Phenotypic Variation and Sexual Size Dimorphism in Dichroplus elongatus (Orthoptera: Acrididae).

Patterns of body size evolution are of particular interest because body size can affect virtually all the physiological and life history traits of an organism. Sexual size dimorphism (SSD), a difference in body size between males and females, is a widespread phenomenon in insects. Much of the variation in SSD is genetically based and likely due to differential selection acting on males and females. The importance of environmental variables and evolutionary processes affecting phenotypeic variation in both sexes may be useful to gain insights into insect ecology and evolution. Dichroplus elongatus Giglio-Tos is a South American grasshopper widely distributed throughout Argentina, Uruguay, most of Chile, and southern Brazil. In this study, we analyzed 122 adult females of D. elongatus collected in eight natural populations from central-east Argentina. Females show large body size variation among the analyzed populations and this variation exhibits a strong relationship with fecundity. Our results have shown that larger females were more fecund than smaller ones. We found that ovariole number varied along a latitudinal gradient, with higher ovariole numbers in populations from warmer locations. A considerable female-biased SSD was detected. SSD for three analyzed morphometric traits scaled isometrically. However, SSD for thorax length displayed a considerable variation across the studied area, indicating a larger relative increase in female size than in male size in warmer environmental conditions.

Latitudinal clines in the grasshopper Dichroplus elongatus: coevolution of the A genome and B chromosomes?

Argentine populations of Dichroplus elongatus (Orthoptera: Acrididae) are polymorphic for B chromosomes. Previous studies showed that B chromosomes affect body size and some fitness components in Northwestern populations. We studied phenotype and B's variation patterns along a latitudinal cline as well as the relationship between karyotype and body size related traits in 17 populations from East. Body size related traits showed a 'saw tooth' pattern of variation being small at low and high latitudes and large at intermediate latitudes in most of the analysed populations. Analyses of variance and principal components demonstrated that in most analysed populations B carrier males are associated with a decrease in body size related traits with respect to individuals with standard karyotype. Accordingly with the relationship between karyotype and body size, an opposite pattern of latitudinal variation in the frequencies of B's with respect to body size variation was observed in this area. i.e. smaller individuals tend to have a higher frequency of B chromosomes. The comparison of the differentiation of both karyotype and body size traits with molecular neutral markers demonstrated the relative importance of selection moulding chromosome and phenotype variation. The observed pattern of phenotypic variation is likely to be the result of local adaptation to season length along the latitudinal gradient. The observed contrary pattern of B's clinal variation may reflect the population ability to maintain this chromosome in relation to the local adaptation. The available evidence indicates that the distribution of B chromosome frequency was shaped by selective factors.

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.

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.

Chromosomal polymorphism, morphological traits and male mating success in Leptysma argentina (Orthoptera).

Leptysma argentina (Acrididae: Orthoptera) is polymorphic for a centric fusion between pairs 3 and 6 of its basic chromosomal complement. With the aim of carrying out an analysis of selection components in this species, male mating success, in relation to karyotype and three morphometric traits, was assessed by using experimental contests with four males and one female per mating cage. Karyotype was assessed in terms of fused chromosome 3/6 dosage and coded as "0", "1" and "2" for unfused homozygotes, heterozygotes and fusion homozygotes, respectively. The individual ANOVA showed significant differences between "successful" and "unsuccessful" males in all four measured characters. Fusion carriers showed a higher relative fitness. Since the phenotypic correlations among traits were significant, the selective effects were assessed by means of a multivariate analysis, which demonstrated that selection acted significantly on femur length only, selection on karyotype being indirect. Given that the fusion frequency has been constant since 1985, the presence of selective trade-offs cannot be ruled out.

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.