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The number of mutations selected during adaptation in a laboratory population of Saccharomyces cerevisiae.

There is currently limited empirical and theoretical support for the prevailing view that adaptation typically results from the fixation of many mutations, each with small phenotypic effects. Recent theoretical work suggests that, on the contrary, most of the phenotypic change during an episode of adaptation can result from the selection of a few mutations with relatively large effects. I studied the genetics of adaptation by populations of budding yeast to a culture regime of daily hundredfold dilution and transfer in a glucose-limited minimal liquid medium. A single haploid genotype isolated after 2000 generations showed a 76% fitness increase over its ancestor. This evolved haploid was crossed with its ancestor, and tetrad dissections were used to isolate a complete series of six meiotic tetrads. The Castle-Wright estimator of the number of loci at which adaptive mutations had been selected, modified to account for linkage and variation among mutations in the size of their effect, is 4.4. The estimate for a second haploid genotype, isolated from a separate population and with a fitness gain of 60%, was 2.7 loci. Backcrosses to the ancestor with the first evolved genotype support the inference that adaptation resulted primarily from two to five mutations. These backcrosses also indicated that deleterious mutations had hitchhiked with adaptive mutations in this evolved genotype.

Acclimatization↗

Assessing quality in the cardiac catheterization laboratory.

Quality assurance and improvement have increasingly been the focus of health care providers, third-party payers, and patients. Because cardiovascular procedures are common, easily identifiable with claims data, and account for a relatively large proportion of health care expenditures, particular attention has been paid to quality assurance in the setting of the diagnostic and interventional cardiac catheterization laboratory. The structure, process, and outcomes domains of quality measurement in the interventional laboratory involve the maintenance of volume standards, the availability of surgical backup, consistent tracking of procedural outcomes and complications so they can be compared with national standards, and the application of evidence-based therapy. Quality assurance i the diagnostic laboratory revolves around the clinical proficiency of the operators, the maintenance and management of catheterization laboratory equipment, and the presence of a continuous quality improvement program. The evolution of interventional equipment and techniques along with the establishment of national registries has led to a gradual improvement in the quality of percutaneous coronary intervention. Given the dynamic nature of cardiology, adaptable quality assurance and quality improvement programs will remain the foundation of successful catheterization labs.

Benchmarking↗

The development of genetically inherited resistance to zinc in laboratory-selected generations of the earthworm Eisenia fetida.

The capacity of species to adapt both physiologically and genetically to contaminants may allow populations to persist in polluted environments. Such 'adaptation' can have important implications for risk assessment, since it may mean that prediction based on extrapolation of toxicity studies with naïve populations may prove invalid for long-term contaminated sites. To investigate the evolution of zinc resistance in Eisenia fetida, worms from a previously unexposed population (parent) were selected and reared over two generations (F1, F2) while exposed to zinc in the laboratory. Relative sensitivities of unexposed and selected generations were then compared by exposing parent, F1 and F2 individuals to zinc in contact filter papers tests. Calculation of effect concentrations from this work indicated differences in sensitivity to zinc for successive generations, with higher toxicity values (LC(50), LC(90), LC(99)) found for the selected worms. The increases in resistance found for F1 and F2 worms were confirmed in a discriminating dose study. In addition to comparing the sensitivities of the parent, F1 and F2 generations for zinc, toxicity tests were also conducted with copper to assess if there was evidence of cross-resistance between the two metals. Results indicated similar increases in resistance to copper to those found for zinc. Mechanisms underlying the increased metal resistance were studied in toxicokinetic experiments. Results indicated no clear trends between the three generations indicating that physiological responses, other than differences in kinetic parameters, are responsible for the increased resistance found in the selected worms.

Journal Article↗

Genome variation and evolution of the malaria parasite Plasmodium falciparum.

Infections with the malaria parasite Plasmodium falciparum result in more than 1 million deaths each year worldwide. Deciphering the evolutionary history and genetic variation of P. falciparum is critical for understanding the evolution of drug resistance, identifying potential vaccine candidates and appreciating the effect of parasite variation on prevalence and severity of malaria in humans. Most studies of natural variation in P. falciparum have been either in depth over small genomic regions (up to the size of a small chromosome) or genome wide but only at low resolution. In an effort to complement these studies with genome-wide data, we undertook shotgun sequencing of a Ghanaian clinical isolate (with fivefold coverage), the IT laboratory isolate (with onefold coverage) and the chimpanzee parasite P. reichenowi (with twofold coverage). We compared these sequences with the fully sequenced P. falciparum 3D7 isolate genome. We describe the most salient features of P. falciparum polymorphism and adaptive evolution with relation to gene function, transcript and protein expression and cellular localization. This analysis uncovers the primary evolutionary changes that have occurred since the P. falciparum-P. reichenowi speciation and changes that are occurring within P. falciparum.

Animals↗

The contribution of ancestry, chance, and past and ongoing selection to adaptive evolution.

The relative contributions of ancestry, chance, and past and ongoing selection to variation in one adaptive (larval feeding rate) and one seemingly nonadaptive (pupation height) trait were determined in populations of Drosophila melanogaster adapting to either low or high larval densities in the laboratory. Larval feeding rates increased rapidly in response to high density, and the effects of ancestry, past selection and chance were ameliorated by ongoing selection within 15-20 generations. Similarly, in populations previously kept at high larval density, and then switched to low larval density, the decline of larval feeding rate to ancestral levels was rapid (15-20 generations) and complete, providing support for a previously stated hypothesis regarding the costs of faster feeding in Drosophila larvae. Variation among individuals was the major contributor to variation in pupation height, a trait that would superficially appear to be nonadaptive in the environmental context of the populations used in this study because it did not diverge between sets of populations kept at low versus high larval density for many generations. However, the degree of divergence among populations (F(ST)) for pupation height was significantly less than expected for a selectively neutral trait, and we integrate results from previous studies to suggest that the variation for pupation height among populations is constrained by stabilizing selection, with a flat, plateau-like fitness function that, consequently, allows for substantial phenotypic variation within populations. Our results support the view that the genetic imprints of history (ancestry and past selection) in outbreeding sexual populations are typically likely to be transient in the face of ongoing selection and recombination. The results also illustrate the heuristic point that different forms of selection-for example directional versus stabilizing selection-acting on a trait in different populations may often not be due to differently shaped fitness functions, but rather due to differences in how the fitness function maps onto the actual distribution of phenotypes in a given population. We discuss these results in the light of previous work on reverse evolution, and the role of ancestry, chance, and past and ongoing selection in adaptive evolution.

Adaptation, Physiological↗

Habitat-specific pigmentation in a freshwater isopod: adaptive evolution over a small spatiotemporal scale.

Pigmentation in the freshwater isopod Asellus aquaticus (Crustacea) differed between habitats in two Swedish lakes. In both lakes, isopods had lighter pigmentation in stands of submerged vegetation, consisting of stoneworts (Chara spp.), than in nearby stands of reed (Phragmites australis). Experimental crossings of light and dark isopods in a common environment showed that pigmentation had a genetic basis and that genetic variance was additive. Environmental effects of diet or chromatophore adjustment to the background had minor influence on pigmentation, as shown by laboratory rearing of isopods on stonewort or reed substrates, as well as analyses of stable isotope ratios for isopods collected in the field. In both study lakes, the average phenotype became lighter with time (across generations) in recently established stonewort stands. Taken together, these results indicate that altered phenotype pigmentation result from evolutionary responses to local differences in natural selection. Based on the assumption of two generations per year, the evolutionary rate of change in pigmentation was 0.08 standard deviations per generation (haldanes) over 20 generations in one lake and 0.22 haldanes over two generations in the other lake. This genetic change occurred during an episode of population growth in a novel habitat, a situation known to promote adaptive evolution. In addition, stonewort stands constitute large and persistent patches, characteristics that tend to preserve local adaptations produced by natural selection. Results from studies on selective forces behind the adaptive divergence suggest that selective predation from visually oriented predators is a possible selective agent. We found no indications of phenotype-specific movements between habitats. Mating within stonewort stands was random with respect to pigmentation, but on a whole-lake scale it is likely that mating is assortative, as a result of local differences in phenotype distribution.

Adaptation, Physiological↗

Genetic variability and adaptive evolution in parthenogenetic root-knot nematodes.

Root-knot nematodes (RKN) of the genus Meloidogyne are biotrophic plant parasites of major agricultural importance, which exhibit very variable modes of reproduction, from classical amphimixis to mitotic parthenogenesis. This review focuses on those RKN species that reproduce exclusively by mitotic parthenogenesis (apomixis), in contrast to those that have meiotic/amphimitic events in their life cycle. Although populations of clonal organisms are often represented as being ecologically isolated and evolutionary inert, a considerable volume of literature provides evidence that asexual RKN are neither: they are widely distributed, extremely polyphagous, and amenable to selection and adaptive variation. The ancestors of the genus are unknown, but it is assumed that the parthenogenetic RKN have evolved from amphimictic species through hybridization and subsequent aneuploidization and polyploidization events. Molecular studies have indeed confirmed that the phylogenetic divergence between meiotic and mitotic RKN lineages occurred early, and have revealed an unexpected level of clonal diversity among populations within apomictic species. Laboratory experiments have shown that asexual RKN can rapidly adapt to new environmental constraints (eg host resistance), although with some fitness costs. Lastly, the molecular and chromosomal mechanisms that could contribute to genome plasticity leading to persistent genetic variation and adaptive evolution in apomictic RKN are discussed. It is concluded that RKN provide an excellent model system in which to study the dynamic nature and adaptive potential of clonal genomes.

Adaptation, Physiological↗

Evolution of high mutation rates in experimental populations of E. coli.

Most mutations are likely to be deleterious, and so the spontaneous mutation rate is generally held at a very low value. Nonetheless, evolutionary theory predicts that high mutation rates can evolve under certain circumstances. Empirical observations have previously been limited to short-term studies of the fates of mutator strains deliberately introduced into laboratory populations of Escherichia coli, and to the effects of intense selective events on mutator frequencies in E. coli. Here we report the rise of spontaneously originated mutators in populations of E. coli undergoing long-term adaptation to a new environment. Our results corroborate computer simulations of mutator evolution in adapting clonal populations, and may help to explain observations that associate high mutation rates with emerging pathogens and with certain cancers.

Adaptation, Physiological↗

"Protoplasm...is soft wax in our hands": Paul Kammerer and the art of biological transformation.

Paul Kammerer's career ended in scandal in 1926 over tampering with his evidence for "Lamarckian" evolution--the infamous midwife toad. But although Kammerer's conclusions proved false, his evidence was probably genuine. In any case his arguments were not simply for Lamarckism and against Darwinism, as the theories are understood today. If we look beyond the scandal, the Kammerer story shows us a great deal about early 20th-century biology: the range of new ideas about heredity and variation, competing theories of biological and cultural evolution and their applications in eugenics, new kinds of laboratories and professional roles for biologists, and changing standards for documenting experimental results.

Adaptation, Physiological↗

Stress-directed adaptive mutations and evolution.

Comparative biochemistry demonstrates that the metabolites, complex biochemical networks, enzymes and regulatory mechanisms essential to all living cells are conserved in amazing detail throughout evolution. Thus, in order to evolve, an organism must overcome new adverse conditions without creating different but equally dangerous alterations in its ongoing successful metabolic relationship with its environment. Evidence suggests that stable long-term acquisitive evolution results from minor increases in mutation rates of genes related to a particular stress, with minimal disturbance to the balanced and resilient metabolism critical for responding to an unpredictable environment. Microorganisms have evolved specific biochemical feedback mechanisms that direct mutations to genes derepressed by starvation or other stressors in their environment. Transcription of the activated genes creates localized supercoiling and DNA secondary structures with unpaired bases vulnerable to mutation. The resulting mutants provide appropriate variants for selection by the stress involved, thus accelerating evolution with minimal random damage to the genome. This model has successfully predicted mutation frequencies in genes of E. coli and humans. Stressed cells observed in the laboratory over hundreds of generations accumulate mutations that also arise by this mechanism. When this occurs in repair-deficient mutator strains with high rates of random mutation, the specific stress-directed mutations are also enhanced.

Adaptation, Biological↗

Molecular evolution of human immunodeficiency virus env in humans and monkeys: similar patterns occur during natural disease progression or rapid virus passage.

Neonatal rhesus macaque 95-3 was inoculated with nonpassaged simian-human immunodeficiency virus strain SHIV-vpu(+), which encodes env of the laboratory-adapted human immunodeficiency virus (HIV) strain IIIB and is considered nonpathogenic. CD4(+) T-cell counts dropped to <200 cells/microl within 4.6 years, and monkey 95-3 died with opportunistic infections 5.9 years postinoculation. Transfer of blood from 95-3 to two naive adult macaques resulted in high peak viral loads and rapid, persistent T-cell depletion. Progeny virus evolved in 95-3 despite high SHIV-vpu(+) neutralizing antibody titers and still used CXCR4 but, in contrast to parental SHIV-vpu(+), productively infected macrophages and resisted neutralization. Sequence analysis revealed three new potential glycosylation sites in gp120; another two were lost. Strikingly similar mutations were detected in a laboratory worker who progressed to AIDS after accidental HIV-IIIB infection (T. Beaumont et al., J. Virol. 75:2246-2252, 2001), thus supporting the SHIV-vpu(+)/rhesus macaque system as a relevant model. Similar mutations were also described after rapid passage of chimeric viruses encoding IIIB env in rhesus and pig-tailed macaques (M. Cayabyab et al., J. Virol. 73:976-984, 1999; Z. Q. Liu et al., Virology 260:295-307, 1999; S. V. Narayan et al., Virology 256:54-63, 1999; R. Raghavan et al., Brain Pathol. 7:851-861, 1997; E. B. Stephens et al., Virology 231:313-321, 1997). Thus, HIV-IIIB env evolved similarly in three different species; this selection occurred in chronically infected individuals during disease progression as well as after rapid virus passage. We postulate that evolutionary pressure led to the outgrowth of more aggressive viral variants in all three species.

Amino Acid Sequence↗

Biological space experiments for the simulation of Martian conditions: UV radiation and Martian soil analogues.

The survivability of resistant terrestrial microbes, bacterial spores of Bacillus subtilis, was investigated in the BIOPAN facility of the European Space Agency onboard of Russian Earth-orbiting FOTON satellites (BIOPAN I -III missions). The spores were exposed to different subsets of the extreme environmental parameters in space (vacuum, extraterrestrial solar UV, shielding by protecting materials like artificial meteorites). The results of the three space experiments confirmed the deleterious effects of extraterrestrial solar UV radiation which, in contrast to the UV radiation reaching the surface of the Earth, also contains the very energy-rich, short wavelength UVB and UVC radiation. Thin layers of clay, rock or meteorite material were shown to be only successful in UV-shielding, if they are in direct contact with the spores. On Mars the UV radiation climate is similar to that of the early Earth before the development of a protective ozone layer in the atmosphere by the appearance of the first aerobic photosynthetic bacteria. The interference of Martian soil components and the intense and nearly unfiltered Martian solar UV radiation with spores of B. subtilis will be tested with a new BIOPAN experiment, MARSTOX. Different types of Mars soil analogues will be used to determine on one hand their potential toxicity alone or in combination with solar UV (phototoxicity) and on the other hand their UV protection capability. Two sets of samples will be placed under different cut-off filters used to simulate the UV radiation climate of Mars and Earth. After exposure in space the survival of and mutation induction in the spores will be analyzed at the DLR, together with parallel samples from the corresponding ground control experiment performed in the laboratory. This experiment will provide new insights into the principal limits of life and its adaptation to environmental extremes on Earth or other planets which and will also have implications for the potential for the evolution and distribution of life.

Bacillus subtilis↗

Life at body temperatures below 0 degrees C: the physiology and biochemistry of Antarctic fishes.

Fishes of the Southern Ocean surrounding Antarctica are dominated by species of the suborder Notothenoidei. For approximately 14MY, these highly successful fishes have evolved under stable thermal conditions that result in a body temperature of ca. 0 degrees C throughout their life histories. Evolution in this chronically cold environment has led to unusual physiological and biochemical characteristics. In some cases, these characteristics are essential to survival and normal biological function at cold body temperature (e.g., development of antifreeze glycoproteins, structural modification of enzymes, cold-stable microtubules, and cardiovascular adaptations). In other instances, mutations that probably would have been lethal in warmer, less oxygen-rich environments than the Southern Ocean have been retained in Antarctic fishes (e.g., loss of hemoglobin production and variable expression of myoglobin in one notothenioid family, the Channichthyidae). These unique animals offer opportunities for insight into evolutionary processes leading to physiological and biochemical characteristics that either arise from strong selective pressure or persist through relaxation of selective pressure. After briefly describing the Antarctic marine environment, I discuss several unique aspects of the physiology and biochemistry of Antarctic fishes, specifically emphasizing our laboratory's recent studies of an unusual pattern of myoglobin expression in the Channichthyid icefishes.

Adaptation, Biological↗

Experimental tests of the adaptive significance of sexual recombination.

Numerous theories have been proposed to explain the advantages of sexual recombination the exchange of hereditary material between different genomes or homologous chromosomes. Many of these candidate benefits have been evaluated in controlled laboratory experiments, which, collectively, strongly indicate that sexual recombination provides important long-term advantages.

Adaptation, Psychological↗

The effects of sex and mutation rate on adaptation in test tubes and to mouse hosts by Saccharomyces cerevisiae.

Some hypotheses for the evolution of sex focus on adaptation to changing or heterogeneous environments, but these hypotheses have rarely been tested. We tested for advantages of sex and of increased mutation rates in yeast strains in two contrasting environments: a standard and relatively homogeneous laboratory environment of minimal medium in test tubes, and the variable environment of a mouse brain experienced by pathogenic strains. Evolving populations were founded as equal mixtures of sexual and obligately asexual genotypes. In the sexuals, cycles of sporulation, meiosis, and mating were induced approximately every 50 mitotic generations, with the asexuals undergoing sporulation but not ploidy cycles or recombination. In both environments, replicate negative control populations established with the same pair of strains were propagated with neither mating nor meiosis. In test tubes with no sex induced, sexuals were fixed in all five replicates within 250 mitotic generations, whereas in mice with no sex induced, asexuals were fixed in all four replicates by 170 generations. Inducing sex altered these outcomes in opposite directions in test tubes and mice, decreasing the fixation frequencies of sexuals in test tubes but increasing them in mice. These contrasts with asexual controls suggest an advantage for sex in mice but not in test tubes, although there was no difference between test tubes and mice in the numbers of populations fixed-for sexuals. In analogous experiments testing for an advantage of increased mutation rates, wild-type genotypes became fixed at the expense of mutators in every replicate of both test tube and mouse populations, indicating a disadvantage for mutators in both environments. Increased rates of point mutation do not appear to accelerate adaptation.

Adaptation, Biological↗

Genetic constraints and the adaptive evolution of rabies virus in nature.

We used a molecular evolutionary approach to investigate the species adaptation of rabies virus in nature. A maximum likelihood analysis of selection pressures revealed that the nucleoprotein (N) and glycoprotein (G) genes of natural viral isolates were highly constrained, especially at nonsynonymous sites, in contrast to the higher rates of nonsynonymous evolution observed in viruses subject to laboratory passage. Positive selection was only found at a single amino acid site--position 183 in the ectodomain of the G gene. The low rate of nonsynonymous evolution in natural isolates of rabies virus may be due to constraints imposed by the need to replicate in multiple cell types within the host, which in turn facilitates cross-species transmission, or because viral proteins are not subject to immune selection. Using known dates in the epidemiologic history of European viral isolates, we estimated that overall rates of nucleotide substitution in rabies virus were similar to those observed in other RNA viruses. Assuming that the average rate of synonymous change does not vary among species, we estimated that the current genetic diversity in lyssavirus genotype 1 may have arisen only during the last 500 years.

Adaptation, Physiological↗

Population genetics of Drosophila amylase. IV. Selection in laboratory populations maintained on different carbohydrates.

Two polymorphic systems impinging on alpha-amylase in Drosophila pseudoobscura have been studied in laboratory populations maintained on medium in which the only carbohydrate source was starch (the substrate of amylase) and replicas maintained on medium in which the only carbohydrate source was maltose (the product of amylase). The two polymorphic systems were alleles at the structural gene (Amy) coding for the enzyme (allozymes) and variation in the tissue-specific expression along the adult midgut controlled by several genes. In the seven populations on maltose medium little consistent change was noted in either system. In the seven populations on starch medium, both polymorphisms exhibited selective changes. A midgut pattern of very limited expression of amylase rose in frequency in all starch populations, as did the frequency of the "fast" (1.00) Amy allele. The overall specific amylase activity did not differ between starch-adapted and maltose-adapted flies.--The results, along with previous studies, indicate that when a gene-enzyme system is specifically stressed in laboratory populations, allozymes often exhibit selective differences. Such results make the selectionist hypothesis at least tenable. Furthermore, the fact that both types of polymorphisms responded to selection indicates the role of structural gene vs. gene regulation changes in adaptive evolution is not an either/or question but one of relative roles and interactions.

Alleles↗

Genomewide evolutionary rates in laboratory and wild yeast.

As wild organisms adapt to the laboratory environment, they become less relevant as biological models. It has been suggested that a commonly used S. cerevisiae strain has rapidly accumulated mutations in the lab. We report a low-to-intermediate rate of protein evolution in this strain relative to wild isolates.

Evolution, Molecular↗