Edward Pfeiler

Phylogenetic relationships of elopomorph fishes inferred from mitochondrial ribosomal DNA sequences.

The superorder Elopomorpha, a grouping which includes all teleost fishes that possess a specialized leptocephalous larva [true eels (Anguilliformes), gulpers and bobtail snipe eels (Saccopharyngiformes), bonefishes, spiny eels, and halosaurs (Albuliformes, including Notacanthiformes), ladyfishes and tarpons (Elopiformes, including Megalopiformes)] comprises >800 species for which phylogenetic relationships are poorly understood. In the present study, we analyzed mitochondrial DNA sequences in segments of the 12S and 16S rRNA genes in 33 elopomorph taxa encompassing all of the previously proposed orders, and 9 of the 15 currently recognized families of the Anguilliformes, as well as outgroup representatives from the superorders Osteoglossomorpha (nine species) and Clupeomorpha (three species), to develop phylogenetic hypotheses based on distance and parsimony methods. Both methods failed to support the monophyly of the Elopomorpha, casting doubt on the validity of the leptocephalus as an elopomorph synapomorphy. The orders Elopiformes, Albuliformes, and Anguilliformes, however, were resolved as monophyletic assemblages. Parsimony analysis supported the separation of the Anguilliformes into two groups (primitive and advanced) based on the presence of divided versus fused frontal bones. In addition, the molecular data indicated a close affinity of the anguilliform Thalassenchelys coheni (incertae sedis), known only from the leptocephalus, with the family Serrivomeridae. The implications of these data as regards the evolution of the elopomorph assemblage are discussed.

FLIES ACROSS THE WATER: GENETIC DIFFERENTIATION AND REPRODUCTIVE ISOLATION IN ALLOPATRIC DESERT DROSOPHILA

Abstract Between sister species of Drosophila, both pre‐ and postzygotic reproductive isolation commonly appear by the time a Neis genetic distance of 0.5 is observed. The degree of genetic differentiation present when allopatric populations of the same Drosophila species exhibit incipient reproductive isolation has not been systematically investigated. Here we compare the relationship between genetic differentiation and pre‐ and postzygotic isolation among allopatric populations of three cactophilic desert Drosophila: D. mettleri, D. nigrospiracula, and D. mojavensis. The range of all three is interrupted by the Gulf of California, while two species, D. mettleri and D. mojavensis, have additional allopatric populations residing on distant Santa Catalina Island, off the coast of southern California. Significant population structure exists within all three species, but only for allopatric populations of D. mojavensis is significant isolation at the prezygotic level observed. The genetic distances for the relevant populations ofD. mojavensis were in the range of 0.12, similar to that for D. mettleri whose greatest D = 0.11 was unassociated with any form of isolation. These observations suggest further investigations of Drosophila populations with genetic distances in this range be undertaken to identify any potential patterns in the relationship between degree of genetic differentiation and the appearance of pre‐ and/or postzygotic isolation.

Mitochondrial DNA evidence for deep genetic divergences in allopatric populations of the rocky intertidal isopod Ligia occidentalis from the eastern Pacific.

Nucleotide sequences from the cytochrome c oxidase subunit I (COI) gene were used to test for genetic differentiation in the rocky intertidal isopod crustacean, Ligia occidentalis (Ligiidae), from the eastern Pacific. Phylogenetic analyses showed that individuals of L. occidentalis from southern California, USA to Manzanillo, Colima, Mexico partitioned into 15 highly-divergent clades. Mean Kimura 2-parameter (K2P) genetic distances among clades ranged from 13.2% to 26.7%. These values are similar to interspecific genetic distances found in a wide variety of crustaceans, including Ligia spp., suggesting that the taxon L. occidentalis represents a complex of cryptic species.

Identification, structural analysis and function of hyaluronan in developing fish larvae (leptocephali)

Hyaluronan (HA) has been identified as the principal glycosaminoglycan (CAG) in the highly hydrated, extracellular body matrix of the larval stage (leptocephalus) of seven species of true eels (Teleostei: Elopomorpha: Anguilliformes) and the ladyfish Elops saurus (Elopiformes), and was found as a minor GAG component in the bonefish Albula sp. (Albuliformes). Identification was based on: (1) HPLC separation of unsaturated disaccharides derived from chondroitinase ABC digests of whole-body GAG extracts; (2) 1H NMR analyses of native GAG polymers; and (3) degradation of GAG extracts by Streptomyces hyaluronan lyase. The unsaturated disaccharide 2-acetamido-2-deoxy-3-O-(beta-D-gluco-4-enepyranosyluronic acid)-D-glucose (DeltaDi-HA) accounted for 92.4-99.8% of the total disaccharides in chondroitinase digests. Trace amounts of unsaturated disaccharides of chondroitin sulfate were also present. Two-dimensional gCOSY spectra of the native HA polymer were similar for all species. Proton assignments for the HA disaccharide repeat (GlcAbeta1-3GlcNAcbeta1-4) in D(2)O, based on gCOSY, DQF-COSY and TOCSY analyses for the eel Ahlia egmontis, were concordant with published chemical shifts for HA oligosaccharides. In addition to its presumed role in maintaining the structural integrity and hydration of the gelatinous body of the leptocephalus, HA is postulated to function as a storage polysaccharide in those species in which it is the predominant GAG.

Phylogeography of the Cactophilic Drosophila and Other Arthropods Associated with Cactus Necroses in the Sonoran Desert

Studies on the population genetics, phylogenetic relationships, systematics and evolution of arthropods that inhabit necrotic tissue of cacti in the Sonoran Desert of North America are reviewed. These studies have focused upon several species of insects (orders Diptera and Coleoptera) and arachnids (order Pseudoscorpiones). For most taxa studied, little genetic structure and high dispersal ability are found in populations inhabiting the mainland and Baja California peninsula regions of the Sonoran Desert, consistent with the availability of the rotting cactus microhabitat which is patchily distributed and ephemeral. There is evidence, however, that the Gulf of California, which bisects the Sonoran Desert, has played a role in limiting gene flow and promoting speciation in several taxa, including histerid beetles, whereas other taxa, especially Drosophila nigrospiracula and D. mettleri, apparently are able to freely cross the Gulf, probably by taking advantage of the Midriff Islands in the northern Gulf as dispersal “stepping stones”. Genetic evidence has also been found for historical population expansions dating to the Pleistocene and late Pliocene in several taxa. Overall, these studies have provided important insights into how arthropods with different life history traits, but generally restricted to a necrotic cactus microhabitat, have evolved in an environmentally harsh and tectonically active region. In addition, they suggest some taxa for further, and more detailed, hypothesis driven studies of speciation.

Genetic diversification and demographic history of the cactophilic pseudoscorpion Dinocheirus arizonensis from the Sonoran Desert.

Sequence data from a segment of the mitochondrial cytochrome c oxidase subunit I (COI) gene were used to examine phylogenetic relationships, estimate gene flow and infer demographic history of the cactophilic chernetid pseudoscorpion, Dinocheirus arizonensis (Banks), from the Sonoran Desert. Phylogenetic trees resolved two clades of D. arizonensis, one from mainland Sonora, Mexico and southern Arizona (clade I) and the other from the Baja California peninsula and southern Arizona (clade II). The two clades were separated by a mean genetic distance (d) of approximately 2.6%. Hierarchical analysis of molecular variance indicated highly significant population structuring in D. arizonensis (overall Phi(ST)=0.860; P<0.0001), with 80% of the genetic variation distributed among the two clades. Most pairwise comparisons of Phi(ST) among populations within each clade, however, were not significant. The results suggest that phoretic dispersal on vagile cactophilic insects such as the neriid cactus fly Odontoloxozus longicornis (Coquillett) provides sufficient gene flow to offset the accumulation of unique haplotypes within each clade of the non-vagile pseudoscorpion. Preliminary results on dispersal capability of O. longicornis were consistent with this conclusion. Tests designed to reconstruct demographic history from sequence data indicated that both clades of D. arizonensis, as well as O. longicornis, have experienced historical population expansions. Potential barriers to gene flow that may have led to genetic isolation and diversification in clades I and II of D. arizonensis are discussed.

Acidic Glycosaminoglycans in Marine Teleost Larvae: Evidence for a Relationship Between Composition and Negative Charge Density in Elopomorph Leptocephali

Glycosaminoglycan (GAG) composition was determined in larvae of 10 species of true eels (subdivision Elopomorpha; order Anguilliformes; families Chlopsidae, Congridae, Moringuidae, Muraenidae and Ophichthidae), two species from the subdivision Clupeomorpha (order Clupeiformes; families Engraulidae and Clupeidae) and one species from the subdivision Euteleostei (order Aulopiformes; family Synodontidae). Using a combination of specific GAG-degrading enzymes, cellulose acetate electrophoresis with known GAG standards and relative degree of staining with Alcian Blue, the predominant GAG in anguilliform leptocephali was identified as undersulfated chondroitin sulfate (uCS); the only exception was Bathycongrus macrurus (Congridae), where heparan sulfate was tentatively identified together with uCS. In representatives of the Clupeomorpha and Euteleostei, however, the predominant GAG was chondroitin sulfate; uCS was not detected. These results, combined with those from previous studies, provide data on GAG composition in 21 species from three of the four currently recognized elopomorph orders (Albuliformes, Elopiformes and Anguilliformes). The combined data suggest that although several different body matrix GAGs have evolved in different groups of leptocephali, selection has favored GAGs (mainly uCS and keratan sulfate) with relatively low average negative charge densities (average of about one negative charge per disaccharide repeat of the acidic GAG chain). The presence of normally sulfated CS (average per disaccharide negative charge density of 2) as the major GAG in the three species of non-elopomorph larvae further suggests that some aspect of the unique developmental pattern in elopomorph leptocephali may place constraints on the maximum negative charge density permitted in the body matrix GAGs. The presence of GAGs with low negative charge density is interpreted as an additional elopomorph synapomorphy, offering further support for the view that the Elopomorpha represents a monophyletic group of fishes. Possible scenarios for the evolution of GAGs in leptocephali are presented.

Phylogenetic relationships of Sonoran Desert cactus beetles in the tribe Hololeptini (Coleoptera: Histeridae: Histerinae), with comments on the taxonomic status of Iliotona beyeri.

Nucleotide sequences from 16S rRNA and cytochrome c oxidase subunit I (COI) were used to examine phylogenetic relationships and evolution of beetles from the tribe Hololeptini (Coleoptera: Histeridae: Histerinae) that inhabit necrotic tissue of columnar cacti in the Sonoran Desert. Phylogenetic and morphological analyses revealed the presence of seven separate lineages, three representing species in the genus Iliotona, including I. beyeri stat. nov., and four species belonging to the genus Hololepta (sensu lato). The possible roles of historical vicariance and host plant associations on the evolution of the Hololeptini from the Sonoran Desert are discussed.

Systematic Status of Bonefishes (Albula spp.) From the Eastern Pacific Ocean Inferred from Analyses of Allozymes and Mitochondrial DNA

Recent molecular evidence suggests that at least eight species of bonefishes (Albuliformes: Albulidae: Albula) are found worldwide (Colborn et al. 2001). Adults of most of these species, including two that are restricted to the eastern Pacific Ocean (Albula sp. A from the Gulf of California and Albula sp. C from the Gulf of Panama), have not been formally described. Because cryptic species of bonefishes are known to occur sympatrically, population genetics data provide an important tool for taxonomic studies on this group. Isozyme analyses conducted on larval, juvenile and adult bonefish from the Guaymas region of the central Gulf of California confirmed that only a single species (Albula sp. A) was present there. In addition, analysis of a segment of the cytochrome b gene from three adult Albula collected from coastal waters of southern California revealed high sequence homology with Albula sp. A, suggesting that the southern California specimens were conspecific with those from Guaymas. However, the southern distributional limit of Albula sp. A, and whether there are regions where Albula sp. A and C occur sympatrically, are unknown. The historical background of the available name Atopichthysesunculus Garman, 1899 [= Albula esuncula (Garman, 1899)], and problems associated with the nomenclature of the two species of eastern Pacific bonefishes, are summarized. The systematic status of the related shafted bonefish, Dixonina (= Albula) nemoptera Fowler, 1911, is also reviewed.

Genetic Diversity and Population Genetics of Mosquitoes (Diptera: Culicidae: Culex spp.) from the Sonoran Desert of North America

The population genetics and phylogenetic relationships of Culex mosquitoes inhabiting the Sonoran Desert region of North America were studied using mitochondrial DNA and microsatellite molecular markers. Phylogenetic analyses of mitochondrial cytochrome c oxidase subunit I (COI) from mosquitoes collected over a wide geographic area, including the Baja California peninsula, and mainland localities in southern Arizona, USA and Sonora, Mexico, showed several well-supported partitions corresponding to Cx. quinquefasciatus, Cx. tarsalis, and two unidentified species, Culex sp. 1 and sp. 2. Culex quinquefasciatus was found at all localities and was the most abundant species collected. Culex tarsalis was collected only at Tucson, Arizona and Guaymas, Sonora. The two unidentified species of Culex were most abundant at Navojoa in southern Sonora. Haplotype and nucleotide diversities in the COI gene segment were substantially lower in Cx. quinquefasciatus compared with the other three species. Analysis of molecular variance revealed little structure among seven populations of Cx. quinquefasciatus, whereas significant structure was found between the two populations of Cx. tarsalis. Evidence for an historical population expansion beginning in the Pleistocene was found for Cx. tarsalis. Possible explanations for the large differences in genetic diversity between Cx. quinquefasciatus and the other species of Culex are presented.