Numerous insects have independently evolved the ability to feed on plants that produce toxic secondary compounds called cardenolides and can sequester these compounds for use in their defense. We surveyed the protein target for cardenolides, the alpha subunit of the sodium pump, Na+,K+-ATPase (ATPα), in 14 species that feed on cardenolide-producing plants and 15 outgroups spanning three insect orders. Despite the large number of potential targets for modulating cardenolide sensitivity, amino acid substitutions associated with host-plant specialization are highly clustered, with many parallel substitutions. Additionally, we document four independent duplications of ATPα with convergent tissue-specific expression patterns. We find that unique substitutions are disproportionately associated with recent duplications relative to parallel substitutions. Together, these findings support the hypothesis that adaptation tends to take evolutionary paths that minimize negative pleiotropy.

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Summary• Low temperature represents a form of abiotic stress that varies predictably with latitude and altitude and to which organisms have evolved multiple physiological responses. Plants provide an especially useful experimental system for investigating the ecological and evolutionary dynamics of tolerance to low temperature because of their sessile lifestyle and inability to escape ambient atmospheric conditions.• Here, intraspecific variation in freezing tolerance was investigated in Arabidopsis thaliana by conducting freezing tolerance assays on 71 accessions collected from across the native range of the species. Assays were performed at multiple minimum temperatures and on both cold-acclimated and non-cold-acclimated individuals.• Considerable variation in freezing tolerance was observed among accessions both with and without a prior cold-acclimation treatment, suggesting that differences among accessions in cold-acclimation capacity as well as differences in intrinsic physiology contribute to variation in this phenotype. A highly significant positive relationship was observed between freezing tolerance and latitude of origin of accessions, consistent with a major role for natural selection in shaping variation in this phenotype.• Clinal variation in freezing tolerance in A. thaliana coupled with considerable knowledge of the underlying genetics and physiology of this phenotype should allow evolutionary genetic analysis at multiple levels.

Numerous animal lineages have expanded and diversified the opsin-based photoreceptors in their eyes underlying color vision behavior. However, the selective pressures giving rise to new photoreceptors and their spectral tuning remain mostly obscure. Previously, we identified a violet receptor (UV2) that is the result of a UV opsin gene duplication specific to Heliconius butterflies. At the same time the violet receptor evolved, Heliconius evolved UV-yellow coloration on their wings, due to the pigment 3-hydroxykynurenine (3-OHK) and the nanostructure architecture of the scale cells. In order to better understand the selective pressures giving rise to the violet receptor, we characterized opsin expression patterns using immunostaining (14 species) and RNA-Seq (18 species), and reconstructed evolutionary histories of visual traits in five major lineages within Heliconius and one species from the genus Eueides. Opsin expression patterns are hyperdiverse within Heliconius. We identified six unique retinal mosaics and three distinct forms of sexual dimorphism based on ommatidial types within the genus Heliconius. Additionally, phylogenetic analysis revealed independent losses of opsin expression, pseudogenization events, and relaxation of selection on UVRh2 in one lineage. Despite this diversity, the newly evolved violet receptor is retained across most species and sexes surveyed. Discriminability modeling of behaviorally preferred 3-OHK yellow wing coloration suggests that the violet receptor may facilitate Heliconius color vision in the context of conspecific recognition. Our observations give insights into the selective pressures underlying the origins of new visual receptors.

Clostridioides difficile toxins (TcdA and TcdB) are major exotoxins responsible for C. difficile infection (CDI) associated diseases. The previously reported TcdB variants showed distinct biological features, immunoactivities, and potential pathogenicity in disease progression. Here, we performed global comparisons of amino acid sequences of both TcdA and TcdB from 3,269 C. difficile genomes and clustered them according to the evolutionary relatedness. We found that TcdB was much diverse and could be divided into eight subtypes, of which four were first described. Further analysis indicates that the tcdB gene undergoes accelerated evolution to maximize diversity. By tracing TcdB subtypes back to their original isolates, we found that the distribution of TcdB subtypes was not completely aligned with the phylogeny of C. difficile. These findings suggest that the tcdB genes not only frequently mutate, but also continuously transfer and exchange among C. difficile strains.

Elucidating the molecular basis of adaptive phenotypic variation represents a central aim in evolutionary biology. Traits exhibiting patterns of clinal variation represent excellent models for studies of molecular adaptation, especially when variation in phenotype can be linked to organismal fitness in different environments. Natural accessions of the model plant species Arabidopsis thaliana exhibit clinal variation in freezing tolerance that follows a gradient of temperature variability across the species' native range (Zhen Y, Ungerer MC. 2008. Clinal variation in freezing tolerance among natural accessions of A. thaliana. New Phytol. 177:419-427). Here, we report that this pattern of variation is attributable, at least in part, to relaxed purifying selection on members of a small family of transcriptional activators (the CBF/DREB1s) in the species' southern range. These regulatory genes play a critical role in the ability of A. thaliana plants to undergo cold acclimation and thereby achieve maximum freezing tolerance. Relative to accessions from northern regions, accessions of A. thaliana from the southern part of their geographic range exhibit levels of nonsynonymous nucleotide polymorphism that are approximately 2.8-fold higher across this small gene subfamily. Relaxed selection on the CBF/DREB1s in southern accessions also has resulted in multiple mutations in regulatory regions resulting in abrogated expression of particular subfamily members in particular accessions. These coding-region and regulatory mutations compromise the ability of these genes to act as efficient transcriptional activators during the cold acclimation process, as determined by reductions in rates of induction and maximum levels of expression in the downstream genes they regulate. This study highlights the potential role of regulatory genes in underlying adaptive phenotypic variation in nature.

Activation of transposable elements in species' genomes represents an important mechanism of new mutation and of potential rapid change in genome size. Thus, it is increasingly recognized that transposable elements likely have played a significant role in shaping species' evolution. In an earlier report, we showed that the genomes of three sunflower species of ancient hybrid origin have experienced large-scale proliferation events of sequences within the Ty3/ gypsy-like superfamily of long terminal repeat (LTR) retrotransposons. In this report, we investigate whether another superfamily of LTR retrotransposon (Ty1/copia-like elements) have experienced similar derepression and proliferation events in the genomes of these sunflower hybrid taxa. We show that Ty1/copia-like elements also have undergone copy number increases following or associated with the origins of these species, although the scale of proliferation is less than that for Ty3/gypsy-like elements. Surveys of sequence heterogeneity of Ty1/copia-like elements in the genomes of the three hybrid and two parental species' genomes reveal that a single sub-lineage of these elements exhibits characteristics of recent amplification, and likely served as the proliferative source lineage. These findings indicate that the genomic and/or environmental conditions associated with the origins of these sunflower hybrid taxa were conducive to derepression of at least two major groups of transposable elements.

BackgroundThe objective of this meta-analysis was to compare the clinical safety and efficacy of robotic right colectomy (RRC) with conventional laparoscopic right colectomy (LRC).MethodsA literature search was performed for comparative studies reporting perioperative outcomes of RRC and LRC. The methodological quality of the selected studies was assessed. Depending on statistical heterogeneity, the fixed effects model or the random effects model were used for the meta-analysis. Operative time, estimated blood loss, length of hospital stay, conversion rates to open surgery, postoperative complications, and related outcomes were evaluated.ResultsSeven studies, including 234 RRC cases and 415 conventional LRC cases, were analyzed. The meta-analysis showed that RRC had longer operative times (P < 0.00001), lower estimated blood losses (P = 0.0002), lower postoperative overall complications (P = 0.02), and significantly faster bowel function recovery (P < 0.00001). There were no differences in the length of hospital stay (P = 0.12), conversion rates to open surgery (P = 0.48), postoperative ileus (P = 0.08), anastomosis leakage (P = 0.28), and bleeding (P = 0.95).ConclusionsCompared to LRC, RRC was associated with reduced estimated blood losses, reduced postoperative complications, longer operative times, and a significantly faster recovery of bowel function. Other perioperative outcomes were equivalent.