Geographical and sexual variation of wing size in the monophagous flower-breeding species Drosophila flavopilosa

Flower samples of Cestrum parqui (Solanaceae) containing preadult forms of Drosophila flavopilosa were collected at four localities from the Maipo Valley in the Central zone of Chile (Lat. 33°40'S), one at sea level and the others at 300 m, 780 m and 1365 m of altitude. Only the adults which emerged after 18 days of permanency of the flowers in a chamber at 22°C were employed for wing analysis. Nine different measurements were made in the wings of 100 males and 100 females from each population. In males, a clear tendency of increasing wing length and width according to elevation above sea level was found. Females, however, exhibited a quite different type of variation; the larger and wider wings corresponding to the populations originated in the laces of intermediate elevation. As a result of the different pattern of wing variation in males and females, the sexual dimorphism was found to be significantly different between the studied populations. The results support the idea that males and females react in a different way to selective pressures affecting wing development.     

Mitochondrial gene divergence of Colombian Drosophila pseudoobscura

Isolated populations of drosophila pseudoobscura, separated from North American populations by about 2,400 km, were found in Colombia in 1960. We compared for sequences of the small ribosomal RNA (srRNA) gene on the mitochondria between North American and Colombian D. pseudoobscura in order to clarify the age of the Colombian isolates. The North American populations were not genetically different from each other but were genetically different from the Colombian populations. The Mexican strains represent the area from which the Colombian founders might have come. The estimated net nucleotide divergence between Mexican and Colombian D. pseudoobscura indicates that the Colombian population is not an ancient lineage. Phylogenies using both distance and parsimony methodologies reinforced this conclusion. The Colombian samples group together with both methods but, according to the bootstrap analysis, not significantly. It appears that the populations have not been separated long enough for their DNA sequences to show much divergence.      

Isozyme Frequency Patterns in DROSOPHILA PAVANI Associated with Geographical and Seasonal Variables

Fourteen population samples of Drosophila pavani were obtained from a number of localities in Chile. The populations sampled were dispersed over 7 degrees of latitude and 1800 meters of elevation, and were drawn at three different times. Sixteen electrophoretic loci were assayed for each population; eight of the loci were analyzed statistically for geographic variation; the other eight were essentially monomorphic. For all eight variable loci, variation in allelic frequencies among populations was highly significant. In all cases, a significant portion of the variation among populations was associated with variation in gross environmental variables (latitude, elevation, month of collection). The implications of the evidence were discussed, and the authors concluded that there was suggestive evidence for selection.     

Chromosomal structure of populations of drosophila flavopilosa studied in larvae collected in their natural breeding sites

Summary D. flavopilosa Frey, is a neotropical species of Drosophila living in Chile that, together with 13 other species, has been included by Wheeler, Takada und Brncic (1962) in the flavopilosa species group. Despite the fact that it has not been possible to breed this species in the laboratory, the circumstance that its habitat is well known, has allowed the study of the chromosomal structure of some natural populations of this species, analyzing larvae directly taken to the laboratory from their natural breeding sites: the flowers of the solanacean Cestrum parqui L'Héritier.The present paper describes the mitetic and salivary gland chromosomes of D. flavopilosa, and gives a composite map of the Standard gene arrangement. In central Chile, natural populations of this species are polymorphic with respect to the gene orders in their chromosomes, due to the presence of four independent inversions, all located in one of the six chromosomes (the right arm of the V-chromosome). Quantitative data on the distribution of the inversions shows that there is an altitudinal gradient in the frequencies of two of the four inversions. Heterozygotes for Inversion A, are more frequent at high altitudes than at sea level. On the contrary, heterozygotes for inversion B, are abundant at sea level, but practically disappear at high altitudes. These altitudinal clines have been observed in two valleys near Santiago, Chile, which run from the Andes mountains to the Pacific coast.The work reported in this article has been carried out under Contract AT (30-1) 2465 US Atomic Energy Commission, and partially supported by Grants from the University of Chile, Faculty of Medicine and the Rockefeller Foundation under a joint program.     

Development of a cost-effective high-throughput process of microsatellite analysis involving miniaturized multiplexed PCR amplification and automated allele identification

Background

Microsatellites are nucleotide sequences of tandem repeats occurring throughout the genome, which have been widely used in genetic linkage analysis, studies of loss of heterozygosity, determination of lineage and clonality, and the measurement of genome instability or the emergence of drug resistance reflective of mismatch repair deficiency. Such analyses may involve the parallel evaluation of many microsatellite loci, which are often limited by sample DNA, are labor intensive, and require large data processing.

Results

To overcome these challenges, we developed a cost-effective high-throughput approach of microsatellite analysis, in which the amplifications of microsatellites are performed in miniaturized, multiplexed polymerase chain reaction (PCR) adaptable to 96 or 384 well plates, and accurate automated allele identification has been optimized with a collective reference dataset of 5,508 alleles using the GeneMapper software.

Conclusions

In this investigation, we have documented our experience with the optimization of multiplex PCR conditions and automated allele identification, and have generated a unique body of data that provide a starting point for a cost-effective, high-throughput process of microsatellite analysis using the studied markers.