The correlation between demography and metabolic rate: a test using the beach vole (Microtus breweri) and the meadow vole (Microtus pennsylvanicus)

Summary This study examines the hypothesis that mammalian species with wide fluctuations in population size will have greater metabolic rates than species with smaller population fluctuations. We tested this hypothesis using two microtine rodents — the beach vole (Microtus breweri) and the meadow vole (M. pennsylvanicus). Although these species experience similar climatic regimes, eat similar foods, and have a very close phylogenetic relationship, they show marked differences in demography. Microtus pennsylvanicus is prone to large supraannual fluctuations in population size, while M. breweri is essentially acyclical. Metabolic rate (oxygen consumption) of each species was measured using open-flow respirometry at ambient temperatures ranging from 2 to 34° C. Basal metabolic rate of M. pennsylvanicus (1.81 ml O₂ g^–1 h^–1) was significantly greater than that of M. breweri (1.39 ml O₂ g^–1 h^–1). The lower critical temperature, estimated by continuous two-phase regression, was 28.9° C for M. pennsylvanicus and 29.8° C for M. breweri. Regression lines below thermoneutrality did not differ in slope, but the elevation for M. pennsylvanicus was significantly higher. Thus, M. pennsylvanicus has a higher metabolic rate at all temperatures examined. These results support the hypothesis that metabolic rate is positively correlated with the extent of population fluctuation. We suggest that further evidence for, or against, this hypothesis should be found by comparing closely matched species pairs, rather than resorting to confounded allometric comparisons of ecologically and phylogenetically diverse taxa.     

Molecular phylogeny of the speciose vole genus Microtus (Arvicolinae, Rodentia) inferred from mitochondrial DNA sequences

Voles of the genus Microtus represent one of the most speciose mammalian genera in the Holarctic. We established a molecular phylogeny for Microtus to resolve contentious issues of systematic relationships and evolutionary history in this genus. A total of 81 specimens representing ten Microtus species endemic to Europe as well as eight Eurasian, six Asian and one Holarctic species were sequenced for the entire cytochrome b gene (1140 bp). A further 25 sequences were retrieved from GenBank, providing data on an additional 23, mainly Nearctic, Microtus species. Phylogenetic analysis of these 48 species generated four well-supported monophyletic lineages. The genus Chionomys, snow voles, formed a distinct and well-supported lineage separate from the genus Microtus. The subgenus Microtus formed the strongest supported lineage with two sublineages displaying a close relationship between the arvalis species group (common voles) and the socialis species group (social voles). Monophyly of the Palearctic pitymyid voles, subgenus Terricola, was supported, and this subgenus was also subdivided into two monophyletic species groups. Together, these groupings clarify long-standing taxonomic uncertainties in Microtus. In addition, the "Asian" and the Nearctic lineages reported previously were identified although the latter group was not supported. However, relationships among the main Microtus branches were not resolved, suggesting a rapid and potentially simultaneous radiation of a widespread ancestor early in the history of the genus. This and subsequent radiations discernible in the cytochrome b phylogeny, show the considerable potential of Microtus for analysis of historical and ecological determinants of speciation in small mammals. It is evident that speciation is an ongoing process in the genus and that the molecular data provides a vital insight into current species limits as well as cladogenic events of the past.     

Non-concerted evolution of the RET76 satellite DNA family in Reticulitermes taxa (Insecta, Isoptera)

The evolutionary dynamics of satellite DNA is most often studied in canonical mating systems, where bisexuality and panmixis are the rule. In eusocial termites, the limited number of reproducers starting a new colony and the maintenance of the colony through few neotenics act as bottle-necks both in space and time. No data on repetitive DNA are available for Isoptera and for their peculiar reproductive strategy. Here we present the first satellite DNA family isolated in European Reticulitermes. RET76 is a G+C rich satellite embodying two sub-families with a 76 bp monomer. RET76 sequences are highly variable (sequence homology is lower than 80% within sub-families and lower than 68% in the entire family) and this variability is equally distributed among the eight analysed taxa, thus depicting a pattern of non-concerted evolution. The absence of variant fixation – together with the strict monomer length conservation – may be explained at the molecular level as due to functional constraints acting on these sequences, and/or at the organismic level by considering the involvement of eusociality in preventing or greatly reducing variant fixation, somehow mimicking an unisexual strategy.     

Accumulation of Y-specific satellite DNAs during the evolution of <Emphasis Type="Italic">Rumex acetosa</Emphasis> sex chromosomes

The study of the molecular structure of young heteromorphic sex chromosomes of plants has shed light on the evolutionary forces that control the differentiation of the X and Y during the earlier stages of their evolution. We have used the model plant Rumex acetosa, a dioecious species with multiple sex chromosomes, 2n = 12 + XX female and 2n = 12 + XY₁Y₂ male, to analyse the significance of repetitive DNA accumulation during the differentiation of the Y. A bulk segregant analysis (BSA) approach allowed us to identify and isolate random amplified polymorphic DNA (RAPD) markers linked to the sex chromosomes. From a total of 86 RAPD markers in the parents, 6 markers were found to be linked to the Ys and 1 to the X. Two of the Y-linked markers represent two AT-rich satellite DNAs (satDNAs), named RAYSII and RAYSIII, that share about 80% homology, as well as with RAYSI, another satDNA of R. acetosa. Fluorescent in situ hybridisation demonstrated that RAYSII is specific for Y₁, whilst RAYSIII is located in different clusters along Y₁ and Y₂. The two satDNAs were only detected in the genome of the dioecious species with XX/XY₁Y₂ multiple sex chromosome systems in the subgenus Acetosa, but were absent from other dioecious species with an XX/XY system of the subgenera Acetosa or Acetosella, as well as in gynodioecious or hermaphrodite species of the subgenera Acetosa, Rumex and Platypodium. Phylogenetic analysis with different cloned monomers of RAYSII and RAYSIII from both R. acetosa and R. papillaris indicate that these two satDNAs are completely separated from each other, and from RAYSI, in both species. The three Y-specific satDNAs, however, evolved from an ancestral satDNA with repeating units of 120 bp, through intermediate satDNAs of 360 bp. The data therefore support the idea that Y-chromosome differentiation and heterochromatinisation in the Rumex species having a multiple sex chromosome system have occurred by different amplification events from a common ancestral satDNA. Since dioecious species with multiple XX/XY₁Y₂ sex chromosome systems of the section Acetosa appear to have evolved from dioecious species with an XX/XY system, the amplification of tandemly repetitive elements in the Ys of the section Acetosa is a recent evolutionary process that has contributed to an increase in the size and differentiation of the already non-recombining Y chromosomes.     

Closely relatedAllium species (Alliaceae) share a very similar satellite sequence

A satellite sequence repeat ofAllium cepa was tested by fluorescent in situ hybridization (FISH) for cross-hybridization to chromosomes of 27 species (in 37 accessions) belonging to 14 sections of four subgenera ofAllium. All investigated species of sect.Cepa, with the two subsects.Cepa andPhyllodolon, revealed clear satellite-specific hybridization signals mainly at their chromosome termini. The tested species belonging to other sections/subgenera revealed no hybridization signals. An exception wasA. roylei, assigned to sect.Oreiprason. Its chromosomes also showed strong terminal hybridization signals. This and other features suggest a close relationship ofA. roylei to the species of sect.Cepa in spite of deviating morphological characters. The divergence between the satellite repeats to species to which theA. cepa repeat cross-hybridized was determined and revealed high degrees of similarity. Therefore, we conclude that this satellite sequence had evolved already in progenitor forms of sect.Cepa and remained unusually well conserved during speciation. This might indicate selection pressure exerted on a secondarily acquired telomere function of the satellite sequence.Dedicated to Dr habil.Peter Hanelt on the occasion of his 65th birthday.     

Distribution and evolution of two satellite DNAs in the genus Beta

Summary EcoRI monomers of a highly repetitive DNA family of Beta vulgaris have been cloned. Sequence analysis revealed that the repeat length varies between 157–160 bp. The percentage of AT-residues is 62% on average. The basic repeat does not show significant homology to the BamHI sequence family of B. vulgaris that was analyzed by us earlier. Both the EcoRI and BamHI sequences are investigated and compared to each other with respect to their genomic organization in the genus Beta. Both repeats were found to be tandemly arranged in the genome of B. vulgaris in a satellite-like manner. The EcoRI satellite DNA is present in three sections (Beta, Corollinae and Nanae) of the genus, whereas the BamHI satellite DNA exists only in the section Beta. The distribution of the EcoRI and BamHI satellite families in the genus is discussed with respect to their evolution.     

Genetic control of enzyme polymorphisms in the California vole,Microtus californicus

Inheritance of 15 polymorphic isozymes was investigated in captive Microtus californicus. Eleven of the isozymes show patterns consistent with a Mendelian model of inheritance: glycerol-3-phosphate dehydrogenase (GPD), lactate dehydrogenases A and B (LDH-A and LDH-B), malic enzyme 2 (ME-2), isocitrate dehydrogenase 1 (ICD-1), phosphogluconate dehydrogenase (PGD), glutamate-oxaloacetate transaminase 1 (GOT-1) phosphoglucomutase 2 (PGM-2), leucine aminopeptidase (LAP), glucosephosphate isomerase (GPI), and esterase 2 (ES-2, from kidney). four of the isozymes show patterns that cannot be interpreted by a simple genetic model: esterases 1 and 4 (ES-1, ES-4, from hemolysate), esterase 3 (ES-3, from plasma), and protein 1 (PT-1). The following pairs of loci are assorting independently: LAP and PGD, LAP and PGM-2, GOT-1 and PGD, GOT-1 and GPD, LAP and GPD, GPD and PGD, GPI and PGD. Data from one test cross mating indicate that GPD and PGM-2 are loosely linked with recombination about 30%. Additional data are needed to confirm this relationship.This study was conducted while the first author was the recipient of an NIH Traineeship. The Departments of Genetics and Zoology provided financial support for the maintenance of the animal colony. This work was supported in part by an NIH-Biomed grant (3-S05-RR-07006-08S1) to W. Z. Lidicker.     

Different AT-rich satellite DNAs in Cucurbita pepo and Cucurbita maxima

Summary The AT-rich highly repeated satellite DNA of Cucurbita pepo (zucchini) and Cucurbita maxima (pumpkin) were cloned and their DNA structure was investigated. DNA sequencing revealed that the repeat length of satellite DNA in Cucurbita pepo is 349–352 base pairs. The percentage of AT-base pairs is about 61%. This satellite is highly conserved in restriction enzyme pattern and DNA sequence; sequence heterogeneity is about 10%. In contrast, the satellite DNA of Cucurbita maxima has a repeat length of 168–169 base pairs. This satellite is also rich in AT-base pairs (64%), existing in at least three different variants as revealed by restriction enzyme analysis and DNA sequencing. The sequence heterogeneity between these variants is about 15%. The two satellite DNAs showed no cross-hybridization to each other and sequence homology is only limited. Nevertheless, we found in the C. pepo genome a high amount of sequences resembling the satellite of C. maxima. In contrast, the satellite repeat of C. pepo is found in the C. maxima DNA only in a few copies. These observations were discussed with respect to satellite DNA evolution and compared to the data received from monocotyledonous species.     

Insertion and amplification of a DNA sequence in satellite DNA of Cucumis sativus L. (cucumber)

Summary Another satellite DNA repeat (type IV) in the genome of Cucumis sativus (cucumber) was found and investigated with respect to DNA sequence, methylation, and evolution. This satellite shows a repeat length of 360 bp and a GC-content of 47%. The repeats of type IV are highly conserved among each other. Evidence for CG and CNG methylation is presented. By comparison to the previously described satellites (type I/II and type III) from cucumber, it is evident that this repeat is created by an insertion of a 180 bp DNA sequence similar to type I–III into another DNA sequence (or vice versa), and subsequent amplification forming a new satellite repeat. The different satellites of the type I/II, type III, and the 180 bp insert of type IV show a sequence homology of 60%–70%, indicating that the complex satellite DNA of cucumber is originated from a common progenitor by mutation, additional insertion, and amplification events. Copies of a sequence similar to a part of type IV are present in the genome of the related species Cucumis melo (melon).     

A high amount of satellite DNA in the genome of Lupinus angustifolius L.

Embryo DNA, isolated from ungerminated seeds of Lupinus angustifolius L., contains an exceptionally high amount of guanine-cytosine-rich satellite DNA. The thermal denaturation curve of total embryo DNA is biphasic with an inflexion point at 62% denaturation, indicating the presence of satellite DNA. The satellite fraction could be separated from the mainband DNA by three successive preparative CsCl-gradient centrifugations. The densities of the DNA fractions are 1.7045 g cm^-3 and 1.6925 g cm^-3, respectively. The percentages of guanine-cytosine calculated from these densities are comparable to the percentages of GC calculated from the melting temperatures. Finally, ressociation studies prove that foldback DNA and highly repeated sequences are much more frequent in the satellite DNA fraction than in the mainband DNA.Abbreviation C _o t the product of the DNA concentration (mol nucleotides l^-1) and the time (s) of incubation in a DNA reassociation reaction - GC guanine-cytosine - np nucleotide parirs - T temperature interval between 16 and 84% denaturation     

Wide distribution of related satellite DNA families within the genus <Emphasis Type="Italic">Pimelia</Emphasis> (Tenebrionidae)

Major satellites of species in the genus Pimelia comprise large portions of their genomes and belong to seven major satellite families which all originate from a common ancestral sequence. Here we present the results of comprehensive screening of 26 Pimelia species belonging to three distinct geographic groups (Ibero-Balearic, African and Canary Islands) for the presence of different Pimelia satellite families in their genomes. Dot-blot hybridization experiments suggest that together with one dominant, highly abundant satellite family, other families are also present in genomes of the majority of examined Pimelia species, but as low-copy number repeats. The estimated abundance of these underrepresented repeats is about 4,000 copies per haploid genome. Signals of highly abundant satellite family from P. scabrosa (PSCA) in examined congeneric species, obtained after PCR amplification and Southern hybridization under high stringency conditions, corroborate sequence preservation of low-copy representatives of satellite families. PRINS localized low-copy repeats within the pericentromeric regions of all chromosomes. These results point to the existence of an extensive library of repetitive DNAs that was already present in the genome of the common ancestor of extant Pimelia taxa, and shifts the period of diversification of Pimelia satellites far in the history of this genus.     

Nucleotide sequence and genomic organization of a tandem satellite array from the rock vole Microtus chrotorrhinus (Rodentia)

A tandem satellite array (herein named MSAT-160) has been isolated and characterized from the rodent Microtus chrotorrhinus. Sequence data from 15 partial or complete monomers revealed a repeat unit length of 160 bp. This unit length was apparently derived from two shorter sub-motifs, one a tetramer (GAAA), the other a hexamer (CTTTCT), through polymerase slippage and mutation. Collectively, perfect or imperfect variants of these two motifs comprise nearly 60% of the component. Southern blot analyses of genomic DNA digested with 14 different restriction endonucleases indicated that most enzymes yielded either classical type A or type B restriction patterns, while RsaI yielded a pattern that combined features of both the A and B types, and BamHI appeared to lack sites altogether in MSAT-160. An examination of restriction patterns from 16 individuals with three enzymes failed to identify intraspecific variation, while a related study compared 11 species and documented interspecific distinctiveness (Modi, submitted). Fluorescence in situ hybridization indicated that the satellite DNA was located at the centromeres of several autosomes and at sex chromosome heterochromatin (GenBank accession No. M86843).     

Characterization of the satellite DNA Msat-160 from the species <Emphasis Type="Italic">Chionomys</Emphasis><Emphasis Type="Italic">nivalis</Emphasis> (Rodentia,Arvicolinae)

The satellite DNA Msat-160 has been previously characterized in several species of the genus Microtus. Here we present the characterization of Msat-160 from Chionomys nivalis, a species with a very primitive karyotype. As in other Microtus species analyzed, C. nivalis Msat-160 is AT rich, has a monomer length of 160 bp, is undermethylated and is mainly located in all the pericentromeric heterochromatin of all autosomes and the X chromosome, but is completely absent from the Y chromosome. Hence, our results support the hypothesis that Msat-160 was initially distributed in the pericentromeric heterochromatin of all autosomes and the X chromosome. The taxonomic status of the genus Chionomys in relation to the genus Microtus is a very interesting issue, so we constructed phylogenetic dendrograms using Msat-160 sequences from several Microtus species. Although the results were not informative about this issue, the presence of Msat-160 in C. nivalis and Microtus species suggested that both genera are closely related and that this satellite DNA was present in the common ancestor. Studies of Msat-160 in different arvicoline species could help to determine the origin of this satellite and, perhaps, to establish the phylogenetic relationships of some arvicoline groups.     

Quantitative nuclear DNA differences associated with genome evolution in Guizotia (Compositae)

The present communication deals with 2C nuclear genome size variation in a fairly small genus Guizotia. Twenty-four accessions belonging to six species, out of seven known, were analysed in order to elucidate the extent of DNA variation both at an intra—as well as interspecific level. At the intraspecific level none of the species exhibited significant differences in their genome size. Between the species, the 2C DNA amounts ranged from 3.61 pg in G. reptans to 11.37 pg in G. zavattarii; over three-fold DNA variation is evident. Apparently these interspecific DNA differences have been achieved independent of the numerical chromosomal change(s), as all the Guizotias share a common chromosome number 2n=2x=30. The cultivated oilseed crop, G. abyssinica (7.57 pg), has accommodated nearly 78% extra DNA in its chromosome complement during the evolutionary time scale of its origin and domestication from the wild progenitor G. schimperi (4.25 pg). The extent of genomic DNA difference(s) between the species has been discussed in the light of their interrelationships and diversity.     

Microsatellite markers for the common vole (<Emphasis Type="Italic">Microtus arvalis</Emphasis>) and their cross-species utility

Fragmentation of natural habitats of the common vole (Microtus arvalis) provides an excellent model system to study the consequences of restricted gene flow and small population sizes for isolated populations. Here we describe the isolation and characteristics of 10 autosomal and one X-linked microsatellite marker. These new markers were tested in 24 voles from a natural population in eastern Germany. Loci were highly polymorphic with numbers of alleles per locus ranging from three to 26 and expected heterozygosities from 0.51 to 0.97. All loci except for the X-linked locus Mar105 followed Hardy–Weinberg expectations. Cross-species amplifications revealed that most loci were polymorphic as well in M. agrestis, M. thomasi, and M. pennsylvanicus.     

Ubiquitin genes as a paradigm of concerted evolution of tandem repeats

Summary Ubiquitin is remarkable for its ubiquitous distribution and its extreme protein sequence conservation. Ubiquitin genes comprise direct repeats of the ubiquitin coding unit with no spacers. The nucleotide sequences of several ubiquitin repeats from each of humans, chicken,Xenopus, Drosophila, barley, and yeast have recently been determined. By analysis of these data we show that ubiquitin is evolving more slowly than any other known protein, and that this (together with its gene organization) contributes to an ideal situation for the occurrence of concerted evolution of tandem repeats. By contrast, there is little evidence of between-cluster concerted evolution. We deduce that in ubiquitin genes, concerted evolution involves both unequal crossover and gene conversion, and that the average time since two repeated units within the polyubiquitin locus most recently shared a common ancestor is approximately 38 million years (Myr) in mammals, but perhaps only 11 Myr inDrosophila. The extreme conservatism of ubiquitin evolution also allows the inference that certain synonymous serine codons differing at the first two positions were probably mutated at single steps.     

A major satellite DNA of soybean is a 92-base pairs tandem repeat

We report the cloning, sequencing and analysis of the major repetitive DNA of soybean (Glycine max). The repeat, SB92, was cloned as several monomers and trimers produced by digestion with XhoI. The deduced consensus sequence of the repeat is 92 base pairs long. Genomic sequences do not fluctuate in length. Their average homology to the consensus sequence is 92%. The consensus of SB92 contains slightly degenerated homologies for several 6-cutters. Therefore, many of them generate a ladder of 92-bp oligomers. The distribution of bands seems to be random, but the occurrence of sites for different 6-cutters varies widely. There is no obvious correlation between the sequences of the neighboring units of SB92 in cloned trimers. Also, there are none of the internal repetitive blocks reported for many satellite DNAs from other species. The SB92 repeat makes up 0.7% of total soybean DNA. This is equivalent to 8×10⁴ copies, or 7 megabases. The repeat is organized in giant tandem blocks over 1 Mb in length, and there are fewer blocks than chromosomes. The polymorphism of these blocks is extremely high. The SB92 repeat is present in identical arrangement and number of copies in the ancestral subspecies Glycine soja. There are 10 times fewer copies of the repeat in a related species Vigna unguiculata (cowpea), and no homologies in several other more distant leguminous plants studied.     

Nuclear DNA changes,genome differentiation and evolution inNicotiana (Solanaceae)

A survey of 51 species fromNicotiana subgg.Tabacum, Rustica andPetunioides has shown that evolution was accompanied by a five-fold variation in nuclear DNA amounts. This variation, however, was not directly correlated with the changes in chromosome number. Drastic rearrangement of karyotypes is characteristic for the evolution ofNicotiana spp. Significant gain or loss in nuclear DNA has often accompanied such changes, but DNA variation has also occurred without significant changes in karyotype arrangements.—The distribution of nuclear DNA is discontinuous inNicotiana, species cluster into DNA groups with consistently regular increments in the mean DNA amounts. The discontinuities are viewed as steady states in terms of genomic balance and biological fitness.—Changes in the amount of nuclear DNA and in the heterochromatin are compared with the morphological, chromosomal and adaptive changes which accompanied speciation in 14 subgeneric sections. The evolutionary significance of DNA variation is discussed.