Carlos Juan

Colonization and diversification: towards a phylogeographic synthesis for the Canary Islands.

Recently, the Canary Islands have become a focus for studies of the colonization and the diversification of different organisms. Some authors have considered Canarian endemisms as relicts of Tertiary origin, but new molecular data suggest a general pattern of continental dispersion followed by in situ speciation. Recent phylogeographic studies are revealing variants of the simple stepping-stone colonization model that seems to hold for many Hawaiian groups. Many factors can generate deviations from such a pattern: the stochastic nature of colonization, competitive exclusion, phylogenetic constraints on adaptive evolution and extinction. An understanding of island colonization and diversification can best be developed from an ecosystem level synthesis as more data for the Canarian archipelago come to hand.

Mitochondrial DNA Phylogeny and Sequential Colonization of Canary Islands by Darkling Beetles of the Genus Pimelia (Tenebrionidae)

Fifteen species of the darkling beetle genus Pimelia (Tenebrionidae: Coleoptera) have been sequenced for a 365 b. p. portion of the mitochondrial Cytochrome Oxidase I gene. Thirteen of these are endemic species inhabiting the Canarian archipelago in the Atlantic and the other two are continental conspecifics. This data set has been phylogenetically analysed by maximum parsimony and distance approaches and the resulting trees used to deduce sequential interisland colonization. This suggests patterns compatible with the geological dating of the islands, but with increasing uncertainty when older events are considered. A colonization sequence from Fuerteventura to Tenerife followed by Tenerife to Gran Canaria and La Gomera, and from the latter to La Palma and then to El Hierro is proposed for the genus. A relatively recent secondary colonization from Gran Canaria to Gomera is deduced.

Phylogeny of the genus Hegeter (Tenebrionidae, Coleoptera) and its colonization of the Canary Islands deduced from Cytochrome Oxidase I mitochondrial DNA sequences

The genus Hegeter comprises 23 species of darkling beetles (Tenebrionidae) endemic to the Macaronesian archipelagos, with 21 of them exclusive to the Canary Islands. We have sequenced 438 bp of the mitochondrial Cytochrome Oxidase I gene in 17 species (24 taxa) of Canarian Hegeter. Estimates of nucleotide composition, transition/transversion ratios and nucleotide change frequencies are very similar to those found in another tenebrionid Canarian genus Pimelia, indicating that similar molecular mechanisms are driving the sequence evolution. The sequence variation found allows phylogenetic analyses of the genus and the deduction of colonization patterns. These involve sequential island invasion with more rapid establishment and radiation than found in the related beetles of the genus Pimelia.

Nested cladistic analysis, phylogeography and speciation in the Timarcha goettingensis complex (Coleoptera, Chrysomelidae).

The Timarcha goettingensis complex is a monophyletic assemblage of closely related leaf beetles (Chrysomelidae), distributed from the north half of the Iberian Peninsula to Central Europe. Oligophagy, mountainous habitat and apterism are factors which are assumed to promote speciation in these beetles. We have used cytochrome oxidase subunit II mitochondrial DNA genealogies obtained from 31 sampling localities and a nested geographical distance analysis to assess the population structure and demographic factors explaining the geographical distributions of the mtDNA haplotypes in the T. goettingensis complex. The results show that there is a significant association between genetic structuring and geography. Inferences about the historical population processes in the species complex are discussed, being in general in accordance with contiguous range expansions and past fragmentations. The use of the cohesion species concept approach suggests the existence of several systematic ranks among the different T. goettingensis populations, which is in part supported by ecological traits such as trophic selection and altitudinal distribution.

Localization of tandemly repeated DNA sequences in beetle chromosomes by fluorescentin situ hybridization

In situ hybridization to chromosomes and nuclei ofTenebrio molitor shows the massive presence of a species-specific satellite DNA in all chromosomes and six sites of rDNA in mitotic chromosomes. These sites are located in two autosomal pairs and in the X and Y chromosomes. In a related species,Misolampus goudoti, in which two different families of highly repetitive DNA have been previously characterized, one family is located in centromeric regions of all chromosomes with the exception of chromosome Y, while the other repeated DNA family is present both in centromeric and distal regions of all chromosomes. rRNA genes in this species are present in a medium-sized autosomal pair only. These results show that molecular cytogenetics can be applied to coleopteran chromosomes and open the way for a physical mapping of DNA sequences in these organisms. The results also provide insights into the type of meiotic association of the X and Y chromosomes in Coleoptera and the distribution of repeated DNAs within the genome of these insects.

Mitochondrial DNA sequence variation and phylogeography of Pimelia darkling beetles on the Island of Tenerife (Canary Islands)

Four morphological taxa of the beetle genus Pimelia (Coleoptera, Tenebrionidae) are known to exist on the Atlantic island of Tenerife. We have obtained DNA sequences for 61 individuals from these taxa across the island for a 200 bp long fragment of the mitochondrial COI gene. In addition, a restriction site polymorphism in the nuclear rRNA ITS-1 sequence was identified and screened in a sample of these individuals using the enzyme Kpn2I. The results were analysed using approaches which allow inferences to be made about the population genetic structure and the mitochondrial genealogy of these closely related beetles. The mtDNA haplotype distribution and the estimates of sequence divergence revealed the presence of two ancient mtDNA lineages which coincide with the disjunct volcanic evolution of the island. The ITS-1 polymorphism was found to be diagnostic of these two lineages. However, the morphological and mitochondrial phylogenies were found to be discordant. We argue that this is possibly the result of rapid morphological change, produced by selection in different habitats, which has been recently superimposed on an older mitochondrial DNA divergence.

Complex structural features of satellite DNA sequences in the genus Pimelia (Coleoptera: Tenebrionidae): random differential amplification from a common 'satellite DNA library'.

The major satellites of the nine species of the subgenera Pimelia s. str. and Amblyptera characterised in this paper are composed of longer monomers (500 and 700 bp) than those described previously in 26 Pimelia s. str. taxa (357 bp, a sequence called PIM357). Sequence analysis reveals partial similarity among these satellites and with the PIM357 monomers. The discrepancy between the phylogeny obtained based on three mitochondrial and two nuclear markers and that deduced from satellite DNA (stDNA) sequences suggests that the different Pimelia satellites were already present in a common ancestor forming what has been called a ‘satellite DNA library’. Thus, the satellite profiles in the living species result from a random amplification of sequences from that ‘library’ during diversification of the species. However, species-specific turnover in the sequences has occurred at different rates. They have included abrupt replacements, a gradual divergence and, in other cases, no apparent change in sequence composition over a considerable evolutionary time. The results also suggest a common evolutionary origin of all these Pimelia satellite sequences, involving several rearrangements. We propose that the repeat unit of about 500 bp has originated from the insertion of a DNA fragment of 141 bp into the PIM357 unit. The 705-bp repeats have originated from a 32-bp direct duplication and the insertion of a 141-bp fragment in inverted orientation relative to a basic structure of 533 bp.

Under the volcano: phylogeography and evolution of the cave-dwelling Palmorchestia hypogaea (Amphipoda, Crustacea) at La Palma (Canary Islands).

BackgroundThe amphipod crustacean Palmorchestia hypogaea occurs only in La Palma (Canary Islands) and is one of the few terrestrial amphipods in the world that have adapted to a strictly troglobitic life in volcanic cave habitats. A surface-dwelling closely related species (Palmorchestia epigaea) lives in the humid laurel forest on the same island. Previous studies have suggested that an ancestral littoral Orchestia species colonized the humid forests of La Palma and that subsequent drought episodes in the Canaries reduced the distribution of P. epigaea favouring the colonization of lava tubes through an adaptive shift. This was followed by dispersal via the hypogean crevicular system.ResultsP. hypogaea and P. epigaea did not form reciprocally monophyletic mitochondrial DNA clades. They showed geographically highly structured and genetically divergent populations with current gene flow limited to geographically close surface locations. Coalescence times using Bayesian estimations assuming a non-correlated relaxed clock with a normal prior distribution of the age of La Palma, together with the lack of association of habitat type with ancestral and recent haplotypes, suggest that their adaptation to cave life is relatively ancient.ConclusionThe data gathered here provide evidence for multiple invasions of the volcanic cave systems that have acted as refuges. A re-evaluation of the taxonomic status of the extant species of Palmorchestia is needed, as the division of the two species by habitat and ecology is unnatural. The information obtained here, and that from previous studies on hypogean fauna, shows the importance of factors such as the uncoupling of morphological and genetic evolution, the role of climatic change and regressive evolution as key processes in leading to subterranean biodiversity.

Genetic structure, phylogeography and demography of two ground-beetle species endemic to the Tenerife laurel forest (Canary Islands)

The volcanic island of Tenerife (Canary archipelago) was formerly covered at 600–1200 m above sea level on most of its northern side by a cloud forest holding much of the endemic insect fauna. In the most significant surviving patches of this laurel forest at the eastern and western tips of the island occur two forest‐specialist, closely related species of Eutrichopus (Coleoptera, Carabidae); here we present data on mitochondrial DNA variation among populations of these species. In total, 116 individuals from 16 localities were sampled and a 638 bp fragment of the cytochrome oxidase subunit II gene was sequenced, obtaining evidence for two distinct evolutionary lineages, in accordance with morphological and biogeographical data. Volcanic events at ~0.7 Ma might be responsible for vicariance and the fragmentation of the geographical range of an ancestral species, causing the establishment of two matrilineal lineages. Using nested clade and historical demography analyses we infer past cycles of demographic bottlenecks followed by population expansion, mostly in agreement with the geological timescale of volcanic events. Recent trends, however, refer to fragmentation of the cloud forest due to human intervention.

Phylogeography of the endangered darkling beetle species of Pimelia endemic to Gran Canaria (Canary Islands)

Phylogenetic and geographical nested clade analysis (NCA) methods were applied to mitochondrial DNA sequences of Pimelia darkling beetles (Coleoptera, Tenebrionidae) endemic to Gran Canaria, an island in the Canary archipelago. The three species P. granulicollis, P. estevezi and P. sparsa occur on the island, the latter with three recognized subspecies. Another species, P. fernandezlopezi (endemic to the island of La Gomera) is a close relative of P. granulicollis based on partial Cytochrome Oxidase I mtDNA sequences obtained in a previous study. Some of these beetles are endangered, so phylogeographical structure within species and populations can help to define conservation priorities. A total of about 700 bp of Cytochrome Oxidase II were examined in 18 populations and up to 75 individuals excluding outgroups. Among them, 22 haplotypes were exclusive to P. granulicollis and P. estevezi and 31 were from P. sparsa. Phylogenetic analysis points to the paraphyly of Gran Canarian Pimelia, as the La Gomera P. fernandezlopezi haplotypes are included in them, and reciprocal monophyly of two species groups: one constituted by P. granulicollis, P. estevezi and P. fernandezlopezi (subgenus Aphanaspis), and the other by P. sparsa‘sensu lato’. The two species groups show a remarkably high mtDNA divergence. Within P. sparsa, different analyses all reveal a common result, i.e. conflict between current subspecific taxonomic designations and evolutionary units, while P. estevezi and P. fernandezlopezi are very close to P. granulicollis measured at the mtDNA level. Geographical NCA identifies several cases of nonrandom associations between haplotypes and geography that may be caused by allopatric fragmentation of populations with some cases of restriction of gene flow or range expansion. Analyses of molecular variance and geographical NCA allow definition of evolutionary units for conservation purposes in both species‐groups and suggest scenarios in which vicariance caused by geological history of the island may have shaped the pattern of the mitochondrial genetic diversity of these beetles.