Game of clones: Is Wolbachia inducing speciation in a weevil with a mixed reproductive mode?

Parthenogenesis is widely distributed in Metazoa but it is especially frequent in weevils (Coleoptera, Curculionidae) with one fifth of all known cases. Previous studies have shown that in the tribe Naupactini parthenogenetic reproduction most likely originated with an infection of the endoparasitic bacterium Wolbachia pipientis. In particular, Pantomorus postfasciatus possess a mixed reproductive mode: some populations have males while in others they are absent, and females produce clones by thelytoky. To better understand this scenario, we studied the population structure and infection status in 64 individuals of P. postfasciatus from Argentina and Brazil. We sequenced two mitochondrial (COI and COII) and one nuclear (ITS-1) fragments and obtained two very divergent haplogroups, one corresponding to the sexual populations uninfected with Wolbachia, and another conforming a monophyletic parthenogenetic (or presumptively parthenogenetic) and infected clade. Each of these haplogroups was identified as an independently evolutionary unit by all species delimitation analyses accomplished: multilocus *BEAST and BP&P, and single locus GMYC and K/θ rule. Additionally, present evidence suggests that Wolbachia infection occurred at least twice in all-female populations of P. postfasciatus with two different bacterial strains. Speciation mediated by Wolbachia is a recently described phenomenon and the case of P. postfasciatus is the first known case in a diplo-diploid insect. A model that describes how thelytoky-inducing phenotypes of Wolbachia could generate new lineages is discussed.

Evolutionary history of the little fire ant Wasmannia auropunctata before global invasion: inferring dispersal patterns, niche requirements and past and present distribution within its native range.

The evolutionary history of invasive species within their native range may involve key processes that allow them to colonize new habitats. Therefore, phylogeographic studies of invasive species within their native ranges are useful to understand invasion biology in an evolutionary context. Here we integrated classical and Bayesian phylogeographic methods using mitochondrial and nuclear DNA markers with a palaeodistribution modelling approach, to infer the phylogeographic history of the invasive ant Wasmannia auropunctata across its native distribution in South America. We discuss our results in the context of the recent establishment of this mostly tropical species in the Mediterranean region. Our Bayesian phylogeographic analysis suggests that the common ancestor of the two main clades of W. auropunctata occurred in central Brazil during the Pliocene. Clade A would have differentiated northward and clade B southward, followed by a secondary contact beginning about 380,000 years ago in central South America. There were differences in the most suitable habitats among clades when considering three distinct climatic periods, suggesting that genetic differentiation was accompanied by changes in niche requirements, clade A being a tropical lineage and clade B a subtropical and temperate lineage. Only clade B reached more southern latitudes, with a colder climate than that of northern South America. This is concordant with the adaptation of this originally tropical ant species to temperate climates prior to its successful establishment in the Mediterranean region. This study highlights the usefulness of exploring the evolutionary history of invasive species within their native ranges to better understand biological invasions.

Speciation in the asexual realm: is the parthenogenetic weevil Naupactus cervinus a complex of species in statu nascendi?

Population genetic theory shows that asexual organisms may evolve into species, which behave as independent evolutionary units. As a result, they form genotypic clusters separated by deep gaps due to geographic isolation and/or divergent selection. Identification of several genetically divergent groups of weevils embodied in the nominal species Naupactus cervinus deserves further study, in order to test if these lineages are evolving independently. In the present paper we tested if the parthenogenetic weevil N. cervinus, native to South America and broadly distributed throughout the world, contains more than one evolutionary unit. For this purpose, we applied three different approaches, a multilocus phylogenetic analysis, the GMYC approach and the K/θ method. We accomplished these analyses through a survey of mitochondrial (COI and COII genes) and nuclear (ITS1 sequence) genetic variation and morphometric analysis in a sample which included individuals from different locations within the native geographic range of N. cervinus. In addition, we compared the divergence accumulated in this species with that in another weevil of the same tribe (Naupactini) showing identical reproductive mode to see if similar levels of morphological variation matches similar levels of genetic divergence. We report the presence of two independent evolutionary units living in sympatry in forest areas. The incongruence between mitochondrial and nuclear datasets analyzed herein reflects incomplete lineage sorting of the nuclear marker and different evolutionary rates between genomes. Ecological divergence driven by natural selection (sympatry) or secondary contact after geographic isolation (allopatry) might explain the deep gaps in mitochondrial phylogenies. Instead, Wolbachia infection was ruled out as a causal factor for such differentiation. We conclude that N. cervinus is probably a species complex with at least two well differentiated lineages that would represent a cluster of species in statu nascendi.

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.

The genome organization and diversification of maize and its allied species revisited: evidences from classical and FISH-GISH cytogenetic analysis.

The present review summarizes our classical and molecular cytogenetic investigations in the genus Zea. The results obtained from the meiotic behavior analysis of Zea species and hybrids, confirm the amphiploid nature of all species in the genus, with a basic number of x = 5 chromosomes. All species with 2n = 20 are diploidized allotetraploids, whereas Z. perennis (2n = 40) is an allooctoploid with four genomes somewhat divergent from one another. These analyses also revealed the existence of postzygotic reproductive isolation among Zea species. Our studies using genomic in situ hybridization (GISH) provide evidence about the evolutionary relationships among maize and its allied species, and reveal remarkable genomic divergences. Particularly, knob sequences were not completely shared between taxa previously considered to be closely related. Our data strongly suggest that the teosinte Z. mays parviglumis is not the only progenitor of cultivated maize. Introgression of Tripsacum into cultivated maize cannot be discarded.

Cytogeography and the evolutionary significance of B chromosomes in relation to inverted rearrangements in a grasshopper species.

The analysis of geographic distribution of polymorphic cytological markers, briefly termed as cytogeography, can be considered an important tool to be applied when studying the evolutionary significance of chromosome variability within a species, either to unravel the adaptive significance of chromosome polymorphisms or to investigate the parasitic nature of some genomic elements. In this article we review cytogeographical studies in Trimerotropis pallidipennis, a grasshopper species whose South American populations display geographical patterns of distribution of inversion and B chromosome polymorphisms. Several lines of evidence that issue from the analysis of the geographic distribution of polymorphic markers suggest that inverted chromosomes are special sequences that are maintained by deterministic forces. On the other hand, the pattern of distribution of B chromosome polymorphism clearly demonstrates its selfish nature, being more frequent in those populations in central environments. We also present the analysis of 272 individuals of T. pallidipennis from Uspallata, and demonstrate that Bs in this population have some influence on body size, enlarging many of the morphometric characters of individuals and we propose it could be the consequence of its genotypic disequilibrium with one inversion. These investigations are finally discussed with regard to the models proposed for the maintenance of B chromosomes in natural populations and in relation to the possible interactions with chromosome inversions.

DMD and BMD in the same family due to distinct mutations.

We report on a family with a boy affected by Duchenne muscular dystrophy (DMD) and an asymptomatic cousin with a Becker-type dystrophin abnormality, diagnosed by chance. Dystrophin gene analysis showed that these conditions were caused by two distinct deletions with breakpoints in different exons. In Xp21 families, DNA analysis and dystrophin testing of asymptomatic males with high CK plasma levels might detect different dystrophin mutations in separate haplotypes as in our family, although we stress there should be clear clinical or familial indications for such testing.

In vivo modulation of myelin gene expression by human recombinant IL-2.

We treated adult mice with human recombinant interleukin-2 (IL-2) and determined the expression of the genes encoding for the major central and peripheral myelin proteins. In the CNS, myelin basic protein (MBP) and myelin-associated glycoprotein (MAG) mRNA levels were the same both in IL-2-treated and in control mice. Proteolipid protein (PLP) transcript was decreased in IL-2-treated animals when compared to controls. In the PNS, the messages for the glycoprotein P0 and for MBP were markedly increased in IL-2-treated animals when compared to controls.

Dystrophin characterization in BMD patients: correlation of abnormal protein with clinical phenotype.

We have investigated protein expression and genotype in 59 Becker muscular dystrophy (BMD) patients. The aim was to identify possible causes of the marked variability in phenotype in patients with similar deletions/mutations. The patients were examined neurologically and functionally and underwent Manual Muscle Testing. Dystrophin expression was analysed by immunohistochemistry and western blot using antibodies against six different segments of the protein. DNA mutations were investigated by PCR amplification of 30 exons. Based on dystrophin expression at the sarcolemma, two groups of patients were identified: group A (29 patients) with the classic patchy distribution of dystrophin and group B (30 patients) with absence or reduction of one or more dystrophin portions and variable, although mostly normal, expression of the other portions of the protein. Dystrophin molecular weight was normal or slightly reduced in group A and was variably reduced, generally conspicuously so, in group B. The quantity of dystrophin expressed varied markedly in both groups. The pattern of immunohistochemical staining in group B patients correlated with milder clinical phenotype, suggesting that small dystrophin molecules lacking a portion in the N-terminus or in the rod domain, are more functional than proteins with normal or slightly reduced molecular weight that display the BMD-typical patchy distribution at the sarcolemma.

Potential geographic distributions and successful invasions of parthenogenetic broad-nosed weevils (Coleoptera: Curculionidae) native to South America.

Ten species of parthenogenetic broad-nosed weevils (Coleoptera: Curculionidae: Entiminae) native to Argentina, southern Brazil, and Uruguay were selected for niche modeling analysis based on climatic data and altitude, to evaluate their potential range expansion inside and outside South America. The selected species belong to five genera of the tribe Naupactini affecting economically important crops. Until present, five of the 10 species analyzed here have invaded prairies and steppes of countries outside South America (Australia, New Zealand, Mexico, United States, and South Africa): Aramigus tessellatus (Say), Atrichonotus sordidus (Hustache), Atrichonotus taeniatulus (Berg), Naupactus leucoloma Boheman, and Naupactus peregrinus (Buchanan). Our niche modeling analyses performed with MAXENT demonstrated that these areas would be also suitable for Aramigus conirostris (Hustache), Eurymetopus fallax (Boheman), Pantomorus auripes Hustache, Pantomorus ruizi (Brèthes), and Pantomorus viridisquamosus (Boheman), consequently, they also have the potential to invade areas outside their native ranges, mainly in southeastern United States, some European countries (e.g., Portugal, France, and southern England), South Africa, New Zealand, and southeastern Australia. All the studied species share similar environmental requirements, the most important variables being the Mean Temperature of Driest Quarter, the Annual Mean Temperature and Isothermality. Long distance dispersal through commercial trade, and parthenogenetic reproduction would increase the threat of these weevils to crop production worldwide.

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.

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.

Population structure of the boll weevil in cotton fields and subtropical forests of South America: a bayesian approach.

The main goal of this contribution is to investigate the genetic structure of boll weevil populations from South America (Argentina and Brazil) and to make further comparisons with a putative source population from USA. Samples were collected in a Paranaense forest under reserve protection, cotton fields and non-cultivated areas. Data from anonymous molecular markers were analysed using both traditional methods of population genetics and Bayesian approaches. Results help to support a previous hypothesis on the presence of two lineages of boll weevil populations in South America: one with characteristics of recent invaders and the other with characteristics of ancient populations. The sample from Urugua-í Provincial Park (Misiones, Argentina) shows the highest percentage of polymorphic loci, the highest values of mean heterozigosity, and the largest number of population-specific alleles, all being typical features of ancient populations. Furthermore, the Urugua-í sample shows two gene pools occurring in sympatry, probably as a consequence of a secondary contact. The remaining samples reveal not only lower percentages of polymorphic loci and heterozygosity values, but also an almost negligible presence of specific alleles. Bayesian methods also suggest the occasional migration of some individuals of ancient lineages from their natural habitats in fragments of the Paranaense forest into cotton fields, and vice versa.

Genomic affinities between maize and Zea perennis using classical and molecular cytogenetic methods (GISH-FISH).

In this study we have analysed and compared the genomic composition, meiotic behaviour, and meiotic affinities of Zea perennis and Zea mays ssp. mays. To do so we studied the parental taxa and the interspecific hybrid Zea perennis x Zea mays ssp. mays, using classical cytogenetic methods, as well as GISH and FISH. GISH enabled us to recognize the genomic source of each chromosome involved in the meiotic configurations of this hybrid, and established the genomic affinities between their parental species. The results obtained here reinforce the hypothesis of the amphiploid origin of Zea perennis and, together with previous research, indicate that the chromosomes with divergent repetitive sequences in maize and Zea luxurians could be the remnants of a relict parental genome not shared with Zea perennis.

Inversion polymorphisms in Trimerotropis pallidipennis (Orthoptera): clinical variation along an altitudinal gradient.

Six populations of Trimerotropis pallidipennis located along an altitudinal gradient in the Antinaco-Los Colorados valley (La Rioja, Argentina), were cytologically analysed. The male karyotype consists of 23 chromosomes (22 + XO) with three long pairs submetacentric, the X-chromosome metacentric and the remaining ones basically acrocentric. Populations from La Rioja were polymorphic for seven pericentric inversions. Correlations between chromosome frequencies and altitude were statistically tested. In most cases, variables were either positively or negatively related in a significant fashion. As a consequence, mean populational heterozygosis (measured as the mean number of heteromorphic bivalents per individual per population) was also positively correlated with altitude. These results suggest that altitude (or other correlated environmental variable) may exert a differential selective pressure on coadapted gene blocks in the mutually inverted sequences. The possible relation of these results with the central-marginal model is discussed.

Dispersal of the cotton boll weevil (Coleoptera: Curculionidae) in South America: evidence of RAPD analysis.

RAPD technique provides useful information on the geographic origin and dispersal of the boll weevil Anthonomus grandis in South America. Nine populations from Argentina, Brazil, Paraguay, Mexico and USA were analyzed. Weevils were captured on native plants (Misiones province, Argentina) and on cotton cultures, except the sample from the United States (USDA laboratory-reared colony). A sample of the 'Peruvian square weevil', A. vestitus, from Ecuador, was included in the analysis in order to compare interspecific variation. The four primers used in the analysis revealed 41 'anonymous loci'. The neighbor-joining tree based on Nei's distances and values of Nm (migrants per generation), indicate that genetic similarity between samples from Tecomán (Mexico) and Puerto Iguazú (Argentina), is higher than among remaining South American populations. This result supports an hypothesis of natural occurrence of the boll weevil in South America, prior to extensive cotton cultivation. Population outbreaks of the species would be associated with increase of agricultural lands.

GISHGenomic in situ hybridization reveals cryptic genetic differences between maize and its putative wild progenitor Zea mays subsp. parviglumis.

The aim of this paper is to test with genomic in situ hybridization the genomic affinities between maize and its putative progenitor Zea mays subsp. parviglumis. Blocking procedures were applied for the purpose of improving discrimination among chromosome regions. Unlabeled genomic DNA from Z. mays subsp. parviglumis as a blocking agent and labeled genomic DNA from maize were hybridized on maize chromosomes. On the other hand, mitotic metaphases from Z. mays subsp. parviglumis were blocked with unlabeled genomic DNA of maize and hybridized with labeled genomic DNA from Z. mays subsp. parviglumis. Both experiments showed that either maize or Z. mays subsp. parviglumis chromosomes have their own unique sequences. This means an unexpected degree of divergence if Z. mays subsp. parviglumis is the only progenitor of maize, a result that is discussed in relation to our previous genomic in situ hybridization observations and to the different scenarios proposed about the origin of maize.

Molecular phylogeny of Larrea and its allies (Zygophyllaceae): reticulate evolution and the probable time of creosote bush arrival to North America.

Nucleotide sequences of Rubisco Large Subunit (rbcL) and the internal transcribed spacers (ITS) of nrDNA were obtained for the five species of Larrea and one species each of Bulnesia (ITS only) and Plectrocarpa (rbcL only). Parsimony analyses were conducted, including sequences from seven genera of Zygophyllaceae reported by other authors-Kallstroemia, Zygophyllum, Augea, Fagonia, Pintoa, Guaiacum, and Porlieria. The main conclusions of the present study are (1) the Argentine endemic Plectrocarpa tetracantha belongs to the subfamily Larreoideae (New World Clade); (2) all three phylogenies obtained from rbcL, ITS, and combined data sets show a close relationship between the tetraploid L. cuneifolia (sect. Bifolium) and the diploid multifoliolate pair L. nitida-L. ameghinoi (sect. Larrea), which could result from a possible intersectional hybrid origin of the tetraploid; (3) L. divaricata (sect. Bifolium) and L. tridentata (sect. Bifolium) form a highly supported monophyletic group, which agrees with previous cytogenetic and molecular evidence; and (4) the rate of nucleotide substitution of rbcL was estimated based on geological and fossil records. Under the molecular clock hypothesis, nucleotide sequence divergence between L. divaricata and L. tridentata suggests a Late Neogene (8.4 to 4.2 mybp) time of arrival of the diploid ancestors of L. tridentata to North American deserts.