Rapid Speciation Research Articles

Evolution of interspecies unilateral incompatibility in the relatives of Arabidopsis thaliana.

The evolutionary concurrence of intraspecies self-incompatibility (SI) and explosive angiosperm radiation in the Cretaceous have led to the hypothesis that SI was one of the predominant drivers of rapid speciation in angiosperms. Interspecies unilateral incompatibility (UI) usually occurs when pollen from a self-compatible (SC) species is rejected by the pistils of a SI species, while the reciprocal pollination is compatible (UC). Although this SI×SC type UI is most prevalent and viewed as a prezygotic isolation barrier to promote incipient speciation of angiosperms, comparative evidence to support such a role is lacking. We show that SI×SI type UI in SI species pairs is also common in the well-characterized accessions representing the four major lineages of the Arabidopsis genus and is developmentally regulated. This allowed us to reveal a strong correlation between UI strength and species divergence in these representative accessions. In addition, analyses of a SC accession and the pseudo-self-compatible (psc) spontaneous mutant of Arabidopsis lyrata indicate that UI shares, at least, common pollen rejection pathway with SI. Furthermore, genetic and genomic analyses of SI×SI type UI in A.lyrata×A.arenosa species pair showed that two major-effect quantitative trait loci are the stigma and pollen-side determinant of UI, respectively, which could be involved in heterospecies pollen discrimination. By revealing a close link between UI and SI pathway, particularly between UI and species divergence in these representative accessions, our findings establish a connection between SI and speciation. Thus, the pre-existence of SI system would have facilitated the evolution of UI and accordingly promote speciation.

Mobilization of retrotransposons as a cause of chromosomal diversification and rapid speciation: the case for the Antarctic teleost genus Trematomus

BackgroundThe importance of transposable elements (TEs) in the genomic remodeling and chromosomal rearrangements that accompany lineage diversification in vertebrates remains the subject of debate. The major impediment to understanding the roles of TEs in genome evolution is the lack of comparative and integrative analyses on complete taxonomic groups. To help overcome this problem, we have focused on the Antarctic teleost genus Trematomus (Notothenioidei: Nototheniidae), as they experienced rapid speciation accompanied by dramatic chromosomal diversity. Here we apply a multi-strategy approach to determine the role of large-scale TE mobilization in chromosomal diversification within Trematomus species.ResultsDespite the extensive chromosomal rearrangements observed in Trematomus species, our measurements revealed strong interspecific genome size conservation. After identifying the DIRS1, Gypsy and Copia retrotransposon superfamilies in genomes of 13 nototheniid species, we evaluated their diversity, abundance (copy numbers) and chromosomal distribution. Four families of DIRS1, nine of Gypsy, and two of Copia were highly conserved in these genomes; DIRS1 being the most represented within Trematomus genomes. Fluorescence in situ hybridization mapping showed preferential accumulation of DIRS1 in centromeric and pericentromeric regions, both in Trematomus and other nototheniid species, but not in outgroups: species of the Sub-Antarctic notothenioid families Bovichtidae and Eleginopsidae, and the non-notothenioid family Percidae.ConclusionsIn contrast to the outgroups, High-Antarctic notothenioid species, including the genus Trematomus, were subjected to strong environmental stresses involving repeated bouts of warming above the freezing point of seawater and cooling to sub-zero temperatures on the Antarctic continental shelf during the past 40 millions of years (My). As a consequence of these repetitive environmental changes, including thermal shocks; a breakdown of epigenetic regulation that normally represses TE activity may have led to sequential waves of TE activation within their genomes. The predominance of DIRS1 in Trematomus species, their transposition mechanism, and their strategic location in “hot spots” of insertion on chromosomes are likely to have facilitated nonhomologous recombination, thereby increasing genomic rearrangements. The resulting centric and tandem fusions and fissions would favor the rapid lineage diversification, characteristic of the nototheniid adaptive radiation.

Molecular Phylogeny ofAstragalussect.Ammodendron(Fabaceae) Inferred from Chloroplastycf1 Gene

Species of Astragalus section Ammodendron (Fabaceae) are restricted to desert areas in central Asia, Afghanistan, and Iran. In the present study, the variable plastid gene ycf1 was used to reconstruct its phylogeny using parsimony, Bayesian and likelihood methods as well as network analysis. Phylogenetic analyses of ycf1 sequences strongly supported monophyly of the section Ammodendron. The ycf1 phylogeny was congruent with the combined nrDNA ITS, trnH-psbA, trnT-Y and ycf1 data sets. The group assembly in neighbor-net splits graph corresponded to the clades retrieved in the phylogenetic analyses and offered a graphical visualization of relationships between species. Poor phylogenetic resolution within the sect. Ammodendron s. str. may be due to rapid speciation and lower rates of nucleotide evolution. In conclusion, the close relationships among several species can provide pieces of evidence for further investigations on the molecular and morphological delimitation of this section.

Skin and Soft Tissue Infections Due to Nontuberculous Mycobacteria.

This review describes recent trends in the epidemiology of nontuberculous mycobacteria (NTM), emerging pathogens, new insights into NTM pathogenesis, and advances in diagnosis and treatment. Emerging pathogens include Mycobacterium chimaera and drug-resistant subspecies of Mycobacterium abscessus. Important virulence mechanisms of pathogenic NTM include the ability to alter the macrophage's permissiveness to intracellular bacterial growth. New diagnostic tools consist of DNA probes, gene sequencing, and matrix-assisted laser desorption ionization-time of flight. These methods allow rapid speciation of NTM species, in some cases directly from patient samples. There are few novel agents available to treat NTM, although some repurposed drugs show excellent activity. The incidence of NTM infections appears to be increasing in a number of regions around the world. Molecular methods are now the diagnostic tools of choice. Discovery of novel effective agents and/or drug combinations with greater likelihood of cure, shorter treatment duration, and fewer side effects are research priorities.

A new species of Buenia (Perciformes: Gobiidae) from the western Mediterranean slope bottoms, the redescription of Buenia jeffreysi and the first Balearic record of Buenia affinis.

A new gobiid species, Buenia lombartei sp. nov. (Teleostei: Gobiidae) is described from the continental slope off the Balearic Islands, western Mediterranean. The Atlantic species Buenia jeffreysii (Günther, 1867) is redescribed and the first record of Buenia affinis Iljin, 1930 is reported for the islands. A key for the species of genus Buenia is provided. The new species can be distinguished from congeneric species by morphological characters: anterior oculoscapular canal semi-closed with pores σ, λ, κ, α, ρ and additional pores and open furrows; suborbital row c of 6 papillae; anal fin I/7-I/8; scales in lateral series 25-27; scales in transverse series 6; pectoral fin rays 18; the second spine of the first dorsal fin longest, backwards reachingto middle second dorsal fin in males when folded down; pelvic fin anterior membrane reduced to less than one sixth of spinous ray in midline depth; tongue well developed and bilobed and several morphometric characters. The new species is also characterized by its reduced colouration and unique depth range and occurrence on upper slope muddy bottoms. Contrary to this, only a small number of gobiid species in the Mediterranean reach beyond the circalittoral to the deep shelf, and just a few have records below the shelf break. Phylogenetic analysis of their mitochondrial COI sequence grouped the two specimens within the same clade as B. jeffreysii, but one apomorphic transversion diagnoses the B. lombartei subclade. A hypothesis about rapid speciation of B. lombartei as well as alternative hypotheses are discussed. During the Last Glacial Maximum, boreal fish species such as cold-water adapted gadids, now restricted to northwestern Europe, entered the Mediterranean. This may have also been the case of the boreal population of B. jeffreysii. At the end of the glacial period, increasingly warming Mediterranean waters might restricted B. jeffreysii to deeper coldwater mud habitats of the upper slope, where it finally adapted to bathyal conditions.

Rapid sexual and genomic isolation in sympatric Drosophila without reproductive character displacement.

The rapid evolution of sexual isolation in sympatry has long been associated with reinforcement (i.e., selection to avoid maladaptive hybridization). However, there are many species pairs in sympatry that have evolved rapid sexual isolation without known costs to hybridization. A major unresolved question is what evolutionary processes are involved in driving rapid speciation in such cases. Here, we focus on one such system; the Drosophila athabasca species complex, which is composed of three partially sympatric and interfertile semispecies: WN, EA, and EB. To study speciation in this species complex, we assayed sexual and genomic isolation within and between these semispecies in both sympatric and allopatric populations. First, we found no evidence of reproductive character displacement (RCD) in sympatric zones compared to distant allopatry. Instead, semispecies were virtually completely sexually isolated from each other across their entire ranges. Moreover, using spatial approaches and coalescent demographic simulations, we detected either zero or only weak heterospecific gene flow in sympatry. In contrast, within each semispecies we found only random mating and little population genetic structure, except between highly geographically distant populations. Finally, we determined that speciation in this system is at least an order of magnitude older than previously assumed, with WN diverging first, around 200K years ago, and EA and EB diverging 100K years ago. In total, these results suggest that these semispecies should be given full species status and we adopt new nomenclature: WN—D. athabasca, EA—D. mahican, and EB—D. lenape. While the lack of RCD in sympatry and interfertility do not support reinforcement, we discuss what additional evidence is needed to further decipher the mechanisms that caused rapid speciation in this species complex.

Establishing Taxonomic Identity and Selecting Genetically Diverse Populations for Conservation of Threatened Plants Using Molecular Markers

The extent of genetic diversity within a species is an important determinant of successful adaptation to adverse environmental conditions. Assessment of extent of genetic diversity/variability is also important to monitor genetic erosion within a species. In threatened plant species, genetic diversity assessment helps in selection of genetically diverse populations to enrich the genetically impoverished populations, thus minimizing the probability of genetic drift. Confirming taxonomic identity of threatened species, particularly those belonging to species complexes with dispute identity, is another essential task in the conservation of threatened species, which is best resolved through molecular approaches. The present study estimated the genetic variability within and among the populations of four threatened species, viz. Justicia beddomei (C.B. Clarke) Bennet (Acanthaceae), Embelia ribes Burm. f. (Myrsinaceae), Madhuca insignis (Radlk.) H.J. Lam (Sapotaceae) and Cycas beddomei Dyer (Cycadaceae) using Inter Simple Sequence Repeat (ISSR) and Simple Sequence Repeat (SSR) markers for selecting the genetically diverse populations. The phylogeny was analysed through ITS (nrDNA) and matK (cpDNA) sequences to confirm the species identity. The phylogenetic analyses confirmed four distinct species of Justicia , which also revealed that J. beddomei and J. adhatoda were sister groups with a common ancestor showing rapid parallel speciation with J. gendarussa in one clade and J. betonica in another. Madhuca insignis with extremely small population in the Western Ghats (Karnataka to Kerala) might have undergone either extensive hybridization or incipient speciation. In case of Embelia species, a greater evolutionary closeness between E. subcoraceae and E. floribunda was revealed, while E. ribes had a distinct clad. Both ISSR and SSR markers distinguished various genotypes of Cycas beddomei .

Evolutionary history of the angiosperm flora of China.

High species diversity may result from recent rapid speciation in a 'cradle' and/or the gradual accumulation and preservation of species over time in a 'museum'. China harbours nearly 10% of angiosperm species worldwide and has long been considered as both a museum, owing to the presence of many species with hypothesized ancient origins, and a cradle, as many lineages have originated as recent topographic changes and climatic shifts-such as the formation of the Qinghai-Tibetan Plateau and the development of the monsoon-provided new habitats that promoted remarkable radiation. However, no detailed phylogenetic study has addressed when and how the major components of the Chinese angiosperm flora assembled to form the present-day vegetation. Here we investigate the spatio-temporal divergence patterns of the Chinese flora using a dated phylogeny of 92% of the angiosperm genera for the region, a nearly complete species-level tree comprising 26,978 species and detailed spatial distribution data. We found that 66% of the angiosperm genera in China did not originate until early in the Miocene epoch (23 million years ago (Mya)). The flora of eastern China bears a signature of older divergence (mean divergence times of 22.04-25.39 Mya), phylogenetic overdispersion (spatial co-occurrence of distant relatives) and higher phylogenetic diversity. In western China, the flora shows more recent divergence (mean divergence times of 15.29-18.86 Mya), pronounced phylogenetic clustering (co-occurrence of close relatives) and lower phylogenetic diversity. Analyses of species-level phylogenetic diversity using simulated branch lengths yielded results similar to genus-level patterns. Our analyses indicate that eastern China represents a floristic museum, and western China an evolutionary cradle, for herbaceous genera; eastern China has served as both a museum and a cradle for woody genera. These results identify areas of high species richness and phylogenetic diversity, and provide a foundation on which to build conservation efforts in China.

Evolution of nectar spur length in a clade of Linaria reflects changes in cell division rather than in cell expansion.

Background and AimsNectar spurs (tubular outgrowths of a floral organ which contain, or give the appearance of containing, nectar) are hypothesized to be a ‘key innovation’ which can lead to rapid speciation within a lineage, because they are involved in pollinator specificity. Despite the ecological importance of nectar spurs, relatively little is known about their development. We used a comparative approach to investigate variation in nectar spur length in a clade of eight Iberian toadflaxes.MethodsSpur growth was measured at the macroscopic level over time in all eight species, and growth rate and growth duration compared. Evolution of growth rate was reconstructed across the phylogeny. Within the clade we then focused on Linaria becerrae and Linaria clementei, a pair of sister species which have extremely long and short spurs, respectively. Characterization at a micromorphological level was performed across a range of key developmental stages to determine whether the difference in spur length is due to differential cell expansion or cell division.Key ResultsWe detected a significant difference in the evolved growth rates, while developmental timing of both the initiation and the end of spur growth remained similar. Cell number is three times higher in the long spurred L. becerrae compared with L. clementei, whereas cell length is only 1.3 times greater. In addition, overall anisotropy of mature cells is not significantly different between the two species.ConclusionsWe found that changes in cell number and therefore in cell division largely explain evolution of spur length. This contrasts with previous studies in Aquilegia which have found that variation in nectar spur length is due to directed cell expansion (anisotropy) over variable time frames. Our study adds to knowledge about nectar spur development in a comparative context and indicates that different systems may have evolved nectar spurs using disparate mechanisms.

Species richness of Eurasian Zephyrus hairstreaks (Lepidoptera: Lycaenidae: Theclini) with implications on historical biogeography: An NDM/VNDM approach.

AimA database based on distributional records of Eurasian Zephyrus hairstreaks (Lepidoptera: Lycaenidae: Theclini) was compiled to analyse their areas of endemism (AoEs), species richness and distribution patterns, to explore their locations of past glacial refugia and dispersal routes.MethodsOver 2000 Zephyrus hairstreaks occurrences are analysed using the NDM/VNDM algorithm, for the recognition of AoEs. Species richness was calculated by using the option ‘Number of different classes’ to count the different classes of a variable presented in each 3.0°×3.0° grid cell, and GIS software was used to visualize distribution patterns of endemic species.ResultsCentres of species richness of Zephyrus hairstreaks are situated in the eastern Qinghai-Tibet Plateau (EQTP), Hengduan Mountain Region (HDMR) and the Qinling Mountain Region (QLMR). Latitudinal gradients in species richness show normal distribution with the peak between 25° N and 35° N in the temperate zone, gradually decreasing towards the poles. Moreover, most parts of central and southern China, especially the area of QLMR-EQTP-HDMR, were identified as AoEs that may have played a significant role as refugia during Quaternary global cooling. There are four major distributional patterns of Zephyrus hairstreaks in Eurasia: Sino-Japanese, Sino-Himalayan, high-mountain and a combined distribution covering all three patterns.ConclusionsZephyrus hairstreaks probably originated at least 23–24 Myr ago in E. Asia between 25° N to 35° N in the temperate zone. Cenozoic orogenies caused rapid speciation of this tribe and extrusion of the Indochina block resulted in vicariance between the Sino-Japanese and the Sino-Himalayan patterns. The four distribution patterns provided two possible dispersal directions: Sino-Japanese dispersal and Sino-Himalayan dispersal.

Genetic evidence for panmixia in a colony-breeding crater lake cichlid fish

Fine-scaled genetic structuring, as seen for example in many lacustrine fish, typically relates to the patterns of migration, habitat use, mating system or other ecological factors. Because the same processes can also affect the propensity of population differentiation and divergence, assessments of species from rapidly speciating clades, or with particularly interesting ecological traits, can be especially insightful. For this study, we assessed the spatial genetic relationships, including the genetic evidence for sex-biased dispersal, in a colony-breeding cichlid fish, Amphilophus astorquii, endemic to Crater Lake Apoyo in Nicaragua, using 11 polymorphic microsatellite loci (n = 123 individuals from three colonies). We found no population structure in A. astorquii either within colonies (no spatial genetic autocorrelation, r ~0), or at the lake-wide level (pairwise population differentiation FST = 0–0.013 and no clustering), and there was no sex-bias (male and female AIc values bounded 0) to this lack of genetic structure. These patterns may be driven by the colony-breeding reproductive behaviour of A. astorquii. The results suggest that strong philopatry or spatial assortative mating are unlikely to explain the rapid speciation processes associated with the history of this species in Lake Apoyo.

Phylogenetic Affinities of Indian Apple Snails: An Insight into the Tibetan Tectonic Terranes

Gastropods belong to a hyper-diverse clade of molluscs and encompass limpets, slugs and snails. Interestingly, gastropods have effectively colonized land, freshwaters and marine habitats. However, the relationships among and within its constituent taxa remained in flux for decades due to delimitation in their morphological and anatomical features and hence molecular approach has taken a lead. An understanding into the history of Tibetan tectonic terranes is attempted in this article to derive through the phylogeny of Pila. In our study, we have re-evaluated the relationships within Ampullariidae by collecting the species of Pila globosa and Pila virens (India) and analyzed them comparing with 18S rDNA sequences from public domain. Obtained sequences are evaluated through Maximum Likelihood method in MEGA v 5 which clearly established a single cluster for the three individual taxons namely P. globosa with high bootstrap support. Pila virens showed a paraphyletic cluster with Pila ovata. A low bootstrap value of a few species of Pila possibly inferred rapid speciation among them. The novel observation in the present study is that P. virens forming a secondary cluster with the species from Vietnam namely Pila polita, Pila conica and Pila ampullacea has authenticated that the south India and Vietnam are in the same latitude namely 9–20°N, supporting the paleomagnetic data which unravelled that India collided with Asia around 57 mya in the Coenozoic era, whereas ampullariids must have been in co-existence since the Cambrian Period indicating their long evolutionary affinity and zoo-geographical relationship.

Rapid hybrid speciation in Darwin's finches

Hybrid Speciation Galapagos finches have driven hypotheses of how speciation occurs. Most commonly, it is assumed that natural selection separates species originating from a single population on the basis of variation in traits that confer advantages for survival and reproduction. Lamichhaney et al

Limited dispersal and geographic barriers cause population differentiation and structuring in Begonia maxwelliana at both large and small scales

ABSTRACTBackground: Genetic divergence is one of the key processes in speciation. In the Begoniaceae, genetic divergence caused by limited gene flow may explain its high species diversity and endemicity. This hypothesis has been supported by past genetic work but there is a lack of empirical studies on the causes of limited gene flow.Aim: To identify the causes of limited gene flow in Begonia.Methods: We examined the genetic structure among the populations of Begonia maxwelliana at the macro- and micro-spatial scales using microsatellites, measured seed dispersal range and observed flowering phenology.Results: Population differentiation and structuring were detected at both the macro- and micro-scales. Dispersal range was short, and all populations showed similar reproductive behaviour.Conclusions: The strong population differentiation and structuring among the populations studied imply that they are evolutionarily significant units and possible candidates for speciation. Geographical barriers and limited seed dispersal restrict gene flow in the populations, and these factors may be responsible for the rapid speciation and large diversity in the family.

DNA binding in combination with capillary electrophoresis and inductively coupled plasma mass spectrometry for the rapid speciation analysis of mercury

AbstractWe herein developed a method for the rapid speciation analysis of Hg2+ and CH3Hg+ by using specific DNA sequences to bind Hg2+ and CH3Hg+ together with capillary electrophoresis and inductively coupled plasma mass spectrometry. The specific DNA sequences can combine with Hg2+ and CH3Hg+ with different binding force, which makes the formed DNA‐Hg2+ and DNA‐CH3Hg+ conjugates possess different negative charges, and thus make Hg2+ and CH3Hg+ can be more easily and rapidly separated by capillary electrophoresis under reverse mode. Under the optimal conditions, the Hg2+ and CH3Hg+ can be baseline separated and detected by capillary electrophoresis‐inductively coupled plasma mass spectrometry within 11 min with a detection limit of 0.12 and 0.10 ng/mL, respectively. With the help of the method, we have successfully determined Hg2+ and CH3Hg+ in dried fish muscle samples with a recovery of 95–101% and a relative standard deviation (n = 6) <5%. In comparison with previous capillary electrophoresis and inductively coupled plasma mass spectrometry methods, the proposed method has shorter separation time and higher sensitivity under the same conditions of inductively coupled plasma mass spectrometry. Higher sensitivity and shorter separation time make our method a valuable technique to the speciation analysis of trace Hg2+ and CH3Hg+ in the environmental and food samples.

Genome-wide single-nucleotide polymorphism data reveal cryptic species within cryptic freshwater snail species-The case of the Ancylus fluviatilis species complex.

DNA barcoding utilizes short standardized DNA sequences to identify species and is increasingly used in biodiversity assessments. The technique has unveiled an unforeseeably high number of morphologically cryptic species. However, if speciation has occurred relatively recently and rapidly, the use of single gene markers, and especially the exclusive use of mitochondrial markers, will presumably fail in delimitating species. Therefore, the true number of biological species might be even higher. One mechanism that can result in rapid speciation is hybridization of different species in combination with polyploidization, that is, allopolyploid speciation. In this study, we analyzed the population genetic structure of the polyploid freshwater snail Ancylus fluviatilis, for which allopolyploidization was postulated as a speciation mechanism. DNA barcoding has already revealed four cryptic species within A. fluviatilis (i.e., A. fluviatilis s. str., Ancylus sp. A–C), but early allozyme data even hint at the presence of additional cryptic lineages in Central Europe. We combined COI sequencing with high‐resolution genome‐wide SNP data (ddRAD data) to analyze the genetic structure of A. fluviatilis populations in a Central German low mountain range (Sauerland). The ddRAD data results indicate the presence of three cryptic species within A. fluviatilis s. str. occurring in sympatry and even syntopy, whereas mitochondrial sequence data only support the existence of one species, with shared haplotypes between species. Our study hence points to the limitations of DNA barcoding when dealing with organismal groups where speciation is assumed to have occurred rapidly, for example, through the process of allopolyploidization. We therefore emphasize that single marker DNA barcoding can underestimate the true species diversity and argue in strong favor of using genome‐wide data for species delimitation in such groups.

Molecular investigation of genetic assimilation during the rapid adaptive radiations of East African cichlid fishes.

Adaptive radiations are characterized by adaptive diversification intertwined with rapid speciation within a lineage resulting in many ecologically specialized, phenotypically diverse species. It has been proposed that adaptive radiations can originate from ancestral lineages with pronounced phenotypic plasticity in adaptive traits, facilitating ecologically driven phenotypic diversification that is ultimately fixed through genetic assimilation of gene regulatory regions. This study aimed to investigate how phenotypic plasticity is reflected in gene expression patterns in the trophic apparatus of several lineages of East African cichlid fishes, and whether the observed patterns support genetic assimilation. This investigation used a split brood experimental design to compare adaptive plasticity in species from within and outside of adaptive radiations. The plastic response was induced in the crushing pharyngeal jaws through feeding individuals either a hard or soft diet. We find that nonradiating, basal lineages show higher levels of adaptive morphological plasticity than the derived, radiated lineages, suggesting that these differences have become partially genetically fixed during the formation of the adaptive radiations. Two candidate genes that may have undergone genetic assimilation, gif and alas1, were identified, in addition to alterations in the wiring of LPJ patterning networks. Taken together, our results suggest that genetic assimilation may have dampened the inducibility of plasticity related genes during the adaptive radiations of East African cichlids, flattening the reaction norms and canalizing their feeding phenotypes, driving adaptation to progressively more narrow ecological niches.

Rapid hybrid speciation in Darwin's finches.

Homoploid hybrid speciation in animals has been inferred frequently from patterns of variation, but few examples have withstood critical scrutiny. Here we report a directly documented example, from its origin to reproductive isolation. An immigrant Darwin's finch to Daphne Major in the Galápagos archipelago initiated a new genetic lineage by breeding with a resident finch (Geospiza fortis). Genome sequencing of the immigrant identified it as a G. conirostris male that originated on Española >100 kilometers from Daphne Major. From the second generation onward, the lineage bred endogamously and, despite intense inbreeding, was ecologically successful and showed transgressive segregation of bill morphology. This example shows that reproductive isolation, which typically develops over hundreds of generations, can be established in only three.

Ion-pairing reversed-phase chromatography coupled to inductively coupled plasma mass spectrometry as a tool to determine mercurial species in freshwater fish

Most of analytical community is focused on reversed phase high performance liquid chromatography (RP-HPLC) for mercury speciation by employing mobile phases comprising of high salts and moderate amounts of organic solvents. This study aims at rapid mercury speciation analysis by ion-pairing RP-HPLC with inductively coupled plasma mass spectrometry (ICP-MS) detection only using low salts for the sake of green analytical chemistry. Two ion-pairing HPLC methods were developed on individual usage of positively and negatively charged ion-pairing reagents (tetrabutylammonium hydroxide –TBAH and sodium dodecylbenzene sulfonate –SDBS), where sodium 3-mercapto-1-propysulfonate (MPS) and l-cysteine (Cys) were individually added in mobile phases to transform mercury species into negative and positive Hg-complexes for good resolution. Addition of phenylalanine was also utilized for rapid baseline separation in combination of short C18 guard columns. Optimum mobile phases of 2.0mM SDBS+2.0mM Cys+1.0mM Phe (pH 3.0) and 4.0mM TBAH+2.0mM MPS+2.0mM Phe (pH 6.0) both achieved baseline separation of inorganic mercury (Hg2+), methylmercury (MeHg), ethylmercury (EtHg) and phenylmercury (PhHg) on two consecutive 12.5-mm C18 columns. The former mobile phase was selected for mercury speciation in freshwater fish because of short separation time (3.0min). Detection limits of 0.015 for Hg2+, 0.014 for MeHg, 0.028 for EtHg and 0.042μgL−1 for PhHg were obtained along with satisfactory precisions of peak height and area (1.0-2.8% for 5.0μgL−1 Hg-mixture standard). Good accordance of determined values of MeHg and total mercury in certified reference materials of fish tissue (GBW 10029) and tuna fish (BCR-463) with certified values as well as good recoveries (91–106%) proved good accuracy of the proposed method. An example application to freshwater fish indicated its potential in routine analysis, where MeHg was presented at 3.7–20.3μgkg−1 as the dominate species.

Admixture mapping in a hybrid zone reveals loci associated with avian feather coloration.

Identifying the genetic bases for colour patterns has provided important insights into the control and expression of pigmentation and how these characteristics influence fitness. However, much more is known about the genetic bases for traits based on melanin pigments than for traits based on another major class of pigments, carotenoids. Here, we use natural admixture in a hybrid zone between Audubon's and myrtle warblers (Setophaga coronata auduboni/S. c. coronata) to identify genomic regions associated with both types of pigmentation. Warblers are known for rapid speciation and dramatic differences in plumage. For each of five plumage coloration traits, we found highly significant associations with multiple single-nucleotide polymorphisms (SNPs) across the genome and these were clustered in discrete regions. Regions near significantly associated SNPs were enriched for genes associated with keratin filaments, fibrils that make up feathers. A carotenoid-based trait that differs between the taxa-throat colour-had more than a dozen genomic regions of association. One cluster of SNPs for this trait overlaps the Scavenger Receptor Class F Member 2 (SCARF2) gene. Other scavenger receptors are presumed to be expressed at target tissues and involved in the selective movement of carotenoids into the target cells, making SCARF2 a plausible new candidate for carotenoid processing. In addition, two melanin-based plumage traits-colours of the eye line and eye spot-show very strong associations with a single genomic region mapping to chromosome 20 in the zebra finch. These findings indicate that only a subset of the genomic regions differentiated between these two warblers are associated with the plumage differences between them and demonstrate the utility of reduced-representation genomic scans in hybrid zones.