Fine Scale Microevolutionary and Demographic Processes Shaping a Wild Metapopulation Dynamics of the South American Fruit Fly Anastrepha fraterculus.

Anastrepha fraterculus (Wiedmann) is an important American pest species. Knowledge of its population dynamics is of particular interest for ecology, evolutionary biology, and management programs. In the present study, phenotypic, genotypic, and spatial data were combined, within the frame of landscape genetics, to uncover the spatial population genetic structure (SGS) and demographic processes of an Argentinian local population from the Yungas ecoregion. Eight simple sequence repeats (SSR) loci and six morphometric traits were analysed considering the hierarchical levels: tree/fruit/individual. Genetic variability estimates were high (HE = 0.72, RA = 4.39). Multivariate analyses of phenotypic data showed that in average 52.81% of variance is explained by the tree level, followed by between individuals 28.37%. Spatial analysis of morphological traits revealed a negative autocorrelation in all cases. SGS analysis and isolation by distance based on SSR showed no significant autocorrelation for molecular coancestry. The comparison between phenotypic (PST) and molecular (FST) differentiation identified positive selection in different fruits for all traits. Bayesian analysis revealed a cryptic structure within the population, with three clusters spatially separated. The results of this study showed a metapopulation dynamics. The genetic background of the components of this metapopulation is expected to change through time due to seasonality, repopulation activities, and high gene flow, with an estimated dispersal ability of at least 10 km. Effective population size (Ne) of the metapopulation was estimated in around 800 flies, and within subpopulations (clusters) Ne was associated with the levels of genetic drift experienced by the founding lineages.

Signatures of natural selection in morphological quantitative traits in Argentinean populations of Senegalia gilliesii (Fabaceae).

In order to elucidate the role of evolutionary forces in shaping the variation of quantitative traits in Senegalia gilliesii we evaluate seven phenotypic traits in three Argentinean populations, two of them sharing environmental and vegetation type conditions, and a third one ecologically differentiated from the former. The phenotypic traits were compared with molecular markers. Here, we search for signatures of selection by means of the comparison PST-FST . We assessed if the averages of the seven phenotypic traits were different among populations by means of ANOVA and we performed discriminant analysis of principal components (DAPC) for both morphological and molecular data. The ANOVA showed significant results only for two traits. For all foliar traits and two spine traits, the PST-FST comparison suggested the occurrence of stabilizing selection. The DAPC obtained from AFLP data showed three well defined groups of populations; when the same analysis was conducted with morphological data the scatterplot showed high overlapping among individuals and could not separate the populations. Overall, our findings suggest a prominent role of stabilizing selection in all foliar traits and stipular spine length. These results could be extrapolated to other tropical and subtropical acacias. Further studies are needed to analyse the mechanisms underlying genetic differentiation in natural populations of S. gilliesii, find its relationship with eco-geographical variables.

EcoGenetics: An R package for the management and exploratory analysis of spatial data in landscape genetics.

The integration of ecology and genetics has become established in recent decades, in hand with the development of new technologies, whose implementation is allowing an improvement of the tools used for data analysis. In a landscape genetics context, integrative management of population information from different sources can make spatial studies involving phenotypic, genotypic and environmental data simpler, more accessible and faster. Tools for exploratory analysis of autocorrelation can help to uncover the spatial genetic structure of populations and generate appropriate hypotheses in searching for possible causes and consequences of their spatial processes. This study presents EcoGenetics, an R package with tools for multisource management and exploratory analysis in landscape genetics.

Isolation by distance and stochastic migration in a fragmented population of Prosopis alba.

PREMISE OF THE STUDY: Gene flow through dispersal of seeds and pollen is a fundamental determinant of spatial genetic structure (SGS) in natural populations of trees at different spatial scales. Within continuous populations, restrictions to gene flow should be manifested in a process of local genetic differentiation, known as isolation by distance. The present work examines the SGS of a Prosopis alba population in a patchy region where urban, forest, and agricultural areas coexist. The analysis discussed here expands our knowledge about the processes affecting the distribution of the genetic variability in populations of disturbed landscapes. METHODS: Three sites with different landscape and demographic characteristics were analyzed. Seven highly variable microsatellite markers were used to survey the relevance of both isolation by distance and stochastic migration in the SGS of the population. KEY RESULTS: The analyses showed that (1) the genetic similarity declined with increasing geographic distance, (2) the population may be conceived as a single genetically continuous unit showing spatial differentiation as consequence of isolation by distance, rather than a structured population following the island model, and (3) there is evidence supporting a past immigration event into one of the study sites, which promoted a local pattern of genetic structure. CONCLUSIONS: These results indicate that in spite of the population fragmentation produced by land-use changes, P. alba maintains the genetic cohesion and a continuous genetic structure in the analyzed area.

Genetics and biology of Anastrepha fraterculus: research supporting the use of the sterile insect technique (SIT) to control this pest in Argentina.

Two species of true fruit flies (taxonomic family Tephritidae) are considered pests of fruit and vegetable production in Argentina: the cosmopolitan Mediterranean fruit fly (Ceratitis capitata Wiedemann) and the new world South American fruit fly (Anastrepha fraterculus Wiedemann). The distribution of these two species in Argentina overlaps north of the capital, Buenos Aires. Regarding the control of these two pests, the varied geographical fruit producing regions in Argentina are in different fly control situations. One part is under a programme using the sterile insect technique (SIT) for the eradication of C. capitata, because A. fraterculus is not present in this area. The application of the SIT to control C. capitata north of the present line with the possibility of A. fraterculus occupying the niche left vacant by C. capitata becomes a cause of much concern. Only initial steps have been taken to investigate the genetics and biology of A. fraterculus. Consequently, only fragmentary information has been recorded in the literature regarding the use of SIT to control this species. For these reasons, the research to develop a SIT protocol to control A. fraterculus is greatly needed. In recent years, research groups have been building a network in Argentina in order to address particular aspects of the development of the SIT for Anastrepha fraterculus. The problems being addressed by these groups include improvement of artificial diets, facilitation of insect mass rearing, radiation doses and conditions for insect sterilisation, basic knowledge supporting the development of males-only strains, reduction of male maturation time to facilitate releases, identification and isolation of chemical communication signals, and a good deal of population genetic studies. This paper is the product of a concerted effort to gather all this knowledge scattered in numerous and often hard-to-access reports and papers and summarize their basic conclusions in a single publication.

Analysis of genetic population structure in Acacia caven (Leguminosae, Mimosoideae), comparing one exploratory and two Bayesian-model-based methods.

Bayesian clustering as implemented in STRUCTURE or GENELAND software is widely used to form genetic groups of populations or individuals. On the other hand, in order to satisfy the need for less computer-intensive approaches, multivariate analyses are specifically devoted to extracting information from large datasets. In this paper, we report the use of a dataset of AFLP markers belonging to 15 sampling sites of Acacia caven for studying the genetic structure and comparing the consistency of three methods: STRUCTURE, GENELAND and DAPC. Of these methods, DAPC was the fastest one and showed accuracy in inferring the K number of populations (K = 12 using the find.clusters option and K = 15 with a priori information of populations). GENELAND in turn, provides information on the area of membership probabilities for individuals or populations in the space, when coordinates are specified (K = 12). STRUCTURE also inferred the number of K populations and the membership probabilities of individuals based on ancestry, presenting the result K = 11 without prior information of populations and K = 15 using the LOCPRIOR option. Finally, in this work all three methods showed high consistency in estimating the population structure, inferring similar numbers of populations and the membership probabilities of individuals to each group, with a high correlation between each other.

Effect of host plant chemistry on genetic differentiation and reduction of gene flow among Anastrepha fraterculus (Diptera: Tephritidae) populations exploiting sympatric, synchronic hosts.

Herbivore host specialization includes changes in behavior, driven by locally induced adaptations to specific plants. These adaptations often result in sexual isolation that can be gauged through detection of reduced gene flow between host associated populations. Hypothetically, reduced gene flow can be mediated both by differential response to specific plant kairomones and by the influence of larval diet on some adult traits such as pheromone composition. These hypotheses could serve as a model to explain rapid radiation of phytophagous tephritid fruit flies, a group that includes several complexes of cryptic species. The South American Fruit Fly Anastrepha fraterculus (Wiedemann) is a complex of at least seven cryptic species among which pheromone mediated sexual isolation resulted in rapid differentiation. Cryptic species also exhibit differences in host affiliation. In search of a model explaining rapid radiation in this group, we studied host plant chemical composition and genetic structure of three host associated sympatric populations of A. fraterculus. Chemical composition among host plant fruit varied widely both for nutrient and potentially toxic secondary metabolite content. Adaptation to plant chemistry appears to have produced population differentiation. We found host mediated differentiation to be stronger between populations exploiting sympatric synchronic hosts differing in chemical composition, than between populations that exploit hosts that fruit in succession. Gene flow among such host associated populations was extremely low. We propose as a working hypothesis for future research, that for those differences to persist over time, isolating mechanisms such as male produced sex pheromones and female preferences resulting from adaptation to different larval diets should evolve.

New microsatellite loci for Prosopis alba and P. chilensis (Fabaceae).

PREMISE OF THE STUDY: As only six useful microsatellite loci that exhibit broad cross-amplification are so far available for Prosopis species, it is necessary to develop a larger number of codominant markers for population genetic studies. Simple sequence repeat (SSR) markers obtained for Prosopis species from a 454 pyrosequencing run were optimized and characterized for studies in P. alba and P. chilensis. • METHODS AND RESULTS: Twelve markers that were successfully amplified showed polymorphism in P. alba and P. chilensis. The number of alleles per locus ranged between two and seven and heterozygosity estimates ranged from 0.2 to 0.8. Most of these loci cross-amplify in P. ruscifolia, P. flexuosa, P. kuntzei, P. glandulosa, and P. pallida. • CONCLUSIONS: These loci will enable genetic diversity studies of P. alba and P. chilensis and contribute to fine-scale population structure, indirect estimation of relatedness among individuals, and marker-assisted selection.

Host association of Spodoptera frugiperda (Lepidoptera: Noctuidae) corn and rice strains in Argentina, Brazil, and Paraguay.

Spodoptera frugiperda (J.E. Smith) is composed of two genetically distinct strains, the so-called corn strain and the rice strain. Whether the two strains differ in their host use is unclear, because laboratory experiments have not been able to show consistent host performance or preference differences between them, and field studies showed high rates of hybridization, as well as some degree asymmetric host use. To determine the distribution of the two strains and their association with host plants, we collected fall armyworm larvae from different crops (corn, rice, alfalfa, and sorghum) and grasses in 15 different localities over 4 yr in Argentina, Brazil, and Paraguay. The strain identity was analyzed using two polymorphisms in the mitochondrial cytochrome oxidase subunit I gene. We identified the corn and rice haplotypes and three types of populations were characterized based on the frequencies of the individuals that belonged to any of these haplotypes: in 44% of populations the corn haplotype predominated, in 44% of populations the rice haplotype was the most frequent, and 11% of populations showed both haplotypes at similar proportions. In total, eight populations (47%) showed the expected pattern, two populations (12%) were polymorphic within the same field, and seven populations (41%) showed the inverse pattern. Taken together, there was no consistent pattern of host association between the two sympatric genotypes and their respective host plants. This investigation supports the need for additional studies to determine which other forces keep the genotypes separate, and what is the degree of genetic differentiation between these populations.

Consistency between marker- and genealogy-based heritability estimates in an experimental stand of Prosopis alba (Leguminosae).

Prosopis represents a valuable forest resource in arid and semiarid regions. Management of promising species requires information about genetic parameters, mainly the heritability (h(2)) of quantitative profitable traits. This parameter is traditionally estimated from progeny tests or half-sib analysis conducted in experimental stands. Such an approach estimates h(2) from the ratio of between-family/total phenotypic variance. These analyses are difficult to apply to natural populations of species with a long life cycle, overlapping generations, and a mixed mating system, without genealogical information. A promising alternative is the use of molecular marker information to infer relatedness between individuals and to estimate h(2) from the regression of phenotypic similarity on inferred relatedness. In the current study we compared h(2) of 13 quantitative traits estimated by these two methods in an experimental stand of P. alba, where genealogical information was available. We inferred pairwise relatedness by Ritland's method using six microsatellite loci. Relatedness and heritability estimates from molecular information were highly correlated to the values obtained from genealogical data. Although Ritland's method yields lower h(2) estimates and tends to overestimate genetic correlations between traits, this approach is useful to predict the expected relative gain of different quantitative traits under selection without genealogical information.

Phylogeographic studies on natural populations of the South American fruit fly, Anastrepha fraterculus (Diptera: Tephritidae).

Anastrepha fraterculus is an important pest of commercial fruits in South America. The variability observed for morphological and behavioural traits as well as genetic markers suggests that A. fraterculus represents a complex of synmorphic species rather than a single biological species. We studied the correlation between geographical distribution and genetic variation in natural populations from Argentina and south Brazil. Fragments of the mitochondrial gene COII were sequenced in 28 individuals. The matrix of aligned sequences was phylogenetically analysed by parsimony, yielding a cladogram of haplotypes. Based on Templeton's nested method, no clade showed any geographic pattern for the gene COII, indicating lack of significant association between haplotypic variability and geographic distribution. The analysis of nucleotide substitution distances by Neighbour-Joining algorithm showed that geographically distant populations exhibit low genetic distances. The corresponding trees clustered the populations without showing any geographic pattern. This result suggests that the populations studied are not reproductively isolated.

Consistency of population genetics parameters estimated from isozyme and RAPDs dataset in species of genus Prosopis (Leguminosae, Mimosoideae).

Genetic variability, population structure and differentiation among 17 populations of 5 species and 2 natural interspecific hybrids of section Algarobia of genus Prosopis were analyzed from data of 23 isozyme and 28 RAPD loci. Both markers indicated that the studied populations are highly variable. P. alba populations in average showed lower values of genetic variability estimates from isozyme data, but this trend was not observed for RAPD markers. The hierarchical analyses of the distribution of genetic variability showed that the highest proportion of variation occurred within populations, the differentiation among species was intermediate and the lowest component was observed among populations within species. The consistency between results from both dataset implies that they are not biased and reflect the actual genetic structure of the populations analyzed. The matrices of Euclidean distances obtained from the two sets of markers were highly correlated according to Mantel test. In both cases the corresponding phenogram and MDS plot tended to cluster conspecific populations while hybrid populations were not intermediate between putative parents. Some disagreements between isozyme and RAPD phenograms were observed mainly in the affinities of hybrid populations. Such inconsistencies might result from reticular rather than dichotomic evolutionary relationships. The phenetic associations retrieved gave no support to the division of the section Algarobia into series.

Phylogenetic relationships in Dichroplus Stål (Orthoptera: Acrididae: Melanoplinae) inferred from molecular and morphological data: testing karyotype diversification.

The neotropical genus Dichroplus and related genera are characterized by a relatively uniform external morphology and a remarkably divergent male genitalia and hence its taxonomy is controversial. It also shows an extreme karyotypic diversification. In this study we used molecular and morphological characters to test the monophyly of the genus and to evaluate chromosome evolution. Twenty-seven species from Dichroplus and related genera were included in the analysis. Morphological characters refer to the general morphology, male genitalia and female structures. Molecular studies were performed, sequencing part of two mitochondrial genes, cytochrome oxidase I and II. Independent and combined phylogenetic analyses of the data were performed under maximum parsimony. The karyotypic characters (rearrangements) were either mapped onto the combined topology or combined with the other data sets. While the molecular analysis confirms some results attained with morphology, some others do not. All point towards the paraphyly of the genus. Our results show the relevance of morphological data in phylogenetic studies because morphology and molecules supply complementary evidence. The mapping of chromosome characters on the combined tree shows that the most extreme karyotype, in D. silveiraguidoi, is a derived condition, probably reached through several centric fusions, and that X-autosome centric fusions were recurrently fixed during the evolution of the group.

First report on RAPDs patterns able to differentiate some Argentinean species of section Algarobia (Prosopis, Leguminosae).

Prosopis species constitute a very important resource in arid and semiarid regions. Some species of section Algarobia hybridise and introgress naturally in areas of sympatry. According to previous isoenzymatic studies these species have high variability within populations. However, the genetic differentiation among species was very low, and these markers failed to provide diagnostic loci for species recognition. Here we analysed by the RAPD technique natural populations of Prosopis alba, P. ruscifolia, P. nigra, P. flexuosa, and P. vinalillo with the purpose of obtaining markers for species and hybrid identification, by analysing the distribution of genetic diversity within and among species. Genetic variability (H = 0.12-0.26) was similar in all populations. Genetic differentiation among populations (FST = 0.39) was highly significant. Hierarchical analysis of genetic structure performed by Wright (1978) method and analysis of molecular variance (AMOVA) indicated that the diversity among populations within species is low (4-13%) and most genetic diversity (54-61%) occurs within populations. The differentiation among species is intermediate (26-42%) between the previous components but highly significant. Five bands provided a tool for identifying any of the species studied, with the exception of P. vinalillo. The difficulty in diagnosing this species is discussed in reference to the hypothesis of its hybrid origin.

First comparative phenetic studies of Argentinean species of Acacia (Fabaceae), using morphometric, isozymal, and RAPD approaches.

Morphological and genetic diversity among Acacia aroma, A. macracantha, A. caven, and A. furcatispina were studied with morphometric, isozymal, and RAPD approaches. The analysis of seven isozyme systems revealed 21 loci, and RAPD analysis showed 34 loci. Most of these loci allowed us to differentiate the species, with the exception of A. aroma and A. macracantha, the two most similar species. The levels of genetic variability estimated by isozymes were higher than those obtained from RAPD analyses. Morphometric characters showed highly significant differences among the species, although A. aroma and A. macracantha are differentiated only by thorn length. The phenogram obtained from isozyme data is consistent with morphological data. The RAPD phenogram based on allelic frequencies showed agreement with morphological and isozymal approaches only at the intraspecific levels, while the RAPD phenogram based on Nei and Li's similarity measures agreed with the phenograms constructed from isozyme and morphological data. High similarities and high indirect gene flow were found between A. aroma and A. macracantha, results that call the relationship between them into question.