Hybridization has many and varied impacts on the process of speciation. Hybridization may slow or reverse differentiation by allowing gene flow and recombination. It may accelerate speciation via adaptive introgression or cause near-instantaneous speciation by allopolyploidization. It may have multiple effects at different stages and in different spatial contexts within a single speciation event. We offer a perspective on the context and evolutionary significance of hybridization during speciation, highlighting issues of current interest and debate. In secondary contact zones, it is uncertain if barriers to gene flow will be strengthened or broken down due to recombination and gene flow. Theory and empirical evidence suggest the latter is more likely, except within and around strongly selected genomic regions. Hybridization may contribute to speciation through the formation of new hybrid taxa, whereas introgression of a few loci may promote adaptive divergence and so facilitate speciation. Gene regulatory networks, epigenetic effects and the evolution of selfish genetic material in the genome suggest that the Dobzhansky-Muller model of hybrid incompatibilities requires a broader interpretation. Finally, although the incidence of reinforcement remains uncertain, this and other interactions in areas of sympatry may have knock-on effects on speciation both within and outside regions of hybridization.

The fire-bellied toads Bombina bombina and B. variegata differ extensively in biochemistry, morphology, and behavior. We use a survey of five diagnostic enzyme loci across the hybrid zone near Cracow in Southern Poland to estimate the dispersal rate, selection pressures, and numbers of loci which maintain this zone. The enzyme clines coincide closely with each other and with morphological and mitochondrial DNA clines. Although the zone lies on a broad transition between environments suitable for bombina and variegata, the close concordance of diverse characters, together with increased aberrations and mortality in hybrids, suggest that the zone is maintained largely by selection against hybrids. There are strong "linkage disequilibria" between each pair of (unlinked) enzyme loci (R̄ = 0.129 [2-unit support limits: 0.119-0.139]). These are probably caused by gene flow into the zone, and they give an estimate of dispersal (σ = 890 [790-940] m gen ). The clines are sharply stepped, with most of the change occurring within 6.15 (5.45-6.45) km, but with long tails of introgression on either side. This implies that the effective selection pressure on each enzyme marker (due largely to disequilibrium with other loci) is s* = 0.17 (0.159-0.181) at the center but that the selection acting directly on the enzyme loci is weak or zero (s < 0.0038). The stepped pattern implies a barrier to gene flow of 220 (48-415) km. This would substantially delay neutral introgression but would have little effect on advantageous alleles; the two taxa need not evolve independently. Strong selection is needed to maintain such a barrier: hybrid populations must have their mean fitness reduced by a factor of 0.65 (0.60-0.77). This selection must be spread over a large number of loci to account for the concordant patterns and the observed cline widths (N = 300 [80-2,000]).

Insect mitochondrial cytochrome oxidase I (COI) genes are used as a model to examine the within-gene heterogeneity of evolutionary rate and its implications for evolutionary analyses. The complete sequence (1537 bp) of the meadow grasshopper (Chorthippus parallelus) COI gene has been determined, and compared with eight other insect COI genes at both the DNA and amino acid sequence levels. This reveals that different regions evolve at different rates, and the patterns of sequence variability seems associated with functional constraints on the protein. The COOH-terminal was found to be significantly more variable than internal loops (I), external loops (E), transmembrane helices (M) or the NH2 terminal. The central region of COI (M5-M8) has lower levels of sequence variability, which is related to several important functional domains in this region. Highly conserved primers which amplify regions of different variabilities have been designed to cover the entire insect COI gene. These primers have been shown to amplify COI in a wide range of species, representing all the major insect groups; some even in an arachnid. Implications of the observed evolutionary pattern for phylogenetic analysis are discussed, with particular regard to the choice of regions of suitable variability for specific phylogenetic projects.

We compare the pattern of morphological and electrophoretic variation in the hybrid zone between Bombina bombina and B. variegata across two transects: one near Cracow and one 200 km away, near Przemyśl in southeastern Poland. Morphological variation across the Przemyśl transect had been surveyed more than 50 years ago; though we found a significant shift at one site, there is no evidence for gross movement over this period. Morphological and electrophoretic changes coincide, and the average shape of the clines is the same across both transects. At the center, most of the change in frequency of six diagnostic allozymes occurs within w = 6.05 km (2‐unit support limits 5.56–6.54 km). These steep gradients are generated not by selection on the allozymes themselves, but by associations with other loci: though these markers are unlinked, they are in strong linkage disequilibrium with each other [R = D/√pquv = 0.22 (0.15–0.29) at the center]. Disequilibria are broken up as alleles diffuse away from the zone and flow into the new genetic background. The net barrier to the flow of genes from bombina into variegata, which is generated by these disequilibria, is B = 51 (22–81) km. The fitness of hybrids must be substantially reduced to produce such a barrier [W̄H/W̄P = 0.58 (0.54–0.68)], and this selection must be spread over many loci [N = 55 (26–88)]. Alleles introgress significantly less far than would be expected from the age of the zone and the estimated dispersal rate [σ = 0.99 (0.82–1.14) km gen.1/2]: this implies selection of se = 0.37 (0.15–0.58)% on the enzymes themselves. There is weak but significant linkage disequilibrium well away from the center of the zone; this, together with the presence of parental and F1 genotypes, suggests some long‐range migration. However, such migration is not likely to cause significant introgression.

The control regions of mitochondrial DNA of two insects, Schistocerca gregaria and Chorthippus parallelus, have been isolated and sequenced. Their sizes are 752 bp and 1,512 bp, respectively, with the presence of a tandem repeat in C. parallelus. (The sequences of the two repeats are highly conserved, having a homology of 97.5%.) Comparison of their nucleotide sequences revealed the presence of several conserved sequence blocks dispersed through the whole control region, showing a different evolutionary pattern of this region in these insects as compared to that in Drosophila. A highly conserved secondary structure, located in the 3' region near the small rRNA gene, has been identified. Sequences immediately flanking this hairpin structure rather than the sequences of this structure themselves are conserved between S. gregaria/C. parallelus and Drosophila, having a sequence consensus of "TATA" at 5' and "GAA(A)T" at 3'. The motif "G(A)nT" is also present in the 3' flanking sequences of mammalian, amphibian, and fish mitochondrial L-strand replication origins and a potential plant mitochondrial second-strand-replication origin, indicating its universal conservation and functional importance related to replication origins. The stem-and-loop structure in S. gregaria/C. parallelus appears to be closely related to that found in Drosophila despite occupying a different position, and may be potentially associated with a second-strand-replication origin. This in turn suggests that such a secondary structure might be widely conserved across invertebrates while their location in the control region may be variable. We have looked for such a conserved structure in the control regions of two other insects, G. firmus and A. mellifera, whose DNA sequences have been published, and their possible presence is discussed. Mitochondrial control regions characterized to date in five different insect taxa (Drosophila, G. firmus, A. mellifera, S. gregaria, and C. parallelus) may be classed into two distinct groups having different evolutionary patterns. It is observed that tandem repetition of regions containing a probable replication origin occurred in some species from disjunct lineages in both groups, which would be the result of convergent evolution. We also discuss the possibility of a mechanism of "parahomologous recombination by unequal crossing-over" in mitochondria, which can explain the generation of such tandemly repeated sequences (especially the first critical repetition) in the control region of mtDNA, and also their convergent evolution in disjunct biological lineages during evolution.