Parallel evolution in the major haemoglobin genes of eight species of Andean waterfowl

Theory predicts that parallel evolution should be common when the number of beneficial mutations is limited by selective constraints on protein structure. However, confirmation is scarce in natural populations. Here we studied the major haemoglobin genes of eight Andean duck lineages and compared them to 115 other waterfowl species, including the bar-headed goose ( Anser indicus ) and Abyssinian blue-winged goose ( Cyanochen cyanopterus ), two additional species living at high altitude. One to five amino acid replacements were significantly overrepresented or derived in each highland population, and parallel substitutions were more common than in simulated sequences evolved under a neutral model. Two substitutions evolved in parallel in the αA subunit of two (Ala-α8) and five (Thr-α77) taxa, and five identical βA subunit substitutions were observed in two (Ser-β4, Glu-β94, Met-β133) or three (Ser-β13, Ser-β116) taxa. Substitutions at adjacent sites within the same functional protein region were also observed. Five such replacements were in exterior, solvent-accessible positions on the A helix and AB corner of the αA subunit. Five others were in close proximity to inositolpentaphosphate binding sites, and two pairs of independent replacements occurred at two different α¹β¹ intersubunit contacts. More than half of the substitutions in highland lineages resulted in the acquisition of serine or threonine (18 gains vs. 2 losses), both of which possess a hydroxyl group that can hydrogen bond to a variety of polar substrates. The patterns of parallel evolution observed in these waterfowl suggest that adaptation to high-altitude hypoxia has resulted from selection on unique but overlapping sets of one to five amino acid substitutions in each lineage.     

Multilocus phylogeography of the Patagonian lizard complex Liolaemus kriegi (Iguania: Liolaemini)

This study presents a detailed phylogeographical analysis of one of the most conspicuous groups of lizards in northwestern Patagonia, the Liolaemus kriegi complex. This region is geographically very complex as a result of Andean orogeny and subsequent volcanism coupled with a long history of glaciations and climatic changes. For 247 individuals we sequenced one mitochondrial gene (cytochrome b) and for a subset we sequenced another mitochondrial gene [12S ribosomal RNA (12S)] and two nuclear fragments [kinesin family member 24 (KIF24) and BA3 ribosomal RNA (BA3)]. We obtained gene trees and mitochondrial and nuclear haploytpe networks, and estimated genetic distances between the main lineages and basic molecular diversity indices. We also performed spatial analysis of molecular variance (SAMOVA) and Bayesian Skyline Plot (BSP) analyses, and concordant patterns from different lines of evidence permitted delimitation of seven lineages: two described species, Liolaemus buergeri and Liolaemus tregenzai; four candidate species, Liolaemus sp. A, Liolaemus sp. B, Liolaemus sp. C, and Liolaemus sp. D; and one lineage that includes all individuals from the geographical range of Liolaemus ceii and L. kriegi, referred to as L. kriegi + L. ceii. We discuss the evolutionary processes that may contribute to the origin of these lineages and their taxonomic and conservation implications. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 113 , 256–269.     

Palaeolimnology of Lake Hess (Patagonia,Argentina): multi-proxy analyses of short sediment cores

In contrast with the extensive palaeolimnological studies carried out in North America and Europe, relatively few studies have described the anthropogenic and/or climate impacts in Patagonian lakes. We addressed these issues by analysing geochemistry, lithology, pigments and chironomid remains from sediment cores collected from Lake Hess (41°22′20″S, 71°44′0″W) located in the Nahuel Huapi National Park in northern Patagonia. The aim of this study is to provide a palaeoenvironmental and climate reconstruction of the past ca. three centuries for this cold oligotrophic, quasi-pristine lake which receives meltwaters from the Tronador ice cap. Chronology was based on ¹³⁷Cs and ²¹⁰Pb measurements of the upper sediments, and the inferred sedimentation rate of 23.2 mg cm⁻² y⁻¹ (0.15 cm y⁻¹) was consistent with both sets of measurements. The sediment from Lake Hess was rich in tephra deposits particularly evident in the lower part of the cores. Tephras are valuable to use for core correlation and can be traced through peaks in the magnetic susceptibility (MS) profiles. Results from the multiproxy analyses in the longest core (83 cm) identify three main phases of change. From the bottom up to 42 cm (ca. ad 1800), the sediment is composed of light-grey organically rich clays. Both pigments and chironomids suggest variable trends in productivity and precipitation regime. At the end of the Little Ice Age chronozone (ad 1770–1850), pigment concentrations were very low. From 42 cm to ca. 25 cm (ad 1800–1940), the sedimentary record is composed of alternating black and dark organic-matter rich mud with variable amounts of macrophyte remains. Pigment concentrations and chironomid head capsule counts were also very low. These facies are composed of very fine plastic sediments with some faintly laminated intervals and an organic matter composition gradually decreasing towards the top of the zone. A sharp change occurs at 25 cm (ca. ad 1940) showing a strong increase in organic matter content, algal nutrients and plant pigments together with a change in the chironomid assemblages. This might document a change in the trophic condition of the lake associated with changes in erosion/deposition rates. Although there are records of human impact in the area studied, involving the use of fires, most of the observed chemical and biological changes in Lake Hess sediment sequence were interpreted in terms of climate changes, especially to changes in moisture balance brought about by variations in the strength of the westerlies. Guest editors: K. Buczkó, J. Korponai, J. Padisák & S. W. Starratt Palaeolimnological Proxies as Tools of Environmental Reconstruction in Fresh Water     

Phylogeny and biogeography of Valerianaceae (Dipsacales) with special reference to the South American valerians

Species of Valerianaceae are a common component of the alpine flora throughout the Northern Hemisphere as well as the Andes of South America. Sequence data from three chloroplast markers (psbA-trnH intron, trnK-matK intron, and the trnL-F region) along with the internal transcribed spacer region (ITS) of nuclear ribosomal DNA were used to infer relationships within Valerianaceae. Both genomes, as well as a combined data set, provide support for the major clades within the group and do not support a monophyletic Valeriana. In addition, these data indicate that Plectritis is nested within South American Valeriana, as opposed to being sister to Centhranthus as previously hypothesized. Valerianaceae appear to have originated in Asia, probably in the Himalayas, and subsequently to have dispersed several times to Europe and to the New World. Our results imply that Valerianaceae colonized South America on multiple occasions from the north. In one of these cases there appears to have been a substantial and rapid radiation, primarily in the high elevation paramo habitat. A variety of methods were used to estimate divergence times to determine when Valerianaceae might have colonized South America. Regardless of the method and fossil constraints applied, our estimates suggest that Valerianaceae colonized South America prior to the formation of the Isthmus of Panama.     

Late Pleistocene divergence between eastern and western populations of wood ducks (Aix sponsa) inferred by the 'isolation with migration' coalescent method

During the Late Pleistocene, glaciers sundered many species into multiple glacial refugia where populations diverged in allopatry. Although deeply divergent mitochondrial DNA (mtDNA) lineages often reflect the number of refugia occupied, it is unlikely that populations that split during the recent Wisconsin glaciations will have reached reciprocal monophyly. We examined mtDNA control region sequences from eastern and western populations of wood ducks (Aix sponsa) to determine whether their current, disjunct distribution is consistent with the occupancy of two glacial refugia. We used the 'isolation with migration' coalescent method (im) to simultaneously estimate effective population sizes, maternal gene flow, and time since divergence. We found 24 unique haplotypes, none of which were shared between the eastern and western populations, but we did not find diagnostic monophyletic lineages suggestive of long-term isolation in multiple glacial refugia. However, a high Phi ST (0.31) indicates that eastern and western populations are well differentiated in mtDNA, and results from im suggest that these populations have been diverging, without extensive gene flow, for 10,000 to 124,000 years. Results from im further suggest that these populations most likely split about 34,000 years ago, and this time of divergence is consistent with the occupancy of multiple glacial refugia during the Late Wisconsin glaciation. Eastern wood ducks are characterized by high genetic diversity, a large effective population size, and a recent population expansion, while western wood ducks have much less genetic diversity, a smaller population size, and have not undergone a recent population expansion.     

Nucleotide sequence data confirm diagnosis and local endemism of variable morphospecies of Andean astroblepid catfishes (Siluriformes: Astroblepidae)

Phylogenetic analysis based on nuclear and mitochondrial DNA sequences was used to test the validity of morphospecies of catfishes of the family Astroblepidae inhabiting the southern‐most limit of their Andean distribution in the upper Ucayali and upper Madre de Dios river basins. Population samples of morphospecies designated a priori on the basis of morphological features were further diagnosed by the presence of unique and unreversed molecular synapomorphies, thereby confirming species validity for seven of nine cases. Although each are distinguished by unique combinations of morphological features, two morphospecies (designated F and H) cannot be diagnosed on the basis of apomorphic changes in molecular sequence that did not also occur in other astroblepid morphospecies or outgroup taxa. Further, one morphospecies (species G) was recovered as nested within the assemblage of populations sampled from morphospecies F, whose morphological diagnosis does not involve unique or apomorphic characters. In contrast, the absence of corroborating molecular apomorphies for species H, otherwise recognized by distinctive and uniquely derived morphological characters, suggests a history of rapid divergence and insufficient time for fixation of genetic differences. Species sharing syntopic distributions were not recovered as sister groups, and in some cases species distributed in adjacent river drainage basins were not more closely related to one another than to species distributed in more distant drainages. Three independent instances were observed of sister‐group relationships involving species distributed in both the Apurimac and Urubamba rivers (Ucayali drainage). These observations combine to suggest that the current distribution of astroblepid species in the southern region may have arisen via a complex history involving both divergence between and dispersal amongst drainage basins that is probably repeated numerous times throughout the Andean distribution of the group. © 2011 The Linnean Society of London, Zoological Journal of the Linnean Society, 2011, 162 , 90–102.     

Phylogeography and population differentiation in terrestrial island populations of Banksia arborea (Proteaceae)

The Banded Iron Formations (BIFs) of south‐western Australia are terrestrial islands characterized by high species richness and endemism. Regional endemics occur across multiple formations without inhabiting the intervening landscape matrix. We investigated whether the occurrence on BIF terrestrial islands has led to genetic differentiation among the eight known populations of the regional endemic, Banksia arborea. Genetic structure was assessed using three chloroplast DNA sequence markers and 11 nuclear microsatellite loci. Phylogenetic relationships were assessed with statistical parsimony and Bayesian methods. Dates of haplotype divergence were estimated using the time to most recent common ancestor of B. arborea and Banksia purdieana, as well as a conservative angiosperm chloroplast (cp)DNA mutation rate. Population genetic diversity and structure was assessed amongst and within populations by genotyping 24 geographically clustered individuals from each BIF and three subpopulations within the Die Hardy Range BIF. Indirect gene flow estimates were determined using a method based on the frequency of private alleles. Banksia arborea showed low genetic diversity in (cp)DNA and a complex structural pattern, with genetic differentiation of some BIF populations and an absence of differentiation amongst others, reflecting either retention of ancestral polymorphism across northern BIF populations or more recent connectivity of these populations. There was little evidence of pollen dispersal both between BIFs and within the large BIF known as Die Hardy Range. © 2015 The Linnean Society of London, Biological Journal of the Linnean Society, 2015, 114 , 860–872.     

Genetic differentiation among Atlantic cod in south and south-east Icelandic waters: synaptophysin (Syp I) and haemoglobin (HbI) variation

Significant differences were found at the synaptophysin ( Syp I) locus between two groups of Icelandic cod Gadus morhua ; Loftstaõahraun (spawning ground), Reykjanesgrunn and Eyrabakkabugur (feeding grounds) on the one hand and Kantur (spawning ground) and Austfjarõadjúp (feeding ground) on the other. There was also a considerable genetic heterogeneity within the former group. The results indicate that the cod in south and south-east Icelandic waters do not belong to one panmictic population.     

Revision of the rhinoceros beetle genus Oryctophileurus Kolbe with description of a new species,the male of O. varicosus Prell,and notes on biogeography (Scarabaeoidea,Dynastinae, Phileurini)

The genus Oryctophileurus is reviewed and its validity is supported by a combination of the following apomorphic characters: a single cephalic horn with lateral carina, pronotal cavity with ocellate punctures and two teeth or tubercles close behind the anterior pronotal margin. The male of Oryctophileurus varicosus Prell, 1934, is described for the first time. A new species, Oryctophileurus guerrai Perger & Grossi sp. n., from subhumid Tucuman-Bolivian forest in the Southern Bolivian Andes is described. The new species is distinguished from its closest relative, O. armicollis Prell, 1911, by a narrower distance between the inner teeth of the dorsal pronotal protuberances and a reduced area of weakly developed ocellate punctures above the posterolateral pronotal margin. The occurrence of Oryctophileurus species in areas of endemism along the eastern slope of the tropical Andes suggests that these populations represent biogeographic “relicts”, and the discovery of Oryctophileurus guerrai sp. n. in the southern Bolivian Andes suggests that this area is underrated with respect to insect diversity and endemism.     

Contrasting genetic metrics and patterns among naturalized rainbow trout (Oncorhynchus mykiss) in two Patagonian lakes differentially impacted by trout aquaculture

Different pathways of propagation and dispersal of non‐native species into new environments may have contrasting demographic and genetic impacts on established populations. Repeated introductions of rainbow trout (Oncorhynchus mykiss) to Chile in South America, initially through stocking and later through aquaculture escapes, provide a unique setting to contrast these two pathways. Using a panel of single nucleotide polymorphisms, we found contrasting genetic metrics and patterns among naturalized trout in Lake Llanquihue, Chile's largest producer of salmonid smolts for nearly 50 years, and Lake Todos Los Santos (TLS), a reference lake where aquaculture has been prohibited by law. Trout from Lake Llanquihue showed higher genetic diversity, weaker genetic structure, and larger estimates for the effective number of breeders (N_b) than trout from Lake TLS. Trout from Lake TLS were divergent from Lake Llanquihue and showed marked genetic structure and a significant isolation‐by‐distance pattern consistent with secondary contact between documented and undocumented stocking events in opposite shores of the lake. Multiple factors, including differences in propagule pressure, origin of donor populations, lake geomorphology, habitat quality or quantity, and life history, may help explain contrasting genetic metrics and patterns for trout between lakes. We contend that high propagule pressure from aquaculture may not only increase genetic diversity and N_b via demographic effects and admixture, but also may impact the evolution of genetic structure and increase gene flow, consistent with findings from artificially propagated salmonid populations in their native and naturalized ranges.     

Congeneric phylogeography: hypothesizing species limits and evolutionary processes in Patagonian lizards of the Liolaemus boulengeri group (Squamata: Liolaemini)

In poorly known groups for which data are insufficient to develop biologically plausible model‐based approaches to phylogeographical analyses, a ‘first hypotheses’ protocol is suggested as offering the best way to generate hypotheses for subsequent model‐based tests. Preliminary hypotheses are formulated about species boundaries and population processes in three species complexes of the Liolaemus boulengeri group in the context of mtDNA ‘congeneric phylogeography’. The temperate South American Liolaemus provides a model with ancient and recent allopatric divergence across ecologically and geologically complex landscapes, incipient speciation, secondary contact, and discordance between molecular and morphological patterns of variation. Moderately dense sampling of widely distributed ‘inertial’ species in the Patagonian Steppe has revealed hidden genetic and probably species diversity, and also hinted at demographic and historical processes that may have shaped the histories of these taxa. Five of the seven focal species of the present study were paraphyletic for mtDNA genealogies, suggesting that they represent complexes of species, and nested clade phylogeographical analysis (NCPA) analyses suggest that different historical and demographic processes have shaped the observed patterns. Introgression and incomplete lineage sorting are hypothesized as being the cause of some of the observed paraphyly. Provisional delimitations of species are proposed and NCPA is used to generate hypotheses of population history, all of which are subject to further testing. Multi‐faceted studies, involving phylogenetic assessments of independent molecular markers and morphological variation across codistributed taxa with estimates of niche breadths in a landscape context, will likely yield the most promising returns for cross‐validation of hypotheses of population and speciation histories. © 2006 The Linnean Society of London, Biological Journal of the Linnean Society, 2006, 89 , 241–275.     

Genetic structure in northeastern populations of the Alpine newt (Triturus alpestris): evidence for post-Pleistocene differentiation

Genetic variation in 13 populations of the Alpine newt, Triturus alpestris, was assessed at the northeastern margin of its range (southern Poland). Variation at six microsatellite loci was scored in 354 newts, and two mitochondrial DNA fragments (c. 2000 bp) were sequenced in a subset of 27 individuals. Significant differences in allele frequencies and the presence of private alleles determined genetic units corresponding to three separate mountain ranges, i.e. the Carpathian, Sudetes and Holy Cross Mountains. F(ST)'s were three times greater in among than in within mountain range pairwise comparisons. An assignment test and pairwise F(ST)'s suggested relatively high levels of gene flow at the local level, although the Sudetes populations revealed some subtle structuring. Genetic variation was lower in the Carpathians and Holy Cross Mountains. The geographic pattern of mitochondrial DNA variation indicated that these newt populations originated from a single glacial refugium/founder population, and that the colonization of southern Poland took place in an easterly direction. The data show that substantial neutral variation and between group divergence has accumulated relatively quickly in these low-vagility organisms. The Alpine newt case exemplifies species history as a factor determining patterns of genetic diversity in marginal populations.     

Population differentiation in the marginal populations of the great tit (Paridae: Parus major)

The major subspecies group of the great tit, Parus major , has experienced demographic and spatial expansions during the last century in several sites at the edges of its distribution range. These expansions, although temporarily very even, have resulted in dissimilar patterns of molecular diversity. Populations locating at regions of contact to other subspecies groups (in Amur, Kirghizia–Kazakhstan, and Iran) show divergence from central population by nuclear and mitochondrial markers. In Amur, gene flow from minor group could be detected based on the existence of private minor alleles in the major population. In Kirghizia and Kazakhstan, the bokharensis and major groups share almost all the microsatellite alleles detected though frequencies differ. In Iran, three geographically close populations are distinct according to the mitochondrial sequences but also indications of present or recent admixture is detected. Populations, which have expanded to regions previously unoccupied by the species (northern UK and Finland), show divergence only by one of the markers. The variability in molecular differentiation may be due to dissimilar expansions, depending on whether the colonized regions have previously been occupied by another subspecies or not, on the amount of colonizing birds, and on the amount of past and present gene flow.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 90 , 201–210.     

中国甜菜夜蛾地理种群的遗传分化与基因流

【目的】为了明确我国甜菜夜蛾Spodoptera exigua地理种群间的遗传分化及基因流,阐明该种害虫在我国的种群历史动态。【方法】本研究对采自我国20个地理种群的529头甜菜夜蛾样品进行线粒体COI基因序列测定与分析,利用DnaSP 5.0和Arlequin 3.11软件分析种群间遗传多样性、遗传分化、基因流水平及分子变异,构建了单倍型系统发育树与单倍型网络图。【结果】在所分析的所有529个序列样本中,共检测出10个单倍型,其中Hap_1为所有种群所共享。总群体遗传多样性较低（Hd=0.257±0.025,Pi=0.0007±0.0001,Kxy=0.323）,群体间遗传分化较小（F_ST=0.211）,基因流水平较高（N_m=1.870）。AMOVA分析表明,甜菜夜蛾遗传变异主要来自种群内,种群间变异水平较低。各种群间遗传分化程度与地理距离无显著相关性（R²=0.005,P>0.05）。各单倍型相互散布在不同种群中,未形成明显系统地理结构。中性检验（Tajima’s D=-2.177, P<0.05; Fu’s F_S=-8.629, P<0.05)与错配分布分析表明,我国甜菜夜蛾种群曾经历种群近期扩张。【结论】研究结果揭示,甜菜夜蛾各种群间的基因交流并未受到地理距离的影响,验证了甜菜夜蛾具有高度的迁飞能力。     

Small body size increases the regional differentiation of populations of tropical mantellid frogs (Anura: Mantellidae)

The processes affecting species diversification may also exert an influence on patterns of genetic variability within species. We evaluated the contributions of five variables potentially influencing clade diversification (body size, reproductive mode, range size, microhabitat and skin texture) on mtDNA divergence and polymorphism among populations of 40 species of frogs (Mantellidae) from two rainforest communities in Madagascar. We report an inverse association between body size and nucleotide divergence between populations but find no influence of other variables on genetic variation. Body size explained ca. 11% of the variation in nucleotide divergence between populations and was coupled with high F_ST levels and an absence of haplotype sharing in small‐bodied and medium‐sized frogs. Low dispersal ability is likely the proximate mechanism producing higher population differentiation in small mantellids. The lack of genetic cohesion among populations establishes regional genetic fragmentation which in turn has the potential to accelerate rates of allopatric speciation in small frogs relative to large species. However, there is little evidence of increased speciation rates in these or other small‐bodied organisms. We reconcile these contradictory observations by suggesting that lower dispersal ability also curbs colonization of new areas, decelerating diversification in weak dispersers. Our results imply that the intermediate dispersal model also applies to amphibians and may explain inconsistent previous results on the correlation of body size and speciation rate.     

A cladistic analysis of the Solatium sessile species group (section Geminata pro parte: Solanaceae)

The Solanum sessile species group is one of eight putatively monophyletic subgroups of the large and diverse Solanum section Geminata. The species group comprises ten species of mostly northern South American trees and shrubs. Characters and character states peculiar to Solanum are discussed in detail. A cladistic analysis of the sessile group yielded four equally parsimonious trees which represent two basic tree topologies. The consensus cladogram is composed of three main clades. Biogeographic analysis using the consensus tree revealed no components in common to the three clades. Possible geographical scenarios are discussed with reference to the geological history of northern South America.     

Genetic Variation in Fragmented Forest Stands of the Andean Oak Quercus humboldtii Bonpl. (Fagaceae)1

Quercus humboldtii is a montane forest dominant species in Colombia, which has experienced significant habitat loss. Using three microsatellite loci, we compared the genetic diversity of adults and seedlings in fragments of small and large size. Results show high genetic diversity, comparable to other temperate oak species (H_o= 0.813, H_e= 0.780, and f=?0.044). However, allelic richness reduction in seedlings of the most fragmented part of the landscape, suggested restricted gene flow and risk of future genetic bottlenecks, if larger tracts of forest disappear.     

Population differentiation and species cohesion in two closely related plants adapted to neotropical high-altitude 'inselbergs', Alcantarea imperialis and Alcantarea geniculata (Bromeliaceae)

Isolated granitic rock outcrops or 'inselbergs' may provide a window into the molecular ecology and genetics of continental radiations under simplified conditions, in analogy to the use of oceanic islands in studies of species radiations. Patterns of variability and gene flow in inselberg species have never been thoroughly evaluated in comparison to related taxa with more continuous distribution ranges, or to other species in the same kingdom in general. We use nuclear microsatellites to study population differentiation and gene flow in two diploid, perennial plants adapted to high-altitude neotropical inselbergs, Alcantarea imperialis and Alcantarea geniculata (Bromeliaceae). Population differentiation is pronounced in both taxa, especially in A. imperialis. Gene flow in this species is considerably lower than expected from the literature on plants in general and Bromeliaceae in particular, and too low to prevent differentiation due to drift (N(e)m < 1), unless selection coefficients/effect sizes of favourable alleles are great enough to maintain species cohesion. Low gene flow in A. imperialis indicates that the ability of pollinating bats to promote gene exchange between inselbergs is smaller than previously assumed. Population subdivision in one inselberg population of A. imperialis appears to be associated with the presence of two colour morphs that differ in the coloration of rosettes and bracts. Our results indicate a high potential for inselbergs as venues for studies of the molecular ecology and genetics of continental radiations, such as the one that gave rise to the extraordinary diversity of adaptive strategies and phenotypes seen in Bromeliaceae.     

Growth in four populations of Leporinus friderici (Bloch, 1794) (Anostomidae, Teleostei) in French Guiana

The growth rates of Leporinus friderici (Bloch, 1794) in four populations from four rivers of French Guiana are compared. According to a statistical analysis of growth curves using the method of maximum likelihood with the Gauss–Markardt algorithm, a marked difference is observed in the growth of the different samples which is attributed to the year of capture rather than to the geographical origin of fishes. It is demonstrated that the main factor affecting growth performances is the length of the rainy season, which corresponds for this species to the feeding period.     

Genetic differentiation among the sympatric brown trout (Salmo trutta) populations of Lough Melvin, Ireland

Three morphotypes of brown trout have been described from Lough Melvin in north-west Ireland: gillaroo; sonaghen; ferox. The extensive genetic differences among the three types indicate that they are reproductively isolated, i.e. separate stocks of trout. For example, only gillaroo possess the Ldh -1(n) allele. Ferox show a high frequency of the Ldh -5(100) allele and this allele does not exceed a frequency of 0.02 in the other two types. Sonaghen are characterized by relatively much higher frequencies for Ck -2(115) and Gpi- 2(135) alleles. Sampling over a period of 7 years and experiments with artificial stocks of gillaroo and sonaghen have demonstrated that these differences are temporally stable. On the basis of observation of mature adults and the genetic composition of fry samples, it has been shown that the three types maintain their genetic integrity as the result of their distinctive spawning habits with gillaroo spawning in the lake and outflowing river, sonaghen in the smaller inflowing rivers and ferox in the deep downstream section of the largest inflowing river. The ferox is a relict population of an early post-glacial trout colonist while gillaroo and sonaghen are representative of a more recent independent colonization. However, it cannot be determined at present whether or not gillaroo and sonaghen were already differentiated prior to colonizing Melvin although the weight of evidence favours a sympatric split. It is proposed that the three types of trout be designated as separate subspecies to highlight the need for independent management and urgent requirement for conservation action.