The complete sequence (15,455 bp) of the mitochondrial DNA of the mosquito Anopheles quadrimaculatus species A is reported. This genome is compact and very A+T rich (77.4% A+T). It contains genes for 2 ribosomal RNAs (rRNAs), 22 transfer RNAs (tRNAs), and 13 subunits of the mitochondrial inner membrane respiratory complexes. The gene arrangement is the same as in Drosophila yakuba, except that the positions of two contiguous tRNAs are reversed and a third tRNA is transcribed from the complementary strand. Protein-coding genes, rRNAs, and most tRNAs were similar to D. yakuba. Two tRNAs had nonstandard secondary structures comparable with those of nematode mitochondrial tRNAs. The very small putative control region (625 bp) contains no sequence motifs similar to those used in vertebrates and other insects for initiation of transcription and replication.

Two Hsp82 genes were isolated from the malaria vector Anopheles albimanus in a single lambda phage clone. The two genes are in a head-to-head arrangement separated by approx. 0.9 kbp. Northern hybridizations and 5' RACE demonstrate that both genes are transcribed, have moderate levels of constitutive transcription, and are also heat-inducible with maximum transcript accumulation occurring after 40 degrees C heat shocks. Both genes have typical heat-shock promoters and conserved intron boundaries in the untranslated leaders. The open reading frames are 99.6% identical differing in only nine silent nucleotide positions in the 2166 bp ORFs. However, precisely outside the ORFs, the flanking DNA of the two genes shows no evidence of common derivation. The high degree of identity between the two ORFs appears to be a result of gene conversion occurring by a process similar to that previously suspected in the A. albimanus Hsp70 genes and several D. melanogaster genes arranged as palindromes. This process probably involves a stem-loop intermediate and is restricted in extent by flanking sequence divergence. These Hsp82 genes clearly demonstrate the extreme precision with which gene conversion can lead to protein-coding-region homogeneity yet allow flanking DNA divergence.

Samples of Anopheles rangeli Gabaldon, Cova Garcia & Lopez, An. trinkae Causey, and An. nuneztovari Gabaldon from Venezuela, Ecuador, Brazil, and Bolivia were analyzed for genetic variability at 24 enzyme loci. Estimates of genetic variability for An. rangeli and An. trinkae from Ecuador and for An. nuneztovari in Venezuela had the following ranges: 46-58% polymorphic loci, 1.7-2.0 (SEM = 0.1-0.3) mean number of alleles per locus, and 0.069-0.113 (SEM = 0.03-0.04) expected mean heterozygosity. Genetic variability estimates of An. rangeli from Bolivia were 20.8-29.2% polymorphic loci, 1.2-1.6 (SEM = 0.1-0.2) mean number of alleles per locus, and 0.037-0.054 (SEM = 0.02-0.03) expected mean heterozygosity. The estimated genetic distance between An. rangeli and An. trinkae ranged from 0.149 to 0.197. The genetic distance between these 2 species and An. nuneztovari ranged from 0.319 to 0.440. Although there were allele frequency differences at some loci between samples of An. nuneztovari sampled from either side of the Andes Mountains in Venezuela, there were no diagnostic loci and the estimated genetic distance was only 0.023. Seven enzyme loci were diagnostic between An. nuneztovari and one or both of its sister species: Acon-2, Ao, Hk-1, Idh-2, Me, Pgi, and Pgm. The diagnostic loci Hk-1 and Acon-2 were not polymorphic in any species. An. rangeli and An. trinkae can be distinguished by the diagnostic loci Ao, Idh-2, and Me-1, and with a 97% probability by Pgm. Distance Wagner and unweighted pair-group method with arithmetic averaging analyses support a close phylogenetic relationship between An. trinkae and An. rangeli.

Sequence variation of the ribosomal DNA internal transcribed spacer 2 (ITS2) was examined for populations of the malaria vector Anopheles nuneztovari collected in Colombia, Venezuela, Bolivia, Suriname, and Brazil. Mosquitoes from Colombia and Venezuela had identical ITS2 sequences and were distinguished from sequences in other populations by three insertion/deletion events (indels) and by one transversion. The length of the ITS2 was 363-369 bp, and it had a G+C content of 55.3%-55.7%. Variation in the length of the ITS2 between and within populations was due to indels in simple repeats. ITS2 consensus sequences were similar or identical for samples from the following three groups: (1) Colombia, Bolivia, and Venezuela; (2) Suriname and northern Brazil; and (3) eastern and central Brazil. The presence of two different consensus sequences from a single location near Manaus, Brazil, suggests that populations from eastern Brazil and those from Suriname converge in this region of the Amazon Basin. These data show that putative cryptic species of An. nuneztovari are distinguished by very minor differences in DNA sequence of the ITS2 region.

Recent field experiments demonstrated the possibility of using the sterile male method for the control of Anopheles albimanus Wiedemann, the most important vector of human malaria in Central America. Until now there was no practical method for excluding females from the releases of sterile males. A genetic method was developed for the preferential elimination of females during any of the four life stages. This genetic sexing system utilizes propoxur ( o -isopropoxyphenyl methyl-carbamate) susceptibility as a recessive conditional lethal, a T(Y:2R) translocation, and an In(2R)inversion. The propoxur resistance allele (dominant) was linked to the Y chromosome via a radiation-induced translocation, and genetic recombination was suppressed by inversions. In one of the strains produced, 99.7 percent of the females are eliminated when treated with propoxur, without male loss.