Biochemical Polymorphisms and Genetic Relationships in Rodents of the Genera Oryzomys and Oligoryzomys (Sigmodontinae) from Brazil

An electrophoretic analysis of 12 allozyme systems (14 loci, 40 alleles) was performed in the rodent genera Oryzomys and Oligoryzomys, in order to determine the levels of genetic variability within and among populations and species. One hundred and fifty-five individuals from 16 populations of Oryzomys russatus, Or. angouya, Oligoryzomys flavescens, and O1. nigripes species were trapped in nine Brazilian localities. Genetic divergence among populations, as well as the interpopulational gene diversity, was higher in Oryzomys than in Oligoryzomys. The dendrogram of the phenetic relationships among the 16 populations of these four species displays three clusters: the first one joins the O1. flavescens populations with those of O1. nigripes; the second groups the populations of Or. russatus, and the last assembles the populations of Or. angouya. The genetic parameters analyzed reveal that the species belonging to the genus Oryzomys are genetically more structured than those of Oligoryzomys, the latter presenting lower levels of intrademe genetic differentiation (G(ST)')     

Nectomys squamipes microsatellites and homologous loci in sigmodontine rodents

Three monomorphic and four highly polymorphic microsatellites of Nectomys squamipes were isolated and characterised in a sample of 141 specimens from eight different Brazilian localities. These seven microsatellites and four others previously described in this species were tested in seven other nonfocus sigmodontine species. At least three loci were successfully amplified in every species, but none was amplified in all species. All sequenced products in nonfocus species showed (GT)(n) motifs as in N. squamipes. Several loci were amplified in Nectomys rattus and Oligoryzomys nigripes, while absence of PCR products was observed more frequently in Oxymycterus dasythricus and Akodon cursor. Two of three monomorphic loci in N. squamipes were polymorphic in other species.     

Phylogenetic relationships among species of the genus Oligoryzomys (Rodentia,Cricetidae) from Central and South America

González‐Ittig, R. E., Salazar‐Bravo, J., Barquez, R. M. & Gardenal, C. N. (2010). Phylogenetic relationships among species of the genus Oligoryzomys (Rodentia, Cricetidae) from Central and South America. —Zoologica Scripta, 39, 511–526. The genus Oligoryzomys includes several species very similar in external morphology, which has resulted in a confusing specific taxonomy. Accurate species identification is particularly important because several species of Oligoryzomys act as natural hosts of hantaviruses affecting humans. Here, we assign specific status to individuals from a wide geographical area of Argentina and Chile using sequences of the mtDNA control region. We also compare cytochrome b sequences of 14 species recognized from Central and South America to infer the phylogenetic relationships within the genus. In addition, the results were analysed using available data on chromosome numbers, and the host–parasite relationships reported for the genus Hantavirus. We confirm the geographical distribution of Oligoryzomys longicaudatus (Argentina, Chile), Oligoryzomys nigripes (Argentina, Paraguay, Brazil), Oligoryzomys chacoensis (Argentina, Bolivia, Paraguay), Oligoryzomys fornesi (Argentina, Paraguay), Oligoryzomys destructor (Argentina, Bolivia) and Oligoryzomys microtis (Bolivia, Brazil). Oligoryzomys longicaudatus is strongly related to the Oligoryzomys flavescens complex, which comprises four clades; O. nigripes is closely related to Oligoryzomys stramineus, and Oligoryzomys vegetus, to Oligoryzomys fulvescens from Central America. Oligoryzomys chacoensis, O. destructor, O. fornesi, O. longicaudatus, O. microtis, O. nigripes, O. stramineus, Oligoryzomys moojeni, Oligoryzomys rupestris, O. fulvescens and O. vegetus are confirmed as valid species, whereas O. flavescens, Oligoryzomys magellanicus, Oligoryzomys griseolus, Oligoryzomys victus, Oligoryzomys andinus and Oligoryzomys arenalis need exhaustive revision. The sister species to all the remaining entities of the genus was O. microtis, suggesting an Amazonian origin for the genus.     

A new karyotype of Oligoryzomys (Sigmodontinae, Rodentia) from central Brazil

A new karyotype of the genus Oligoryzomys was described for specimens collected in Brazilian Cerrado in Tocantins and Goiás States. Conventional staining, G-, C-banding, and Ag-NOR staining techniques were used for describe this karyotype with 2n=70, AN=74 or 76. The chromosome complement, with the highest diploid number known among Oligoryzomys species, differs from all others previously reported. This small sized Oligoryzomys species can be differentiated from other Brazilian Oligoryzomys not only by chromosomal complement, but also in some morphological attributes. The new species is apparently restricted in distribution and endemic of Brazilian Cerrado, occurring only in Rio Tocantins basin.     

Isolation and characterization of macaroni penguin (Eudyptes chrysolophus) microsatellite loci and their utility in other penguin species (Spheniscidae, AVES)

We report the characterization of 25 microsatellite loci isolated from the macaroni penguin (Eudyptes chrysolophus). Thirteen loci were arranged into four multiplex sets for future genetic studies of macaroni penguin populations. All 25 loci were tested separately in each of four other penguin species [Adélie penguin (Pygoscelis adeliae), chinstrap penguin (Pygoscelis antarctica), gentoo penguin (Pygoscelis papua) and king penguin (Aptenodytes patagonicus)]. Between eight and 12 loci were polymorphic per species. These loci are expected to be useful for studies of population genetic structure in a range of penguin species.     

The molecular phylogenetics of the genus Oligoryzomys (Rodentia: Cricetidae) clarifies rodent host–hantavirus associations

Several species of the genus Oligoryzomys are natural hosts of different hantavirus genotypes affecting humans. The systematics of the genus is confusing, which complicates the identification of the rodent host and hence the potential endemic areas of hantavirus pulmonary syndrome. In this study, we analyse molecular data to infer phylogenetic relationships among Central and South American specimens of Oligoryzomys, and compare our results with previously published data on karyotypic, geographic distribution and host–virus associations to solve contradictory taxonomic reports. We identified 25 clades, each one corresponding to a different putative species. The phylogenetic trees show that Oligoryzomys longicaudatus is strongly related to the Oligoryzomys flavescens complex, which comprises four clades; Oligoryzomys nigripes is related to Oligoryzomys stramineus, Oligoryzomys vegetus is related to Oligoryzomys fulvescens from Central America, and Oligoryzomys brendae is the sister species of Oligoryzomys aff. destructor. We identified the following rodent host–hantavirus genotype relationships: O. longicaudatus–Andes; O. flavescens ‘West'–Bermejo; O. flavescens ‘East'–Lechiguanas; O. nigripes–Juquitiba; Oligoryzomys microtis–Rio Mamore and Rio Mamore‐3; Oligoryzomys chacoensis–Oran; Oligoryzomys costaricencis–Choclo; Oligoryzomys delicatus–Maporal; Oligoryzomys utiaritensis–Castelo dos Sonhos; Oligoryzomys sp. RT2012–Rio Mamore‐4; Oligoryzomys sp. (and not Oligoryzomys fornesi)–Anajatuba. This work, besides contributing to the development of prevention programmes for hantavirus epidemiology in Latin America, represents a comprehensive update of the systematics of the genus Oligoryzomys. © 2014 The Linnean Society of London     

Autosomal and sex-linked microsatellite loci in the green oak leaf roller Tortrix viridana L. (Lepidoptera, Tortricidae)

Eight microsatellite markers were developed for the lepidopteran species Tortrix viridana using an enrichment protocol. The loci were highly variable with number of alleles ranging from four to 38. Six of the eight loci were in Hardy-Weinberg equilibrium. The other two were linked to the Z-chromosome. Values of observed heterozygosity ranged for the autosomal loci from 0.510 to 0.957. All loci will be useful to study dispersal and the autosomal loci, as well for phylogeographical studies.     

Molecular identification and phylogenetic relationships among the species of the genus Oligoryzomys (Rodentia, Cricetidae) present in Argentina, putative reservoirs of hantaviruses

The systematics and geographical distribution of the species of Oligoryzomys present in Argentina are poorly known. From some of the species different hantavirus genotypes have been recovered. In order to contribute to the accurate identification of those species and to infer their phylogenetic relationships, we analysed data of restriction sites and sequences of the mtDNA d -loop region. The samples used represent almost all Oligoryzomys species known to occur in Argentina. The trees obtained with the two types of data were similar, showing high bootstrap values for the majority of the nodes. Our results support the idea that the specific name Oligoryzomys longicaudatus should be applied only to individuals from the south of Argentina and Chile and confirm that O. nigripes and O. delticola are conspecific. Specimens identified as O. flavescens conform three related clades, probably belonging to a species complex. This study also emphasises that the use of a DNA fragment characterised by a high evolutionary rate compare with other mitochondrial segments as the d -loop, was appropriate to infer the phylogenetic relationships in a group originated by a rapid speciation process. We also suggest the following modifications in the rodent–hantavirus relationships: the only viral genotype associated to O. longicaudatus would be Andes Sout, while O. chacoensis would be the natural host of the genotype Oran. One of the forms of the O. flavescens complex would be the natural host of the genotype Bermejo. Our data are consistent with the hypothesis that each hantavirus genotype is associated with a specific rodent host.     

New polymorphic microsatellite loci for the Macedonian crested newt, Triturus macedonicus, and cross-priming testing in four other crested newt species

Eleven microsatellites have been characterized for Triturus macedonicus. Nine loci showed different variation patterns in a sample of 40 individuals from a single breeding pond in Zagori province (Greece), with an average number of 4.2 alleles per locus and an expected heterozygosity ranging from 0.1199 to 0.8079. Distinct cross-priming amplification rates were recovered on four additional crested newt species. Two monomorphic T. macedonicus loci were polymorphic in other Triturus species. The microsatellites developed herein could be a useful intraspecific genetic tool to undertake fine-scale population genetic analyses as well as in the study of contact zones between crested newt species.     

Microsatellite loci in the stable fly,Stomoxys niger niger (Diptera: Muscidae) on La Réunion Island

Six polymorphic microsatellite markers have been isolated from a microsatellite‐enriched DNA library from the stable fly, Stomoxys niger niger. These loci exhibited five to 10 alleles per locus and an expected heterozygosity ranging from 0.57 to 0.81 in the studied populations from La Réunion Island (Indian Ocean). They should therefore be useful for the study of genetic diversity and population structure of S. niger. In addition, cross‐species amplification of four of these six loci in the closely allied species Stomoxys calcitrans produced interpretable results, two of which would be useful for population biology studies.     

Characterization,multiplex conditions,and cross‐species utility of tetranucleotide microsatellite loci for Blanchard's cricket frog (Acris crepitans blanchardi)

We have isolated and characterized 17 tetranucleotide microsatellite loci for Blanchard's cricket frog (Acris crepitans blanchardi), an anuran common in the central USA. Sixteen loci were organized into four multiplex amplification reactions. These loci were highly polymorphic when screened in 55 individuals from two distant populations, with 11–48 alleles per locus (average = 24.8). Observed and expected heterozygosities ranged from 0.18 to 0.97 and from 0.17 to 0.96, respectively. Nine loci were also polymorphic in Acris crepitans crepitans, with seven polymorphic in Acris gryllus. Five loci amplified in all three taxa. These loci will be useful for population‐ and species‐level investigations of this widespread group.     

Thirteen polymorphic microsatellite loci in the Neotropical fish Prochilodus argenteus (Characiformes,Prochilodontidae)

Prochilodus species inhabit the main river systems of South America and usually present commercial value to inland fishing. In the present study, we describe the isolation and characterization of 13 novel microsatellite loci in Prochilodus argenteus. The number of alleles per polymorphic locus varied from four to 22 and the observed heterozygosity ranged from 0.333 to 0.893. Additionally, cross‐species amplification was successful in two other Prochilodus species. These loci will be useful for studies of the population genetic structure in this fish group.     

Development, characterisation, inheritance, and cross-species utility of American lobster (Homarus americanus) microsatellite and mtDNA PCR-RFLP markers.

Effective management of exploited species demands contemporary knowledge of population structure and mating patterns. Genetic markers can prove useful in providing this knowledge. Despite its commercial importance, genetic markers for American lobster (Homarus americanus) are limited. We developed 12 tetra- and 1 trinucleotide microsatellite loci for American lobster that exhibit little stuttering after PCR amplification. Gene diversity of these loci ranged from 0.516 to 0.929. A four-locus multiplex permits rapid genotyping of progeny in parentage experiments with a paternity exclusion probability over the four loci of 97.8%. We examined the loci for conformity to Hardy-Weinberg expectations (HWE) and linkage using individuals from one location and found that four loci deviated from HWE. We also tested inheritance and pairwise linkage using 48 embryos from each of two females. With the exception of two loci that were derived from the same clone and separated by 72 bp, no evidence of linkage was found. We, for the first time, demonstrate the occurrence of multiple paternity in American lobster. We also observed an apparent occurrence of dispermic androgenesis, possibly the first documentation of such an event within a species. Ten of the loci amplified in European lobster (Homarus gammarus), although two were monomorphic and one deviated significantly from HWE. We quantified mitochondrial DNA (mtDNA) sequence variation through the use of PCR amplification of two DNA fragments, followed by digestion with restriction enzymes; eight haplotypes were detected. One of the two fragments amplified in European lobster. Both sets of markers should prove useful for population discrimination purposes, and the microsatellites, in particular the four-locus multiplex, should prove highly amenable to rapidly addressing questions about mating patterns.     

Characterization of microsatellite DNA markers for a mahseer species,Tor tambroides (Cyprinidae) and cross‐amplification in four congeners

This study reports the isolation and characterization of microsatellite DNA markers in a mahseer species, Tor tambroides (Pisces, Cyprinidae). Of a total of 14 loci evaluated, 10 were polymorphic in T. tambroides samples, with an average of 2.86 alleles per locus. Deviations from Hardy–Weinberg equilibrium were observed at one locus and there was no indication of linkage disequilibrium among loci. A high level of cross‐amplification among four congeners was achieved, with 12 loci successfully amplifying and 11 loci showing polymorphism in at least one other species. These markers will be a useful resource for population genetic studies and broodstock management of closely related mahseer species.     

Isolation and characterization of 13 polymorphic microsatellites for the black murex, Hexaplex nigritus

Thirteen polymorphic microsatellite markers have been isolated and characterized for the black murex (Hexaplex nigritus). These loci are moderately to highly variable with seven to 37 alleles within 113 individuals from four populations in the Northern Gulf of California. Expected heterozygosities ranged from 0.43 to 0.98. High variability indicates that these markers will be useful for studying population structure and connectivity in this species.     

Characterization of 14 polymorphic microsatellite loci for the facultatively eusocial sweat bee Halictus rubicundus (Hymenoptera, Halictidae) and their variability in related species

Sweat bees display considerable variation in social organization and a few species, such as Halictus rubicundus, are even facultatively eusocial. Fourteen polymorphic, unlinked microsatellite loci were isolated from H. rubicundus and characterized in 45 females. The number of alleles per locus ranged from two to 18 (mean 10.1), observed heterozygosity ranged from 0.24 to 0.98 (mean 0.71) and expected heterozygosity ranged from 0.24 to 0.98 (mean 0.70). Six or more loci cross-amplified in four other sweat bees. These loci will be useful for the study of social evolution and population genetic structure in H. rubicundus and many other sweat bees.     

High prevalence of multiple paternity within fruits in natural populations of Silene latifolia, as revealed by microsatellite DNA analysis

Data on multiple paternity within broods has been gathered in several animal species, and comparable data in plants would be of great importance to understand the evolution of reproductive traits in a common framework. In this study, we first isolated and characterized six microsatellite loci from the dioecious plant Silene latifolia (Caryophyllaceae). The polymorphism of the loci was assessed in 60 individual females from four different populations. Two of the investigated loci showed a pattern of inheritance consistent with X-linkage. These microsatellite loci were highly polymorphic and therefore useful tools for parentage analysis. We then used four of the markers to determine paternity within naturally pollinated fruits in four European populations. This study revealed widespread multiple paternity in all populations investigated. The minimum number of fathers per fruit varied from one to nine, with population means ranging from 3.4 to 4.9. The number of fathers per fruit was not significantly correlated with offspring sex ratios. High prevalence of multiple paternity within fruits strongly suggest that pollen competition is likely to occur in this species. This may substantially impact male reproductive success and possibly contribute to increase female and offspring fitness, either through postpollination selection or increased genetic diversity. Wide variation in outcrossing rates may be an overlooked aspect of plant mating systems.     

Ten polymorphic microsatellite markers developed in the masson pine moth Dendrolimus punctatus Walker (Lepidoptera: Lasiocampidae)

A set of 10 polymorphic di‐ and trinucleotide microsatellite loci were developed for the forestry pest insect, masson pine moth, Dendrolimus punctatus Walker. The expected heterozygosity at these loci ranges from 0.285 to 0.859, and the observed allele numbers from five to 19. Cross species amplification of these loci in four other congeneric pine moth species indicates variable levels of loci conservation and thus cross‐applicability. Therefore, the microsatellite loci reported here should be useful for population genetic and other related studies in the masson pine moth and other closely related species.     

Isolation, characterization and multiplex polymerase chain reaction of novel microsatellite loci for the avian parasite Philornis downsi (Diptera: Muscidae)

An enrichment technique was used to isolate 11 di-, tri-, and tetra microsatellites for the parasitic fly Philornis downsi (Diptera: Muscidae). These loci were polymerase chain reaction amplified in singleplexes or two-plexes for P. downsi. The loci showed low to moderate polymorphism, exhibited between three and four alleles, and observed heterozygosity ranged from 0.05 to 0.86. These new markers will be useful for population-level and paternity analyses and will provide valuable information about the ecology of this high-impact parasite of vulnerable bird species.     

Twenty-one novel microsatellite DNA loci isolated from the Puget Sound white-crowned sparrow, Zonotrichia leucophrys pugetensis

The white‐crowned sparrow, Zonotrichia leucophrys, has served as a model species for studies of song and reproductive physiology. Here, we describe primers for 21 novel microsatellite loci isolated from the Puget Sound subspecies, Zonotrichia leucophrys pugetensis, which will be useful for parentage and population genetic analyses. Based on genotypes from seven to 22 adult birds from one population, the average number of alleles per locus was 10.9 (four to 21 alleles) and observed heterozygosity varied from 0.50 to 1.00. All loci also amplified products in at least one of three other passerine species tested.