Combining geometric morphometrics with pattern recognition for the investigation of species complexes

The parasitoid Hymenoptera contain a large number of species complexes that are as yet unresolved by traditional taxonomic studies. Evolutionary studies as well as biological control programmes often require further investigations which cannot rely on simple qualitative morphological characters; in many cases, particularly with dried specimens housed in museums, molecular approaches cannot be used. Recent developments in geometric morphometrics and statistical exploratory approaches open new perspectives for the objective evaluation of morphological characters in this taxonomic context. In this study, geometric morphometrics and pattern recognition approaches were applied to the wing shape and venation of two closely related braconid species considered to differ by subtle qualitative morphological head characters. Exploratory analyses such as kernel density estimates and Gaussian mixture analyses were used to explore the structure of the data in the multivariate morphometric space. Discrimination techniques (linear discriminant functions and neural networks combined with cross‐validations) were used to estimate the taxonomic value of qualitative characters. Gaussian mixtures highlighted the existence of two non‐overlapping groups. A good congruence was found between one of the two groups and the a priori defined Bassus tumidulus. The misclassification rate was higher for B. tegularis specimens, which also appeared morphometrically heterogeneous. Discrimination between the two a priori defined species was incomplete with misclassification rates higher than, or equal to, 6%. In most cases, the lack of congruence between species and morphometrically defined subgroups could be related to specimens that exhibited ambiguous qualitative character states. In summary, if two entities are present, they still need to be defined morphologically, while B. tegularis heterogeneity calls for further investigation of specimens of known origin and hosts. © 2003 The Linnean Society of London, Biological Journal of the Linnean Society, 2003, 80, 89–98.     

Combining geometric morphometrics and pattern recognition to identify interspecific patterns of skull variation: case study in sympatric Argentinian species of the genus Calomys (Rodentia: Cricetidae: Sigmodontinae)

Sympatric species of vesper mice Calomys laucha and Calomys musculinus are difficult to discriminate, especially in natural history collections where they are identified by external body measurements and cranial characteristics. Accurate identification of these two species can be important because only one of them, C. musculinus , is a Junin virus reservoir, the aetiological agent of the Argentine Hemorragic Fever. Research has focused into the development of molecular techniques to unambiguously identify these species. We apply statistical procedures from the field of pattern recognition to three-dimensional geometric morphometric data based on skull landmarks to identify sympatric species C. laucha , C. musculinus and Calomys venustus . Pattern recognition techniques correctly identified the three species without any prior information on specimen identity. By contrast to expectations, C. venustus differed from the other two species mainly on the basis of shape and not by its centroid size. The main sources of difference between C. laucha and C. musculinus were of shape, specifically localized at the landmarks defined by: (1) the sutures between the premaxillaries, the nasals and the frontals; (2) the sutures between the parietals, the frontals and the squamosals; and (3) the suture between the parietals and the interparietal. Nevertheless, allometries dominate the patterns of interspecific variation between these latter species and may partly explain past identification difficulties. Morphological evolution is discussed. The need for objective methods to define phenotypic clusters is highlighted with respect to the need for fast and precise biodiversity assessments.  © 2008 The Linnean Society of London, Biological Journal of the Linnean Society , 2008, 94 , 365–378.     

Gill morphometrics of two Antarctic fish species Pleuragramma antarcticum and Notothenia gibberifrons

Summary Gill dimensions of 27 juvenile and adult Pleuragramma antarcticum from the southern Weddell Sea and of 28 juvenile and adult Notothenia gibberifrons from the South Orkney and South Shetland Islands were estimated. The unit gill area (UGA) of P. antarcticum ranged from 75 to 167 mm²/g (mean=105); gill area index (GAI) and water-blood distance (WBD) were found to be 1.38 cm² and 3.3 m, respectively. The exponent d_g in the relationship total gill area to weight was found to be 0.90. It is concluded that P. antarcticum belongs to sluggish species and, although pelagic, its routine energy costs tend to be low. However, the closely packed lamellae (NL/mm=21) indicate more active behaviour in comparison with N. gibberifrons. A preliminary estimation of growth parameters (P, k) is presented. The sluggish behaviour of N. gibberifrons, as expected from its benthic mode of life, is reflected by the gill parameters: the very low UGA ranged from 39 to 118 mm²/g; GAI and d_g were found to be 0.73 cm² and 0.98, respectively. Comparing gill dimensions and general respiration characteristics of fish from antarctic, temperate and tropical waters it is concluded that antarctic fish have increased their scope for activity.     

Combining geometric morphometrics,molecular phylogeny,and micropaleontology to assess evolutionary patterns in Mallomonas (Synurophyceae: Heterokontophyta)

Synurophytes, also known as scaled chrysophytes, are ecologically important algae that produce an array of siliceous structures upon which their taxonomy is based. Despite occupying a key position within the photosynthetic heterokonts, the evolutionary history of synurophytes remains poorly constrained. Here, modern and Middle Eocene siliceous scales of the morphotaxon Mallomonas insignis are used as a model to investigate synurophyte evolutionary patterns. Structural details of scale morphology were examined comparatively with scanning electron microscopy and scored for geometric morphometric analyses to assess the stability of shape characters. Although consistent size differences exist (modern scales are larger than Eocene counterparts), the populations cannot be differentiated on the basis of shape or microstructural detail, implying considerable evolutionary stasis in scale morphology. A time‐calibrated relaxed molecular clock analysis using a three‐gene concatenated data set (27 strains) suggests that the M. insignis lineage predates the available fossil record, having diverged from closest congeneric taxa in the Cretaceous (≥94 Ma). However, the molecular analysis also implies that considerable genetic variability is present within several morphotaxa of Mallomonas, implying that substantial genetic variability has arisen despite the retention of uniform scale morphologies, and resulting in the widespread occurrence of cryptic taxa. Results from the synurophyte lineage are consistent with the notion of protracted ghost ranges (>10 Ma) implied by the molecular phylogenies of other algal groups, together pointing to the paucity of the fossil record of these organisms on these timescales.     

Tracing geographical patterns of population differentiation in a widespread mangrove gastropod: genetic and geometric morphometrics surveys along the eastern African coast

In the present study, we assessed the inter‐ and intrapopulation genetic and morphological variation of Cerithidea decollata along the eastern coast of Africa. The population structure of C. decollata along the latitudinal gradient was examined by sequencing 420 bp of the mitochondrial cytochrome c oxidase I (COI) gene in 172 snails from 29 sites, in a combined analysis with geometric morphometrics in 1799 snails from 32 sites. Analysis of molecular variance and spatial analysis of molecular variance showed a moderate spatial population differentiation from Kenya to the Republic of South Africa, suggesting genetic divergence between the northern, central, and southern regions. This structure appears to be the result of life‐history traits combined with oceanographic features. Haplotype network and mismatch analysis suggest a recent population expansion during the Holsteinian interglacial period in the northern region and several colonization events in the central and southern regions. The morphometric approach suggests that morphological variation in shell shape is somewhat independent of the genetic divergence, revealing an overlap of shape across the latitudinal gradient but significant differences among‐population at a local level. This may indicate that similar ecological pressures are acting along the coast, leading to the occurrence of similar morphological characters. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 107 , 647–663.     

Discrimination of Eubazus (Hymenoptera, Braconidae) sibling species using geometric morphometrics analysis of wing venation

Abstract.  Complexes of sibling and cryptic species are encountered frequently in parasitic Hymenoptera. Geometric morphometrics is a useful tool to detect minimal morphological variations, which often are undetectable by traditional morphological studies and even by classical morphometric approaches. We applied geometric morphometrics to wing venation to assess a complex case of sibling species in the genus Eubazus (Hymenoptera, Braconidae), parasitoids of conifer bark weevils of the genus Pissodes (Coleoptera, Curculionidae). The results and methods were compared with previous taxonomic studies on the same species, involving classical multivariate morphometrics, isoenzyme analyses, cross-mating experiments and biological observations. Geometric morphometrics confirmed the previous division into four distinct species. However, this approach enabled the four species to be separated simultaneously, with a reliability of 98.6% for well-classified females and 93.1% for males. A similar result in previous studies was obtained only by combining isoenzyme analyses and several canonical variate analyses, including many morphometric characters. Furthermore, measurements of wing venation were less time-consuming, more reliable and required less prior knowledge of braconid taxonomy than the measurements needed for the classical morphometrics methods. Geometric morphometrics was used also to test the effect of host species on wing shape. Several female populations of Eubazus semirugosus originating from three different Pissodes spp. were compared. Significant differences were found in wing shape between conspecific Eubazus from different host species. The results are discussed in relation to reproductive isolation and genetic flow between the four species.     

Limited genetic connectivity of Pavona gigantea in the Mexican Pacific

Coral reefs are the most complex and diverse of aquatic ecosystems. Their vulnerability and deterioration in the face of anthropogenic disturbance require the adoption of conservation and restoration efforts to maintain their resilience, for which connectivity is of paramount importance. Dispersal of meroplanktonic larval stages drives the levels of connectivity among coral populations and is influenced by the local current regime, the synchronization of spawning events, and the capacity of larvae to reach recruitment sites. This research aims to quantify the levels of connectivity among Pavona gigantea populations in the Mexican Pacific, using two mitochondrial genes and a nuclear gene. Mitochondrial genes were insufficiently variable to test geographical heterogeneity, whereas the more variable (h ≥ 0.86) nuclear rDNA indicated significant geographic differentiation (Φ _ST  = 0.159, P < 0.001) among five locations along the Mexican Pacific, but no evidence of isolation by distance. Gene flow was limited among most sampled locales, and the largest estimate suggested moderate and unidirectional gene flow from Huatulco Bays to La Paz Bay and Marietas Islands. We found partial agreement between the patterns of connectivity among localities and the general pattern of superficial oceanographic circulation of the region, particularly in reference with the expected influence of the northward flowing West Mexican Current. These results suggest a limited demographic connectivity among Pavona gigantea populations along the Mexican Pacific, mediated by passive larval transport, and highlight the difficulty of predicting connectivity patterns on the basis of highly variable oceanographic regimes and reproductive events. The limited connectivity is of consequence for the viability and vulnerability of local populations and should be considered in the management and conservation strategies in the region.     

Combining genetic and biochemical approaches to identify functional molecular contact points

Protein-protein interactions are required for many viral and cellular functions and are potential targets for novel therapies. Here we detail a series of genetic and biochemical techniques used in combination to find an essential molecular contact point on the duck hepatitis B virus polymerase. These techniques include differential immunoprecipitation, mutagenesis and peptide competition. The strength of these techniques is their ability to identify contact points on intact proteins or protein complexes employing functional assays. This approach can be used to aid identification of putative binding sites on proteins and protein complexes which are resistant to characterization by other methods.     

Advances in insectivore and rodent systematics due to geometric morphometrics

• 1 Morphometrics, the study of the variation and change in form amongst organisms, serves as a basic methodological tool in various fields of biological research, including systematics. Because it includes information about spatial relationships amongst anatomical landmarks, geometric morphometrics is more suitable for analyzing morphometric variation than methods based on distance measurements.
• 2 Geometric morphometrics allows us to answer general ecological and evolutionary questions about shape.
• 3 In this paper, landmark‐based methods are described and illustrated, based on a dataset of measurements from 295 Apodemus mandibles, and the applications of such methods in the systematics of insectivores (Eulipotyphla) and rodents (Rodentia) are summarized.

     

Low genetic structure and diversity of Red-billed Tropicbirds in the Mexican Pacific

Understanding genetic structure and gene flow can elucidate the mechanisms of diversification and adaptation in seabirds and help define conservation and management units. From 2012 to 2016, we collected blood and feather samples from 156 Red-billed Tropicbirds (Phaethon aethereus) from seven colonies distributed along the Gulf of California and Mexican tropical Pacific to estimate genetic diversity, genetic structure, and gene flow using microsatellite markers and mitochondrial DNA (mtDNA; control region) sequences. Nuclear and mtDNA data revealed relatively low or null levels of genetic diversity, respectively, possibly the result of a founder effect in the eastern Pacific followed by a subsequent population expansion. Nuclear data revealed significant genetic structure among the colonies, but the differences were not associated with regional grouping (i.e., Gulf of California vs. Tropical Pacific). Greater gene flow was observed from the tropical Pacific toward the Gulf of California, possibly related to shared dispersal patterns during the non-breeding season (individuals traveling north to reach warm currents with abundant prey). With the exception of one colony in the Mexican tropical Pacific, we found no evidence of recent bottleneck events. Nonetheless, the overall reduced genetic diversity suggests a high intrinsic vulnerability and risk of extinction for this species.     

Multivariate and geometric morphometrics in the analysis of sexual dimorphism variation in Podarcis lizards

Podarcis bocagei and P. carbonelli are two closely related lacertid species, very similar morphologically and ecologically. We investigated sexual dimorphism patterns presented by both species in allopatry and in sympatry. Sexual size and shape dimorphism patterns were analyzed using both multivariate and geometric morphometric techniques. Multivariate morphometrics revealed a marked sexual dimorphism in both species--males being larger with more robust habitus and females presenting a longer trunk. General patterns of sexual size dimorphism are not modified in sympatry, although there is evidence for some morphological change in male head size. The application of geometric morphometrics offered a more detailed image of head shape and revealed that males present a more developed tympanic area than do females, while females have a more rounded head. Differences in the degree of sexual shape dimorphism were detected in sympatry, but no consistent patterns were observed. From the results of the study, and based on previous knowledge on the populations studied, we conclude that the morphological differences observed are probably not caused by exploitative competition between the species, but rather appear attributable to the modification of the relative influence of sexual and natural selection on both sexes.     

基于几何形态测量学的黄蜻翅的雌雄二态性研究

【目的】本研究运用几何形态测量学方法对黄蜻Pantala flavescens Fabricius前翅和后翅的雌雄二态性进行分析,探讨黄蜻雌雄性在翅上的形态差异。【方法】通过对黄蜻的前翅和后翅做数字化标点,获得翅的形态信息。经主成分分析(PCA)和薄片样条法(TPS)获得雌雄性在翅型和翅脉上的差异性。【结果】PCA结果表明,黄蜻雌雄性的前翅和后翅在翅型、翅脉结构上存在着明显差异,而且后翅间的差异更大。经TPS分析可知,雌雄性黄蜻前翅上的差异部位主要发生在亚翅结和三角室,后翅的差异部位主要发生在亚翅结、肘脉域和臀角区域。对黄蜻前后翅的大小分析(CS)可知,雌性黄蜻前后翅较大,雄性黄蜻前后翅较小。【结论】黄蜻雌雄性间的翅型结构差异性表明雌雄性在其各自生活史中履行不同的职责,特定的飞行模式逐渐形成特定的翅型、翅脉结构。     

Integration of cytochrome c oxidase I barcodes and geometric morphometrics to delimit species in the genus Gnopharmia (Lepidoptera: Geometridae,Ennominae)

We tested the hypothesis of species taxonomy in the genus Gnopharmia (Macariini, Ennominae) that was recently established in a review based on discrete morphological characters. For this objective we integrated both DNA‐based and morphometric approaches in order to infer species boundaries. A 658‐bp fragment of the mitochondrial cytochrome c oxidase subunit 1 (CO1) (DNA barcode) was analysed from populations of five species distributed throughout the Middle East to assess their consistency with traditionally defined morphospecies. Signals in the morphological variation of the aedeagus of all relevant populations were evaluated using geometric landmarks. Consistent groupings compatible with the current taxonomic classification were found with both approaches. The results strongly support the distinction of seven closely related species. © 2013 The Linnean Society of London     

Combining the least cost path method with population genetic data and species distribution models to identify landscape connectivity during the late Quaternary in Himalayan hemlock

Himalayan hemlock (Tsuga dumosa) experienced a recolonization event during the Quaternary period; however, the specific dispersal routes are remain unknown. Recently, the least cost path (LCP) calculation coupled with population genetic data and species distribution models has been applied to reveal the landscape connectivity. In this study, we utilized the categorical LCP method, combining species distribution of three periods (the last interglacial, the last glacial maximum, and the current period) and locality with shared chloroplast, mitochondrial, and nuclear haplotypes, to identify the possible dispersal routes of T. dumosa in the late Quaternary. Then, both a coalescent estimate of migration rates among regional groups and establishment of genetic divergence pattern were conducted. After those analyses, we found that the species generally migrated along the southern slope of Himalaya across time periods and genomic makers, and higher degree of dispersal was in the present and mtDNA haplotype. Furthermore, the direction of range shifts and strong level of gene flow also imply the existence of Himalayan dispersal path, and low area of genetic divergence pattern suggests that there are not any obvious barriers against the dispersal pathway. Above all, we inferred that a dispersal route along the Himalaya Mountains could exist, which is an important supplement for the evolutionary history of T. dumosa. Finally, we believed that this integrative genetic and geospatial method would bring new implications for the evolutionary process and conservation priority of species in the Tibetan Plateau.     

Combining gene expression and genetic analyses to identify candidate genes involved in cold responses in pea

Cold stress affects plant growth and development. In order to better understand the responses to cold (chilling or freezing tolerance), we used two contrasted pea lines. Following a chilling period, the Champagne line becomes tolerant to frost whereas the Terese line remains sensitive.