Nuclear Gene Markers Research Articles

Cephalotrichella echinicola, sp. nov. (Palaeonemertea : Cephalotrichellidae), a new nemertean associated with sea urchins from Nha Trang Bay (South China Sea)

About 50 nemertean species have been reported to live in symbiotic relationships with other invertebrates, but only two hoplonemertean species are associated with echinoderms (starfish). The palaeonemertean Cephalotrichella echinicola, sp. nov. is described from samples collected in Nha Trang Bay, Vietnam, South China Sea. The species is the first known nemertean associated with sea urchins (Metalia sternalis and M. spatagus), living on both the oral and the aboral surfaces of the host and freely moving among its spines. The internal morphology of the new species is described based on histological sections and confocal laser scanning microscopy with phalloidin and antibody labelling. Sequences of three nuclear gene markers (18S rRNA, 28S rRNA, and histone H3) and two mitochondrial gene markers (16S rRNA and COI) were compared with those of other palaeonemertean species, and a phylogenetic analysis suggested that C. echinicola is closest to the free-living Cephalotrichella alba Gibson & Sundberg, 1992. Both the morphological data and the phylogenetic analysis provide additional evidence for distinguishing the families Cephalotrichidae and Cephalotrichellidae and support the rejection of Cephalotrichella as a junior synonym of Cephalothrix. A new diagnosis of the genus Cephalotrichella is given. http://zoobank.org/urn:lsid:zoobank.org:pub:091B5D56-71B2-4F4C-9AD8-F666B4610DE2

Phylogenetic analysis of Rhizoclonium (Cladophoraceae, Cladophorales), and the description of Rhizoclonium subtile sp. nov. from China

The genus Rhizoclonium (Cladophoraceae, Cladophorales) accommodates uniserial, unbranched filamentous algae, closely related to Cladophora and Chaetomorpha. Its taxonomy has been problematic for a long time due to the lack of diagnostic morphological characters. To clarify the species diversity and taxonomic relationships of this genus, we collected and analyzed thirteen freshwater Rhizoclonium specimens from China. The morphological traits of these specimens were observed and described in detail. Three nuclear gene markers small subunit ribosomal DNA (SSU), large subunit ribosomal DNA (LSU) and internal transcribed spacer 2 (ITS2) sequences were analyzed to elucidate their phylogenetic relationships. The results revealed that there were at least fifteen molecular species assignable to Rhizoclonium and our thirteen specimens were distributed in four clades. On the basis of morphological and molecular evidence we propose the new species, R. subtile sp. nov.

Diversity and phylogeny of Sargassum (Fucales, Phaeophyceae) in Singapore

Sargassum species play key ecological roles on coral reefs, yet their diversity remains poorly known. Precise identification of Sargassum species, however, is improving with molecular genetic tools, though these have yet to be applied rigorously in Singapore. Historical records list 41 species, but no more than ten were verified based on herbarium vouchers, and even fewer (five species) were confirmed in the field based on a single nuclear gene marker in a previous study. Here, we revised the diversity of Sargassum in Singapore by examining all the morphologically distinct forms collected from the local coral reef environment. A total of six morphotypes, Sargassum aquifolium (Turner) C.Argardh (1820), S. cf. granuliferum C.Argardh (1820), S. ilicifolium (Turner) C.Argardh (1820), S. swartzii C.Argardh (1820), S. polycystum C.Argardh (1824), and an undescribed taxon ‘Sargassum sp.’ (Mattio and Payri 2009), were delineated based on morphological characteristics. The morphotypes were placed in five molecular clades based on phylogenetic analyses of the nuclear ITS-2, chloroplastic partial RuBisCO operon rbcLS, and mitochondrial cox3. Sargassum cf. granuliferum, though morphologically distinct from all other species, is not phylogenetically distinct from S. polycystum. Our results provide a species list for Singapore that will be valuable for future studies on macroalgal biogeography and species-specific ecological relationships with other reef organisms, particularly corals.

Barcoding and multi-locus phylogeography of the globally distributed calcareous tubeworm genus Hydroides Gunnerus, 1768 (Annelida, Polychaeta, Serpulidae)

Hydroides is a large and diverse group of calcareous tubeworms (Serpulidae, Annelida) recognised by a distinctive but variable two-tiered operculum. Despite considerable research using several species of Hydroides as models in ecological and biofouling studies, phylogenetic and biogeographic relationships within the genus are still poorly understood. Using combined mitochondrial (COI, cytochrome b) and nuclear (18S, 28S and ITS) gene markers for 284 individuals of 45 morphospecies of Hydroides, we investigated the global phylogenetic and biogeographic relationships within the genus. Phylogenetic topologies were well supported and indicated high genetic diversity within Hydroides, revealing potential cryptic species. Present results also include the first COI barcoding data enabling rapid and effective species identification of Hydroides on a global scale. Phylogenetic relationships within Hydroides were more concordant with geographical distributions than morphological similarity of their opercula. Molecular divergence estimates suggested the origin and subsequent diversification in the western Tethys Sea followed by a shift of the historical centre of diversity from the Indo-Mediterranean region to the central Indo-Pacific during the last 50 million years. Further studies on population genetics of species consisting of multiple lineages would provide a better understanding on the status of potential cryptic species. Furthermore, paleogeographic studies based on fossil Hydroides tubes would provide evidence to test this biogeographic hypothesis.

Assessing the Ability of Chloroplast and Nuclear DNA Gene Markers to Verify the Geographic Origin of Jatoba (Hymenaea courbaril L.) Timber.

Deforestation-reinforced by illegal logging-is a serious problem in many tropical regions and causes pervasive environmental and economic damage. Existing laws that intend to reduce illegal logging need efficient, fraud resistant control methods. We developed a genetic reference database for Jatoba (Hymenaea courbaril), an important, high value timber species from the Neotropics. The data set can be used for controls on declarations of wood origin. Samples from 308 Hymenaea trees from 12 locations in Brazil, Bolivia, Peru, and French Guiana have been collected and genotyped on 10 nuclear microsatellites (nSSRs), 13 chloroplast SNPs (cpSNP), and 1 chloroplast indel marker. The chloroplast gene markers have been developed using Illumina DNA sequencing. Bayesian cluster analysis divided the individuals based on the nSSRs into 8 genetic groups. Using self-assignment tests, the power of the genetic reference database to judge on declarations on the location has been tested for 3 different assignment methods. We observed a strong genetic differentiation among locations leading to high and reliable self-assignment rates for the locations between 50% to 100% (average of 88%). Although all 3 assignment methods came up with similar mean self-assignment rates, there were differences for some locations linked to the level of genetic diversity, differentiation, and heterozygosity. Our results show that the nuclear and chloroplast gene markers are effective to be used for a genetic certification system and can provide national and international authorities with a robust tool to confirm legality of timber.

Genetic diversity and population structure of penis fish (Urechis unicinctus) based on mitochondrial and nuclear gene markers

Urechis unicinctus is distributed only in Bohai Gulf of China and Korean and Japanese coast. The wild populations of this species have sharply declined in China and Japan. We collected 105 samples from six localities of Bohai Gulf and Korea coast, and investigated genetic diversity and population structure with mitochondrial COI, 16S-rRNA and nuclear 28S-rRNA gene fragments. Genetic diversity of U. unicinctus based on COI sequences was still high (Hd: 0.9595, π: 0.0101), however, 28S-rRNA gene sequences showed low level of genetic diversity (Hd: 0.4084, π: 0.0007). Based on COI sequences, FST values between populations ranged from −0.00204 to 0.05210, and 99.12% genetic diversity was contributed by different individuals within population. Both phylogenetic trees and median-joining network did not show clear geographic cluster, haplotypes from different populations were mixed. Our results indicated low level of genetic divergence among different localities of U. unicinctus, and this species should be treated as a whole population among China, Japan and Korea coast during species conservation.

A new endemic Impatiens species on Mount Gorongosa (Mozambique) demonstrates the conservation importance of montane areas in Africa

A new Impatiens species from Mozambique, Impatiens wuerstenii, is described and illustrated. The material used for the description was initially mistaken for the morphologically similar Impatiens species I. psychadelphoides, a known endemic from Mozambique. However, a combined molecular – morphological study demonstrates that the populations found on Mount Gorongosa do not belong to I. psychadelphoides but are a species on their own. Phylogenetic analyses based on chloroplast and nuclear gene markers demonstrate that I. wuerstenii is sister to the floral morphologically distinct I. salpinx, and that I. psychadelphoides and I. cecilii are sister to that lineage. The presence of a new endemic species on Mount Gorongosa indicates the importance of this mountain area on the overall biodiversity of the region and emphasizes the need for a more thorough conservation and biological exploration of the mountain.

Hybrid origin of Asian aspermic <i>Fasciola</i> flukes is confirmed by analyzing two single-copy genes, <i>pepck</i> and <i>pold</i>

Nuclear gene markers, phosphoenolpyruvate carboxykinase (pepck) and DNA polymerase delta (pold), have been developed for precise discrimination of Fasciola flukes instead of internal transcribed spacer 1. In this study, these two genes of 730 Fasciola flukes from eight Asian countries were analyzed. The results were compared with their mitochondrial NADH dehydrogenase subunit 1 (nad1) lineages for obtaining a definitive evidence of the hybrid origin of aspermic Fasciola flukes. All the flukes categorized into the aspermic nad1 lineages possessed both the fragment patterns of F. hepatica and F. gigantica (mixed types) in pepck and/or pold. These findings provide clear evidence for the hybrid origin of aspermic Fasciola lineages and suggest that “aspermic Fasciola flukes” should hereafter be called “hybrid Fasciola flukes”.

Prevalence and Identity of Taenia multiceps cysts “Coenurus cerebralis” in Sheep in Egypt

Coenurosis is a parasitic disease caused by the larval stage (Coenurus cerebralis) of the canids cestode Taenia multiceps. C. cerebralis particularly infects sheep and goats, and pose a public health concerns. The present study aimed to determine the occurrence and molecular identity of C. cerebralis infecting sheep in Egypt. Infection rate was determined by postmortem inspection of heads of the cases that showed neurological manifestations. Species identification and genetic diversity were analyzed based on PCR-sequence analysis of nuclear ITS1 and mitochondrial cytochrome oxidase (COI) and nicotinamide adenine dinucleotide dehydrogenase (ND1) gene markers. Out of 3668 animals distributed in 50 herds at localities of Ashmoun and El Sadat cities, El Menoufia Province, Egypt, 420 (11.45%) sheep showed neurological disorders. Postmortem examination of these animals after slaughter at local abattoirs indicated to occurrence of C. cerebralis cysts in the brain of 111 out of 420 (26.4%), with overall infection rate 3.03% of the involved sheep population. Molecular analysis of representative samples of coenuri at ITS1 gene marker showed extensive intra- and inter-sequence diversity due to deletions/insertions in the microsatellite regions. On contrast to the nuclear gene marker, considerably low genetic diversity was seen in the analyzed mitochondrial gene markers. Phylogenetic analysis based on COI and ND1 gene sequences indicated that the generated sequences in the present study and the reference sequences in the database clustered in 4 haplogroups, with more or less similar topologies. Clustering pattern of the phylogenetic tree showed no effect for the geographic location or the host species.

First genetic characterization of Fasciola hepatica in Argentina by nuclear and mitochondrial gene markers

Fasciola hepatica is a trematode showing genetic variation among isolates from different regions of the world. The objective of this work was to characterize for the first time F. hepatica isolates circulating in different regions of Argentina. Twenty-two adult flukes were collected from naturally infected bovine livers in different areas from Argentina and used for DNA extraction. We carried out PCR amplification and sequence analysis of the ribosomal internal transcribed spacer 1 (ITS1), mitochondrial nicotinamide adenine dinucleotide dehydrogenase subunits 4 and 5 (nad4 and nad5) and mitochondrial cytochrome c oxidase subunit I (cox1) genes as genetic markers. Phylogenies were reconstructed using maximum parsimony algorithm. A total of 6 haplotypes were found for cox1, 4 haplotypes for nad4 and 3 haplotypes for nad5. The sequenced ITS1 fragment was identical in all samples. The analyzed cox1 gene fragment is the most variable marker and is recommended for future analyses. No geographic association was found in the Argentinean samples.

Molecular phylogeny of Pasiphaeidae (Crustacea, Decapoda, Caridea) reveals systematic incongruence of the current classification

Caridean shrimps constitute one of the most diverse groups of decapod crustaceans, notwithstanding their poorly resolved infraordinal relationships. One of the systematically controversial families in Caridea is the predominantly pelagic Pasiphaeidae, comprises 101 species in seven genera. Pasiphaeidae species exhibit high morphological disparity, as well as ecological niche width, inhabiting shallow to very deep waters (>4000m). The present work presents the first molecular phylogeny of the family, based on a combined dataset of six mitochondrial and nuclear gene markers (12S rDNA, 16S rDNA, histone 3, sodium–potassium ATPase α-subunit, enolase and ATP synthase β-subunit) from 33 species belonged to six genera of Pasiphaeidae with 19 species from 12 other caridean families as outgroup taxa. Maximum likelihood and Bayesian inference analyses conducted on the concatenated dataset of 2265bp suggest the family Pasiphaeidae is not monophyletic, with Psathyrocaris more closely related to other carideans than to the other five pasiphaeid genera included in this analysis. Leptochela occupies a sister position to the remaining genera and is genetically quite distant from them. At the generic level, the analysis supports the monophyly of Pasiphaea, Leptochela and Psathyrocaris, while Eupasiphae is shown to be paraphyletic, closely related to Parapasiphae and Glyphus. The present molecular result strongly implies that certain morphological characters used in the present systematic delineation within Pasiphaeidae may not be synapomorphies and the classification within the family needs to be urgently revised.

Sequence-based species delineation and molecular phylogenetics of the transitional Nearctic–Neotropical grasshopper genus Taeniopoda (Orthoptera, Romaleidae)

Taeniopoda is a genus of grasshoppers currently represented by 12 species distributed from southern USA to Panama, with most of them occurring along the transitional Nearctic–Neotropical region in central and southern Mexico. Despite being a small group of conspicuous, colourful species, the systematics of Taeniopoda has been largely neglected, including its phylogenetic affinity with the morphologically similar, monotypic genus Romalea. Here, we assessed the species limits in 11 of the species of Teniopoda based on two mitochondrial (mt) markers (COI, cyt b). Phylogenetic relationships were reconstructed adding two nuclear gene markers (28S, H3). A relaxed molecular clock analysis was performed based on the mt markers. We detected nuclear mt paralogues (numts) and the probable introgression of T. tamaulipensis mtDNA in specimens of T. eques from central Mexico. Between six and 14 species of Taeniopoda were delimited by the sequence-based approaches performed (COI divergence with thresholds of 1 and 2%; General Mixed Yule-Coalescent (GMYC) model). The GMYC and 1% threshold analyses with COI were more congruent with the currently recognized morphology-based taxonomy with 10 and 11 putative species, respectively. Four of these species were regarded as ‘stable’, since they were supported by at least one of the molecular analyses and by diagnostic morphological features. The species-based phylogeny recovered Taeniopoda as paraphyletic with respect to the monotypic genus Romalea. Three morphologically and geographically congruent major clades were recovered, two with species having a considerably elevated pronotal crest and one with its members having it less elevated. The origin and subsequent diversification of Taeniopoda were estimated to occur from the mid and late Miocene to Pliocene, respectively. The current species diversity in Taeniopoda was estimated to occur during the Pleistocene, which was probably influenced by the climatic oscillations that occurred during this period and the uplift of mountain ranges in Central America.

Complete mitochondrial genomes of prasinophyte algae Pyramimonas parkeae and Cymbomonas tetramitiformis.

Mitochondria are archetypal eukaryotic organelles that were acquired by endosymbiosis of an ancient species of alpha-proteobacteria by the last eukaryotic common ancestor. The genetic information contained within the mitochondrial genome has been an important source of information for resolving relationships among eukaryotic taxa. In this study, we utilized mitochondrial and chloroplast genomes to explore relationships among prasinophytes. Prasinophytes are represented by diverse early-diverging green algae whose physical structures and genomes have the potential to elucidate the traits of the last common ancestor of the Viridiplantae (or Chloroplastida). We constructed de novo mitochondrial genomes for two prasinophyte algal species, Pyramimonas parkeae and Cymbomonas tetramitiformis, representing the prasinophyte clade. Comparisons of genome structure and gene order between these species and to those of other prasinophytes revealed that the mitochondrial genomes of P.parkeae and C.tetramitiformis are more similar to each other than to other prasinophytes, consistent with other molecular inferences of the close relationship between these two species. Phylogenetic analyses using the inferred amino acid sequences of mitochondrial and chloroplast protein-coding genes resolved a clade consisting of P.parkeae and C.tetramitiformis; and this group (representing the prasinophyte clade I) branched with the clade II, consistent with previous studies based on the use of nuclear gene markers.

Genetic Diversity and Structure of Lolium Species Surveyed on Nuclear Simple Sequence Repeat and Cytoplasmic Markers.

To assess the genetic diversity and population structure of Lolium species, we used 32 nuclear simple sequence repeat (SSR) markers and 7 cytoplasmic gene markers to analyze a total of 357 individuals from 162 accessions of 9 Lolium species. This survey revealed a high level of polymorphism, with an average number of alleles per locus of 23.59 and 5.29 and an average PIC-value of 0.83 and 0.54 for nuclear SSR markers and cytoplasmic gene markers, respectively. Analysis of molecular variance (AMOVA) revealed that 16.27 and 16.53% of the total variation was due to differences among species, with the remaining 56.35 and 83.47% due to differences within species and 27.39 and 0% due to differences within individuals in 32 nuclear SSR markers set and 6 chloroplast gene markers set, respectively. The 32 nuclear SSR markers detected three subpopulations among 357 individuals, whereas the 6 chloroplast gene markers revealed three subpopulations among 160 accessions in the STRUCTURE analysis. In the clustering analysis, the three inbred species clustered into a single group, whereas the outbreeding species were clearly divided, especially according to nuclear SSR markers. In addition, almost all Lolium multiflorum populations were clustered into group C4, which could be further divided into three subgroups, whereas Lolium perenne populations primarily clustered into two groups (C2 and C3), with a few lines that instead grouped with L. multiflorum (C4) or Lolium rigidum (C6). Together, these results will useful for the use of Lolium germplasm for improvement and increase the effectiveness of ryegrass breeding.

A novel set of single-copy nuclear gene markers in white oak and implications for species delimitation

Oaks have often been the focus of research on plant evolution owing to their propensity to intercross and their important role in ecology and economy. Compared with traditional molecular markers, such as amplified fragment length polymorphisms (AFLPs) and simple sequence repeats (SSRs), multiple single-copy nuclear genes (SCNGs) are of greater utility in inferring evolutionary processes in oaks. Nineteen primer pairs were developed from expressed sequence tags (ESTs) of Quercus mongolica and Q. robur that could produce orthologous products in Chinese white oaks (section Quercus). These SCNG markers showed a moderate to high level of nucleotide polymorphism in 42 individuals of two closely related white oaks, Q. mongolica and Q. liaotungensis, and demonstrated high transferability across seven white oaks, four oaks from section Cerris, and one oak from section Lobatae. A phylogenetic tree based on these SCNGs provided resolution at deep nodes and robust support for delimiting populations of Q. mongolica and Q. liaotungensis; Bayesian analysis clustered individuals into their respective species with high probability and no admixture. When the same individuals were used, Bayesian clusters based on either 194 AFLPs or 19 SSRs gave comparable results, but one or several individuals respectively were identified as having admixed ancestry. This indicates that the hybridization rate between these two oaks may have been overestimated using SSR markers due to the occurrence of homoplasy. The SCNGs are powerful for species delimitation of white oaks, and these markers could be useful for future phylogenetics and phylogeography research in white oaks.

A first higher-level time-calibrated phylogeny of antlions (Neuroptera: Myrmeleontidae)

In this study, we reconstruct the first time-calibrated phylogeny of the iconic antlion family, the Myrmeleontidae (Neuroptera: Myrmeleontiformia). We use maximum likelihood and Bayesian inference to analyse a molecular dataset based on seven mitochondrial and nuclear gene markers. The dataset encompasses 106 species of Neuroptera, including 94 antlion species. The resulting phylogenetic framework provides support for a myrmeleontid classification distinguishing four subfamilies: Acanthaclisinae, Myrmeleontinae, Palparinae, and Stilbopteryginae. Within Myrmeleontinae, Myrmecaelurini and Nemoleontini are recovered as monophyletic clades; Gepini also appears as a valid tribe, distinct from Myrmecaelurini whereas Myrmecaelurini and Nesoleontini on one hand and Brachynemurini and Dendroleontini on the other hand, appear closely related. Some preliminary information related to generic and specific levels are also implied from our results, such as the paraphyly of several genera. Dating analyses based on thoroughly evaluated fossil calibrations indicate that the antlion family likely originated in the Cretaceous, between 135 and 138 million years ago (depending on the set of fossil calibrations), and that all higher-level lineages appeared during the Early Cretaceous. This first phylogenetic hypothesis will provide a valuable basis to further expand the taxonomic coverage and molecular sampling, and to lay the foundations of future systematic revisions.

Northeast India Helminth Parasite Information Database (NEIHPID): Knowledge Base for Helminth Parasites.

Most metazoan parasites that invade vertebrate hosts belong to three phyla: Platyhelminthes, Nematoda and Acanthocephala. Many of the parasitic members of these phyla are collectively known as helminths and are causative agents of many debilitating, deforming and lethal diseases of humans and animals. The North-East India Helminth Parasite Information Database (NEIHPID) project aimed to document and characterise the spectrum of helminth parasites in the north-eastern region of India, providing host, geographical distribution, diagnostic characters and image data. The morphology-based taxonomic data are supplemented with information on DNA sequences of nuclear, ribosomal and mitochondrial gene marker regions that aid in parasite identification. In addition, the database contains raw next generation sequencing (NGS) data for 3 foodborne trematode parasites, with more to follow. The database will also provide study material for students interested in parasite biology. Users can search the database at various taxonomic levels (phylum, class, order, superfamily, family, genus, and species), or by host, habitat and geographical location. Specimen collection locations are noted as co-ordinates in a MySQL database and can be viewed on Google maps, using Google Maps JavaScript API v3. The NEIHPID database has been made freely available at http://nepiac.nehu.ac.in/index.php

Sergey gen. n., a new doryctine genus from temperate forests of Mexico and Cuba (Hymenoptera, Braconidae).

The new doryctine genus Sergey gen. n. is described with four new species (Sergey cubaensis Zaldívar-Riverón & Martínez, sp. n., Sergey coahuilensis Zaldívar-Riverón & Martínez, sp. n., Sergey tzeltal Martínez & Zalídivar-Riverón, sp. n., Sergey tzotzil Martínez & Zalídivar-Riverón, sp. n.) from temperate forests of Mexico and Cuba. Similar to many other doryctine taxa, the new genus has a considerably elongated, petiolate basal sternal plate of the first metasomal tergite, although it can be distinguished from these by having the mesoscutum sharply declivous anteriorly with sharp anterolateral edges. The described species have been characterised molecularly based on two mitochondrial (COI, cyt b) and one nuclear (28S) gene markers. Based on the mitochondrial gene genealogies reconstructed, the evidence suggests the existence of incomplete lineage sorting or hybridization in the populations from Chiapas and Oaxaca assigned to Sergey tzeltal sp. n.

Genetic variation and phylogenetic relationships among Rehmannia (Scrophulariaceae) species as revealed by a novel set of single-copy nuclear gene markers

Single-copy nuclear genes (SCNGs) are effective tools for phylogenetic analysis in plants. In this study, genetic variation and interspecific relationships in the genus Rehmannia were investigated by a novel set of SCNG markers. In total, 12 pairs of primers were successfully amplified and sequenced in six Rehmannia species after screening 258 candidate genes from the transcriptome datasets of Rehmannia glutinosa. The moderate to high level of nucleotide polymorphisms were identified in these SCNG markers, and nucleotide diversity (π) per locus ranged from 0.00498 to 0.01100, with 6–10 haplotypes. R. chingii and R. piasezkii exhibited a high level of genetic variation, while R. glutinosa and R. solanifolia had no genetic diversity. Phylogenetic analyses suggested that R. glutinosa - R. solanifolia, R. elata - R. piasezkii should be classified as one species, respectively. Molecular dating supported the rapid speciation of Rehmannia in Pleistocene. The single-copy DNA markers developed here would be also helpful for the study on the process of speciation and the population dynamic history of Rehmannia.

Constrained body shape among highly genetically divergent allopatric lineages of the supralittoral isopod Ligia occidentalis (Oniscidea).

Multiple highly divergent lineages have been identified within Ligia occidentalis sensu lato, a rocky supralittoral isopod distributed along a ~3000km latitudinal gradient that encompasses several proposed marine biogeographic provinces and ecoregions in the eastern Pacific. Highly divergent lineages have nonoverlapping geographic distributions, with distributional limits that generally correspond with sharp environmental changes. Crossbreeding experiments suggest postmating reproductive barriers exist among some of them, and surveys of mitochondrial and nuclear gene markers do not show evidence of hybridization. Populations are highly isolated, some of which appear to be very small; thus, the effects of drift are expected to reduce the efficiency of selection. Large genetic divergences among lineages, marked environmental differences in their ranges, reproductive isolation, and/or high isolation of populations may have resulted in morphological differences in L. occidentalis, not detected yet by traditional taxonomy. We used landmark-based geometric morphometric analyses to test for differences in body shape among highly divergent lineages of L. occidentalis, and among populations within these lineages. We analyzed a total of 492 individuals from 53 coastal localities from the southern California Bight to Central Mexico, including the Gulf of California. We conducted discriminant function analyses (DFAs) on body shape morphometrics to assess morphological variation among genetically differentiated lineages and their populations. We also tested for associations between phylogeny and morphological variation, and whether genetic divergence is correlated to multivariate morphological divergence. We detected significant differences in body shape among highly divergent lineages, and among populations within these lineages. Nonetheless, neither lineages nor populations can be discriminated on the basis of body shape, because correct classification rates of cross-validated DFAs were low. Genetic distance and phylogeny had weak to no effect on body shape variation. The supralittoral environment appears to exert strong stabilizing selection and/or strong functional constraints on body shape in L. occidentalis, thereby leading to morphological stasis in this isopod.