Identification of Pathotypes in the Daylily Rust Pathogen Puccinia hemerocallidis

Daylily rust, caused by Puccinia hemerocallidis, has been present in the United States since 2000. In 2003, inoculations with a single isolate of P. hemerocallidis identified daylily cultivars with high levels of resistance to the fungus. The present study was carried out to determine if pathotypes of P. hemerocallidis are present in the south eastern United States. Sixteen isolates of P. hemerocallidis were each inoculated onto leaf segments from 19 daylily cultivars and the resulting disease phenotype assessed. A significant effect of rust isolate on host reaction phenotype was observed for nine of the 19 daylily cultivars. Five of the nine cultivars displayed reaction phenotypes with different isolates of P. hemerocallidis that included at least one susceptible or moderately susceptible and also resistant phenotypes. These results indicate that different pathotypes of the fungus are present in the south east United States. Daylily hybridizers interested in screening for host resistance to P. hemerocallidis will need to include multiple isolates of the fungus to allow for this host specialization.     

The effects of riverine impoundment on genetic structure and gene flow in two stream fishes in the Mobile River basin

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Fat frogs,mobile genes: unexpected phylogeographic patterns for the ornate chorus frog (Pseudacris ornata)

The southeastern coastal plain of the United States is a region marked by extraordinary phylogeographic congruence that is frequently attributed to the changing sea levels that occurred during the glacial‐interglacial cycles of the Pleistocene epoch. A phylogeographic break corresponding to the Apalachicola River has been suggested for many species studied to date that are endemic to this region. Here, we used this pattern of phylogeographic congruence to develop and test explicit hypotheses about the genetic structure in the ornate chorus frog (Pseudacris ornata). Using 1299 bp of mtDNA sequence and seven nuclear microsatellite markers in 13 natural populations of P. ornata, we found three clades corresponding to geographically distinct regions; one spans the Apalachicola River (Southern Clade), one encompasses Georgia and South Carolina (Central Clade) and a third comprises more northerly individuals (Northern Clade). However, it does not appear that typical phylogeographic barriers demarcate these clades. Instead, isolation by distance across the range of the entire species explained the pattern of genetic variation that we observed. We propose that P. ornata was historically widespread in the southeastern United States, and that a balance between genetic drift and migration was the root of the genetic divergence among populations. Additionally, we investigated fine‐scale patterns of genetic structure and found the spatial scale at which there was significant genetic structure varied among the regions studied. Furthermore, we discuss our results in light of other phylogeographic studies of southeastern coastal plain organisms and in relation to amphibian conservation and management.     

Corn pith weevil,Geraeus penicillus (Coleoptera: Curculionidae), in transgenic,insect‐resistant and conventional field corn hybrids

The corn pith weevil Geraeus penicillus (Herbst) is occasionally found boring in corn stalks throughout the eastern and Midwestern United States. Injury caused by G. penicillus is not typically economical, but may be confused with that of the European corn borer Ostrinia nubilalis (Hubner), an important economic pest throughout the United States. During efforts to assess European corn borer infestations, we discovered G. penicillus in field corn in south‐eastern Pennsylvania, including hybrids genetically modified (i.e. Bt hybrids) to control European corn borer among other herbivore species. Our analysis across sites indicated that tunnels of G. penicillus were significantly more abundant in transgenic Bt hybrids than non‐Bt hybrids, but comparisons of Bt hybrids and their near isolines revealed mostly similar numbers of G. penicillus tunnels, suggesting other hybrid features might be affecting the distribution of G. penicillus. Tunnels of G. penicillus were equally distributed among the three transgenic trait packages represented in our study. In plants where we found G. penicillus, tunnels were more abundant in stalks free of European corn borer damage. Our report appears to be the first to note G. penicillus feeding in Bt corn hybrids. These findings are notable because they document insect damage in Bt hybrids that may be mistaken for European corn borer damage and may provide evidence of an insect herbivore proliferating following a mild winter or possibly even moving into competitor‐free space.     

Microsatellite markers for the endangered Roanoke logperch, Percina rex (Percidae) and their potential utility for other darter species

The Roanoke logperch (Percina rex Jordan and Evermann), an endangered fish, occurs in only six watersheds in the Roanoke and Chowan river drainages of Virginia, USA. The species’ population genetic structure is poorly known. We developed 16 microsatellite markers that were reliably scorable and polymorphic P. rex. Markers were also screened in seven other darter species of the genus Percina. Most markers exhibited successful amplification and polymorphism in several species. These markers may therefore prove useful for population genetic studies in other darters, a diverse but highly imperiled group.     

Crossing the uncrossable: novel trans‐valley biogeographic patterns revealed in the genetic history of low‐dispersal mygalomorph spiders (Antrodiaetidae,Antrodiaetus) from California

Antrodiaetus riversi is a dispersal‐limited, habitat‐specialized mygalomorph spider species endemic to mesic woodlands of northern and central California. Here, we build upon prior phylogeographic research using a much larger geographic sample and include additional nuclear genes, providing more detailed biogeographic insights throughout the range of this complex. Of particular interest is the uncovering of unexpected and replicated trans‐valley biogeographic patterns, where in two separate genetic clades western haplotypes in the California south Coast Ranges are phylogenetically closely related to eastern haplotypes from central and northern Sierran foothills. In both instances, these trans‐valley phylogenetic patterns are strongly supported by multiple genes. These western and eastern populations are currently separated by the Central Valley, a well‐recognized modern‐day and historical biogeographic barrier in California. For one clade, the directionality is clearly northeast to southwest, and all available evidence is consistent with a jump dispersal event estimated at 1.2–1.3 Ma. During this time period, paleogeographic data indicate that northern Sierran rivers emptied to the ocean in the south Coast Ranges, rather than at the San Francisco Bay. For the other trans‐valley clade genetic evidence is less conclusive regarding the mechanism and directionality of biogeographic exchange, although the estimated timeframe is similar (approximately 1.8 Ma). Despite the large number of biogeographic studies previously conducted in central California, to the best of our knowledge no prior studies have discussed or revealed a northern Sierran to south Coast Range biogeographic connection. This uniqueness may reflect the low‐dispersal biology of mygalomorph spiders, where ‘post‐event’ gene exchange rarely erases historical biogeographic signal.     

The diatom genus Pseudo‐nitzschia (Bacillariophyceae) in New South Wales,Australia: morphotaxonomy,molecular phylogeny,toxicity, and distribution

Species belonging to the potentially harmful diatom genus Pseudo‐nitzschia, isolated from 16 localities (31 sampling events) in the coastal waters of south‐eastern Australia, were examined. Clonal isolates were characterized by (i) light and transmission electron microscopy; (ii) phylogenies, based on sequencing of nuclear‐encoded ribosomal deoxyribonucleic acid (rDNA) regions and, (iii) domoic acid (DA) production as measured by liquid chromatography–mass spectrometry (LC‐MS/MS). Ten taxa were unequivocally confirmed as Pseudo‐nitzschia americana, P. arenysensis, P. calliantha, P. cuspidata, P. fraudulenta, P. hasleana, P. micropora, P. multiseries, P. multistriata, and P. pungens. An updated taxonomic key for south‐eastern Australian Pseudo‐nitzschia is presented. The occurrence of two toxigenic species, P. multistriata (maximum concentration 11 pg DA per cell) and P. cuspidata (25.4 pg DA per cell), was documented for the first time in Australia. The Australian strains of P. multiseries, a consistent producer of DA in strains throughout the world, were nontoxic. Data from 5,888 water samples, collected from 31 oyster‐growing estuaries (2,000 km coastline) from 2005 to 2009, revealed 310 regulatory exceedances for “Total Pseudo‐nitzschia,” resulting in six toxic episodes. Further examination of high‐risk estuaries revealed that the “P. seriata group” had highest cell densities in the austral summer, autumn, or spring (species dependent), and lowest cell densities in the austral winter, while the “P. delicatissima group” had highest in winter and spring.     

Diversity of the snail‐eating snakes Pareas (Serpentes,Pareatidae) from Taiwan

Pareatidae are a group of mollusc‐eating snakes widely distributed in South‐eastern Asia. Due to their dietary specialization, the asymmetric dentition of pareatids has recently become an interesting issue in evolutionary biology. However, phylogenetic relationships and species diversity of pareatids are still poorly studied. A total of three Pareas species, P. formosanus (Van Denburgh 1909), P. compressus (Oshima 1910) and P. komaii (Maki 1931), have been reported for Taiwan. However, only P. formosanus is currently regarded as a valid species. Using mitochondrial sequence phylogeny, nuclear c‐mos haplotype network, as well as multivariate morphometrics, we re‐evaluated the taxonomic status of Pareas from Taiwan, the Ryukyus and adjacent regions. These lines of evidence showed congruent results for the coexistence of three Pareas species in Taiwan, with prominent genetic and morphological differentiation and differing level of dentition asymmetry. The currently used name P. formosanus should be applied only to the snakes with red iris, comparatively short lower jaw and totally smooth dorsal scales. An examination of the type material indicated that the name P. compressus should be regarded as a junior synonym of P. formosensis sensu stricto. Pareas komaii (Oshima 1910) is confirmed as a valid taxon with yellow iris, elongated lower jaw and strongly keeled dorsals. The third clade is characterized by a yellow iris, elongated lower jaw and weakly keeled dorsals. Despite their sympatric occurrence, every examined individual showed consistent grouping from mitochondrial, nuclear and morphological markers, indicating there is no gene flow among these three clades. Here, we describe the third clade as a new specie, Pareas atayal sp. nov.     

Population genetic structure and intraspecific genetic distance of Periplaneta americana (Blattodea: Blattidae) based on mitochondrial and nuclear DNA markers

The American cockroach (Periplaneta americana) is a globally invasive pest that can cause significant economic loss and threaten human health. Although it is abundant and lives in close proximity to humans, few studies have investigated the genetic diversity of P. americana. Our study analyzed 1,053 P. americana and other Periplaneta species' samples from different locations in China and the United States. A traditional tree‐based method using 17 unique mitochondrial COI haplotypes of P. americana and 20 haplotypes of the other Periplaneta species accurately identified P. americana with a barcoding threshold of 5.1%. To identify the population genetic structure of P. americana, we investigated wingless gene and pooled them with obtained mtDNA data for a combined analysis. Although the genetic diversity of the USA group was relatively higher than the China group, the number of haplotypes and alleles of both groups was small. The analysis of molecular variance (AMOVA), intraspecific phylogeny, and haplotype networks indicated that P. americana had very little global genetic differentiation. The weak geographic genetic structure might reflect the human‐mediated dispersal of P. americana. Despite no apparent phylogeographic assignment of mtDNA and nuclear lineages was observed in both BI trees, the integrated COI sequence data identified four distinct P. americana haplotype groups, showing four ancient maternal lineages of P. americana in China and the United States.     

Holocene vegetation and climate change on the Colorado Great Plains,USA, and the invasion of Colorado piñon (Pinus edulis)

Aim To reconstruct the last c. 7000 years of vegetation and climate change in an unusual region of modern Great Plains grassland and scarp woodland in south‐east Colorado (USA), and to determine the late Holocene biogeography of Colorado piñon (Pinus edulis) at its easternmost extent, using a series of radiocarbon‐dated packrat (Neotoma sp.) middens. Location The West Carrizo Canyon drains the Chaquaqua Plateau, a plateau that projects into the western extent of the southern Great Plains grasslands in south‐eastern Colorado, USA. Elevations of the study sites are 1448 to 1525 m a.s.l. Today the plateau is mostly Juniperus scopulorum–P. edulis woodland. Methods Plant macrofossils and pollen assemblages were analysed from 11 ¹⁴C‐dated packrat middens. Ages ranged from 5990 yr bp (6839 cal. yr bp ) to 280 yr bp (485 cal. yr bp ). Results The results presented here provide information on the establishment and expansion of Juniperus–P. edulis woodland at its eastern limits. The analysis of both plant macrofossils and pollen from the 11 middens documents changes in plant communities over the last 7000 years, and the establishment of P. edulis at its easternmost limit. Though very minor amounts of P. edulis pollen occur as early as the middle Holocene, plant macrofossils were only recovered in middens dating after c. 480 cal. yr bp . Main conclusions Originally, midden research suggested a late glacial refuge to the north‐east of the Carrizo Canyon site, and a middle Holocene expansion of P. edulis. Results reported here are consistent with a late Holocene expansion, here at its eastern limits, but noted elsewhere at its northern and north‐eastern limits. In general, this late Holocene expansion is consistent with pollen data from sediments in Colorado and New Mexico, and suggests that P. edulis is still expanding its range at its present extremes. This has implications for further extension of its range due to changing climatic conditions in the future.     

LIFE-HISTORY VARIATION AND COMPARATIVE PHYLOGEOGRAPHY OF DARTERS (PISCES: PERCIDAE) FROM THE NORTH AMERICAN CENTRAL HIGHLANDS

Phylogeography seeks to evaluate the relationship between genetic variation and geographic distribution of a species to examine the influence of historical events on divergence. Congruent phylogeographic patterns in codistributed species indicate historical association of the taxa being compared, and the uniform action of biogeographic events in shaping genetic variation. We sought to evaluate the congruence of patterns of genetic variation of five closely related fish species across a well-defined biogeographic boundary. We gathered allozymic and mitochondrial DNA (mtDNA) sequence data for five species of darters (Percidae: Etheostomatini) from populations distributed among biogeographic regions in the Ozark and Ouachita highlands of the south-central United States. Comparisons among species revealed noncongruence in the magnitude of genetic divergence in both allozymes and mtDNA sequences. We hypothesized that noncongruence resulted, in part, from differences in life histories of the species in our comparison. To address this hypothesis, we evaluated the association between gene flow (measured by N_em) and variation in body size and fecundity variables because they have been shown to influence gene flow in fishes. Correlation analysis revealed an association between gene flow and fecundity (r = 0.88), but not with body size (r = 0.36) or reproductive investment per individual (r = –0.23). The result was similar when independent contrasts of the original variables were used in correlation analyses. Phylogeographic analysis of mtDNA sequence data indicated the importance of history, evident in gene trees of Percina nasuta and Percina phoxocephala. Divergence rates between these two taxa may differ because of historically persistent differences in population sizes, reflected in present-day abundance and fecundity differences. Conversely, Percina caprodes showed little evidence of divergence in mtDNA sequences and yielded the highest mean N_em values from allozyme data. Comparisons among closely related, codistributed taxa may help discriminate among the possibilities for noncongruence in biogeographic studies by allowing evaluation of the influence of life history on patterns of gene flow.     

The higher‐level phylogenetic relationships of the Eumeninae (Insecta,Hymenoptera, Vespidae), with emphasis on Eumenes sensu lato

Cladistic analyses were carried out to infer the phylogenetic relationships among taxa that were originally part of the large genus Eumenes. Terminals belonging to other eumenine lineages were also included, as well as terminals from other vespid subfamilies. Analyses under equal weights and implied weights were carried out, and better results were obtained with the latter. The results corroborated the monophyly of Eumeninae, and recovered Zethini sensu lato as the sister‐lineage to the remaining eumenines. Eumenes sensu lato as originally recognized is paraphyletic relative to Odynerus sensu lato. A natural classification at the tribal level congruent with the phylogenetic results may be proposed, and the names Zethini, Odynerini, and Eumenini are already available. This is the most comprehensive phylogeny of the Eumeninae to date. A new generic synonymy is Alfieria Giordani Soika, 1934 = Delta de Saussure, 1855.     

Rapid evolution and the genomic consequences of selection against interspecific mating

While few species introduced into a new environment become invasive, those that do provide critical information on ecological mechanisms that determine invasions success and the evolutionary responses that follow invasion. Aedes albopictus (the Asian tiger mosquito) was introduced into the naturalized range of Aedes aegypti (the yellow fever mosquito) in the United States in the mid‐1980s, resulting in the displacement of A. aegypti in much of the south‐eastern United States. The rapid displacement was likely due to the superior competitive ability of A. albopictus as larvae and asymmetric mating interference competition, in which male A. albopictus mate with and sterilize A. aegypti females, a process called “satyrization.” The goal of this study was to examine the genomic responses of a resident species to an invasive species in which the mechanism of character displacement is understood. We used double‐digest restriction enzyme DNA sequencing (ddRADseq) to analyse outlier loci between selected and control lines of laboratory‐reared A. aegypti females from two populations (Tucson, AZ and Key West, Florida, USA), and individual females classified as either “resisted” or “mated with” A. albopictus males via mating trials of wild‐derived females from four populations in Florida. We found significant outlier loci in comparing selected and control lines and between mated and nonmated A. aegypti females in the laboratory and wild‐derived populations, respectively. We found overlap in specific outlier loci between different source populations that support consistent genomic signatures of selection within A. aegypti. Our results point to regions of the A. aegypti genome and potential candidate genes that may be involved in mating behaviour, and specifically in avoiding interspecific mating choices.     

Implications of High Molecular Divergence of Nuclear rRNA and Phylogenetic Structure for the Dinoflagellate Prorocentrum (Dinophyceae,Prorocentrales)

The small and large nuclear subunit molecular phylogeny of the genus Prorocentrum demonstrated that the species are dichotomized into two clades. These two clades were significantly different (one‐factor ANOVA, p < 0.01) with patterns compatible for both small and large subunit Bayesian phylogenetic trees, and for a larger taxon sampled dinoflagellate phylogeny. Evaluation of the molecular divergence levels showed that intraspecies genetic variations were significantly low (t‐test, p < 0.05), than those for interspecies variations (> 2.9% and > 26.8% dissimilarity in the small and large subunit [D1/D2], respectively). Based on the calculated molecular divergence, the genus comprises two genetically distinct groups that should be considered as two separate genera, thereby setting the pace for major systematic changes for the genus Prorocentrum sensu Dodge. Moreover, the information presented in this study would be useful for improving species identification, detection of novel clades from environmental samples.     

Variation of within‐crop microhabitat use by Drosophila suzukii (Diptera: Drosophilidae) in blackberry

Drosophila suzukii Matsumara (Diptera: Drosophilidae) is an invasive vinegar fly that infests ripe and ripening soft skinned fruits. In the south‐eastern United States, blackberry (Rubus spp.) crops are heavily impacted by D. suzukii, and current management tactics rely on the use of broad‐spectrum insecticides targeted to adult populations. An improved understanding of D. suzukii biology and ecology are necessary to create sustainable management options. Knowledge of how D. suzukii interacts with resources will enable targeted management actions in the future. In this present study, we monitored larval infestation throughout the blackberry canopy and found that infestation was highest in the inner portion of the canopy and lower in more exposed locations. We also documented higher humidity within the cane canopy relative to the edge of the field. A difference in abiotic conditions may create within‐crop microhabitats that D. suzukii is able to exploit. Future research will explore how to take advantage of these microhabitats in pest management programs.     

Evolutionary insight into the Peripatopsis balfouri sensu lato species complex (Onychophora: Peripatopsidae) reveals novel lineages and zoogeographic patterning

Evolutionary relationships in the widely distributed velvet worm Peripatopsis balfouri sensu lato species complex were examined using DNA sequence data, gross and SEM morphology. Sequence data were generated for the COI mtDNA and the 18S rRNA loci and analysed using a Bayesian inference approach, maximum likelihood and maximum parsimony. Phylogenetic analyses of the combined DNA sequence data revealed that Peripatopsis clavigera specimens from the southern Cape (clade 1) was sister to P. balfouri sensu lato specimens from the Cederberg Mountains (clade 2). Within the main P. balfouri sensu lato species complex, three addition clades could be discerned (clades 3, 4 and 5). The obligatory troglobitic species Peripatopsis alba was equidistant between the Cape Peninsula and adjacent interior (clade 3) and the two Boland and Hottentots Holland Mountains (clades 4 and 5). On the Cape Peninsula, P. stelliporata specimens nested among the sympatric P. balfouri sensu lato specimens. The Cape Peninsula specimens were sister to specimens from Jonkershoek site 1, Kogelberg and Simonsberg. Two Boland clades were retrieved, comprising Du Toit's Kloof, Bain's Kloof and Mitchell's Pass and sister (in clade 4) to specimens from the Boland and adjacent Hottentots Holland Mountains in clade 5. These results revealed complex biogeographic patterning in the P. balfouri sensu lato species complex. The presence of sympatric, yet genetically discrete species pairs at six of the sample localities (Du Toit's Kloof, Simonsberg, Jonkershoek sites 1 and 2, Kogelberg and Landroskop) suggests that there is reproductive isolation between the lineages. Divergence time estimations suggest a Miocene/Pliocene/Pleistocene cladogenesis. A taxonomic revision of the P. balfouri sensu lato species complex was undertaken to stabilize the taxonomy. P. clavigera is monophyletic and retained for the southern Cape specimens, P. balfouri sensu stricto is now confined to the Cape Peninsula and adjacent interior, while P. stelliporata is regarded as a junior synonym of the latter taxon, P. alba is endemic to the Wynberg Cave systems on the Cape Peninsula. Three novel species (Peripatopsis cederbergiensis, sp. n., Peripatopsis bolandi sp. n. and Peripatopsis purpureus, sp. n.) are described.     

Discordant patterns of introgression across a narrow hybrid zone between two cryptic lineages of an Iberian endemic newt

The study of natural hybrid zones can illuminate aspects of lineage divergence and speciation in morphologically cryptic taxa. We studied a hybrid zone between two highly divergent but morphologically similar lineages (south‐western and south‐eastern) of the Iberian endemic Bosca's newt (Lissotriton boscai) in SW Iberia with a multilocus dataset (microsatellites, nuclear and mitochondrial genes). STRUCTURE and NEWHYBRIDS analyses retrieved few admixed individuals, which classified as backcrosses involving parental individuals of the south‐western lineage. Our results show asymmetric introgression of mtDNA beyond the contact from this lineage into the south‐eastern lineage. Analysis of nongeographic introgression patterns revealed asymmetries in the direction of introgression, but except for mtDNA, we did not find evidence for nonconcordant introgression patterns across nuclear loci. Analysis of a 150‐km transect across the hybrid zone showed broadly coincident cline widths (ca. 3.2–27.9 km), and concordant cline centres across all markers, except for mtDNA that is displaced ca. 60 km northward. Results from ecological niche modelling show that the hybrid zone is in a climatically homogenous area with suitable habitat for the species, suggesting that contact between the two lineages is unlikely to occur further south as their distributions are currently separated by an extensive area of unfavourable habitat. Taken together, our findings suggest the genetic structure of this hybrid zone results from the interplay of historical (biogeographic) and population‐level processes. The narrowness and coincidence of genetic clines can be explained by weak selection against hybrids and reflect a degree of reproductive isolation that is consistent with cryptic speciation.