Identification of a Gene Regulating the Tissue Expression of a Phosphoglucomutase Locus in Rainbow Trout

Nine percent of the rainbow trout (Salmo gairdneri) from a hatchery source have a greater than 100-fold increase in expression of a phosphoglucomutase (PGM) locus, Pgm1, in the liver but have normal expression of this locus in other tissues. The results of genetic crosses are consistent with a single regulatory gene with additive inheritance being responsible for the differences in the amount of PGM activity in the liver.—The allele responsible for the expression of Pgm1 in the liver is apparently a recent mutation. This is supported by its restricted distribution in rainbow trout and the absence of liver Pgm1 expression in closely related species. This genetic system is valuable for future analysis of the control of gene expression and in determining the relative evolutionary importance of genetic variation at structural and regulatory genes.     

HETEROZYGOSITY AND DEVELOPMENTAL RATE IN A STRAIN OF RAINBOW TROUT (SALMO GAIRDNERI)

Rainbow trout (Salmo gairdneri) with greater heterozygosity at enzyme loci also have greater developmental stability, as measured by bilateral symmetry of five meristic traits. Fish with increased amounts of liver phosphoglucomutase activity have greater developmental stability and develop faster than fish with normal activity. These observations suggest that the differences in developmental stability between homozygotes and heterozygotes may be the result of differences in developmental rate. Faster developmental rates are expected to decrease the probability of accidents during critical periods of development, resulting in a more stable or uniform phenotype. As indicated by differences in hatching time, heterozygotes tend to develop more rapidly than homozygotes. This association is not strongly expressed within families at any locus except Pgm1-t. However, heterozygotes for Mdh3,4 and Hex hatched significantly sooner than homozygotes in a population sample. These results suggest that differences in developmental rate between homozygotes and heterozygotes may account for the positive association between developmental stability and heterozygosity in rainbow trout.     

DEVELOPMENTAL INSTABILITY AND HIGH MERISTIC COUNTS IN INTERSPECIFIC HYBRIDS OF SALMONID FISHES

Interspecific hybrids have been proposed to have reduced developmental stability in comparison to their parental species because the parental genomes have not undergone selection for the maintenance of developmental stability when they occur together. We present data from four interspecific hybrids of salmonid fishes that support this view. Natural hybrids of bull trout (Salvelinus confluentus) with brook trout (Salvelinus fontinalis) and laboratory hybrids of rainbow trout (Salmo gairdneri) with Yellowstone (Salmo clarki bouvieri), westslope (S. c. lewisi), and coastal (S. c. clarki) cutthroat trout all have higher levels of fluctuating asymmetry than either of their parental species raised in the same environment. Thus, the hybrids have reduced developmental stability. The hybrids do not have meristic counts intermediate to the counts of the parental species. The hybrids usually have counts as high as the species with the higher count for those characters that differ between the parental species and often have higher counts for those characters that do not differ between the parental species. We suggest that the tendency for interspecific hybrids to have high meristic counts may be related to differences between the species in the length and timing of the developmental periods during which the counts of the characters are determined.     

Evidence of genetic and environmental influences on meristic variation in the rainbow trout,Salmo gairdneri Richardson

Synopsis An investigation of the effect of parentage and temperature on the expression of selected meristic characters of rainbow trout, Salmo gairdneri, reveals statistical differences in the number of vertebrae, dorsal fin rays and anal fin rays among test batches of progeny of different parentage. Batches of fertilized ova of the same stock or parentage, when incubated at 9.5 °C and 14.8 °C, produced lower mean dorsal and anal fin ray counts and a higher mean pectoral fin ray count at the higher temperature. Incubation temperature had no significant effect on the number of vertebrae.Significant statistical differences occurring in various characters among groups of rainbow trout of different parental origin provide convincing evidence of genetic inputs into meristic expression. However, the existence of both environmental and genetic inputs into meristic expression in rainbow trout necessitates further investigation before the meaning of observed meristic plasticity in rainbow trout can be resolved.     

Developmental stability in brown trout: are there any effects of heterozygosity or environmental stress?

We studied the developmental stability of brown trout, Salmo trutta L., in 10 populations (five acidified, five control) in Norway, measured as fluctuating asymmetry (FA) and departure from the morphological norm. We measured four meristic and four morphometric characters, and scored the level of biochemical heterozygosity at 49 loci (20 polymorphic). We reared eggs of a single population in a hatchery using four different water qualities (three replicates of each treatment) to test the effect of acidification stress on developmental instability. There were no significant differences in the level of FA, in departure from the morphological norm between brown trout sampled from lakes with acidified or control water qualities, or in brown trout hatched at different water qualities. There was no correlation between level of heterozygosity and FA or departure from the morphological norm, either when tested within populations or among populations. There were no single-locus effects on developmental stability tested for 11 loci. We conclude that measures of developmental stability or morphological variability are not useful for detecting acidification stress in brown trout. Furthermore, we conclude that developmental stability in our material varies independently of heterozygosity.     

Deviation from morphological intermediacy in interstrain hybrids of rainbow trout,Salmo gairdneri

Synopsis Deviations from morphological intermediacy in six first generation hybrids between three hatchery strains of rainbow trout, raised in a common environment, are reported. Hybrids have higher mean counts of four meristic characters than their maternal parental strain in a significantly greater number of cases (18 out of 24). Furthermore, eight of eleven hybrid indices are not intermediate. These results are discussed in reference to several mechanisms and models proposed to account for observed responses of meristic characters to environmental and genetic influences.     

MITOCHONDRIAL GENOTYPES HAVE NO DETECTABLE EFFECTS ON MERISTIC TRAITS IN CUTTHROAT TROUT HYBRID SWARMS

Efforts to detect effects of cytoplasmic genes are confounded by the problem of partitioning nuclear and cytoplasmic effects. In this study we test for effects of mtDNA haplotype on development in hybrid populations of cutthroat trout (Oncorhynchus clarki) with randomly associated nuclear and mtDNA genotypes. We have previously described several intraspecific hybrid swarms formed by interbreeding of westslope cutthroat trout (O. c. lewisi) and Yellowstone cutthroat trout (O. c. bouvieri). Genetic distance between these subspecies is high (Nei's D = 0.30; mtDNA P = 0.02), and diagnostic alleles at multiple nuclear loci and two distinct mtDNA haplotypes are present in the hybrids. Historical associations between alleles at nuclear loci and between cytotypes and nuclear alleles have largely decayed. We test for differences in meristic characters between fish with alternate mtDNA haplotypes. Counts and fluctuating bilateral asymmetry for these characters have been previously shown to be sensitive indicators of genetic differences that affect development. No differences were found between mtDNA types in meristic counts or fluctuating asymmetry. Therefore, the alternate mtDNA haplotypes had no detectable effect on development as measured by meristic counts in these hybrid populations. However, diagnostic alleles at one nuclear allozyme locus (CK-CI) were associated with several fin ray counts.     

Morphological variation in a secondary contact between divergent lineages of brown trout (Salmo trutta) from the Iberian Peninsula

The aim of this study was to analyze the morphological variation of brown trout (Salmo trutta) in the Duero basin, an Atlantic river basin in the Iberian Peninsula, where a spatial segregation of two divergent lineages was previously reported, based on isozyme, microsatellite and mtDNA data. In these studies, two divergent pure regions (Pisuerga and Lower-course) and several hybrid populations between them were identified. Morphological variation was evaluated in 11 populations representative of the genetic differentiation previously observed in the Duero basin, using multivariate analysis on 12 morphometric and 4 meristic traits. A large differentiation between populations was observed (interpopulation component of variance: 41.8%), similar to that previously detected with allozymes and microsatellites. Morphometric differentiation was also reflected by the high classification success of pure and hybrid individuals to their respective populations, using multivariate discriminant functions (94.1% and 79.0%, respectively). All multivariate and clustering analyses performed demonstrated a strong differentiation between the pure regions. The hybrid populations, though showing large differentiation among them, evidenced an intermediate position between the pure samples. Head and body shape traits were the most discriminant among the morphometric characters, while pectoral rays and gillrakers were the most discriminant among the meristic traits. These results confirmed the high divergence of the brown trout from the Duero basin and suggest some traits on which selection could be acting to explain the spatial segregation observed.     

Stock Transfers in Spanish Brown Trout Populations: A Long-term Assessment

Synopsis Stocking of fish from other populations has been commonly employed for enhancement of wild brown trout, Salmo trutta, populations in north Spain. Young hatchery reared brown trout of central European origin were introduced into some Asturian rivers every year since 1984. Based on variation at the isozyme locus LDH-C1* and at the microsatellite locus BFRO 002, two genetic markers race-specific in Salmo trutta, we detected introgression of foreign genomes into native gene pools in some Spanish trout populations where only pure native individuals were present 10 years ago. We strongly suggest development of alternative management policies for conservation of Spanish natural brown trout populations without endangering the traditional recreational fisheries. Jorge I. Izquierdo, Ana G. F. Castillo: These two authors contributed equally to the article.     

Meristic variation among world hatchery stocks of rainbow trout,Salmo gairdneri Richardson

Synopsis Sixteen meristic characters were examined among 10 stocks of rainbow trout to establish their inter- and intrastock variability and relative discriminating power. The number of vertebrae, dorsal fin rays, scales above and below and lateral line, oblique scale rows and pectoral fin rays were found to be the most useful meristic characters in identifying stocks. Examination of the extent of meristic divergence among 10 hatchery or naturalized stocks from various regions of the world, using analysis of variance, the generalized distance function and per cent overlap, revealed that each stock exhibited highly significant differences in at least one meristic character, thereby demonstrating a high degree of intraspecific variability in meristic expression.In multivariate terms, stocks from Idaho and Alaska were almost totally divergent from all other stocks examined. Multivariate analysis of meristic features proves to be a useful method for identifying the extent of phenotypic variability among and within divergent stocks of rainbow trout. The observed meristic variability in test stocks of hatchery fish is seemingly genetically set within the limits previously described for native populations of rainbow trout.     

The relationship between developmental instability of gudgeon Gobio gobio and abundance or morphology of its ectoparasite Paradiplozoon homoion (Monogenea)

Gudgeon Gobio gobio (Cyprinidae) were collected monthly from the Vlara Stream (Czech Republic) during 2004 and examined for the presence of the ectoparasite Paradiplozoon homoion (Diplozoidae; Monogenea). Over 26% of P. homoion showed abnormalities of the attachment sclerites, indicating either a stressful environment or that the quality of the fish host was suboptimal for parasite development. Here, the hypotheses that parasite abundance and the occurrence of morphological abnormalities are correlated with fluctuating asymmetry (FA, the random deviation from the perfect bilateral symmetry) in its fish host were tested. The FA and morphological abnormality are estimators of developmental instability. To estimate fish developmental instability nine meristic and two morphometric bilateral characters on the right and the left sides of the fish were measured. In general, there was no correlation between mean fish FA and parasite abundance or the proportion of parasites with abnormalities. There was, however, a significant relationship between the proportion of abnormal P. homoion and the number of asymmetric meristic traits per individual fish. This study thus indicates that developmental instability of P. homoion , measured as morphological abnormality of the attachment apparatus, is only weakly dependent on the fish quality as measured by FA.     

Morphological variation in hybrids between Salmo marmoratus, and alien Salmo species in the Volarja stream, Soca River basin, Slovenia

There were significant correlations between colour pattern, LDH -5*, genotype and certain meristic characters in 59 hybrid trout Salmo , sp. from the Volarja stream, Soca River basin, Slovenia. It is concluded that panmixia between native Salmo marmoratus , and introduced S. trutta , of Atlantic, Danubian and Mediterranean origin had not been reached in this zone, despite the long period of introgression. The result is in agreement with other studies dealing with introgression in Salmo , and for management purposes certain morphological characters, especially colour pattern, can be a valuable tool in restoring the marble trout population in the Soca River.     

Testing the relationship between domestication and developmental instability in rainbow trout,Oncorhynchus mykiss (Teleostei,Salmonidae)

Fish domestication is an evolutionary process arising in captivity through genetic and developmental mechanisms, producing organisms performing more poorly than wild conspecifics in the natural environment. Culture conditions could be suboptimal for fish at particular life cycle stages, presenting environmental disturbances leading to developmental instability. The limited size of captive lots, moreover, can result in the loss of genetic variation, and the resulting homozygosity (as well as hybridization and mutation) could have strong harmful effects on developmental stability. Rainbow trout are the most widely‐cultured species in Europe and North America, having been in culture for more than a century. Prolonged artificial selection for desired traits and incidental effects of domestication has led to the development of a ‘farmed type’. Fluctuating asymmetry, variations in meristic counts, and skeletal anomalies were examined in several rainbow trout captive and wild clonal lines as indicators of developmental instability. Differences in developmental stability were identified among lines and correlated with different degrees of exposure to captivity. Some relationship between meristic counts and domestication level was found in the present study, with the number of vertebrae and of dorsal pterygiophores and rays being the strongest predictors of the domestication level. However, the occurrence of skeletal anomalies and fluctuating asymmetry were apparently not related to the level of exposure to captivity. The findings of the present study will facilitate the selection of clonal lines with divergent phenotypes for subsequent quantitative trait loci analyses aimed at identifying genome regions linked with morpho‐anatomical and physiological adaptive responses to captivity. © 2015 The Linnean Society of London, Biological Journal of the Linnean Society, 2015, 114 , 608–628.     

Evidence for co‐dominant allelic expression of a phosphoglucomutase regulatory locus polymorphism in the Atlantic salmon

Starch gel electrophoresis of the phosphoglucomutase isozyme, PGM‐1, in liver tissue in Atlantic salmon Salmo salar shows three phenotypes – full, partial and no expression. The genetic basis of the variation has not been established. Studies of the inheritance of the variation and of liver enzyme levels carried out indicate that the variation, as in rainbow trout Onchorynchus mykiss , reflects co‐dominant allelic variation at a cis ‐acting regulatory locus, designated PGM‐1r *, with one allele promoting and the other suppressing expression.     

Absence of developmental incompatibility in hybrids between rainbow trout and two subspecies of cutthroat trout

We examined the developmental rate of hybrids between rainbow trout (Salmo gairdneri) and two subspecies of cutthroat trout: westslope cutthroat trout (Salmo clarki lewisi) and Yellowstone cutthroat trout (Salmo clarki bouvieri). These taxa show considerable genetic divergence at 42 structural loci encoding enzymes; the mean Nei's d between the rainbow trout and the two species of cutthroat trout is 0.22. We used four measures of developmental rate: time of hatching and yolk resorption, rate of increase in activity of four enzymes, and time of initial detection of seven isozyme loci. The two cutthroat trout subspecies reached hatching and yolk resorption earlier than rainbow trout. Cutthroat trout had higher relative enzyme activities than rainbow trout from deposition of eye pigment to hatching. There was no difference in the rate of increase in enzyme activity or time of initial expression of these loci between these species. Hybrids showed developmental rates intermediate or similar to that of the parental species using all measures. Our results indicate an absence of regulatory and developmental incompatibility between these taxa.This research was supported by NSF Grants ISP-8011449 and BSR-8300039. M.M.F. was supported by a postgraduate scholarship from the Natural Sciences and Engineering Research Council of Canada.     

Morphological differentiation among local trout (Salmo trutta) populations

The trout (Salmo trutta) has been divided into three forms: sea-run trout, lake-run brown trout, and resident brown trout. They differ in their living environment, migratory behaviour, growth and appearance. As local trout populations are often isolated, and gene flow between them is minimal, differentiation between populations can be expected. The morphology of 1-year-old trout from ten populations representing all three forms was studied in a common-garden experiment. The fish were reared under similar environmental conditions, and 20 morphometric characters were measured from each individual fish. Marked morphological differentiation was found, and differences between populations were greater than differences between forms. The results suggest that the differences have a genetic basis, and they are likely to indicate adaptation to local environmental conditions in the native habitat of the trout.     

Reticulate evolution: ancient introgression of the Adriatic brown trout mtDNA in softmouth trout Salmo obtusirostris (Teleostei: Salmonidae)

Two populations of softmouth trout ( Salmo obtusirostris ) from the rivers Neretva (Bosnia and Herzegovina) and Jadro (Croatia), along with two neighbouring populations of brown trout ( Salmo trutta ) were analysed with a suite of genetic markers (two mtDNA genes, two nuclear genes, and nine microsatellites) as well as morphological characters. The Jadro softmouth trout were fixed for a brown trout mtDNA haplotype of the Adriatic lineage, which is 1.7% divergent from a previously described haplotype characteristic for the Neretva softmouth trout. All other genetic markers, as well as morphological analysis, supported the clear distinction of softmouth trout from the rivers Neretva and Jadro from brown trout in neighbouring populations, and thus a mtDNA capture event is assumed. Population specific microsatellite allele profiles, as well as a high number of private alleles for both populations of softmouth trout, support the hybridization between brown trout and the Jadro softmouth trout most likely being of ancient origin, thus leading to a reticulate evolutionary pattern of mtDNA in this taxon.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 90 , 139–152.     

Genetic and morphological characterization of a Lake Ohrid endemic, Salmo (Acantholingua) ohridanus with a comparison to sympatric Salmo trutta

Analysis of both uni‐(two mtDNA gene sequences) and bi‐parentally (seven microsatellite loci) inherited genetic markers, together with analysis of 40 morphological characters, described Salmo ohridanus as a highly divergent member of the genus Salmo . Based on comparative substitution rate differences at the cytochrome b gene, and a rough estimated age of the Salmo trutta complex ( i.e. at least 2 million years), the S . ohridanus and Salmo obtusirostris clade probably split from a common ancestor of brown trout Salmo trutta >4 million years ago, overlapping with minimum age estimates of the formation of Europe's oldest freshwater habitat, Lake Ohrid. Comparative analysis with Lake Ohrid brown trout (known regionally as Salmo letnica ), supported the notion that these fish have more recently colonized the lake and phylogenetically belong to the Adriatic lineage of brown trout. It is further suggested that species‐specific saturation in the mtDNA control region underestimated the divergence between S . ohridanus and S . trutta . Evidence of rare hybridization between S . ohridanus and Lake Ohrid brown trout was seen at both mtDNA and microsatellite markers, but there was no support for extensive introgression.     

Effect of hatchery environment on cranial morphology and developmental stability of Atlantic salmon (Salmo salar L.) from North‐West Russia

The study addresses the effect of hatchery rearing on morphological variation and developmental stability of Atlantic salmon parr from North‐West Russia. Totally, we collected nine samples. Four wild samples were collected from each of the rivers Kola, Umba, Keret’ and Shuia. Five samples of hatchery‐reared parr were the first‐generation progeny of wild adults from these rivers reared separately at the four hatcheries (one hatchery was represented by two samples). Ten meristic and 48 morphometric cranial characters were analysed. We studied the morphological divergence between wild and hatchery fishes of the same river of origin. To analyze developmental stability we used fluctuating asymmetry (random deviations from perfect bilateral symmetry). It was found that hatchery‐reared parr significantly differ from wild parr in both meristic characters and the shape of cranial bones. Different hatcheries caused similar effect on morphological variation in all populations. Fluctuating asymmetry in morphometric characters was significantly higher in hatchery fish than in wild from the Shuia River, indicating a higher level of developmental instability. However, wild parr from the Keret’ River had significantly higher fluctuating asymmetry than cultivated parr of the same origin, possible due to a high infection pressure of the parasite Gyrodactylus salaris Malmberg which has led to significant decline of the wild salmon population in this river, or from genetic changes caused by cultivation. The obtained results indicate a notable effect of hatchery environment on Atlantic salmon’s phenotype.     

Phylogeography and taxonomic status of trout and salmon from the Ponto‐Caspian drainages,with inferences on European Brown Trout evolution and taxonomy

Current taxonomy of western Eurasian trout leaves a number of questions open; it is not clear to what extent some species are distinct genetically and morphologically. The purpose of this paper was to explore phylogeography and species boundaries in freshwater and anadromous trout from the drainages of the Black and the Caspian Seas (Ponto‐Caspian). We studied morphology and mitochondrial phylogeny, combining samples from the western Caucasus within the potential range of five nominal species of trout that are thought to inhabit this region, and using the sequences available from GenBank. Our results suggest that the genetic diversity of trout in the Ponto‐Caspian region is best explained with the fragmentation of catchments. (1) All trout species from Ponto‐Caspian belong to the same mitochondrial clade, separated from the other trout since the Pleistocene; (2) the southeastern Black Sea area is the most likely place of diversification of this clade, which is closely related to the clades from Anatolia; (3) The species from the Black Sea and the Caspian Sea drainages are monophyletic; (4) except for the basal lineage of the Ponto‐Caspian clade, Salmo rizeensis, all the lineages produce anadromous forms; (5) genetic diversification within the Ponto‐Caspian clade is related to Pleistocene glacial waves; (6) the described morphological differences between the species are not fully diagnostic, and some earlier described differences depend on body size; the differences between freshwater and marine forms exceed those between the different lineages. We suggest a conservative taxonomic approach, using the names S. rizeensis and Salmo labrax for trout from the Black Sea basin and Salmo caspius and Salmo ciscaucasicus for the fish from the Caspian basin.