David W. Hall

Nucleotide Polymorphism and Phenotypic Associations Within and Around the phytochrome B2 Locus in European Aspen (Populus tremula, Salicaceae)

We investigated the utility of association mapping to dissect the genetic basis of naturally occurring variation in bud phenology in European aspen (Populus tremula). With this aim, we surveyed nucleotide polymorphism in 13 fragments spanning an 80-kb region surrounding the phytochrome B2 (phyB2) locus. Although polymorphism varies substantially across the phyB2 region, we detected no signs for deviations from neutral expectations. We also identified a total of 41 single nucleotide polymorphisms (SNPs) that were subsequently scored in a mapping population consisting of 120 trees. We identified two nonsynonymous SNPs in the phytochrome B2 gene that were independently associated with variation in the timing of bud set and that explained between 1.5 and 5% of the observed phenotypic variation in bud set. Earlier studies have shown that the frequencies of both these SNPs vary clinally with latitude. Linkage disequilibrium across the region was low, suggesting that the SNPs we identified are strong candidates for being causally linked to variation in bud set in our mapping populations. One of the SNPs (T608N) is located in the “hinge region,” close to the chromophore binding site of the phyB2 protein. The other SNP (L1078P) is located in a region supposed to mediate downstream signaling from the phyB2 locus. The lack of population structure, combined with low levels of linkage disequilibrium, suggests that association mapping is a fruitful method for dissecting naturally occurring variation in Populus tremula.

Abstinence-Only Education and Teen Pregnancy Rates: Why We Need Comprehensive Sex Education in the U.S

The United States ranks first among developed nations in rates of both teenage pregnancy and sexually transmitted diseases. In an effort to reduce these rates, the U.S. government has funded abstinence-only sex education programs for more than a decade. However, a public controversy remains over whether this investment has been successful and whether these programs should be continued. Using the most recent national data (2005) from all U.S. states with information on sex education laws or policies (N = 48), we show that increasing emphasis on abstinence education is positively correlated with teenage pregnancy and birth rates. This trend remains significant after accounting for socioeconomic status, teen educational attainment, ethnic composition of the teen population, and availability of Medicaid waivers for family planning services in each state. These data show clearly that abstinence-only education as a state policy is ineffective in preventing teenage pregnancy and may actually be contributing to the high teenage pregnancy rates in the U.S. In alignment with the new evidence-based Teen Pregnancy Prevention Initiative and the Precaution Adoption Process Model advocated by the National Institutes of Health, we propose the integration of comprehensive sex and STD education into the biology curriculum in middle and high school science classes and a parallel social studies curriculum that addresses risk-aversion behaviors and planning for the future.

Using association mapping to dissect the genetic basis of complex traits in plants

Association or linkage disequilibrium mapping has become a very popular method for dissecting the genetic basis of complex traits in plants. The benefits of association mapping, compared with traditional quantitative trait locus mapping, is, for example, a relatively detailed mapping resolution and that it is far less time consuming since no mapping populations need to be generated. The surge of interest in association mapping has been fueled by recent developments in genomics that allows for rapid identification and scoring of genetic markers which has traditionally limited mapping experiments. With the decreasing cost of genotyping future emphasis will likely focus on phenotyping, which can be both costly and time consuming but which is crucial for obtaining reliable results in association mapping studies. In addition, association mapping studies are prone to the identification of false positives, especially if the experimental design is not rigorously controlled. For example, population structure has long been known to induce many false positives and accounting for population structure has become one of the main issues when implementing association mapping in plants. Also, with increasing numbers of genetic markers used, the problem becomes separating true from false positive and this highlights the need for independent validation of identified association. With these caveats in mind, association mapping nevertheless shows great promise for helping us understand the genetic basis of complex traits of both economic and ecological importance.

Precise estimates of mutation rate and spectrum in yeast.

Significance Spontaneous mutations are rare and difficult to observe in large numbers experimentally. By sequencing the genomes of 145 diploid mutation accumulation (MA) lines of the budding yeast Saccharomyces cerevisiae, we identified nearly 1,000 mutations, a larger number than in any prior eukaryotic MA experiment as far as we are aware. For the first time, to our knowledge, in MA data, we were able to estimate rates of context-dependent single-nucleotide mutations. We were also able to observe mutational classes not seen in earlier yeast MA experiments and infer the rate of strongly deleterious mutations from patterns of missing mutations in each mutational class. Our findings both answer outstanding questions in the field, as well as highlight the need for more studies of spontaneous mutation. Mutation is the ultimate source of genetic variation. The most direct and unbiased method of studying spontaneous mutations is via mutation accumulation (MA) lines. Until recently, MA experiments were limited by the cost of sequencing and thus provided us with small numbers of mutational events and therefore imprecise estimates of rates and patterns of mutation. We used whole-genome sequencing to identify nearly 1,000 spontaneous mutation events accumulated over ∼311,000 generations in 145 diploid MA lines of the budding yeast Saccharomyces cerevisiae. MA experiments are usually assumed to have negligible levels of selection, but even mild selection will remove strongly deleterious events. We take advantage of such patterns of selection and show that mutation classes such as indels and aneuploidies (especially monosomies) are proportionately much more likely to contribute mutations of large effect. We also provide conservative estimates of indel, aneuploidy, environment-dependent dominant lethal, and recessive lethal mutation rates. To our knowledge, for the first time in yeast MA data, we identified a sufficiently large number of single-nucleotide mutations to measure context-dependent mutation rates and were able to (i) confirm strong AT bias of mutation in yeast driven by high rate of mutations from C/G to T/A and (ii) detect a higher rate of mutation at C/G nucleotides in two specific contexts consistent with cytosine methylation in S. cerevisiae.

Genetic differentiation, clinal variation and phenotypic associations with growth cessation across the Populus tremula Photoperiodic Pathway

Perennial plants monitor seasonal changes through changes in environmental conditions such as the quantity and quality of light. To ensure a correct initiation of critical developmental processes, such as the initiation and cessation of growth, plants have adapted to a spatially variable light regime and genes in the photoperiodic pathway have been implicated as likely sources for these adaptations. Here we examine genetic variation in genes from the photoperiodic pathway in Populus tremula (Salicaceae) for signatures diversifying selection in response to varying light regimes across a latitudinal gradient. We fail to identify any loci with unusually high levels of genetic differentiation among populations despite identifying four SNPs that show significant allele frequency clines with latitude. We do, however, observe large covariance in allelic effects across populations for growth cessation, a highly adaptive trait in P. tremula. High covariance in allelic effects is a signature compatible with diversifying selection along an environmental gradient. We also observe significantly higher heterogeneity in genetic differentiation among SNPs from the photoperiod genes than among SNPs from randomly chosen genes. This suggests that spatially variable selection could be affecting genes from the photoperiod pathway even if selection is not strong enough to cause individual loci to be identified as outliers. SNPs from three genes in the photoperiod pathway (PHYB2, LHY1, and LHY2) show significant associations with natural variation in growth cessation. Collectively these SNPs explain 10–15% of the phenotypic variation in growth cessation. Covariances in allelic effects across populations help explain an additional 5–7% of the phenotypic variation in growth cessation.

Molecular evolution and emergence of avian gammacoronaviruses

Abstract Coronaviruses, which are single stranded, positive sense RNA viruses, are responsible for a wide variety of existing and emerging diseases in humans and other animals. The gammacoronaviruses primarily infect avian hosts. Within this genus of coronaviruses, the avian coronavirus infectious bronchitis virus (IBV) causes a highly infectious upper-respiratory tract disease in commercial poultry. IBV shows rapid evolution in chickens, frequently producing new antigenic types, which adds to the multiple serotypes of the virus that do not cross protect. Rapid evolution in IBV is facilitated by strong selection, large population sizes and high genetic diversity within hosts, and transmission bottlenecks between hosts. Genetic diversity within a host arises primarily by mutation, which includes substitutions, insertions and deletions. Mutations are caused both by the high error rate, and limited proof reading capability, of the viral RNA-dependent RNA-polymerase, and by recombination. Recombination also generates new haplotype diversity by recombining existing variants. Rapid evolution of avian coronavirus IBV makes this virus extremely difficult to diagnose and control, but also makes it an excellent model system to study viral genetic diversity and the mechanisms behind the emergence of coronaviruses in their natural host.

Automated reconstruction of ancient languages using probabilistic models of sound change

One of the oldest problems in linguistics is reconstructing the words that appeared in the protolanguages from which modern languages evolved. Identifying the forms of these ancient languages makes it possible to evaluate proposals about the nature of language change and to draw inferences about human history. Protolanguages are typically reconstructed using a painstaking manual process known as the comparative method. We present a family of probabilistic models of sound change as well as algorithms for performing inference in these models. The resulting system automatically and accurately reconstructs protolanguages from modern languages. We apply this system to 637 Austronesian languages, providing an accurate, large-scale automatic reconstruction of a set of protolanguages. Over 85% of the system’s reconstructions are within one character of the manual reconstruction provided by a linguist specializing in Austronesian languages. Being able to automatically reconstruct large numbers of languages provides a useful way to quantitatively explore hypotheses about the factors determining which sounds in a language are likely to change over time. We demonstrate this by showing that the reconstructed Austronesian protolanguages provide compelling support for a hypothesis about the relationship between the function of a sound and its probability of changing that was first proposed in 1955.

Selection Transforms the Landscape of Genetic Variation Interacting with Hsp90

The protein-folding chaperone Hsp90 has been proposed to buffer the phenotypic effects of mutations. The potential for Hsp90 and other putative buffers to increase robustness to mutation has had major impact on disease models, quantitative genetics, and evolutionary theory. But Hsp90 sometimes contradicts expectations for a buffer by potentiating rapid phenotypic changes that would otherwise not occur. Here, we quantify Hsp90’s ability to buffer or potentiate (i.e., diminish or enhance) the effects of genetic variation on single-cell morphological features in budding yeast. We corroborate reports that Hsp90 tends to buffer the effects of standing genetic variation in natural populations. However, we demonstrate that Hsp90 tends to have the opposite effect on genetic variation that has experienced reduced selection pressure. Specifically, Hsp90 tends to enhance, rather than diminish, the effects of spontaneous mutations and recombinations. This result implies that Hsp90 does not make phenotypes more robust to the effects of genetic perturbation. Instead, natural selection preferentially allows buffered alleles to persist and thereby creates the false impression that Hsp90 confers greater robustness.

Adaptive evolution of the Populus tremula photoperiod pathway

Perennial plants monitor seasonal changes through changes in environmental conditions such as the quantity and quality of light and genes in the photoperiodic pathway are known to be involved in controlling these processes. Here, we examine 25 of genes from the photoperiod pathway in Populus tremula (Salicaceae) for signatures of adaptive evolution. Overall, levels of synonymous polymorphism in the 25 genes are lower than at control loci selected randomly from the genome. This appears primarily to be caused by lower levels of synonymous polymorphism in genes associated with the circadian clock. Natural selection appears to play an important role in shaping protein evolution at several of the genes in the photoperiod pathways, which is highlighted by the fact that approximately 40% of the genes from the photoperiod pathway have estimates of selection on nonsynonymous polymorphisms that are significantly different from zero. A surprising observation we make is that circadian clock‐associated genes appear to be over‐represented among the genes showing elevated rates of protein evolution; seven genes are evolving under positive selection and all but one of these genes are involved in the circadian clock of Populus.

Fitness Epistasis among 6 Biosynthetic Loci in the Budding Yeast Saccharomyces cerevisiae

We generated all possible haploid and homozygous diploid genotypes at 6 biosynthetic loci in yeast and scored their fitness to examine whether there was any pattern of weak synergistic epistasis, which is a requirement of the deterministic mutation model for the evolution of sex. We measured 4 components of fitness: haploid growth rate, haploid mating efficiency, diploid growth rate, and diploid sporulation efficiency. We found that in agreement with previous work in yeast, epistasis tended to be small in magnitude and variable in sign, regardless of the fitness component measured. The number of background mutations had either no effect or no consistent effect on epistasis distributions. For every combination of 2 loci in a mutation-free background, we also generated all heterozygous genotypes so that we could partition diploid epistasis into additive x additive, additive x dominance, and dominance x dominance epistasis. Our main interest was in determining whether dominance by dominance epistasis was large and negative, which is a requirement of diploid models with inbreeding to explain high levels of recombination. Dominance by dominance epistasis estimates obtained by partitioning diploid epistasis for growth rates were both positive and negative. With the caveat that our results are based on only 6 biosynthetic loci, epistasis for fitness is not supported as an explanation for the maintenance of sex or the high rate of meiotic recombination in yeast.