Thomas E. Dowling

Evolution of Animal Mitochondrial DNA: Relevance for Population Biology and Systematics

In the past decade, mitochondrial DNA (mtDNA) analysis has become established as a powerful tool for evolutionary studies of animals. These studies, recently reviewed by Avise (8) and Wilson et al (149), have used mtDNA analyses to provide insights into population structure and gene flow, hybridization, biogeography, and phylogenetic relationships. Major advances have been made in our understanding of the molecular biology of animal mtDNA (reviewed in 7, 28). A powerful synergism exists between the two fields of research: Evolutionary studies provide comparative data on mtDNA organization and function, and molecular investigations can, and should, improve the level of sophistication of evolutionary studies that use mtDNA. Here we focus on molecular aspects of animal mtDNA that are especially relevant to its use in evolutionary studies. Thus, we consider the form and frequency of three types of change in mtDNA: base substitution, length variation, and sequence rearrangement. Attention is also given to the nature and possible consequences of interactions between nuclear and mitochondrial (mt) genes. In the concluding sections, we discuss the way in which the knowledge of molecular processes allows informed use of mtDNA variation in evolutionary studies. Only animal mtDNAs are considered. The molecular biology and evolution of chloroplast DNA and nonmetazoan mtDNAs have been reviewed elsewhere (41, 66, 111, 121, 123).

A Conservation Plan for Native Fishes of the Lower Colorado River

Abstract The native fish fauna of the lower Colorado River, in the western United States, includes four “big-river” fishes that are federally listed as endangered. Existing recovery implementation plans are inadequate for these critically imperiled species. We describe a realistic, proactive management program founded on demographic and genetic principles and crafted to avoid potential conflicts with nonnative sport fisheries. In this program, native species would breed and their progeny grow in isolated, protected, off-channel habitats in the absence of nonnative fishes. Panmictic adult populations would reside in the main channel and connected waters, exchanging reproductive adults and repatriated subadults with populations occupying isolated habitats. Implementation of the plan would greatly enhance recovery potential of the four listed fishes.

Evolutionary Relationships of the Plagopterins (Teleostei: Cyprinidae) from Cytochrome b Sequences

Abstract Sequences of cytochrome b (cytb) were used to examine composition and phylogenetic relationships of cyprinid fishes of the tribe Plagopterini, endemic to the Great Basin and Lower Colorado River in southwestern North America. The plagopterin genera, Lepidomeda, Meda, Plagopterus, and Snyderichthys, were most closely affiliated with the chubs Couesius and Margariscus of northern and eastern North America. As indicated by previous morphologic, allozymic, and mtDNA studies, Snyderichthys is intimately related to Lepidomeda. The relationship is paraphyletic, however, according to our molecular data. Snyderichthys from the Snake and Bear River drainages are part of a clade that includes Lepidomeda mollispinis and Lepidomeda albivallis according to the cytb sequence, with Snyderichthys from the central and southern Bonneville basin more divergent. This paraphyly and the complex geographic relationships of mtDNA sequences indicate a complex history of the group and cast doubt on the validity of morphologically diagnosed Snyderichthys. Estimates of divergence time, based on a combination of fossil and molecular data, indicate that the plagopterins are an ancient clade, at least 17 million years old.

REPRODUCTIVE ISOLATION AND INTROGRESSION BETWEEN NOTROPIS CORNUTUS AND NOTROPIS CHRYSOCEPHALUS (FAMILY CYPRINIDAE): COMPARISON OF MORPHOLOGY, ALLOZYMES, AND MITOCHONDRIAL DNA

Hybrid zones in fluvial fishes may be heterogeneous from drainage to drainage. The comparison of data from morphology, allozymes, and mitochondrial DNA (mtDNA) indicates variability in the causes and degree of restriction of gene flow between Notropis cornutus and Notropis chrysocephalus. Allozyme marker loci show frequency‐dependent introgression; i.e., the rarer species, whichever it is at a particular locality, tends to exhibit a higher proportion of introgressed alleles. Unlike allozymes, introgression of mtDNA haplotypes varies geographically. In westward‐flowing Michigan drainages, N. cornutus mtDNA haplotypes are more common in F1 hybrids and backcrosses, independent of parental frequencies. In eastward‐flowing Michigan drainages, N. chrysocephalus mtDNA is more common in F1 hybrids and backcrosses; this pattern may be due to local ecological effects or frequency‐dependent introgression. Morphological data alone are not sufficient to distinguish all classes of hybrids. The lack of concordance of morphological, allozymic, and mtDNA introgression patterns implies operation of one or two factors: 1) geographically variable patterns of selection against different hybrid and backcross combinations or 2) genetic differences between Michigan populations inhabiting eastward‐ and westward‐flowing drainage systems accumulated during historical isolation.

Functional and ecological significance of rDNA intergenic spacer variation in a clonal organism under divergent selection for production rate

It has recently been hypothesized that variation in the intergenic spacer (IGS) of rDNA has considerable developmental, evolutionary and ecological significance through effects on growth rate and body C : N : P stoichiometry resulting from the role of the IGS in production of rRNA. To test these ideas, we assessed changes in size and structure of the repetitive region of the IGS, juvenile growth rate (JGR), RNA and phosphorus (P) contents in clonal lineages of Daphnia pulex derived from a single female and subjected to divergent selection on weight–specific fecundity (WSF). As a result of selection, WSF diverged rapidly, with significant reductions within two generations. Other significant changes accompanying shifts in WSF were that juveniles produced by low–WSF females grew more rapidly and had higher RNA and P contents. An increased predominance of long IGS variants was observed in lineages with elevated JGRs and low WSF. The observed variations in IGS length were related to the number of subrepeat units carrying a promoter sequence in the repetitive region. These results strongly support the hypothesized relationships, indicate a genetic mechanism for the evolution of such associations and demonstrate that Daphnia (and perhaps other parthenogens) possess considerable potential for rapid adaptive change in major life–history traits.

EFFECTS OF INTRINSIC AND EXTRINSIC FACTORS ON POPULATION FRAGMENTATION IN THREE SPECIES OF NORTH AMERICAN MINNOWS (TELEOSTEI: CYPRINIDAE)

Geographic patterns of genetic variation (mitochondrial DNA [mtDNA] and allozymes) were used to examine effects of intrinsic characteristics (e.g., vagility, habitat specificity, and reproductive behaviors) and extrinsic factors (e.g., climatic and geological history) on population fragmentation. The three species of cyprinid fishes examined (Tiaroga cobitis, Meda fulgida, and Agosia chrysogaster) occupied similar historical ranges within the lower Colorado River drainage, but differ in intrinsic characteristics conducive to population fragmentation. Relationships among populations were similar across species, reflecting common historical influences, but results indicate the distribution of variation among species is strongly affected by intrinsic characteristics. Variation within two species (T. cobitis and M. fulgida) is subdivided among populations, suggesting little gene flow among rivers. In contrast, similarity of A. chrysogaster populations throughout the Gila River drainage supports the hypothesis that levels of gene flow are high for this species. Levels of mtDNA divergence were much higher than expected for both T. cobitis and A. chrysogaster suggesting long‐term isolation of geographic regions. These results indicate that both long‐term and short‐term extrinsic factors have shaped basic patterns of variation within these fishes; however, the intrinsic characteristics of each species have strongly affected the population genetic structure of these fishes.

Historical Biogeography of the New-World Pupfish Genus Cyprinodon (Teleostei: Cyprinodontidae)

Abstract Analysis of mtDNA sequence variation (2,548 bp from ND2, cytb, and part of the control region) indicates that the genus Cyprinodon began diverging in the Late Miocene from a common ancestor with Megupsilon, a monotypic genus on the Mesa del Norte of Mexico. The geographic pattern of mtDNA variation, with estimates of divergence time, suggests that by the end of the Miocene Cyprinodon occurred from the Atlantic Coast and West Indies to near the western margin of North America via ancestral Rio Grande and Colorado River systems. Phylogeographic structure within the major mtDNA complexes supports a variety of hypotheses from geology and previous phylogenetic analyses for Late Neogene connections among basins in southwestern North America now separated by formidable barriers to dispersal. Comparison of the mtDNA tree with previous phylogenetic inferences from allozymes indicates that reticulate evolution involving divergent lineages probably was important in the history of Cyprinodon.

Variable microsatellite markers amplify across divergent lineages of cyprinid fishes (subfamily Leusicinae)

Thomas F. Turner1,∗, Thomas E. Dowling2, Richard E. Broughton3 & John R. Gold4 1Department of Biology and Museum of Southwestern Biology, University of New Mexico, Albuquerque, NM 87131; 2Department of Biology, Arizona State University, Tempe, AZ 85287; 3Department of Zoology, University of Oklahoma, Norman, OK 73019; 4Center for Biosystematics and Biodiversity, Texas A&M University, College Station, TX 77843 (∗Corresponding author: E-mail: turnert@unm.edu)

THE EXTENT OF INTROGRESSION OUTSIDE THE CONTACT ZONE BETWEEN NOTROPIS CORNUTUS AND NOTROPIS CHRYSOCEPHALUS (TELEOSTEI: CYPRINIDAE)

The cyprinid fishes, Notropis cornutus and N. chrysocephalus, hybridize in a long, narrow zone in the midwestern United States. To quantify the extent of introgression of genetic markers outside of this zone, samples were collected along transects starting near the region of contact (as defined by morphological characters), followed by samples progressively more distant. Diagnostic allozymic and mitochondrial DNA (mtDNA) restriction site markers were used to estimate the extent of introgression outside of the zone, while polymorphic allozyme and mtDNA markers were used to evaluate the potential for gene flow among populations within transects. Analysis of populations from the northern transect provided evidence for differentiation of populations for some of the markers; however, on average, enough gene flow has occurred to overcome substantial differentiation. Introgressed mtDNA and allozyme haplotypes were rare and found only in the population closest to the contact zone. The rarity of introgressed alleles in the more northern populations is consistent with the recent origin of these populations after the Wisconsin glaciation (less than 12,000 years bp) and/or selection maintaining the northern boundary of the contact zone. Analysis of populations from the southern transect revealed evidence for population subdivision but no evidence for introgression at the diagnostic allozyme loci; however, nearly all individuals from this transect possessed introgressed mtDNA haplotypes, with samples furthest from the contact zone exhibiting the highest frequencies of introgression. Patterns of variation for one of the polymorphic allozyme markers (Est‐A) and introgressed mtDNAs were highly correlated, suggesting that allozymic heterogeneity at this locus is also the result of introgression. The most likely explanation for these data is that these introgressed haplotypes are indicators of a more southern position of the contact zone during the Pleistocene, with the contact zone shifting northward with the recession of the glacial front. Such movement implicates selection in the maintenance of distributional limits of these species, and hence, the width and position of the contact zone.

Influence of hydrogeographic history and hybridization on the distribution of genetic variation in the pupfishes Cyprinodon atrorus and C. bifasciatus

The evolutionary importance of hybridization in animals has been subject of much debate. In this study, we examined the influence of hydrogeographic history and hybridization on the present distribution of nuclear and mitochondrial DNA variation in two pupfish species, Cyprinodon atrorus and Cyprinodon bifasciatus. Results presented here indicate that there has been limited introgression of nuclear genes; however, mtDNA introgression has been substantial, with complete replacement of the C. bifasciatus mitochondrial genome by that of C. atrorus. Subsequent to this replacement, there has been diversification of mitochondrial haplotypes along major geographic regions in the basin. Evidence was also found that mitochondrial replacement follows a predictable, cyclical pattern in this system, with isolation and diversification followed by re‐contact and replacement of C. bifasciatus mitochondrial haplotypes by those of C. atrorus. This pattern is best explained by a combination of a numeric bias towards C. atrorus and mating site selection rather than selection for C. atrorus mitochondrial genome. These results demonstrate the important role hybridization can play in evolution.