Synchronous allelic expression at the glucosephosphate isomerase A and B loci in interspecific sunfish hybrids

Allelic isozymes of glucosephosphate isomerase at the Gpi-A and -B loci were separated by starch gel electrophoresis in the warmouth (Lepomis gulosus) and green sunfish (L. cyanellus). The specific tissue distributions and developmental expressions of the GPI-A₂, -AB, and -B₂ isozymes were not different between these two species. The synchrony of allelic expression in normal intraspecific sunfish crosses was demonstrated by means of an electrophoretic variant at the Gpi-B locus. In embryos formed from warmouth × green sunfish hybrid crosses, the paternal GPI-A₂ isozymes were first expressed at the same time in both reciprocal hybrids, at 21–25 hr after fertilization. The maternal and paternal GPI-B subunits were synchronously expressed in reciprocal hybrids just prior to hatching. The parental allelic isozymes at both loci showed codominant expression in all tissues of the mature F₁ hybrids. These results are consistent with the absence of allelic asynchrony and inhibition in interspecific hybrids formed from more evolutionarily related species.     

The use of floating overhead cover by warmwater stream fishes

Increasing the amount of woody debris in streams has often increased population size of one or more focal species whereas clearing a stream of woody debris has often reduced populations. Alterations of the amount of woody debris change multiple aspects of the habitat simultaneously, so it is very difficult to know what particular stimulus or combination of stimuli evoked the changes in the fish populations. The purpose of this research was to alter habitat by the addition of overhead cover alone, and to see whether or not that single change would affect the distribution of stream-dwelling fishes. The addition of solid, floating cover objects 1.8 m by 2.4 m resulted in increased local populations of bluntnose minnow (Pimphales notatus), creek chub (Semotilus atromaculatus), and longear sunfish (Lepomis megalotis) five to six weeks later, but did not change the distribution of four other species: white sucker (Catostomus commersoni), green sunfish (Lepomis cyanellus), bluegill (Lepomis macrochirus), and largemouth bass (Micropterus salmoides). These results showed that for some species at least, changes in channel morphology, visual isolation, and amelioration of flow produced by adding three-dimensional complex woody debris were not essential for making locally attractive habitat changes in a warmwater stream. The effectiveness of overhead cover as used here would be expected to vary for different species and depend on such things as predation threat, flow, and food levels.     

Confirmation of intraspecific crossing and single and joint segregation of biochemical loci ofVolvariella volvacea

Nineteen lines ofV. volvacea were obtained from worldwide sources and analyzed for allozyme activity. Twelve loci (Aat, Acp-2, Dia, Est-4, Gipi, Lap-1, Pep-GL, Pep-LLL-1, Pep-LLL-2, Pgm, Pgd, andSod) were monomorphic; 5 loci (Ada, Gpt, Mpi, Np, andPep-PAP) were polymorphic. Nine unique genotypes were distinguished among the 19 lines. Multilocus enzyme electrophoresis was used to confirm intraspecific crosses between putative homokaryons. Putative homokaryons were recognized by electrophoresis of single-spore-derived cultures and used as breeding stock. These breeding stocks were grown together in dual culture on agar, and selections were made from the interaction zone. These selections were then transferred to make spawn. Intraspecific crosses were confirmed by the presence of heteromeric allozymes. To determine single and joint segregation of biochemical loci, 84 single-spore-derived offspring from a confirmed hybrid sporocarp with five heterozygous loci were analyzed. Mendelian segregation was verified for the five loci. One linkage (Np withGpt,r=0.0) was found among 10 pairwise comparisons for joint segregation. The basidiospores ofV. volvacea are haploid and homokaryotic isolates may mate to yield heterokaryons.     

Purification of the creatine kinase isozymes of the green sunfish (Lepomis cyanellus) with Blue Sepharose CL-6B.

Three homodimeric creatine kinase isozymes (A₂, B₂, and C₂) of the green sunfish (Lepomis cyanellus) were purified by a combination of affinity chromatography, gel filtration, and preparative starch gel electrophoresis. The final preparations were isozymically pure and were used to elicit antibodies in rabbits. The use of the group-specific adsorbant Blue Sepharose CL-6B (Pharmacia) and specific elution conditions for creatine kinase facilitated purification. Fish creatine kinase isozymes are sensitive to denaturation and cannot be readily purified by procedures routinely used for mammalian creatine kinase isozymes.     

Ecological and morphological differentiation of pumpkinseed sunfish in lakes without bluegill sunfish

Summary Over the last three decades, sunfish of the familyCentrarchidae have become recognized as a model system in which the ecological consequences of species interactions can be observed and tested. The evolutionary consequences of species interactions in sunfish have received less attention. Bluegill (Lepomis machrochirus) and pumpkinseed (Lepomis gibbosus) sunfish are two common and well-studied species that occupy separate ecological niches. Adult bluegill are generalists that feed in the open water on zooplankton during much of the year, while adult pumpkinseeds specialize on crushing hard-bodied prey such as snails. These species coexist over much of their geographical ranges, but bluegill are historically absent from several large drainage basins in the northeastern US. Here we show that pumpkinseeds from an Adirondack lake without bluegills have differentiated into two morphological forms, one of which is planktivorous. Differentiation is independent of sex and occurs over a broad range of sizes. Thus, the ecological diversity that exists between the bluegill and pumpkinseeds in sympatry has been replaced by a comparable degree of diversity within pumpkinseeds in allopatry.     

What Causes Partial F1 Hybrid Viability? Incomplete Penetrance versus Genetic Variation

Background

Interspecific hybrid crosses often produce offspring with reduced but non-zero survivorship. In this paper we ask why such partial inviability occurs. This partial inviability could arise from incomplete penetrance of lethal Dobzhansky-Muller incompatibilities (DMIs) shared by all members of a hybrid cross. Alternatively, siblings may differ with respect to the presence or number of DMIs, leading to genotype-dependent variation in viability and hence non-Mendelian segregation of parental alleles in surviving F1 hybrids.

Methodology/Principal Findings

We used amplified fragment length polymorphisms (AFLPs) to test for segregation distortion in one hybrid cross between green and longear sunfish (Lepomis cyanellus and L. megalotis). Hybrids showed partial viability, and twice as much segregation distortion (36.8%) of AFLPs as an intraspecific control cross (18.8%). Incomplete penetrance of DMIs, which should cause genotype-independent mortality, is insufficient to explain the observed segregation distortion.

Conclusions/Significance

We conclude that F1 hybrid sunfish are polymorphic for DMIs, either due to sex-linked DMI loci (causing Haldane''s Rule), or polymorphic autosomal DMI loci. Because few AFLP markers were sex-linked (2%), the most parsimonious conclusion is that parents may have been heterozygous for loci causing hybrid inviability.     

Inheritance and linkage relationships of morphological and isozyme loci in chickpea (Cicer arietinum L.)

Inheritance and linkage relationships of several morphological and isozyme loci are described in chickpea (Cicer arietinum L.). Segregation data obtained from several F₂ families confirmed the previously observed mode of inheritance for most of the morphological loci. Additional morphological markers in chickpea are also described. Most of the isozyme loci studied showed codominant expression and fit expected Mendelian segregation ratios. However, distorted ratios were also observed for some loci. Linkage was found betweenPgd-c, the locus encoding the cytosolic form of 6-phosphogluconate dehydrogenase, andHg, the locus controlling plant growth habit. These 2 loci were separated by approximately 18 recombinational map units. A similar linkage between comparable loci was previously reported in pea (Pisum sativum L.) (Weeden and Wolko 1990). Linkage was also detected among 3 isozyme loci; the cytosolic form of phosphoglucomutase (Pgm-c), glucose-1-phosphate transferase (Gpt1), and the plastid specific form of 6-phosphogluconate dehydrogenase (Pgd-p). The linkage of 2 loci (Pgm-c andPgd-p) in this cluster is also conserved in pea and lentil (Lens Miller). The linkage between an acid phosphatase locus (Acp3) and the locus specifying the cytosolic form of glucosephosphate isomerase (Gpi-c) in chickpea suggested another linkage group in common with pea. Additionally, other linkages that were not previously observed in chickpea or related genera included the linkage of the cytosolic form of aconitase (Aco-c) with adenylate kinase (Adk1) and fructokinase (Fk3), and the linkage of a locus encoding the mitochondrial specific aconitase (Aco-m) with a seed protein locus (Spr1). The loci determining flower color (P), epicotyl color (Gst), seed coat color (T ³), and seed surface (Rs) were associated with the locus encoding glucose-1-phosphate transferase (Gpt2). These results, along with previous studies, suggest that pea, lentil and chickpea have several common linkage groups consisting of homologous genes. This also indicates that linkages found in one genus can be used to predict similar linkages in related genera in the development of linkage maps.     

Linkage relationships of six enzyme Loci in interspecific sunfish hybrids (genus lepomis)

Backcross hybrids produced from the bluegill, the red-ear sunfish, and their F₁ interspecific hybrid have been analyzed for the inheritance of six enzyme phenotypes. Malate dehydrogenase A and B, tetrazolium oxidase, 6-phosphogluconate dehydrogenase, skeletal muscle esterase, and liver α-glycerophosphate dehydrogenase are all inherited in a mendelian manner as codominant alleles at nuclear loci. 6-phosphogluconate dehydrogenase and α-glycerophosphate dehydrogenase are encoded by linked loci, undergoing recombination at a frequency of 15%-22%. No other case of linkage was observed. The absence of linkage between the homologous malate dehydrogenase loci is of particular interest. These interspecific hybrids appear to be very useful for studies of biochemical genetics.     

Comparative genome analysis of mungbean (Vigna radiata L. Wilczek) and cowpea (V. unguiculata L. Walpers) using RFLP mapping data

Genome relationships between mungbean (Vigna tradiata) and cowpea (V. Unguiculata) based on the linkage arrangement of random genomic restriction fragment length polymorphism (RFLP) markers have been investigated. A common set of probes derived from cowpea, common bean (Phaseolus vulgaris), mungbean, and soybean (Glycine max) PstI genomic libraries were used to construct the genetic linkage maps. In both species, a single F₂ population from a cross between an improved cultivar and a putative wild progenitor species was used to follow the segregation of the RFLP markers. Approximately 90% of the probes hybridized to both mungbean and cowpea DNA, indicating a high degree of similarity in the nucleotide sequences among these species. A higher level of polymorphism was detected in the mungbean population (75.7%) than in the cowpea population (41.2%). Loci exhibiting duplications, null phenotypes, and distorted segregation ratios were detected in both populations. Random genomic DNA RFLP loci account for about 89% of the currently mapped markers with a few cDNA and RAPD markers added. The current mungbean map is comprised of 171 loci/loci clusters distributed in 14 linkage groups spanning a total of 1570cM. On the other hand, 97 markers covered 684 cM and defined 10 linkage groups in the current cowpea map. The mungbean and cowpea genomes were compared on the basis of the copy number and linkage arrangement of 53 markers mapped in common between the two species. Results indicate that nucleotide sequences are conserved, but variation in copy number were detected and several rearrangements in linkage orders appeared to have occurred since the divergence of the two species. Entire linkage groups were not conserved, but several large linkage blocks were maintained in both genomes.     

Predator Generalization Decreases the Effect of Introduced Predators in the San Marcos Salamander,Eurycea nana

The introduction of novel predators into an environment can have detrimental consequences on prey species, especially if these species lack the ability to recognize these predators. One such species that may be negatively affected by introduced predators is the federally threatened San Marcos salamander (Eurycea nana). Previous research found that predator‐naïve (captive‐hatched) salamanders showed decreased activity in response to the chemical cues of both a native fish predator (Micropterus salmoides) and an introduced fish predator (Lepomis auritus), but not to a non‐predatory fish (Gambusia geiseri). We tested the hypothesis that E. nana recognized the introduced Lepomis (and other non‐native Lepomis) because they share chemical cues with other native congeneric Lepomis predators in the San Marcos River. We examined the antipredator response of predator‐naïve E. nana to chemical cues from (1) a sympatric native sunfish (Lepomis cyanellus; Perciformes: Centrarchidae); (2) a sympatric introduced sunfish (L. auritus); (3) an allopatric sunfish (Lepomis gibbosus); (4) a sympatric non‐native, non‐centrarchid cichlid (Herichthys cyanoguttatum; Perciformes: Cichlidae); and (5) a blank water control to determine whether individuals make generalizations about novel predators within a genus and across a family. Exposure to chemical cues from all fish predator treatments caused a reduction in salamander activity (antipredator response). Additionally, there were no differences in the antipredator responses to each predatory fish treatment. The similar responses to all sunfish treatments indicate that E. nana shows predator generalization in response to novel predators that are similar to recognized predators. Additionally, the antipredator response to H. cyanoguttatum indicates that predator generalization can occur among perciform families.     

Development and characterization of 24 polymorphic microsatellite markers for bluegill sunfish, <Emphasis Type="Italic">Lepomis macrochirus</Emphasis>

The bluegill sunfish (Lepomis macrochirus) is a notorious exotic fish, which causes serious damages to freshwater ecosystems in the world. Therefore, it is a matter of urgency to control this species for the conservation of ecosystem. New microsatellite loci were developed for L. macrochirus using high throughput 454 GS-FLX+ pyrosequencing. We selected 24 primer pairs that were tested on 47 individuals from Lake Biwa, which encompasses the oldest and largest population of L. macrochirus in Japan. Allele per locus varied 2–17. Expected and observed heterozygosities ranged from 0.185 to 0.888 and 0.120 to 0.872, respectively. Applied to L. macrochirus in Lake Biwa, these markers could detect genetic differentiation among populations, which showed a pattern of isolation by distance. These markers are expected to be useful in the estimation of population structure and ecology of L. macrochirus.     

2(S),3(S)-3-hydroxy-4-methyleneglutamic Acid from Gleditsia caspica

A new natural product, 2(S),3(S)-3-hydroxy-4-methyleneglutamic acid (G3) has been isolated from seeds of Gleditsia caspica. The structure has been established by chemical and spectroscopic methods. Catalytic reduction of G3 yields 2(S),4(S)-4-methylglutamic acid and a new amino acid, 2(S),3(S),4(S)-3-hydroxy-4-methylglutamic acid. Ozonolysis of G3 followed by oxidation gives 2(S),3(R)-3-hydroxyaspartic acid. The S- (or l-) configurations at C₂ in G3 and in 2(S),3(S),4(S)-3-hydroxy-4-methyglutamic acid and the S-configurations at C₃ for G3 and 2(S),3(S),4(S)-3-hydroxy-4-methylglutamic acid and at C₄ for 2(S),3(S),4(S)-3-hydroxy-4-methylglutamic acid are inferred from the configurations at C₂ in 2(_S),4(_S)-4-methylglutamic acid and at C₂ and C₃ in 2(S),3(R)-3-hydroxyaspartic acid. The seeds also contain appreciable quantities of 2(S),3(S),4(R)-3-hydroxy-4-methylglutami c acid (G1) and 2(S),4(R)-4-methylglutamic acid.     

Inheritance and linkage relationships of isozyme loci in cucumber (Cucumis sativus L.)

Summary Genetic analyses were conducted among 18 provisionary isozyme loci in Cucumis sativus L. Fourteen loci demonstrated simple Mendelian inheritance while observed variation at four loci (Gpi2, Gr2, Pgm3, Skdh2) was determined not to have a predictable genetic basis. Joint segregation analyses among the 14 genetically predictable polymorphic loci resulted in the assignment of 12 loci to four linkage groups. Linkage groups contain the following loci: (1) Gr1, Pgm1, Idh, Pgd1; (2) Pep-pap, Mdh2, Mdh3, Gpi1; (3) Pep-la, Per4; (4) Pgd2, G2dh. Mpi2 and Mdh1 segregated independently. Recombination fractions for linked loci ranged between 0.051 (Pgm1-Idh) to 0.385 (Pep-la-Per4). Some practical applications of isozyme marker loci for cucumer improvement are discussed.     

Effects of a non-native cichlid fish (African jewelfish, Hemichromis letourneuxi Sauvage 1880) on a simulated Everglades aquatic community

In an 8-month mesocosm experiment, we examined how a simulated Everglades aquatic community of small native fishes, snails, and shrimp changed with the addition of either a native predator (dollar sunfish Lepomis marginatus) or a non-native predator (African jewelfish Hemichromis letourneuxi) compared to a no-predator control. Two snail species (Planorbella duryi, Physella cubensis) and the shrimp (Palaemonetes paludosus) displayed the strongest predator-treatment effects, with significantly lower biomasses in tanks with Hemichromis. One small native fish (Heterandria formosa) was significantly less abundant in Hemichromis tanks, but there were no significant treatment effects for Gambusia holbrooki, Jordanella floridae, or Pomacea paludosa (applesnail). Overall, there were few treatment differences between native predator and no-predator control tanks. The results suggest that the potential of Hemichromis to affect basal food-web species that link primary producers with higher-level consumers in the aquatic food web, with unknown consequences for Florida waters.     

Differential sensitivity of CYP1A to 3,3′,4′,4-tetrachlorobiphenyl and benzo(a)pyrene in two Lepomis species

Although Lepomis species are abundant in a wide variety of habitats throughout North America and could serve as potentially valuable biomonitoring tools, few studies have examined the induction of pollutant biomarkers in this genus. We hypothesized that the induction of cytochrome P-450 1A (CYP1A), a sensitive and widely used indicator of response to aquatic contaminants, would serve as an effective biomarker of organic pollutant exposure in Lepomis species. We examined the response of CYP1A and two of the major pollutant-responsive phase II enzymes, glutathione S-transferase (GST), and uridine diphosphate glucuronyltransferase (UDPGT), in Lepomis exposed to organic pollutants under laboratory and field conditions. Two Lepomis species (longear sunfish, Lepomis megalottis and bluegill, Lepomis macrochirus) were exposed in the laboratory via intraperitoneal injection to corn oil (vehicle), benzo(a)pyrene (BaP) (10 and 50 mg/kg), a polynuclear aromatic hydrocarbon (PAH) or 3,4,3′,4′-tetrachlorobiphenyl (PCB 77) (0.1 and 1.0 mg/kg), a dioxin-like planar halogenated aromatic hydrocarbon (HAH), and sacrificed 2 (BaP) or 7 (corn oil, PCB77) days later. Lepomis hepatic CYP1A exhibited differential sensitivity to these two classes of environmental contaminants. CYP1A activity was weakly induced in bluegill exposed to 1.0 mg/kg PCB 77 (3 fold induction over controls) but strongly induced in both bluegill and longear sunfish exposed to 50 mg/kg BaP (37 and 15 fold induction over controls, respectively). In contrast, hepatic GST activity in both species remained unchanged following the treatment with either compound and hepatic UDPGT activity, which was assessed only in BaP-treated longear sunfish, was unaffected by that chemical, indicating these phase II enzymes may not be sensitive pollutant biomarkers in this genus. Further, longear sunfish collected from a PCB contaminated site displayed relatively low levels of CYP1A activity despite PCB body burdens associated with strong induction of CYP1A activity in other fish species. The strong induction of CYP1A by BaP with much weaker CYP1A response to PCB indicates that CYP1A in Lepomis sp. could be an excellent biomarker for PAH pollution, but may not be a reliable indicator of site contamination by halogenated hydrocarbons. We conclude that Lepomis species provide a useful model for examining the regulation and potential consequences of differential pollutant sensitivity, but that CYP1A in these species should be used with caution as an indicator of halogenated contaminants.     

Analysis of genetic mapping in a waxy/dent maize RIL population using SSR and SNP markers

A maize genetic linkage map was generated using SSR and SNP markers in a F_7:8 recombinant inbred line (RIL) population derived from a cross of waxy corn (KW7) and dent corn (Mo17). A total of 465 markers, including 459 SSR and 6 SNP markers, were assigned to 10 linkage groups which spanned 2,656.5 cM with an average genetic distance between markers of 5.7 cM, and the number of loci per linkage group ranged from 39 to 55. The SSR (85.4%) and SNP (83.3%) markers showed Mendelian segregation ratios in the RIL population at a 5% significance threshold. In linkage analysis of six SNP loci associated with kernel starch synthesis genes (ae1, bt2, sh1, sh2, su1, and wx1), all six loci were successfully mapped and are closely linked with SSR markers in chromosomes 3 (sh2), 4 (su1 and bt2), 5 (ae1), and 9 (sh1 and wx1). The SSR markers linked with genes in starch synthesis may be utilized in marker assisted breeding programs. The resulting genetic map will be useful in dissection of quantitative traits and the identification of superior QTLs from the waxy hybrid corn. Additionally, these data support further genetic analysis and development of maize breeding programs.     

Invasive quagga mussels can be attenuated by redear sunfish (Lepomis microlophus) in the Southwestern United States

The degree to which invasive quagga mussels have spread across the Southwestern United States is unprecedented, and until now, none of the controls currently available is risk-free to the environment or human health. Here we demonstrate that stocking high-density (0.42 fish/m³ or 1.90 fish/m²) redear sunfish (Lepomis microlophus) in an infested lake enclosure removed adult quagga mussels efficiently and appeared to suppress their growth and recruitment. However, mussels were not eradicated during the short term experimental course. Long-term study needs to be conducted to determine whether mussels can be completely eliminated by redear sunfish and what optimum density would be required for balancing high efficacy mussel control with cost-effectiveness. Redear sunfish is a species native to the Southeastern United States that has become a popular, stocked sportfish in the Southwest. Targeted stocking of redear sunfish may be an efficient and environmentally friendly approach to mitigate the colonization of invasive quagga mussels in suitable open water ecosystems, especially those with new dreissenid infestations or those threatened by invasion.     

Linkage Analysis of the Multilocus Glucosephosphate Isomerase Isozyme System in Sunfish (Centrarchidae, Teleostii)

The purpose of the present investigation is to determine whether the two duplicated glucosephosphate isomerase (EC 5.3.1.9) loci Gpi-A and Gpi-B reside on the same chromosome in teleostean fishes. Interspecific sunfish hybrids were employed for the cross because of the different species-specific electrophoretic mobilities of the allelic isozymes at each GPI locus and because of their genomic compatibility. F1 sunfish hybrids, formed from a male warmouth (Lepomis gulosus) X female green sunfish (L. cyanellus) cross, were mated to form the F2 generation. The number of each of the nine different isozyme phenotypes, revealed by starch gel electrophoresis, was determined using 256 F2 individuals. The high frequency of recombinant phenotypes in the F2 generation indicated that the two GPI loci are not linked. An excess of F2 individuals heterozygous at both loci was observed and is interpreted as being caused by heterosis. The absence of linkage for the homologous loci encoding GPI subunits and for other multilocus isozyme systems is consistent with the postulate that the genomes of present-day vertebrates arose through one or more polyploidization events early in vertebrate evolution.     

Isolation of Polymorphic RAPD-SSR Markers from Xinjiang Arctic Grayling (<Emphasis Type="Italic">Thymallus arcticus grubei</Emphasis>) and a Test of Cross-Species Amplification

A total of 18 polymorphic microsatellite loci were isolated and characterized from RAPD products in the Xinjiang Arctic Grayling (Thymallus arcticus grubei). The number of alleles (N_a) per locus varied from 2 to 10. Observed (H_o) and expected (H_e) heterozygosities ranged from 0.64 to 0.92, and from 0.63 to 0.88, respectively. Considerable differences were found among HBH, FH and FY populations in the number of alleles, effective number of alleles, number of genotypes at all of these loci. These new RAPD-SSR markers have provided a helpful tool for genetic analyses and resources conservation of T. arcticus grubei. Five additional fish species, Amur grayling (Thymallus grubii), Taimen (Hucho taimen), Sea perch (Lateolabrax japonicus), Lenok (Brachymystax lenok) and Red seam bream (Pagrosomus major) were assessed for cross-species amplification. Three of the five species showed at least one polymorphic locus. In addition, seven loci were found to be polymorphic in at least one species.