Allopaternal care in the tessellated darter,Etheostoma olmstedi (Pisces: Percidae)

Synopsis Large male tessellated darters, Etheostoma olmstedi, defend flat-bottomed rocks, the undersides of which serve as spawning substrates. Because females attach eggs directly onto bare stone, a spawning bout necessarily decreases the surface area available for further breeding at a nest. In apparent response to the female preference of ovipositing on rocks with the most bare surface, fathers abandon nests with their attached offspring to search for rocks with more uncovered surface. Egg abandonment also results after roving fathers intercept and spawn with ripe females and remain at a different rock to care for a new clutch. In essence, large males appear to maximize the number of eggs they fertilize by sequentially monopolizing breeding patches that are temporarily most desirable to females. The costs to a father of abandoning his eggs appear to be low for two reasons: (1) non-territorial males, created by a scarcity of suitable rocks, readily occupy vacant nests, and clean and aerate the abandoned eggs; and (2) after water hardening, eggs are less vulnerable to cannibalism. The maintenance of eggs by nonparental males may be both incidental to scrubbing the ceiling for their own breeding and adaptive in that viable eggs stimulate females to spawn; neither would appear to involve parental investment costs. Thus, allopaternal care in the tessellated darter may have evolved because it is performed in the selfish pursuit of spawning opportunities while entailing little, if any, of the costs normally subsumed within parental investment.     

Life history of the gulf darter,Etheostoma swaini (Pisces: Percidae)

Synopsis Etheostoma swaini, the gulf darter, was collected from the Black Creek drainage in southern Mississippi (February 1978 – April 1979). The gulf darter generally inhabits small- to moderate-size creeks and occurs over a sand or sandy mud bottom, often in association with aquatic vegetation or a layer of organic debris. Larval dipterans were the most important food items, both numerically and volumetrically. Chironomids were found in 71–100% of the stomachs in all except the unusual March 16 collection. The length frequency distribution and the scale annuli analysis indicated there were three year-classes present in the population at any one time. Fifty-one percent of the specimens taken were less than 12 months old. During the mid-February to late March spawning season gulf darters were most often collected over clean gravel or gravelsand substrates. Laboratory observations suggest that the female burrows into the gravel where the demersal, adhesive eggs are deposited. Female gulf darters significantly outnumbered males at a ratio of 59:41.     

History repeated: recent and historical mitochondrial introgression between the current darter Etheostoma uniporum and rainbow darter Etheostoma caeruleum (Teleostei: Percidae)

Incongruence between recognized taxonomy and phylogenetic relationships between two species from a diverse clade (Percidae: Etheostomatinae) of stream fishes was found in a mitochondrial (mt) DNA gene tree. Two darters in subgenus Oligocephalus , Etheostoma uniporum current darter and Etheostoma caeruleum rainbow darter were sampled throughout their sympatric distribution in the Ozark Highlands of the central United States. Sequences from cytochrome (cyt) b and the first intron of the nuclear marker S7 were analysed separately using maximum parsimony and Bayesian methods. Cyt b recovered both species as polyphyletic; E. uniporum haplotypes were interspersed within E. caeruleum . However, both species were monophyletic and non-sister taxa based on S7. The cyt b gene tree pattern is caused by introgressive hybridization resulting in the mtDNA replacement of E. uniporum haplotypes by those of E. caeruleum . Some E. uniporum haplotypes are shared with geographically proximate E. caeruleum , and this is consistent with recent hybridization, while other E. uniporum haplotypes indicate historical sorting of introgressed lineages. The mechanism of introgression is likely asymmetric sneaking behaviour by male E. uniporum , a mating tactic observed in related species. MtDNA replacement may have occurred in E. uniporum due to drift fixation in a historically small female effective population. Additional evidence for darter hybridization will likely be discovered in future molecular genetic surveys of the nearly 200 species in eastern North America.     

Phylogeography of the greenside darter complex, Etheostoma blennioides (Teleostomi: Percidae): a wide-ranging polytypic taxon

The greenside darter, Etheostoma blennioides (Teleostomi: Percidae), is a wide-ranging polytypic taxon that occurs throughout eastern North America. A previous morphological study recognized four subspecies (blennioides, newmanii, gutselli, and pholidotum), several morphological races, and three zones of morphological intergradation. We generated complete cytochrome b (1140bp) sequence data for 51 individuals from across the range of the greenside darter inclusive of all of the currently recognized taxa to assess genetic variation and taxonomic boundaries. Both maximum parsimony and mixed model Bayesian analyses resulted in two strongly supported deeply divergent clades including (1) a Tennessee River drainage clade, and (2) an Ohio River and Great Lakes basins, Interior Highlands, and Atlantic slope clade. Etheostoma blennius, a closely related congener, nested within the Tennessee River clade of E. blennioides, rendering the complex paraphyletic. Test of alternative topologies failed to support the current taxonomic designations. The inclusion of nuclear sequence data from intron 1 of the S7 ribosomal protein (523bp) from a subset of the populations was included to independently test whether the currently recognized taxa conform to distinct evolutionary lineages and also to clarify potential issues associated with ancestral hybridization. Although the nuclear data was less variable than the mitochondrial data, the monophyly of several of the subspecies could not be rejected.     

Incomplete behavioural isolation and asymmetric female preference in darter sister species (Percidae: Etheostoma)

In this study, female preferences and behavioural isolation were estimated in a pair of allopatric sister species, Etheostoma duryi and Etheostoma flavum. Dichotomous mate preference trials were conducted to determine whether females prefer to associate with conspecific over heterospecific males and free‐spawning assays were conducted to determine whether those preferences translated into behavioural isolation. Dichotomous mate choice trials revealed asymmetric female preference, as female E. flavum preferred conspecific males, whereas female E. duryi showed no preference. Free‐spawning assays indicated that behavioural isolation remains incomplete between E. duryi and E. flavum (I_B = 0·19). In addition to female mating preferences, male behaviour also appeared to influence mating outcomes as male E. flavum consistently courted conspecific females more often in free‐spawning assays whereas male E. duryi did not. The data therefore suggest that despite marked divergence in male nuptial colour, divergence in female preferences between these species may not be sufficient to maintain species boundaries upon secondary contact. These results contrast with similar work in a sympatric darter species pair and may be explained by considering the contributions of reinforcement and differences in colour pattern as well as colour value.     

Phylogeography and post-glacial colonization patterns of the rainbow darter, Etheostoma caeruleum (Teleostei: Percidae)

Aim To examine the effects of historical climate change and drainage isolation on the distribution of mitochondrial DNA cytochrome b genetic variation within the rainbow darter, Etheostoma caeruleum (Percidae: Etheostomatinae). Location Eastern North American streams including tributaries to the Mississippi River, Great Lakes, Potomac River and Hudson Bay drainages. Methods Parsimony analyses, Bayesian analyses and haplotype networks of mitochondrial DNA sequences. Results Four major clades were recovered from sampled populations of E. caeruleum. Three of four clades are distributed in the western portion of the species’ range (primarily west of the Mississippi River). Samples from this region do not form a monophyletic group, and sequences often vary greatly between samples from adjacent stream systems (up to 7.2% divergence). A basal clade includes samples from the White River system in the Ozark Highlands. The northern Ozarks–upper Midwest clade includes samples from Missouri River tributaries and the upper Midwest (Hudson Bay, upper Mississippi River, and western Lake Michigan drainage). The eastern clade is composed of individuals from the Ohio River, Great Lakes and Potomac River. The Mississippi River corridor clade includes samples from middle and lower Mississippi River tributaries. Main conclusions The four major clades of E. caeruleum are deep allopatric lineages with well‐defined boundaries and have additional phylogeographical structure within each clade. The Ozark Highlands have the greatest levels of diversity relative to distributional area, with marked cytochrome b subdivisions between adjacent stream systems. Samples from previously glaciated areas do not have a subset of the cytochrome b diversity found in unglaciated areas, but four separate source areas are identified based on phylogenetic analyses. Dispersal into previously glaciated areas followed several known glacial outlets and, based on sequence divergence between populations, may have occurred during different glacial or interglacial stages. The disjunct distribution and cytochrome b pattern of E. caeruleum in the Mississippi River corridor clade is consistent with late Pleistocene and Recent changes in the course and characteristics of the middle and lower Mississippi River. Phylogeographical boundaries between clades of E. caeruleum correspond to independent sources of biogeographical information and provide insight into historical stream drainage relationships, post‐glacial colonization and drainage isolation patterns.     

PERMANENT GENETIC RESOURCES: Microsatellite DNA primers for the candy darter, Etheostoma osburni and variegate darter, Etheostoma variatum, and cross-species amplification in other darters (Percidae)

In order to investigate a potential hybrid zone between the candy darter, Etheostoma osburni, and variegate darter, Etheostoma variatum, and examine population variation within E. osburni, a suite of primers for 15 polymorphic microsatellite loci were developed. The average number of alleles per locus was 5.5 in E. osburni and 7.6 in E. variatum, and the average observed heterozygosities were 62.5% and 71.4%, respectively. There were no deviations from Hardy-Weinberg equilibrium and no observed linkage disequilibrium after Bonferroni correction. The utility of these primers was also tested in 11 species of darters representing all four genera of darters. Success of cross-species amplification was largely consistent with phylogenetic relationships of darters.     

Characterization of eight polymorphic microsatellite DNA markers for the greenside darter,Etheostoma blennioides (Percidae)

The greenside darter, Etheostoma blennioides is a small benthic fish found in fast‐flowing streams in eastern North America. In Canada, this species is native to three, and introduced into one, Great Lakes tributaries in southwestern Ontario. It is currently listed as a species of Special Concern. To characterize population genetic structure and diversity in the Canadian populations of greenside darter, eight polymorphic microsatellite markers were developed for the species. The polymerase chain reaction primers were tested between 32 and 60 individuals from the Sydenham River and yielded a high number of alleles (four to 42 per locus), and observed heterozygosities ranging from 0.14 to 0.82.     

Systematics and population genetics of the coldwater (Etheostoma ditrema) and watercress (Etheostoma nuchale) darters,with comments on the Gulf darter (Etheostoma swaini) (Percidae: subgenus Oligocephalus)

Current taxonomy of the Etheostoma asprigene species group recognizes four species, two of which (Etheostoma ditrema Ramsey and Suttkus and Etheostoma nuchale Howell and Caldwell) are restricted to isolated springs and spring-fed stream systems above the Fall Line of the Mobile Basin of Alabama, Georgia, and Tennessee. Previous studies of morphological and biochemical variation between disjunct populations of E. ditrema support the presence of multiple independent lineages. Unfortunately, the lack of phylogenetic methodology has made it impossible to distinguish historically differentiated non-sister taxa from currently intergrading sister lineages.The current study examines members of the E. asprigene species group, particularly E. ditrema and E. nuchale, from throughout their respective ranges for products of 44 presumptive gene loci. Highly significant levels of genetic subdivision (F_IT) and genetic differentiation (F_ST) are observed both within and between species of this group, supporting the presence of previously unrecognized diversity within this clade. Phylogenetic evaluation of polyallelic loci supports the specific recognition of E. nuchale and three taxa currently under the name E. ditrema: nominal E. ditrema (upper Coosa River system), E. sp. cf. E. ditrema (Coldwater Spring), and E. sp. cf. E. ditrema (central Coosa River system) based on either fixed allelic products or significant allele frequency differences. The Gulf darter, Etheostoma swaini, is also identified as a composite of at least three diagnosable evolutionary species, two of which (Black Warrior River system above and Cahaba River system below the Fall Line) are more closely related to E. nuchale and species of the E. ditrema complex than to remaining E. swaini. It is recommended that future conservation efforts with these highly geographically restricted and imperiled species incorporate these patterns of genetic subdivision into management and recovery plans.     

Microsatellite DNA primers for the endangered vermilion darter,Etheostoma chermocki,and cross‐species amplification in other darters (Percidae: Etheostoma)

The endangered vermilion darter (Etheostoma chermocki) is endemic to the Black Warrior River system in the Mobile Basin in Alabama. Restoration and conservation of this species require an understanding of its population genetic structure, which can be characterized using microsatellite DNA. Nine microsatellite loci were developed; eight loci were polymorphic. Although observed heterozygosity was lower than expected heterozygosity in most polymorphic loci, only one locus showed significant deviation from Hardy–Weinberg equilibrium. These nine markers were tested in an additional 24 species of Etheostoma and appear to have sufficient allelic variation to be useful in studies of population genetic structure.     

Living on the bottom: Kinematics of benthic station‐holding in darter fishes (Percidae: Etheostomatinae)

Darters represent a substantial radiation of freshwater fishes that live in close association with the substrate in North American streams and rivers. A key feature of any darter species is therefore its ability to stay in place or to “hold station” in flowing water. Here, we quantify the station‐holding performance of two morphologically divergent darter species, the fantail darter Etheostoma flabellare and the Missouri saddled darter Etheostoma tetrazonum. We also characterize the primary kinematic responses of the two species when holding station in flow speeds ranging from 4 to 56 cm s^?1 in a flow tank on either plexiglas or small rock substrate. We then present a series of hypotheses about the potential hydrodynamic and functional consequences of the observed postural changes and the links among morphology, posture, and station‐holding performance. On both substrates, E. tetrazonum was able to hold station at higher flow speeds than E. flabellare. On rocks, E. tetrazonum slipped at an average speed of 55.7 cm s^?1 whereas E. flabellare slipped at 40.2 cm s^?1. On plexiglas, E. tetrazonum slipped at an average speed of 24.7 cm s^?1 whereas E. flabellare slipped at 23.1 cm s^?1. We measured body and fin positions of the two species from individual frames of high‐speed video while holding station on rocks and plexiglas. We found that on both substrates, the two species generally exhibited similar kinematic responses to increasing flow: the head was lowered and angled downward, the back became more arched, and the median and caudal fin rays contracted as water flow speed increased. The ventral halves of the pectoral fins were also expanded and the dorsal halves contracted. These changes in posture and fin position likely increase negative lift forces thereby increasing substrate contact forces and reducing the probability of downstream slip. J. Morphol., 2010. © 2009 Wiley‐Liss, Inc.     

Systematics of the subgenus Oligocephalus (Teleostei: Percidae: Etheostoma) with complete subgeneric sampling of the genus Etheostoma

The genus Etheostoma is the most diverse clade of freshwater fishes in North America. While studies have been performed with complete sampling of a single subgenus, none have included representatives of all remaining subgenera. The subgenus Oligocephalus is the largest, consisting of 25-27 species in four species groups, and its monophyly has never been clearly demonstrated. The monophyly of this subgenus and its constituent groups was tested using parsimony and Bayesian analyses of ND2 (mtDNA) and the first intron of S7 (nDNA) with complete species sampling from Oligocephalus and complete subgeneric sampling from Etheostoma. Although the subgenus Oligocephalus was not recovered as a monophyletic group in any analyses, monophyletic E. whipplei, Southwestern Darter, and E. spectabile (in part) species groups were recovered in all analyses. All analyses agree that E. okaloosae and both subspecies of E. hopkinsi are not closely related to other members of the subgenus Oligocephalus. E. exile is, however, presenting the strongest evidence yet that recognition of the subgenus Boleichthys is unwarranted.     

Intrastream variation in growth rates and time of first otolith increment formation for young of the year Etheostoma simoterum (Cope) (Perciformes: Percidae)

Growth rates and otolith-estimated hatching dates of young of the year Etheostoma simoterum were studied at four localities along an elevational gradient in the Little River, Tennessee, U.S.A. during the summer of 1983. Changes in otolith increment width and mean daily air temperature corresponded well with each other, suggesting that increments were laid down daily. Growth rate increased nearly threefold from the highest elevation site to the lowest. Median otolith-estimated hatching dates differed by about 2 weeks between sites, and the pattern was not directly related to elevation. These data indicate that within-stream, life-history variation of a broadly distributed species should be taken into consideration when studying factors regulating stream fish populations along elevational gradients.     

Population structure and genetic diversity in the endangered bluemask darter (Etheostoma akatulo)

Bluemask darters (Etheostoma akatulo) were sampled from the four drainages where extant populations of this narrowly endemic freshwater fish are known to exist. Population genetic diversity and structure were assessed at 10 microsatellite loci. All populations exhibited low levels of genetic variation, with expected heterozygosity ranging from 0.2 to 0.35. Significant population subdivision was found among most tributaries, and genetic divergence was strongly correlated with geographic distance. Bayesian population assignment and pairwise population differentiation measures both identified a lack of differentiation between E. akatulo populations inhabiting Cane Creek and the Caney Fork. This observation reduced the number of distinct breeding populations of this species to three. We also used approximate Bayesian computation to compare three models of demographic history in this species. A constant population size model was favored over models that included historic or recent population reductions. Our results suggest that impoundment of the Caney Fork and its tributaries, by completion of Great Falls Dam in 1916, was not responsible for the reduced genetic diversity in the sampled populations. Given the low levels of genetic diversity within populations and the limited geographic distribution, future conservation efforts should seek to maximize available habitat while simultaneously limiting the influences of anthropogenic stressors in the system.