Morphometric Differentiation Among Anastrepha fraterculus (Diptera: Tephritidae) Exploiting Sympatric Alternate Hosts.

Anastrepha fraterculus (Wiedemann) is currently considered a complex of cryptic species infesting fruits from Mexico to Argentina and represents an interesting biological model for evolutionary studies. Moreover, detecting and quantifying behavioral, morphological, and genetic differentiation among populations is also relevant to the application of environment-friendly control programs. Here, phenotypic differentiation among individuals coexisting in the wild in a Northern region of Argentina was unveiled and associated with host choice. Six morphometric traits were measured in sympatric flies exploiting three different host species. Phenotypic variation was shown to be host-dependent regardless of geographical or temporal overlap. Flies collected from synchronous alternate hosts (peach and walnut) differed from each other despite the lack of geographical isolation. By contrast, flies emerging from guavas that ripen about two months later than peach and walnut showed no significant differentiation in comparison to flies collected from walnuts, but they differ significantly from flies originating from peaches. This result is consistent with the hypothesis that the same population of flies shifts from walnuts to guavas throughout the year, whereas the population of flies that uses peaches as a host is probably exploiting other alternate hosts when peach availability decreases. Further research is needed to study the underlying mechanism. Results are consistent with previous molecular markers (inter-simple sequence repeat-ISSR) research on flies stemming from the same hosts and the same area, suggesting that differentiation among flies emerging from alternative hosts occurs at both genetic and phenotypic levels. The contribution of host preference in long-term genetic differentiation is discussed.

Demonstration Using Field Collections that Argentina Fall Armyworm Populations Exhibit Strain-specific Host Plant Preferences.

Spodoptera frugiperda, the fall armyworm, is a major economic pest throughout the Western Hemisphere of corn (maize), cotton, sorghum, and a variety of agricultural grasses and vegetable crops. Studies in the United States, the Caribbean, and Brazil demonstrated the existence of two subpopulations (previously designated "host strains") that differ in their choice of plant host. Specifically, the corn strain is preferentially found in corn and sorghum, while the rice strain is dominant in rice, turf grass, and alfalfa. However, inconsistent results were reported in surveys of fall armyworm in Argentina, with some indicating that the host plant preferences of the two strains might be compromised or even nonexistent. If correct, this would complicate efforts to control this pest by considerably expanding the range of habitats that would have to be considered as potential sources for fall armyworm infestations in specific crops. A reexamination of Argentine fall armyworm, this time with field collections rather than the laboratory colonies used in previous studies, confirmed the existence of the two strains and their host preferences. Specifically, the corn strain was consistently the majority population infesting corn and was usually so in sorghum, while the rice strain was predominant in pasture/turf grasses and alfalfa. The one outlier was a collection from rice, which had a corn strain majority. Overall, the data were generally consistent with strain behaviors observed in other areas of the Western Hemisphere.

Evidences of local adaptation in quantitative traits in Prosopis alba (Leguminosae).

Signals of selection on quantitative traits can be detected by the comparison between the genetic differentiation of molecular (neutral) markers and quantitative traits, by multivariate extensions of the same model and by the observation of the additive covariance among relatives. We studied, by three different tests, signals of occurrence of selection in Prosopis alba populations over 15 quantitative traits: three economically important life history traits: height, basal diameter and biomass, 11 leaf morphology traits that may be related with heat-tolerance and physiological responses and spine length that is very important from silvicultural purposes. We analyzed 172 G1-generation trees growing in a common garden belonging to 32 open pollinated families from eight sampling sites in Argentina. The multivariate phenotypes differ significantly among origins, and the highest differentiation corresponded to foliar traits. Molecular genetic markers (SSR) exhibited significant differentiation and allowed us to provide convincing evidence that natural selection is responsible for the patterns of morphological differentiation. The heterogeneous selection over phenotypic traits observed suggested different optima in each population and has important implications for gene resource management. The results suggest that the adaptive significance of traits should be considered together with population provenance in breeding program as a crucial point prior to any selecting program, especially in Prosopis where the first steps are under development.

Male courtship behavior of the South American fruit fly, Anastrepha fraterculus, from an Argentinean laboratory strain.

The South American fruit fly Anastrepha fraterculus (Wiedemann) (Diptera: Tephritidae) is a pest of fruit species of warm regions of the Americas, including Argentina. Some authors claim that this taxon includes a group of cryptic species. In order to evaluate possible targets of sexual selection, it is necessary to analyze ethological aspects of male courtship and identify particular steps that strongly influence mating success. A mating test designed to evaluate behavioral differences between insects that achieve copulation (successful males) and those that did not mate (unsuccessful males) could also be relevant for the possible implementation of control programs based on sterile insect technique. Reared insects need to be evaluated periodically, since genetic drift and artificial selection associated with rearing conditions could have a detrimental effect on their ability to compete for matings in nature. In this study, courtship behavior of A. fraterculus males from a laboratory strain was analyzed for the first time through video recordings. Three components for the activities were identified: calling, wing positions, and movements. Also, the time that males spent on each step of the courtship was registered, including the last activities before attempting copulation. Data showed that mating achievement occurs relatively quickly; 65% of the successful males reached copulation within the first ten minutes after the male and female were placed together. Behavioral differences were detected between successful and unsuccessful males. The former group tended to invest more time in activities directly related with mating (Spin, Arrowhead, Attempt); however, as courtship progressed, unsuccessful males increased the time dedicated to activities not directly associated to mating (Call 0, Relax,Stationary). There was not a single sequence of activities leading to success, but the analysis of the last activities performed before mating attempts indicated that the most frequent position before successful attempts was Arrowhead, occurring in 68% of cases, whereas in unsuccessful males this position was observed only 18% of the time before mounting. Although the behavior of the strain analyzed here should be compared with that of natural populations, one would not expect to observe significant differences as compatibility and competitiveness with wild collected flies was previously shown under field cage conditions. Behavioral tests such as those applied here might be important to assess quality of mass reared strains for sterile insect technique implementation programs.

Effects of B chromosomes and supernumerary segments on morphometric traits and adult fitness components in the grasshopper, Dichroplus elongatus (Acrididae).

The South American species, Dichroplus elongatus, is polymorphic for B chromosomes and supernumerary segments in chromosome pairs M6 (SS6), S9 (SS9) and S10 (SS10). Both forms of supernumerary heterochromatin shape chiasma frequency and distribution and B chromosomes also affect male fertility. Here, we analysed the effects of these polymorphisms on morphometric traits (total, 3rd femur, 3rd tibia, thorax and tegmen lengths) and several adult fitness components, including male and female mating success, and female reproductive potential. B chromosomes tend to decrease, and SS6 segments to increase the body size of carriers. The analysis of reproductive potential suggested that B chromosome carrying females have higher numbers of embryos per clutch and ovarioles per ovary. The uni- and multivariate analysis of mating success revealed that sexual selection favours larger individuals of both sexes and males with standard karyotype. B chromosomes may have accumulation mechanisms, which involve preferential transmission of B chromosomes to germ cells or functional gametes. The maintenance of Bs might be explained by interactions among accumulation mechanisms and trade-offs between detrimental and favourable effects on different fitness components.

Mitochondrial DNA variation in the grasshopper Sinipta dalmani: application of long-PCR to the development of a homologous probe.

RFLP analysis of mtDNA in natural populations is a valuable tool for phylogeographic and population genetic studies. The amplification of long DNA fragments using universal primers may contribute to the development of novel homologous probes in species for which no previous genomic information is available. Here we report how we obtained the complete mtDNA genome of Sinipta dalmani (Orthoptera) in 2 fragments (7 and 9 kb) using primers of conserved regions. The specificity of the PCR reactions was ultimately confirmed by several lines of evidence. These fragments were used as a probe for a mtDNA RFLP study in S. dalmani that analyzed the pattern of haplotype distribution and nucleotide diversity within and among chromosomally differentiated natural populations. Our results suggest that the restriction in gene flow detected at the molecular level may explain the chromosome differentiation detected previously and the maintenance of chromosome polymorphism in some areas of S. dalmani geographic distribution.

Mitotically unstable B chromosome polymorphism in the grasshopper Dichroplus elongatus.

Dichroplus elongatus, a widespread South American phytophagous grasshopper, exhibits polymorphisms for supernumerary chromosomes and segments (SS) in natural populations in Argentina. In this paper we review the available information on B chromosome polymorphism in D. elongatus related to geographic distribution, patterns of chromosome variation and influence on sperm formation. In D. elongatus the different forms of supernumerary variants are not independent. The proportion of B-carrying individuals (B prevalence) is negatively correlated with SS10 and positively with SS6 frequencies. The analysis of population structure considering the different supernumerary variants would suggest that the patterns of chromosome variation can not be explained only by random factors. Geographic distribution was analyzed scoring the prevalence of B chromosomes in 13 natural populations collected in three different biogeographical provinces from Northwest (Las Yungas province) and East (Espinal and Pampeana provinces) of Argentina. The detected heterogeneity may be explained by significant differentiation between Northwest and East regions and among populations within Las Yungas and Pampeana provinces. Correlation analysis suggested that B chromosome prevalence is associated with maximum temperature and with latitude. Additional information about the nature of the patterns of B chromosome variation was obtained comparing them with those obtained at the mitochondrial DNA level. The hierarchical analysis of molecular differentiation revealed discrepancy with respect to chromosome differentiation and also suggested that the pattern of B chromosomes may not be explained by historical factors. We also discussed the probable influence on fertility of carriers considering the production of abnormal sperm formation (macro and microspermatids) in relation to the number of Bs per follicle.

Sexual selection on multivariate phenotype in wild and mass-reared Ceratitis capitata (Diptera: Tephritidae).

Males with a larger thorax and narrower face were found to be favoured by directional sexual selection in a previous field cage experiment where Ceratitis capitata males from the Seib 6-96 genetic sexing strain competed with wild males from Alto Valle (Patagonia) for the possession of wild females. Targets of sexual selection, however, might differ between wild and laboratory females as a response to adaptation to mass-rearing conditions. To evaluate possible divergences on the targets of sexual selection as a by-product of adaptation to mass-rearing conditions, field cage tests were performed with both wild and laboratory females. To avoid possible bias due to correlation among the measured traits (eye length [EL], face width [FW], head width [HW], and thorax length [TL]), a multivariate analysis was applied. Consistent with the previous experiment, the results indicated that TL and FW are probable targets of directional sexual selection independently of female strain. However, laboratory females were less selective than wild ones. Additionally, correlational sexual selection was detected acting on the multivariate phenotype. The effects of correlational selection overlap with those of directional selection on each single trait. The analysis of mating pair characteristics showed patterns that do not match the expectations for a random mating system. The current analysis indicates that during mating pair formation two processes overlap. On the one hand, sexual selection favours males with larger size (TL) and narrower faces (FW). This effect occurs in both wild and laboratory females. In addition, assortative mating based on both phenotype and origin was also observed.

Ribosomal DNA variation in the grasshopper, Dichroplus elongatus.

We report an RFLP analysis of ribosomal DNA variation in natural populations of the grasshopper, Dichroplus elongatus, previously analyzed for mitochondrial DNA variation. DNA samples were digested with five restriction enzymes, BamHI, EcoRI, HindIII, PstI, and XbaI. BamHI was the only enzyme that showed no variation. The remaining enzymes showed fragment size variation at both intra- and interpopulation levels. Stepwise regression analysis revealed that the average number of length variants per individual is significantly associated with altitude. Moreover, the same analysis indicated that the frequency of some restriction variants exhibits a significant regression on both geographic and climatic variables. The intra- and interpopulation variability of rDNA was analysed by Lynch's and Hedrick's similarity indices using presence or absence of a fragment and band intensities, respectively. The corresponding neighbour-joining (N-J) trees based on Lynch's and Hedrick's genetic distances resulted in similar topologies. However, these trees were not in agreement with the N-J dendrogram obtained from mtDNA data previously reported by Clemente et al. (2000). The disagreement between mtDNA and rDNA phenograms along with the observed correlation between rDNA variability and geographical and climatic variables suggest some form of selection, besides genetic drift and migration, is involved in the pattern of rDNA variation.

Negative genetic correlation between traits of the Drosophila head, and interspecific divergence in head shape.

For morphological traits that are negatively correlated, the genetic correlation (rg) between them might strongly influence patterns of morphological divergence and shape. Here, the pattern of divergence between two sibling species of cactophilic Drosophila, D. buzzatii and D. koepferae, is examined for two traits that are known to be negatively correlated in other Drosophila species: face width (FW) and width of both eyes (EW). Head width (HW, the sum of FW and EW, i. e. the total width of the head capsule) was also examined. Genetic and phenotypic correlations were estimated in the laboratory G2 generation of a sample of wild D. buzzatii derived from a population where D. koepferae is not present. Phenotypic correlations were also estimated in D. buzzatii and D. koepferae from another, very different, population where the species are sympatric. Consistent with studies in other Drosophila species, rg was negative and significant for the correlation between FW and EW, and positive (but nonsignificant at a matrix-wide P-value of 0.05) for the correlations of HW with both FW and EW. This well-defined correlation pattern was also consistent with the phenotypic correlations in both D. buzzatii and D. koepferae. No significant difference in these traits was detected between D. buzzatii populations, but head shape has diverged between D. buzzatii and D. koepferae. Specifically, the two negatively correlated traits, FW and EW, have evolved in opposite directions in these two species, with HW showing no significant interspecific difference. The overall picture of this divergence pattern shows a striking concordance with the present evidence of negative correlations between FW and EW, and is consistent with the notion of rg-related constraints on the pattern of interspecific differentiation.

Mating system parameters in species of genus Prosopis (Leguminosae).

The section Algarobia of genus Prosopis involves important natural resources in arid and semiarid regions of the world. Their rationale use requires a better knowledge of their biology, genetics and mating system. There are contradictory information about their mating system. Some authors claim they are protogynous and obligate outcrosser. However, some evidence have been shown indicating that they might not be protogynous and that they might be somewhat self-fertile. The current paper analyses genetic structure and mating system parameters in populations of seven species of this section from South and North America based on isozyme data. In all species a significant homozygote excess was found in the offspring population but not in mother plant genotypes. Multilocus and mean single locus outcrossing rates (tm, ts) indicated that about 15% selfing can occur in the studied populations. The heterogeneity between pollen and ovule allele frequencies was low suggesting population structuration, in agreement with the estimates of correlation of tm within progeny (rt) and correlation of outcrossed paternity (rp). The difference of FIS estimates between offspring and mother plants suggest some selection favouring heterozygotes between seedling and adult stages.

Multivariate analysis of male mating success on morphometric traits and chromosome dosage in the grasshopper Sinipta dalmani.

Sinipta dalmani is a South American grasshopper that is chromosomally polymorphic for a pericentric inversion in the M4 pair. The inversion affects morphology, showing a negative average effect on body size. In the present work, male mating success was analysed in competition cages and possible correlations examined between this fitness component, body size and inversion polymorphism. The outcome of the study revealed that mated males were larger and had lower inversion dosage than unmated ones. The analysis of standardized selection differentials (S') demonstrated that directional selection operated on chromosome dosage and 3rd tibia, 3rd femur, thorax and tegmina lengths. The analysis of karyotype frequencies showed that mated males had a higher frequency of the standard homozygote karyotype. The analysis of selection gradient (beta') and the transformed logistic regression (alpha) showed an important effect of selection on 3rd tibia length. Identical conclusions were obtained when linear and logistic multiple regression were performed on the first three components of PCA analyses. Our results indicate that differences in some body size-related traits are determined by the karyotype, and that of these length of 3rd tibia provides the greatest contribution to variation in fitness. The selective effects detected on chromosome dosage and the other morphological traits may be considered as indirect ones caused by correlated effects.

Unexpected low genetic differentiation among allopatric species of section Algarobia of Prosopis (leguminosae).

The Chaqueña Biogeographic Province, in South America, is the main diversity centre of Prosopis. A group of sympatric species of Section Algarobia in this region constitutes a syngameon, characterised by frequent hybridization and introgression. These processes have been postulated as responsible for the low genetic differentiation observed among species within this group. In this study genetic variability and differentiation among geographically isolated species of the same section was analysed through isoenzyme electrophoresis. Variability parameters and fixation indices were estimated to determine the genetic structure of populations. Two Argentinean, P. ruscifolia ('vinal') and P. flexuosa ('algarrobo amarillo'), and one North American species, P. glandulosa (honey mesquite) were studied. All of them showed, similarly to other species of the same section, high genetic variability and exhibit homozygote excess, probably due to population substructure and low rates of selfing. In contrast to our hypothesis, genetic similarity among species is not related to geographic distance. Genetic distances between P. glandulosa and South American species are similar to those observed among species of this subcontinent. The results obtained suggest that the high genetic similarity among the species of the section Algarobia studied is not due to hybridization.

Sexual selection related to developmental stability in Drosophila buzzatii.

Relationships among developmental stability, sexual selection, and body size were examined in the cactophilic fly Drosophila buzzatii. Developmental stability, as measured in terms of fluctuating asymmetry (FA) of wing length, was positively associated to male's copulatory success in a mass-mating experiment with wild-reared flies. Previous studies showed that body size is positively correlated with mating success and other fitness components in this species, but no phenotypic correlation between wing asymmetry and size was found in the present study. Thus, wing asymmetry does not indicate male quality in terms of phenotypic variation in body size. This study suggests that sexual selection against wing asymmetry may be mediated through unknown factors related to developmental stability rather than body size.

Correlations among size-related traits affected by chromosome inversions in Drosophila buzzatii: the comparison within and across environments.

Genetic variation in correlations among size-related traits of head, thorax, and wings was examined in Drosophila buzzatii, by comparing the pattern of the Phenotypic Correlation Matrix (Rp) between inversion karyotypes of the second chromosome. No similarity in Rp was observed between some karyotypes in a natural population. The pattern of Rp in wild-reared heterokaryotypes, but not in homokaryotypes, was similar to the whole population represented by laboratory-reared flies. While phenotypic correlations in wild-reared flies were found to be larger than in laboratory-reared flies, similarity in the pattern of Rp was very high for one homokaryotype reared in both environments: the relatively homogeneous lab environment and the more variable field environment. While no such a similarity across environments was detected between different karyotypes, the pattern of Rp was similar for a same homokaryotype in different populations. Thus, the lack of karyotypic similarity in Rp is, at least partially, genetic. These results indicate that chromosomal inversions are factors affecting genetic correlations among traits known to be phenotypically correlated with adult fitness components in this species.

Genetic and phenotypic correlations among size-related traits, and heritability variation between body parts in Drosophila buzzatii.

Recent studies have shown that body size is a heritable trait phenotypically correlated with several fitness components in wild populations of the cactophilic fly Drosophila buzzatii. To obtain further information on size-related variation, heritabilities as well as genetic and phenotypic correlations among size-related traits of several body parts (head, thorax and wings) were estimated. The study was carried out on an Argentinean natural population in which size-related selection was previously detected. The genetic parameters were estimated using offspring-parent regressions (105 families) in the laboratory G2 generation of a sample of wild flies. The traits were also scored in Wild-Caught Flies (WCF). Laboratory-Reared Flies (LRF) were larger and less variable than WCF. Although heritability estimates were significant for all traits, heritabilities were higher for thorax-wing traits than for head traits. Phenotypic and genetic correlations were all positive. The highest genetic correlations were found between traits which are both functionally and developmentally related. Genetic and phenotypic correlations estimated in the lab show similar correlation pattern (r = 0.49; P = 0.02, Mantel's test). However, phenotypic correlations were found to be typically larger in WCF than in LRF. The genetic correlation matrix estimated in the relatively homogeneous lab environment is not simply a constant multiplicative factor of the phenotypic correlation matrix estimated in WCF.

Correlations among size-related traits are affected by chromosome inversions in an adaptive polymorphism in Drosophila buzzatii.

Genetic variation in correlations among size-related traits of head, thorax and wings was examined in Drosophila buzzatii, by comparing the correlation pattern of the phenotypic correlation matrix (CP-Rp) between inversion karyotypes of the second chromosome. CP-Rp differed between some karyotypes in a natural population. CP-Rp in homokaryotypic classes of wild-reared flies, but not in heterokaryotypes, differed from the whole population represented by laboratory-reared flies. Similarity in CP-Rp was highly significant for a same homokaryotype in two populations. In one of them, the chromosome is polymorphic for four inversions. In the other population, one of the inversions is almost fixed. CP-Rp was significantly similar between these populations, illustrating that similarity of CP-Rp may even occur between populations which have greatly diverged in frequencies of some genotypes affecting correlation patterns. It is suggested that chromosomal inversions are factors affecting genetic correlations among traits known to be phenotypically correlated with fitness components.

An adaptive chromosomal polymorphism affecting size-related traits, and longevity selection in a natural population of Drosophila buzzatii.

Size-related phenotypic variation among second-chromosome karyotypes in Drosophila buzzatii was examined in an Argentinian natural population. For all measured traits (thorax and wing length; wing, head and face width), this inversion polymorphism exhibited a significant and (additive) linear contribution to the phenotypic variance in newly emerged wild flies. The results suggest that only overall body size, and not body shape, is affected, as no karyotypic variation was found for any trait when the effects of differences in within-karyotype size were removed with Burnaby's method. Likewise, in an experiment of longevity selection in the wild, variation in chromosomal frequencies was verified in the direction predicted on the basis of: (i) previous studies on longevity selection for body size in the wild and (ii) the pattern of chromosomal effects we observed on size. The direction of such selection is consistent with a pattern of antagonistic selection detected in previous studies on the inversion polymorphism.

Genetic structure is determined by stochastic factors in a natural population of Drosophila buzzatii in Argentina.

D. buzzatii is a cactophilic species associated with several cactaceae in Argentina. This particular ecological niche implies that this species is faced with a non-uniform environment constituted by discrete and ephemeral breeding sites, which are colonized by a finite number of inseminated females. The genetic consequences of this population structure upon the second chromosome polymorphism were investigated by means of F-statistics in a natural endemic population of Argentina. The present study suggests that differentiation of inversion frequencies in third instar larvae among breeding sites has taken place mainly at random and selection is not operating to determine the structure of this population. The average number of parents breeding on a single pad seems to be similar to the number colonizing Opuntia ficus indica rotting cladodes in Carboneras, a derived population from Spain. There is no significant excess of heterokaryotypes within pads or in the population as a whole. The results obtained in the present study suggest that the potential role of selective versus stochastic factors relative to the among pad heterogeneity in the population here studied is different from that of the Spanish population previously reported. Potential mechanisms responsible for these differences are discussed.

The evolutionary history of Drosophila buzzatii. XXVII. Thorax length is positively correlated with longevity in a natural population from Argentina.

The correlation between body size and longevity was tested in an Argentinian natural population of Drosophila buzzatii. Mean thorax length of flies newly emerging from rotting cladodes of Opuntia vulgaris was significantly smaller than that of two samples of flies caught at baits. The present results which might be interpreted as directional selection for longevity favoring larger flies are in agreement with previous results achieved in a Spanish natural population of D. buzzatii. Flies emerging from different substrates showed significant differences in thorax length, suggesting that an important fraction of phenotypic variance can be attributed to environmental variability. However, laboratory and field work in different populations of D. buzzatii showed a significant genetic component for thorax length variation.