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.     

DEVELOPMENTAL STABILITY OF RAINBOW TROUT HYBRIDS: GENOMIC COADAPTATION OR HETEROZYGOSITY?

I compare the developmental stability of first generation hybrids between hatchery strains of rainbow trout (Salmo gairdneri) to that of the three pure parental strains raised in a common environment. Two of three reciprocal hybrid pairs show significantly less fluctuating asymmetry of four meristic characters than is found in parental strains. In contrast, the third hybrid pair shows reduced but not significantly lower developmental stability compared to pure strains. These hybrids had previously been found to develop slower than their maternal parental strains, indicating divergence of parental regulatory mechanisms controlling early ontogeny. A significant positive association between the degree of relative delay in hybrid developmental rate and their degree of developmental instability supports this view. For example, the only hybrid pair with decreased developmental stability also had the largest relative delay in development time of all hybrids. Neither absolute developmental rate nor enzyme heterozygosity at 42 loci alone can explain the degree of fluctuating asymmetry in these hybrids. The developmental stability of hybrids is apparently a result of the interaction between the developmental divergence between parental strains and their genomic heterozygosity due to hybridization.     

Major morphological effects of a regulatory gene: Pgm1-t in rainbow trout

We have investigated the morphological effects of a genetic locus, Pgm1- t, that affects the expression of a phosphoglucomutase locus (Pgm1) in liver of rainbow trout (Salmo gairdneri). We have previously shown that embryos with liver Pgm1 expression hatch earlier than those without liver Pgm1 expression. We predicted that this difference in developmental rate should cause a reduction in meristic counts in the more rapidly developing fish with liver Pgm1 expression. Eight meristic (countable) characters in nine full-sib groups segregating for the presence or absence of liver Pgm1 expression are in agreement with this prediction. In eight of the nine families, there is a significant difference in the multivariate distribution of the eight meristic counts between full sibs with and without liver Pgm1 expression. This separation in multivariate space is based on a tendency for lower meristic counts in fish with liver Pgm1 expression. The magnitude of these morphological differences is similar to that between two subspecies of cutthroat trout (Salmo clarki) that show substantial genetic divergence at structural loci encoding enzymes (Nei's D = 0.34). These data support the view that small changes in the developmental process caused by genetic differences at regulatory genes can have large effects on morphology.      

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.     

Erosion of interspecific reproductive barriers resulting from hatchery supplementation of rainbow trout sympatric with cutthroat trout

The frequency of hybridization between cutthroat (Onchorhynchus clarki clarki) and rainbow (O. mykiss irideus) trout from coastal habitats in British Columbia, Canada, was examined in seven populations where the two species are sympatric with no history of rainbow trout stocking and compared with areas where native rainbow trout populations have been supplemented with hatchery fish (three populations). Four nuclear markers were used to identify each species and interspecific hybrids and one mitochondrial marker showed the direction of gene exchange between species. The frequency of hybrids was significantly higher (Fisher exact test, P < 0.001) in river systems where hatchery rainbow trout have been introduced (50.6% hybrids) than in populations where the two species naturally co-occur without supplementation (9.9% hybrids).     

Linkage Relationships Reflecting Ancestral Tetraploidy in Salmonid Fish

Fifteen classical linkage groups were identified in two salmonid species (Salmo trutta and Salmo gairdneri) and three fertile, interspecific hybrids (S. gairdneri X Salmo clarki, Salvelinus fontinalis X Salvelinus namaycush and S. fontinalis X Salvelinus alpinus) by backcrossing multiply heterozygous individuals. These linkage relationships of electrophoretically detected, protein coding loci were highly conserved among species. The loci encoding the enzymes appeared to be randomly distributed among the salmonid chromosomes. Recombination frequencies were generally greater in females than in males. In males, certain linkage groups were pseudolinked with other linkage groups, presumably because of facultative multivalent pairing and directed disjunction of chromosomes. Five such pseudolinkage groups were identified and they also appeared to be common among species and hybrids. Duplicate loci were never classically linked with each other, although some exhibited pseudolinkage and some showed evidence of exchanging alleles. Gene-centromere recombination frequencies estimated from genotypic distributions of gynogenetic offspring were consistent with map locations inferred from female intergenic recombination frequencies. These linkage relationships support the contention that all extant salmonids arose from a common tetraploid progenitor and that this progenitor may have been a segmental allotetraploid.     

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.     

Failure of interspecies androgenesis in salmonids

Androgenetic development of salmonid embryos was induced in recipient oocytes from the same or other species (intra- or interspecies androgenesis). Parameters for induced androgenesis were investigated in brown trout Salmo trutta and brook trout Salvelinus fontinalis . Reciprocal androgenetic and control crosses were conducted among fishes from three genera: Oncorhynchus (rainbow trout, O. mykiss ), Salmo (brown trout) and Salvelinus (brook trout), and within two genera: Salmo (brown trout and Atlantic salmon, S. salar ) and Salvelinus (brook trout and Arctic charr, S. alpinus ). Live hatched androgenetic progenies were obtained in all intraspecies variants, where oocytes and sperm originated from the same species. Interspecies androgenesis resulted in no viable larvae, despite the fact that most hybrid controls and intraspecies androgenetic individuals were viable. When recipient oocytes originated from other genera (interspecific intergeneric androgenesis), embryonic development ceased in early developmental stages, except for haploid controls of brook trout produced in eggs of brown trout. Survival of embryos to the eyed-egg stage was relatively high in the intrageneric androgenesis experiment. Nevertheless, none of these embryos survived to hatching. Some of the presumed Atlantic salmon individuals developing in brown trout eggs contained maternal DNA, questioning the accuracy of enucleation using irradiation. The inability to induce interspecific androgenesis among the examined salmonid species may have been the result of substantial kariotypical and developmental differences between spermatozoal donors and oocyte recipients, causing an incompatibility between maternal cytoplasmic regulatory factors and the paternal nuclear genome.     

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.     

Genetic variation,inheritance, and quaternary structure of malic enzyme in brook trout (Salvelinus fontinalis)

Electrophoretic variation is described for malic enzyme (ME) for the first time in brook trout (Salvelinus fontinalis). Since the quaternary structure of ME was not clear from examination of banding patterns in brook trout alone, ME phenotypes in rainbow trout (Salmo gairdneri) × brook trout hybrids as well as in esocid species demonstrated that ME is tetrameric. A model of two duplicated loci is proposed to account for the observed variation. One locus (ME-2) is fixed and one locus (ME-1) is variable with three electrophoretically distinct alleles; the protein products of ME-1 are reduced in activity relative to the protein products of ME-2. Joint segregation was examined between ME-1 and ten other biochemical loci in brook trout, and between ME-1, ME-2, and nine other biochemical loci in a splake—lake trout (Salvelinus namaycush) × brook trout hybrid—backcross. All pairwise examinations showed random assortment except ME-2 with an isocitrate dehydrogenase locus (IDH-3), which showed complete linkage in the splake backcross. This may be due to a chromosomal aberration.Authorized for publication as Paper No. 5599 in the Journal Series of The Pennsylvania Agricultural Experiment Station, University Park, Pennsylvania, in cooperation with the Benner Spring Fish Research Station, The Pennsylvania Fish Commission, Bellefonte, Pennsylvania. M.S. was supported by an NSF Graduate Fellowship.     

Persistence of maternal isoenzyme patterns of the lactate dehydrogenase and phosphoglucomutase system during early development of hybrid trout

Developmental changes in the isoenzyme patterns of LDH and PGM during early embryogenesis (until hatching) in two trout species (Salmo trutta and Salmo irideus) as well as in their reciprocal interspecific hybrids are reported, completing a previous study which had commenced from hatching onward (Hitzeroth et al., 1968). Isoenzymes of both systems occur in unfertilized eggs and in embryos from fertilization onward. Until about the Augenpunkt stage, the isoenzyme patterns remain unchanged and are identical to the patterns of unfertilized eggs; the reciprocal hybrids correspond to the respective maternal species. Additional isoenzymes appearing for the first time at about the Augenpunkt stage in the interspecific hybrids show the same electrophoretic mobility as those in the animals of the respective maternal species at this developmental stage. It is discussed whether the appearance of additional isoenzymes characterizes the activation of maternally derived alleles of the embryonic genome, or whether maternally derived cytoplasmic factors are responsible for the occurrence of these additional bands.Supported by the Deutsche Forschungsgemeinschaft.     

Evidence for interspecific hybridization between native white-spotted charr Salvelinus leucomaenis and non-native brown trout Salmo trutta on Hokkaido Island, Japan

Hybrids between native white-spotted charr Salvelinus leucomaenis and non-native brown trout Salmo trutta were identified in streams of Hokkaido, Japan, using both appearance and genetic characters. The DNA analyses indicated that the specimens were hybrids between female S. leucomaenis and male S. trutta . Occurrence of such hybrids implies increased mating opportunities between these species in wild streams.     

Host-related life history traits in interspecific hybrids of cactophilic Drosophila

In the genus Drosophila (Diptera: Drosophilidae), interspecific hybridization is a rare phenomenon. However, recent evidence suggests a certain degree of introgression between the cactophilic siblings Drosophila buzzatii Patterson & Wheeler and Drosophila koepferae Fontdevila & Wasserman. In this article, we analyzed larval viability and developmental time of hybrids between males of D. buzzatii and females of D. koepferae, raised in media prepared with fermenting tissues of natural host plants that these species utilize in nature as breeding sites. In all cases, developmental time and larval viability in hybrids was not significantly different from parental lines and, depending on the cross, hybrids developed faster than both parental species or than the slowest species. When data of wing length were included in a discriminant function analysis, we observed that both species can be clearly differentiated, while hybrids fell in two categories, one intermediate between parental species and the other consisting of extreme phenotypes. Thus, our results point out that hybrid fitness, as measured by developmental time and viability, is not lower than in the parental species.     

Comparison of competitive ability between native and introduced salmonids: evidence from pairwise contests

Brown trout, Salmo trutta, and rainbow trout, Oncorhynchus mykiss, have been introduced to freshwaters in Hokkaido, Japan. Today, it is recognized that these introduced salmonids have negative impacts on native salmonids such as white-spotted charr, Salvelinus leucomaenis, and masu salmon, O. masou. In particular, interspecific competition may be an important mechanism that could contribute to the exclusion for native salmonids. In this study, experimental pairwise contests were conducted to compare interference competitive ability between native and introduced salmonids. We demonstrated that brown trout were competitively superior to white-spotted charr and masu salmon whereas rainbow trout were superior to white-spotted charr. We suggest that introduced brown trout negatively impact both white-spotted charr and masu salmon, and introduced rainbow trout negatively impact white-spotted charr.     

An experimental test of the effect of predation upon behaviour and trait correlations in the threespine stickleback

Benthic and limnetic threespine stickleback (Gasterosteus aculeatus) are a classic example of ecological speciation. Behavioural and armour divergence between these species has been predicted to be the result of divergent selection driven in part by differential predation from cutthroat trout (Oncorhynchus clarki). To experimentally test this prediction, we reared split families of benthic–limnetic hybrids in the presence or absence of trout predation. Our results show that the presence of trout had little effect upon stickleback behaviour. We then compared performance in behavioural assays among stickleback that varied in armour to test if armour morphology correlates with behaviour. Our measurements revealed trait correlations between several behaviours and components of armour morphology. Trout predation did not result in an increased correlation between traits, therefore differential trout predation between benthics and limnetics is unlikely to be the cause of these correlations. The presence of trait correlations in advanced generation hybrids suggests that pleiotropy or linkage between genes underlying behaviour and armour morphology may be greater than previously appreciated.     

Genetic and biochemical aspects of lactate dehydrogenase isozymes in the salmonid eye

Polyacrylamide gel electrophoresis reveals similar differences between the retinal-specific lactate dehydrogenase (LDH) isozymes of the salmon (Salmo salar L.) and the sea-trout (Salmo trutta forma trutta L.) as those previously described for the salmon and the brown trout (Salmo trutta L.). F₁ salmon × sea-trout hybrids give a classic hybrid isozyme pattern, but the F₂ hybrids all possess the parental sea-trout type pattern. Loss of part of the salmon genome in these latter hybrids is the most likely explanation. It was observed that when the individual eye isozymes of the salmon, the sea-trout, and the rainbow trout (Salmo gairdneri Richardson) were eluted from preparative polyacrylamide gels and re-electrophoresed, an apparent interconversion of certain isozyme bands occurred. This phenomenon was also evident using starch gel. However, the major cathodally migrating isozyme in each case (presumably the E4 isozyme) re-electrophoresed pure. The reasons for these interconversions are, as yet, unclear. Attempts to produce in vitro hybridization between the various isolated individual isozymes were unsuccessful. K_m pyruvate values for the different salmon isozymes were of the order expected from results already published for other teleosts.     

Short-term toxicity of oil-based drilling fluid to the brackish-water shrimp Palaemonetes africanus

Thirty-nine morphometric and five meristic comparisons were carried out on 12-month-old interspecific and intergeneric hybrids of the African clariid catfishes Heterobranchus longifilis, Clarias gariepinus and Clarias anguillaris from experimental earthen ponds in Nigeria. Canonical discriminant analysis accounted for 89.4% of the variations, with eigenvalues of 151.69 and 9.06, respectively, the highest loading in the two axes coming from only 12 variables. A re-analysis based on those 12 variables revealed four major groups among the nine genetic combinations from a plot of axes 1 and 2, which accounted for 95.2% of the variations. The Mahalanobis square distance (D ²) between the offspring of the various mating combinations also revealed the varying levels of relationship between them based on their morphometric and meristic characters. The interspecific hybrids showed some level of heterosis only in the inheritance of the length of the frontal fontanelles compared to that of the putative Clarias species. The study shows the level of similarities in many of the morphometric and merisitic features, especially among the hybrids, and the distinguishing features that can be used to identify the hybrids from their putative parents.     

Interspecific association of brown trout (Salmo trutta) with non‐native brook trout (Salvelinus fontinalis) at the fry stage

The introduction of non‐native brook trout (Salvelinus fontinalis) in Europe has led to displacement and decreasing populations of native brown trout (Salmo trutta). Some studies have found that brown trout shift to a diet niche similar to brook trout when the two species live in sympatry, which conflicts with the competitive exclusion principle. A change in feeding niche may be a sign of early interspecific association and social learning, leading to behavioral changes. As a first step to address this possibility, it is essential to assess the interspecific association between the species during the early ontogenetic life stages. In this study, we therefore assess whether juvenile brown trout associate with non‐native juvenile brook trout to the same extent as with conspecifics by setting up two experiments: (i) a binomial choice test allowing visual and chemical cues to estimate the species specificity of group preference, and (ii) an association test without physical barriers to estimate the degree of association of a focal brown trout with a group of either conspecifics or heterospecifics. In experiment (1), we found that focal juvenile brown trout preferred to associate with the stimuli groups and did not discriminate either against conspecific or heterospecific groups. Furthermore, more active individuals showed stronger preference for the stimuli group than less active ones, regardless of species. In experiment (2), we found that brook trout groups had a tighter group structure than brown trout groups, and that focal brown trout showed stronger association with brook trout than with brown trout. These results indicate that brown trout may associate with brook trout at an early life stage, which would allow for interspecific social learning to occur. Future studies should look closer into causes and consequences of interspecific association and social learning, including potential effects on the phenotype selection in brown trout populations.     

Morphologic comparisons of hatchery-reared specimens of Scaphirhynchus albus, Scaphirhynchus platorynchus, and S. albus × S. platorynchus hybrids (Acipenseriformes: Acipenseridae)

Extensive habitat modifications within the Mississippi and Missouri rivers have presumably interfered with the reproductive isolating mechanisms between the endangered pallid sturgeon, Scaphirhynchus albus, and the sympatric shovelnose sturgeon, Scaphirhynchus platorynchus, causing hybridization between these two species. Several character indices were developed to assist fisheries biologists in identifying specimens of S. albus, S. platorynchus, and their putative hybrids. The indices have numerous assumptions, including that pure strains of both parental species are within the sample analyzed and that hybrids are morphologically intermediate relative to their parents. If these indices have produced inaccurate identifications, then previous work on Scaphirhynchus studies in the Mississippi and Missouri rivers are questionable, including status surveys, captive propagation efforts, or the harvesting of tissues for genetic studies. In this study we tested indices by examining progeny of ‘known’ pallid, shovelnose, and hybrid sturgeon propagated, raised, and preserved at hatcheries. These 60 specimens [78–600 mm standard length (SL)] were propagated with breeding stock from the upper Missouri River drainage, where hybridization between these two species presumably does not occur. Existing indices did not correctly identify small (<250 mm SL) or a combination of small and large (>250 mm SL) sizes of S. albus, S. platorynchus, and their hybrids. Indices worked fairly well in identifying large S. platorynchus, but not in differentiating large S. albus from hybrids. We used principal components analysis (PCA) as an alternative approach to character indices. No a priori knowledge of the identity of the specimen is required with this multivariate technique, which avoids potential circular reasoning present in indices. We were able to completely or almost completely separate both sturgeon species and their hybrids by extracting principal components from a correlation matrix of 13 meristic characters in a standard PCA and extracting size‐corrected principal components from a covariance matrix of 51 morphometric variables using a sheared PCA. Additionally, we demonstrated that first generation hybrids were intermediate with respect to their parental species. Multivariate analyses with a reduced character set of six meristic and 12 morphometric variables also led to accurate and reliable specimen identification. Head spines and numerous qualitative characters are also extremely useful in differentiating between Scaphirhynchus species and their hybrids. In addition to all morphometric characters, some meristic characters and the degree of head spine fusion vary significantly with SL of sturgeons. Recording appropriate data from released specimens, including photovouchers, and making this information available is essential for researchers to have any scientific or legal basis for genetic or any other studies involving these sturgeons.