Viviana A. Confalonieri

Are flightless Galapaganus weevils older than the Galápagos Islands they inhabit

The 15 species in the weevil genus Galapaganus Lanteri 1992 (Entiminae: Curculionidae: Coleoptera) are distributed on coastal Perú and Ecuador and include 10 flightless species endemic to the Galápagos islands. These beetles thus provide a promising system through which to investigate the patterns and processes of evolution on Darwin’s archipelago. Sequences of the mtDNA locus encoding cytochrome oxidase subunit I (COI) were obtained from samples of seven species occurring in different ecological zones of the oldest south-eastern islands: San Cristóbal, Española and Floreana, and the central island Santa Cruz. The single most parsimonious tree obtained shows two well-supported clades that correspond to the species groups previously defined by morphological characters. Based on a mtDNA clock calibrated for arthropods, the initial speciation separating the oldest species, G. galapagoensis (Linell) on the oldest island, San Cristóbal, from the remaining species in the Galápagos occurred about 7.2 Ma. This estimate exceeds geological ages of the extant emerged islands, although it agrees well with molecular dating of endemic Galápagos iguanas, geckos and lizards. An apparent explanation for the disagreement between geological and molecular time-frames is that about 7 Ma there were emerged islands which subsequently disappeared under ocean waters. This hypothesis has gained support from the recent findings of 11-Myr-old submarine seamounts (sunken islands), south-east of the present location of the archipelago. Some species within the darwini group may have differentiated on the extant islands, 1–5 Ma.

Microsatellite typing of ancient maize: insights into the history of agriculture in southern South America.

Archaeological maize specimens from Andean sites of southern South America, dating from 400 to 1400 years before present, were tested for the presence of ancient DNA and three microsatellite loci were typed in the specimens that gave positive results. Genotypes were also obtained for 146 individuals corresponding to modern landraces currently cultivated in the same areas and for 21 plants from Argentinian lowland races. Sequence analysis of cloned ancient DNA products revealed a high incidence of substitutions appearing in only one clone, with transitions prevalent. In the archaeological specimens, there was no evidence of polymorphism at any one of the three microsatellite loci: each exhibited a single allelic variant, identical to the most frequent allele found in contemporary populations belonging to races Amarillo Chico, Amarillo Grande, Blanco and Altiplano. Affiliation between ancient specimens and a set of races from the Andean complex was further supported by assignment tests. The striking genetic uniformity displayed by the ancient specimens and their close relationship with the Andean complex suggest that the latter gene pool has predominated in the western regions of southern South America for at least the past 1400 years. The results support hypotheses suggesting that maize cultivation initially spread into South America via a highland route, rather than through the lowlands.

Molecular evidence for a novel encapsulated genotype of Trichinella from Patagonia, Argentina

At present, Trichinella spiralis is the only species of this genus reported from South America. Herein, we detail a molecular analysis of a new encapsulated isolate of muscle larvae of Trichinella, found in a mountain lion (Puma concolor) coming from the Patagonia, Argentina. We studied three DNA regions previously probed to be useful for the identification of all eleven recognized Trichinella genotypes: expansion segment 5 (ES5), cytochrome c-oxidase subunit I (COI) and 5S ribosomal DNA intergenic spacer region (5S ISR). BLAST searches with these DNA sequences showed that the mitochondrial and nuclear ribosomal regions most closely resemble other Trichinella sequences available in GenBank. However, they did not exactly match any of the eleven recognized genotypes. The phylogenetic analysis from COI and 5S ISR sequences showed that the mountain lion isolate is grouped with encapsulated members, in concordance with morphological data. Furthermore, this new isolate was located at the base of the encapsulated genotypes, signifying that it is an old genotype that could have emerged earliest in this group. These data strongly suggest that this isolate from the Patagonia represents the twelfth genotype (T12) described in the genus Trichinella. Nevertheless, further studies are necessary to adequately establish this isolate as a unique genotype.

Trichinella patagoniensis n. sp. (Nematoda), a new encapsulated species infecting carnivorous mammals in South America ☆

Until a few years ago, Trichinella spiralis was the only taxon of the genus Trichinella detected in both domestic and wild animals of South America. Recently, a new genotype, named Trichinella T12, was identified in cougars (Puma concolor) from Argentina, on the basis of molecular studies using mitochondrial and nuclear ribosomal markers. In the present study, cross-breeding experiments indicated that Trichinella T12 is reproductively isolated from all other encapsulated Trichinella spp. and suggested that it is biologically more similar to Trichinella britovi and Trichinella murrelli than to the other encapsulated species/genotypes. Biological assays revealed that the reproductive capacity index of Trichinella T12 was ~4 and >2000 times lower than those of T. spiralis in mice and rats, respectively. The reproductive capacity index of Trichinella T12 in domestic pigs ranged from 0.0 to 0.05. Larvae parasitising the muscles of carnivores were infective to mice after freezing at -5°C for 3 months, but they lost infectivity after freezing at -18°C for 1 week. The region within the rDNA, known as the expansion segment V, showed a unique sequence which differs from those of all other known Trichinella spp./genotypes. The biological, geographical and molecular data support the classification of the genotype Trichinella T12 as a new species widespread in the Neotropical region, for which we propose the name Trichinella patagoniensis n. sp.

Complex mutational patterns and size homoplasy at maize microsatellite loci

Microsatellite markers have become one of the most popular tools for germplasm characterization, population genetics and evolutionary studies. To investigate the mutational mechanisms of maize microsatellites, nucleotide sequence information was obtained for ten loci. In addition, Single-Strand Conformation Polymorphism (SSCP) analysis was conducted to assess the occurrence of size homoplasy. Sequence analysis of 54 alleles revealed a complex pattern of mutation at 8/10 loci, with only 2 loci showing allele variation strictly consistent with stepwise mutations. The overall allelic diversity resulted from changes in the number of repeat units, base substitutions, and indels within repetitive and non-repetitive segments. Thirty-one electromorphs sampled from six maize landraces were considered for SSCP analysis. The number of conformers per electromorph ranged from 1 to 7, with 74.2% of the electromorphs showing more than one conformer. Size homoplasy was apparent within landraces and populations. Variation in the amount of size homoplasy was observed within and between loci, although no differences were detected among populations. The results of the present study provide useful information on the interpretation of genetic data derived from microsatellite markers. Further efforts are still needed to determine the impact of these findings on the estimation of population parameters and on the inference of phylogenetic relationships in maize investigations.

Partial host fidelity in nest selection by the shiny cowbird (Molothrus bonariensis), a highly generalist avian brood parasite

Obligate avian brood parasites can be host specialists or host generalists. In turn, individual females within generalist brood parasites may themselves be host specialists or generalists. The shiny cowbird Molothrus bonariensis is an extreme generalist, but little is known about individual female host fidelity. We examined variation in mitochondrial control region sequences from cowbird chicks found in nests of four common Argentinean hosts. Haplotype frequency distributions differed among cowbird chicks from nests of these hosts, primarily because eggs laid in nests of house wrens Troglodytes aedon differed genetically from those laid in nests of the other three hosts (chalk‐browed mockingbird Mimus saturninus, brown‐and‐yellow marshbird Pseudoleistes virescens, and rufous‐collared sparrow Zonotrichia capensis). These differences in a maternally inherited marker indicate the presence of a nonrandom laying behaviour in the females of this otherwise generalist brood parasite, which may be guided by choice for nest type, as house wrens nest in cavities whereas the other three species are open cup nesters.

Microsatellite variation in maize landraces from Northwestern Argentina: genetic diversity, population structure and racial affiliations.

The highland region or Northwestern Argentina (NWA) is one of the southernmost areas of native maize cultivation and constitutes an expansion of the peruvian Andes sphere of influence. To examine the genetic diversity and racial affiliations of the landraces cultivated in this area, 18 microsatellite markers were used to characterize 147 individuals from 6 maize races representative of traditional materials. For the whole data set, a total of 184 alleles were found, with an average of 10.2 alleles per locus. The average gene diversity was 0.571. The observed patterns of genetic differentiation suggest that historical association is probably the main factor in shaping population structure for the landraces studied here. In agreement with morphological and cytogenetic data, Bayesian analysis of NWA landraces revealed the occurrence of three main gene pools. Assessment of racial affiliations using a combined dataset including previous data on American landraces showed a clear relationship between one of these gene pools and typical Andean races, whereas the remaining two gene pools exhibited a closer association to Caribbean accessions and native germplasm from the United States, respectively. These results highlight the importance of integrating regional genetic studies if a deeper understanding of maize diversification and dispersal is to be achieved.

Diversity of Boll Weevil Populations in South America: A Phylogeographic Approach

A phylogeographic approach was conducted to assess the geographic structure and genetic variation in populations of the boll weevil Anthonomus grandis, which is the most harmful insect pest of cotton in the Americas. COI and COII mitochondrial gene sequences were analyzed to test a former hypothesis on the origin of the boll weevil in Argentina, Brazil and Paraguay, using samples from Mexico and USA as putative source populations. The analysis of variability suggests that populations from South American cotton fields and nearby disturbed areas form a phylogroup with a central haplotype herein called A, which is the most common and widespread in USA and South America. The population from Texas has the A haplotype as the most frequent and gathers in the same group as the South American populations associated with cotton. The sample from Tecomán (México) shows high values of within-nucleotide divergence, shares no haplotype in common with the South American samples, and forms a phylogroup separated by several mutational steps. The sample from Iguazú National Park (Misiones Province, Argentina) has similar characteristics, with highly divergent haplotypes forming a phylogroup closer to the samples from cotton fields, than to the Mexican group. We propose that in South America there are: populations with characteristics of recent invaders, which would be remnants of “bottlenecks” that occurred after single or multiple colonization events, probably from the United States, and ancient populations associated with native forests, partially isolated by events of historical fragmentation.

Wolbachia infection in the tribe Naupactini (Coleoptera, Curculionidae): association between thelytokous parthenogenesis and infection status.

Several parthenogenetic species of broad‐nosed weevils exist, some of them of economic importance because of their pest status. Screening of the maternally inherited Wolbachia bacterium in 29 weevils of the tribe Naupactini, using multilocus sequence typing allowed us to assess a significant correlation between asexuality and infection, and suggests an involvement of Wolbachia in the origin of this reproductive mode. The nine Wolbachia strains retrieved from the Naupactini belong to the B supergroup. Phylogenetic analysis of these strains, along with other 23 strains obtained from arthropods and nematodes, supports previous hypotheses that horizontal transfer of Wolbachia amongst species from unrelated taxa has been pervasive.

Eggshell spotting in brood parasitic shiny cowbirds (Molothrus bonariensis) is not linked to the female sex chromosome

For avian brood parasites in which individual females are host-specialists, the arms race between hosts and parasites has favored egg color polymorphism in the parasite, with female lineages laying mimetic eggs that resemble those of the host species they parasitize. Female sex-linked inheritance of egg color fosters evolutionary stability of egg polymorphism if female lineages show both consistent eggshell color and host use. This co-evolutionary relationship is unlikely to occur if individual brood parasites use different hosts or if egg color is not maternally inherited. The shiny cowbird (Molothrus bonariensis) is an extreme generalist brood parasite that shows a very high degree of egg polymorphism. We tested whether egg spotting in this species has female sex-linked inheritance. If genetic factors controlling the expression of egg spotting were present on the female-specific W chromosome, we expected co-segregation between spotting patterns and mtDNA haplotypes, as both W and mtDNA are maternally inherited. In contrast to the known maternal inheritance of spotting patterns in great tits, we found no associations between eggshell spotting and mtDNA haplotypes, which suggests that eggshell spotting is not maternally inherited in this cowbird species.