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Biomedical subjects

G M Hewitt

Publications and source records attributed to G M Hewitt.

At least 55 records · Page 3Linked to original sources

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.

Animals↗

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.

Animals↗

The sequence and structure of the meadow grasshopper (Chorthippus parallelus) mitochondrial srRNA, ND2, COI, COII ATPase8 and 9 tRNA genes.

The nucleotide sequence of the mitochondrial ND2, COI, COII, ATPase8, srRNA and nine tRNA genes have been sequenced from two individual of the meadow grasshopper Chorthippus parallelus. Comparisons are made to other insects for which the same regions are completely sequenced. Percentage A + T is found to be relatively low in C. parallelus though consistent with that of the other Orthopteran, Locusta migratoria. The relative number of substitutions observed in the different protein-coding genes was analysed between pairs of insect species sharing different levels of relatedness. A clear change in this rate was observed between the within-genus and between-genera comparisons. This change is interpreted in terms of the functional constraints acting on these four different genes. The patterns seem to result from an early saturation of COI and COII genes with synonymous substitutions, and a tolerance of ND2 and ATPase8 function to high levels of amino acid replacements. This analysis highlights a need for further sequence studies and comparisons between taxa of different levels of divergence in order to understand the patterns of mtDNA evolution on which many evolutionary investigations are based.

Adenosine Triphosphatases↗

The insect cytochrome oxidase I gene: evolutionary patterns and conserved primers for phylogenetic studies.

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.

Amino Acid Sequence↗

Highly conserved nuclear copies of the mitochondrial control region in the desert locust Schistocerca gregaria: some implications for population studies.

Animal mitochondrial DNA has proved a valuable marker in intraspecific systematic studies. However, if nucleotide sequence heterogeneity exists at the individual level, its usefulness will be much reduced. The study demonstrates that the presence of highly conserved non-coding mitochondrial sequences in the nuclear genome of Schistocerca gregaria greatly impairs the use of mtDNA in population genetic studies. Caution is called for in other organisms; and it seems necessary to check for conserved nuclear copies of mitochondrial sequences before launching into a large scale analysis of populations using mtDNA as a genetic marker. Experimental procedures are suggested for this purpose.

Animals↗

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 inhibiting 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 Canarian 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.

Animals↗

Evolution and structural conservation of the control region of insect mitochondrial DNA.

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.

Animals↗

Cloning and sequence analysis of an extremely homogeneous tandemly repeated DNA in the grasshopper Eyprepocnemis plorans.

Digestion of total nuclear DNA of the grasshopper Eyprepocnemis plorans with seven different restriction endonucleases (REs), and subsequent agarose gel electrophoresis, has shown the presence of highly repetitive DNA yielding the typical ladder-like banding pattern. The most clear pattern was produced by DraI, the monomer being some 180 bp. This repeat unit was subsequently cloned and sequenced. Bidirectional sequencing of five randomly chosen clones showed exactly the same nucleotides in all 180 positions. The possible explanations for such an extreme homogeneity of this tandem repeat are discussed in the light of current hypotheses on repetitive DNA function and molecular drive mechanisms.

Animals↗

Phylogeny of the Coleoptera based on mitochondrial cytochrome oxidase I sequence data.

A 400 base pair region of the mitochondrial cytochrome oxidase I gene (COI) was sequenced for thirty-seven species of beetle, representing fifteen families. The sequence was found to be highly variable, with 12-18% divergence within families, and up to 27% divergence between families. Phylogenetic analysis using the neighbour-joining method shows the Carabidae (Adephaga) as a distinct clade, but also shows that all other beetles (Polyphaga) diverged soon after the Adephaga/Polyphaga split. Whilst some species are grouped within their respective families, others are too diverged for easy resolution. The main disagreement with trees constructed using comparative morphology and the fossil record is the position of the Coccinellidae. Whilst COI may be an informative gene for molecular systematics at lower taxonomic levels, or in other insects, improved resolution of this particular phylogeny will require a more highly conserved sequence.

Amino Acid Sequence↗

Postglacial expansion and genome subdivision in the European grasshopper Chorthippus parallelus.

A noncoding nuclear DNA marker sequence (Cpn1-1) was used to investigate subdivision in the grasshopper Chorthippus parallelus and deduce postglacial expansion patterns across its species range in Europe. Investigation of the spatial distribution of 71 Cpn1-1 haplotypes and estimation of levels of genetic differentiation (KST values) between populations and geographic regions provided evidence for subdivision of C. parallelus into at least five major geographic regions and indicated that the French form of C. parallelus originated after range expansion from a Balkan refugium. Further evidence for subdivision of C. parallelus between Italy and northern Europe suggests that the Alps may have formed a significant barrier to gene flow in this grasshopper.

Animals↗

Natal dispersal and genetic structure in a population of the European wild rabbit (Oryctolagus cuniculus).

A combination of behavioural observation, DNA fingerprinting, and allozyme analysis were used to examine natal dispersal in a wild rabbit population. Rabbits lived in territorial, warren based social groups. Over a 6-year period, significantly more male than female rabbits moved to a new social group before the start of their first breeding season. This pattern of female philopatry and male dispersal was reflected in the genetic structure of the population. DNA fingerprint band-sharing coefficients were significantly higher for females within the same group than for females between groups, while this was not the case for males. Wright's inbreeding coefficients were calculated from fingerprint band-sharing values and compared to those obtained from allozyme data. There was little correlation between the relative magnitudes of the F-statistics calculated using the two techniques for comparisons between different social groups. In contrast, two alternative methods for calculating FST from DNA fingerprints gave reasonably concordant values although those based on band-sharing were consistently lower than those calculated by an 'allele' frequency approach. A negative FIS value was obtained from allozyme data. Such excess heterozygosity within social groups is expected even under random mating given the social structure and sex-biased dispersal but it is argued that the possibility of behavioural avoidance of inbreeding should not be discounted in this species. Estimates of genetic differentiation obtained from allozyme and DNA fingerprint data agreed closely with reported estimates for the yellow-bellied marmot, a species with a very similar social structure to the European rabbit.

Analysis of Variance↗

Using chloroplast DNA to trace postglacial migration routes of oaks into Britain.

Postglacial migration is a major factor responsible for the patterns of genetic variation we see in natural populations. Fossil pollen data indicate that early postglacial colonists such as oak, were able to take both western and eastern migration routes into Britain. Analysis at a finer level is now permitted by the use of modern molecular techniques. A 13-bp duplication in the chloroplast tRNA(Leu1) intron occurs in natural populations of East Anglian oaks, but is not found in other parts of Britain or from mainland Europe. The distribution of this marker suggests that the mutation occurred either in southern England, or during migration from the mainland, and became fixed in a source population from which East Anglia was colonized. Planting of non-native trees for roadside boundaries and in the grounds of old houses and estates, explains the absence of the marker from some East Anglian oaks.

Biological Evolution↗

Possible origin of a B chromosome deduced from its DNA composition using double FISH technique.

Double fluorescent in situ hybridization (FISH) with two DNA probes (a 180 bp tandemly repeated DNA and ribosomal DNA) was performed in embryo cells of the grasshopper Eyprepocnemis plorans. Repetitive DNA was present in most standard chromosomes (excepting 7, 8 and 10) and in the proximal two-thirds of the B chromosome, which was its major location in the complement. Ribosomal DNA was present distally on the B, and in the active nucleolar organizer regions (NORs) of the X, 9, 10 and 11 chromosomes. A small number of rRNA gene clusters was also observed in the pericentromeric regions of chromosomes 1-8. The double FISH technique showed that the B chromosome (B2 type) is mainly composed of a 180 bp tandem repeat and ribosomal DNA, the minute short arm being the only region that does not hybridize with them. The location and order of the centromere and both the DNA sequences on the B chromosome coincide only with those in the X chromosome, indicating that the B most probably derives from the X.

Animals↗

Nuclear DNA introgression across a Pyrenean hybrid zone between parapatric subspecies of the grasshopper Chorthippus parallelus.

Two parapatric subspecies of the European grasshopper Chorthippus parallelus meet along the Pyrenees and form a hybrid zone. A nuclear DNA sequence marker (cpnl-1), involving the presence or absence of a 5 bp insertion, was found to differentiate between the two subspecies along either side of the High Pyrenees but further electrophoretic and sequence analyses revealed that considerable mixing of the subspecific genomes had occurred towards the western and eastern ends of the Pyrenees. The cline for this marker was relatively narrow in two adjoining western central high cols (Peyrelue: 9.7 km and Portalet: 13.3 km) but was significantly wider in another central high col towards the east (Quillane: 42.3 km), indicating that a different combination of forces has been operating on this locus in different regions of the Pyrenees.

Animals↗

Presence of highly repetitive DNA sequences in Tribolium flour-beetles.

Digestion of genomic DNA from seven species of Tribolium (Coleoptera) with Sau3AI, TaqI and ClaI restriction enzymes shows the presence of remarkable amounts of highly repetitive DNA sequences in these species. In Tribolium freemani the sequences are tandemly repeated with a satellite monomer of 166 bp, A-T rich (70.5 per cent), representing 31 per cent of the total genome and located in centromeric chromosome areas as demonstrated by in situ hybridization. The sequence has the potential to form secondary structures such as stems or cruciforms due to the presence of frequent inverted repeats. Tribolium castaneum, T. anaphe and T. madens show homologous sequences to T. freemani satellite DNA but T. confusum, T. audax, T. brevicornis and other tenebrionid beetles, such as Tenebrio molitor and Misolampus goudoti, do not. A phylogenetic dendrogram, based on the homology and abundance of highly repetitive sequences deduced by dot-blot hybridization, chemotaxonomic and karyological characters, is proposed for the seven studied species of Tribolium.

Animals↗

Nuclear DNA sequence divergence between parapatric subspecies of the grasshopper Chorthippus parallelus.

Sequence data from a non-coding nuclear DNA segment (Cpnl-1) was used to investigate the divergence between Spanish and French subspecies of the grasshopper Chorthippus parallus which form a hybrid zone in the Pyrenees. The spatial distribution of eighteen Cpnl-1 sequence haplotypes from eighteen French and fifteen Spanish populations and phylogenetic analyses using both distance and parsimony procedures provide strong evidence that the Pyrenean hybrid zone was formed by postglacial secondary contact and that the French subspecies arose by range expansion from a Balkan or Italian refugium. In addition, the data provide evidence for subdivision between grasshoppers from Northern Spain and those from Central and Pyrenean Spanish populations.

Animals↗