cockroach gut microbiomes. The M. darwiniensis Blattabacterium genome may reflect relaxed selection for some of its encoded functions, and the loss of this endosymbiont in all remaining termite genera may result from its replacement by a functionally complementary gut microbiota.     

Symbiosis, morphology, and phylogeny of Hoplonymphidae (Parabasalia) of the wood-feeding roach Cryptocercus punctulatus

Anaerobic cellulolytic flagellate protists of the hindguts of lower termites and the wood-feeding cockroach Cryptocercus are essential to their host's ability to digest lignocellulose. Many have bacteria associated with their surfaces and within cytoplasmic vesicles-likely important symbioses as suggested by molecular and other data. Some of the most striking examples of these symbioses are in the parabasalid family Hoplonymphidae, but little or no data exist on the structural aspects of their symbioses, their relationships with bacteria through different life-cycle stages, or their diversity and phylogenetic relationships in Cryptocercus. We investigated these areas in the hoplonymphid genera Barbulanympha and Urinympha from Cryptocercus punctulatus using light and electron microscopy, and analysis of small subunit rRNA. Microscopy reveals variation in density of bacterial surface symbionts related to life-cycle stage, a glyococalyx possibly important in bacterial adhesion and/or metabolite exchange, and putative viruses associated with bacterial surface symbionts. Patterning of surface bacteria suggests protists emerging from the resistant (dormant) stage are colonized by a small population of bacterial cells, which then divide to cover their surface. Additionally, cytoplasmic protrusions from the protist are covered by bacteria. Phylogenetic analysis rejects the monophyly of Hoplonymphidae, suggesting multiple origins or losses of these bacterial symbioses.     

A case for ancestral transfer of symbionts between cockroaches and termites

Living species of the cockroach family Cryptocercidae have intestinal symbionts that are congeneric with some of the gut protozoa found in Isoptera. Presence of such closely related symbionts in cryptocercids and in termites has been frequently interpreted as a uniquely derived homologous character shared between the two xylophagous groups. This may not be the most parsimonious interpretation. Cryptocercus nymphs placed into Zootermopsis (dampwood termite) colonies were killed and eaten by the termites. Termites placed into a Cryptocercus nest box were also fully consumed. Modern Cryptocercus punctulatus and Zootermopsis are often found in the same decaying logs in the Pacific Northwest of the U.S.A., and it is likely that their ancestors also cohabited in at least a portion of their ranges. By occasionally killing and consuming an intruder from the other group, gut protozoa could have been acquired and exchanged between termites and Cryptocercus or their ancestors, under natural conditions and before the life histories of the protozoa became specialized within the host orders. Implications for assessing the phylogeny of the two dictyopteroid groups are also discussed.     

Metabolic stasis in an ancient symbiosis: genome-scale metabolic networks from two <Emphasis Type="Italic">Blattabacterium cuenoti</Emphasis> strains,primary endosymbionts of cockroaches

BACKGROUND: Cockroaches are terrestrial insects that strikingly eliminate waste nitrogen as ammonia instead of uric acid. Blattabacterium cuenoti (Mercier 1906) strains Bge and Pam are the obligate primary endosymbionts of the cockroaches Blattella germanica and Periplaneta americana, respectively. The genomes of both bacterial endosymbionts have recently been sequenced, making possible a genome-scale constraint-based reconstruction of their metabolic networks. The mathematical expression of a metabolic network and the subsequent quantitative studies of phenotypic features by Flux Balance Analysis (FBA) represent an efficient functional approach to these uncultivable bacteria. RESULTS: We report the metabolic models of Blattabacterium strains Bge (iCG238) and Pam (iCG230), comprising 296 and 289 biochemical reactions, associated with 238 and 230 genes, and 364 and 358 metabolites, respectively. Both models reflect both the striking similarities and the singularities of these microorganisms. FBA was used to analyze the properties, potential and limits of the models, assuming some environmental constraints such as aerobic conditions and the net production of ammonia from these bacterial systems, as has been experimentally observed. In addition, in silico simulations with the iCG238 model have enabled a set of carbon and nitrogen sources to be defined, which would also support a viable phenotype in terms of biomass production in the strain Pam, which lacks the first three steps of the tricarboxylic acid cycle. FBA reveals a metabolic condition that renders these enzymatic steps dispensable, thus offering a possible evolutionary explanation for their elimination. We also confirm, by computational simulations, the fragility of the metabolic networks and their host dependence. CONCLUSIONS: The minimized Blattabacterium metabolic networks are surprisingly similar in strains Bge and Pam, after 140 million years of evolution of these endosymbionts in separate cockroach lineages. FBA performed on the reconstructed networks from the two bacteria helps to refine the functional analysis of the genomes enabling us to postulate how slightly different host metabolic contexts drove their parallel evolution.     

Genetic and morphological evidence for substantial hidden biodiversity in a freshwater crayfish species complex

North America is a center of biodiversity for freshwater crayfish, with >300 described species, mostly in the family Cambaridae. However, the phylogenetic and phylogeographic relationships among these taxa are poorly understood, despite their ubiquitous occurrence in North American freshwater habitats. Here, we target the geographically widespread Orconectesvirilis and its sister taxa for genetic and morphological investigation. We used sequence data from two mitochondrial genes (16S rRNA and cytochrome oxidase I) and from one nuclear gene (glyceraldehyde-3-phosphate dehydrogenase), as well as morphological data from the male gonopod, to gain insights into phylogenetic relationships among individuals collected in the northeastern United States, where this taxon is considered to be invasive, and from Kansas. Our data reveal evidence for three cryptic species in our collections, in addition to O.virilis sensu stricto; all four clades are separated by both mitochondrial and nuclear sequence data. In addition, two of the newly discovered species, O.sp. nov. A and O.sp. nov. B, are distinguishable from O.virilis by significant differences in gonopod morphology (no data on gonopod morphology were available for O.sp. nov. C). O.sp. nov. A was collected at numerous sites in Massachusetts; additional sampling is required to determine if this species is native to New England. Two other taxa, O.sp. nov. B and O.sp. nov. C, were both collected from sites in eastern Kansas, and historically have probably been considered to be O.nais. In summary, this investigation indicates that this species complex may harbor a great deal of unknown biodiversity over the rest of its described range, and suggests that further attention to the phylogenetics and phylogeography of North American crayfishes may yield interesting insights into the evolution of biodiversity.     

Phylogeny of Cryptocercus species (Blattodea: Cryptocercidae) inferred from nuclear ribosomal DNA

The wood-feeding cockroaches of the genus Cryptocercus occur in temperate forests. Of the seven known species, five occur in the United States and two in Eurasia. Until 1997, all populations in the United States were considered a single species. Populations in the western United States were elevated to a species status based on variation in DNA sequence and morphology. In 1999, three new species were described from the eastern United States based on variation in chromosome number and mitochondrial DNA, bringing the number of species in the United States to five. The objective of this study was to determine if the DNA sequence of nuclear rRNA also signals the existence of four species in the eastern United States and to compare the inferred relationships with those proposed based on mitochondrial sequences. We obtained the DNA sequence from a portion of the 5.8S and 28S rRNA genes and the entire ITS2 region from 38 individuals and 30 additional clones to assess intraindividual, intraspecific, and interspecific variation. We found extensive sequence variation among the various species and little or no intraindividual and intraspecific variation. Phylogenetic analysis indicated the existence of monophyletic lineages among the eastern United States samples, which largely corresponded to the four species previously described. The inferred relationships were well-supported by bootstrap analysis and decay indices. Although the nuclear rRNA sequences resulted in a coherent phylogenetic tree, the ITS2 region contained many insertions and deletions, which may introduce homoplasy and ambiguity in alignment as more taxa are added to the data set.     

Molecular phylogeny and biogeography of the Korean woodroaches Cryptocercus spp

Phylogenetic relationships and biogeography of Northeast Asian Cryptocercus were inferred based on the DNA sequences of mitochondrial COII and 16S rRNA genes and nuclear 18S rRNA gene. The results suggest that two clades exist in Korean populations. The southwestern population (Cryptocercus from Jiri-san) was more closely related to the populations from Northeast China and eastern Russia than to all the other Korean Cryptocercus. According to molecular-based estimated divergence times, the divergence event occurred between Cryptocercus in Jiri-san, Northeast China and eastern Russia and those in the remaining South Korea during the Miocene (7.5-17.4Myr ago), and then the divergence event between Cryptocercus in Jiri-san and those in Northeast China and eastern Russia occurred 0.8-1.9Myr ago. In the Korean Peninsula, Jiri-san is located in the most southwestern region among the high mountains surveyed. The location is the farthest from Northeast China and eastern Russia among sampling localities in South Korea. Thus, it was unexpected that the southwestern populations are more closely related to those from Northeast China and eastern Russia rather than to the other Korean Cryptocercus. Based on Korean topography and estimated divergence times, possible scenarios are proposed for the current geographical distribution of Korean Cryptocercus.     

Development, phylogeny, and taxonomy of Bostrycapulus (Caenogastropoda: Calyptraeidae), an ancient cryptic radiation

Calyptraeid gastropods are well know for the taxonomic difficulties caused by their simple, phenotypically variable shells. In this paper I demonstrate that what was previously considered to be a single species, Crepidula aculeata , is an ancient (3–15 Myr) cryptic species complex made up of at least eight species, and that this group should be placed in the genus Bostrycapulus . Despite the difficulty in finding diagnostic adult shell and anatomical features upon which species can be unambiguously identified, DNA sequences, protoconch morphology, embryonic morphology and developmental characters clearly differentiate these eight species. A single species with direct development and nurse eggs is present in the South Atlantic, and a species with planktotrophic development occurs in the equatorial Pacific. The species from Japan, Australia, Florida, the Pacific coasts of Mexico and Central America, and the Cape Verde Islands all have direct development. Most of these species are separated by > 15% divergence in COI sequence data. The fossil record of Bostrycapulus goes back to the Miocene, which agrees with genetic estimates of divergences within the genus ranging from 3 to 15 Mya. Surprisingly, these ancient species differ only slightly in morphology from each other and genetic differentiation does not correlate with geographical distance. I revise the genus Bostrycapulus on the basis of differences in adult morphology, embryonic morphology, mode of development, protoconch morphology, and DNA sequence data. I also describe four new species ( B. pritzkeri sp. nov., B. odites sp. nov., B. latebrus sp. nov. and B. urraca sp. nov. ) and remove three others ( B. gravispinosus , B. calyptraeformis , and B.  cf. tegulicius ) from synonymy with B. aculeatus .  © 2005 The Linnean Society of London, Zoological Journal of the Linnean Society , 2005, 144 , 75−101.