Context-dependent concordance between physiological divergence and phenotypic selection in sister taxa with contrasting phenology and mating systems.

PREMISE: The study of phenotypic divergence of, and selection on, functional traits in closely related taxa provides the opportunity to detect the role of natural selection in driving diversification. If the strength or direction of selection in field populations differs between taxa in a pattern that is consistent with the phenotypic difference between them, then natural selection reinforces the divergence. Few studies have sought evidence for such concordance for physiological traits. METHODS: Herbarium specimen records were used to detect phenological differences between sister taxa independent of the effects on flowering time of long-term variation in the climate across collection sites. In the field, physiological divergence in photosynthetic rate, transpiration rate, and instantaneous water-use efficiency were recorded during vegetative growth and flowering in 13 field populations of two taxon pairs of Clarkia, each comprising a self-pollinating and a outcrossing taxon. RESULTS: Historically, each selfing taxon flowered earlier than its outcrossing sister taxon, independent of the effects of local long-term climatic conditions. Sister taxa differed in all focal traits, but the degree and (in one case) the direction of divergence depended on life stage. In general, self-pollinating taxa had higher gas exchange rates, consistent with their earlier maturation. In 6 of 18 comparisons, patterns of selection were concordant with the phenotypic divergence (or lack thereof) between sister taxa. CONCLUSIONS: Patterns of selection on physiological traits measured in heterogeneous conditions do not reliably reflect divergence between sister taxa, underscoring the need for replicated studies of the direction of selection within and among taxa.

Could seasonally deteriorating environments favour the evolution of autogamous selfing and a drought escape physiology through indirect selection? A test of the time limitation hypothesis using artificial selection in Clarkia.

Background and Aims: The evolution of selfing from outcrossing may be the most common transition in plant reproductive systems and is associated with a variety of ecological circumstances and life history strategies. The most widely discussed explanation for these associations is the reproductive assurance hypothesis - the proposition that selfing is favoured because it increases female fitness when outcross pollen receipt is limited. Here an alternative explanation, the time limitation hypothesis, is addressed, one scenario of which proposes that selfing may evolve as a correlated response to selection for a faster life cycle in seasonally deteriorating environments. Methods: Artificial selection for faster maturation (early flowering) or for low herkogamy was performed on Clarkia unguiculata (Onagraceae), a largely outcrossing species whose closest relative, C. exilis, has evolved higher levels of autogamous selfing. Direct responses to selection and correlated evolutionary changes in these traits were measured under greenhouse conditions. Direct responses to selection on early flowering and correlated evolutionary changes in the node of the first flower, herkogamy, dichogamy, gas exchange rates and water use efficiency (WUE) were measured under field conditions. Key Results: Lines selected for early flowering and for low herkogamy showed consistent, statistically significant responses to direct selection. However, there was little or no evidence of correlated evolutionary changes in flowering date, floral traits, gas exchange rates or WUE. Conclusions: These results suggest that the maturation rate and mating system have evolved independently in Clarkia and that the time limitation hypothesis does not explain the repeated evolution of selfing in this genus, at least through its indirect selection scenario. They also suggest that the life history and physiological components of drought escape are not genetically correlated in Clarkia, and that differences in gas exchange physiology between C. unguiculata and C. exilis have evolved independently of differences in mating system and life history.

Outcrossing and photosynthetic rates vary independently within two Clarkia species: implications for the joint evolution of drought escape physiology and mating system.

Background and Aims Mating systems of plants are diverse and evolutionarily labile. Abiotic environmental factors, such as seasonal drought, may impose selection on physiological traits that could lead to transitions in mating system if physiological traits are genetically correlated with traits that influence mating system. Within Clarkia, self-fertilizing taxa have higher photosynthetic rates, earlier flowering phenology, faster individual floral development and more compressed flowering periods than their outcrossing sister taxa, potentially reducing the selfing taxa's exposure to drought. In theory, this contrast in trait combinations between sister taxa could have arisen via correlated evolution due to pleiotropy or genetic linkage. Alternatively, each trait may evolve independently as part of a life history that is adaptive in seasonally dry environments. Methods To evaluate these hypotheses, we examined relationships between photosynthetic rates (adjusted for plant height and leaf node position) and outcrossing rates (estimated by allozyme variation in progeny arrays) during two consecutive years in multiple wild populations of two mixed-mating Clarkia taxa, each of which is sister to a derived selfing taxon. If the negative association between photosynthetic rate and outcrossing previously observed between sister taxa reflects correlated evolution due to a strong negative genetic correlation between these traits, then a similarly negative relationship would be observed within populations of each taxon. By contrast, if the combination of elevated photosynthetic rates and reduced outcrossing evolved independently within taxa, we predicted no consistent relationship between photosynthetic rate and outcrossing rate. Key Results We found no significant difference in outcrossing rates within populations between groups of plants with high versus low photosynthetic rates. Conclusions Overall, these results provide support for the hypothesis that the joint divergence in photosynthetic rate and mating system observed between Clarkia sister taxa is the result of independent evolutionary transitions.

Interoceptive awareness and its relationship to hippocampal dependent processes.

Many neuropsychological and animal lesion studies point to the hippocampus as being critical for mediating interoceptive awareness, while neuroimaging studies have been used to argue for the importance of the insula and anterior cingulate cortex. Here, using healthy young adults - as with the neuroimaging data - we tested for an association between performance on a hippocampal dependent learning and memory (HDLM) measure (logical memory percent retention) and interoceptive awareness assessed on three tasks - heart rate tracking, water loading and the Multidimensional Assessment of Interoceptive Awareness questionnaire (MAIA). After controlling for other relevant potentially confounding variables, we found significant associations between both the water loading and MAIA measures (which were both correlated) and HDLM performance. These findings imply that hippocampal dependent processes are involved in interoceptive awareness in healthy young adults. More tentatively, they suggest that medial temporal lobe structures may mediate interoceptive tasks that involve ingestion and/or integration of past and current state-based information.

Seasonal changes in physiological performance in wild Clarkia xantiana populations: Implications for the evolution of a compressed life cycle and self-fertilization.

PREMISE OF THE STUDY: One explanation for the evolution of selfing, the drought escape hypothesis, proposes that self-fertilization may evolve under conditions of intensifying seasonal drought as part of a suite of traits that enable plants to accelerate the completion of their life cycle, thereby escaping late-season drought. Here, we test two fundamental assumptions of this hypothesis in Clarkia xantiana: (1) that a seasonal decline in precipitation causes an increase in drought stress and (2) that this results in changes in physiological performance, reflecting these deteriorating conditions. METHODS: We examined seasonal and interannual variation in abiotic environmental conditions (estimated by ambient temperature, relative humidity, predawn leaf water potentials, and carbon isotope ratios) and physiological traits (photosynthesis, conductance, transpiration, instantaneous water-use efficiency, ascorbate peroxidase and glutathione reductase activities, quantum yield of photosystem II, PSII potential efficiency) in field populations of C. xantiana in 2009 and 2010. KEY RESULTS: In both years, plants experienced intensifying drought across the growing season. Gas exchange rates decreased over the growing season and were lower in 2009 (a relatively dry year) than in 2010, suggesting that the temporal changes from early to late spring were directly linked to the deteriorating environmental conditions. CONCLUSIONS: Seasonal declines in transpiration rate may have increased survival by protecting plants from desiccation. Concomitant declines in photosynthetic rate likely reduced the availability of resources for seed production late in the season. Thus, the physiological patterns observed are consistent with the conditions required for the drought escape hypothesis.

Temperature-driven flower longevity in a high-alpine species of Oxalis influences reproductive assurance.

How high-alpine plants confront stochastic conditions for animal pollination is a critical question. We investigated the effect of temperature on potential flower longevity (FL) measured in pollinator-excluded flowers and actual FL measured in pollinated flowers in self-incompatible Oxalis compacta and evaluated if plastically prolonged potential FL can ameliorate slow pollination under cool conditions. Pollinator-excluded and hand-pollinated flowers were experimentally warmed with open-top chambers (OTCs) on a site at 3470 m above sea level (asl). Flower-specific temperatures, and pollinator-excluded and open-pollination flower life-spans were measured at six alpine sites between 3100 and 3470 m asl. Fruit set was analyzed in relation to inferred pollination time. Warming reduced potential FL. Variable thermal conditions across the alpine landscape predicted potential and actual FL; flower senescence was pollination-regulated. Actual FL and potential FL were coupled. Prolonged potential FL generally increased fruit set under cooler conditions. Plastic responses permit virgin flowers of O. compacta to remain open longer under cooler temperatures, thereby ameliorating slow pollination, and to close earlier when pollination tends to be faster under warmer conditions. Plastic potential FL provides adaptive advantages in the cold, thermally variable alpine habitat, and has important implications for reproductive success in alpine plants in a warming world.

Goals, preferences, and concerns of patients with acute myeloid leukemia at time of treatment decision.

INTRODUCTION: Current cancer value-based models require documentation of patient goals of care and an evidence-based treatment course commensurate with patient goals. This feasibility study assessed the utility of an electronic tablet-based questionnaire to elicit patient goals, preferences, and concerns at a treatment decision making time point in patients with acute myeloid leukemia. MATERIALS AND METHODS: Seventy-seven patients were recruited from three institutions prior to seeing the physician for treatment decision-making visit. Questionnaires included demographics, patient beliefs, and decision-making preferences. Analyses included standard descriptive statistics appropriate for the level of measurement. RESULTS: Median age was 71 (range = 61-88), 64.9% female, 87.0% white, and 48.6% college educated. On average, patients completed the surveys unassisted in 16.24 min and providers reviewed the dashboard in 3.5 min. All but one patient completed the survey prior to starting treatment (98.7%). Providers reviewed the survey results prior to seeing the patient 97.4% of the time. When asked their goals of care, 57 (74.0%) patients agreed with the statement "my cancer is curable" and 75 (97.4%) agreed that the treatment goal was to get rid of all cancer. Seventy-seven (100%) agreed the goal of care is to feel better and 76 (98.7%) agreed the goal of care is live longer. Forty-one (53.9%) indicated they wanted to make treatment decisions together with the provider. The top two concerns were understanding treatment options (n = 24; 31.2%) and making the right decision (n = 22; 28.6%). DISCUSSION: This pilot demonstrated the feasibility of using technology for decision-making at the point of care. Eliciting patient goals of care, treatment outcomes expectations, decision-making preferences, and top concerns may provide clinicians with information to inform the treatment discussion. A simple electronic tool may provide valuable insight into patient understanding of disease to better tailor patient-provider discussion and treatment decision-making.

Integrating electronic geriatric assessment and frailty screening for adults with acute myeloid leukemia to drive personalized treatment decisions.

PURPOSE: Evidenced based guidelines for patients with Acute Myeloid Leukemia (AML) acknowledge increasing importance of frailty assessment when deciding on treatment, yet comprehensive geriatric assessment (GA) results are not easily incorporated into clinic workflows and the electronic health record. This study assessed the feasibility of electronic GA use in a real-world environment. METHODS: Patients with AML, ≥ 60 years and at a treatment decision-making point were recruited at three academic institutions. An electronic GA (eGA) was completed by patients prior to starting treatment. Results were immediately available on a dashboard. Data on feasibility, useability and acceptability of the intervention were collected immediately after the clinical visit. Patients completed follow up surveys at 3 months and chart reviews were done to capture treatment and toxicities. RESULTS: 77 patients were enrolled with a median age of 71 years (range=61-88). The eGA results were 25 fit (31.0 %), 22 (32.0 %) intermediate, and 23 (31.0 %) frail. There was 62.7 % (n = 47) provider concordance with the eGA result and 27 (36.0 %) post visit reports indicated that the eGA results influenced the treatment decision. On average, patients completed the surveys unassisted in 16.24 min and providers reviewed the dashboard in 3.5 min. CONCLUSION: Patients easily completed an eGA prior to starting treatment. Results were reviewed by the physician and considered in the decision for optimal treatment. One third of physician reports indicated the results were used to inform the treatment decision. Feasibility of completing the eGA in the clinic without workflow disruption and utility of the results was demonstrated.

Physiological performance and mating system in Clarkia (Onagraceae): does phenotypic selection predict divergence between sister species?

PREMISE OF THE STUDY: The evolution of self-fertilization often occurs in association with other floral, life history, and fitness-related traits. A previous study found that field populations of Clarkia exilis (a predominantly autogamous selfer) and its sister species, Clarkia unguiculata (a facultative outcrosser) differ in mean photosynthetic rates and instantaneous water use efficiency (WUE(i)). Here, we investigate the strength and direction of selection on these traits in multiple populations of each taxon to determine whether natural selection may contribute to the phenotypic differences between them. METHODS: In spring 2008, we measured instantaneous gas exchange rates in nine populations during vegetative growth (Early) and/or during flowering (Late). We conducted selection gradient analyses and estimated selection differentials within populations and across pooled conspecific populations to evaluate the strength, direction, and consistency of selection on each trait early and late in the season. KEY RESULTS: The direction and relative strength of selection on photosynthetic rates in these taxa corresponds to the phenotypic difference between them; C. exilis has higher photosynthetic rates than C. unguiculata, as well as stronger, more consistent selection favoring rapid photosynthesis throughout the growing season. Patterns of selection on transpiration, WUE(i), and the timing of flowering progression are less consistent with phenotypic differences (or lack thereof) between taxa. CONCLUSIONS: We detected several examples where selection was consistent with the phenotypic divergence between sister taxa, but there were also numerous instances that were equivocal or in which selection did not predict the realized phenotypic difference between taxa.

Stability of pollen-ovule ratios in pollinator-dependent versus autogamous Clarkia sister taxa: testing evolutionary predictions.

It has been proposed that natural selection should favor distinct temporal patterns of sex allocation in selfing vs pollinator-dependent taxa. In autogamous selfers in which pollen receipt is highly reliable, selection should favor genotypes that maintain low and stable pollen to ovule (P : O) ratios throughout flowering. By contrast, in outcrossers the optimum P : O ratio of an individual's flowers will depend on pollinator abundances and mating opportunities, both of which may vary over time. In this case, selection may favor temporal variation among flowers in the P : O ratio. An opposing prediction is that selfing taxa will be developmentally more unstable than outcrossers because of lower homeostasis caused by high homozygosity. We compared temporal changes in the P : O ratio in two pairs of sister taxa in the genus Clarkia. We examined hundreds of glasshouse-raised maternal families representing three wild populations each of the outcrossing, insect-pollinated Clarkia unguiculata, the facultatively autogamous Clarkia exilis and the outcrossing and selfing subspecies of Clarkia xantiana: ssp. xantiana and parviflora, respectively. Temporal change in the P : O ratio was significantly greater in both outcrossers than in their selfing sister taxa, although the proportional changes in the P : O ratio (relative to the first bud produced) did not differ significantly between sister taxa (0.07 < P < 0.10). Our results provide partial support for the hypothesis that the P : O ratio is more stable in selfing than in outcrossing taxa and reject the hypothesis that selfers are less stable.

Functional role of long-lived flowers in preventing pollen limitation in a high elevation outcrossing species.

Low pollinator visitation in harsh environments may lead to pollen limitation which can threaten population persistence. Consequently, avoidance of pollen limitation is expected in outcrossing species subjected to habitually low pollinator service. The elevational decline in visitation rates on many high mountains provides an outstanding opportunity for addressing this question. According to a recent meta-analysis, levels of pollen limitation in alpine and lowland species do not differ. If parallel trends are manifested among populations of alpine species with wide elevational ranges, how do their uppermost populations contend with lower visitation? We investigated visitation rates and pollen limitation in high Andean Rhodolirium montanum. We test the hypothesis that lower visitation rates at high elevations are compensated for by the possession of long-lived flowers. Visitation rates decreased markedly over elevation as temperature decreased. Pollen limitation was absent at the low elevation site but did occur at the high elevation site. While initiation of stigmatic pollen deposition at high elevations was not delayed, rates of pollen arrival were lower, and cessation of pollination, as reflected by realized flower longevity, occurred later in the flower lifespan. Comparison of the elevational visitation decline and levels of pollen limitation indicates that flower longevity partially compensates for the lower visitation rates at high elevation. The functional role of flower longevity, however, was strongly masked by qualitative pollen limitation arising from higher abortion levels attributable to transference of genetically low-quality pollen in large clones. Stronger clonal growth at high elevations could counterbalance the negative fitness consequences of residual pollen limitation due to low visitation rates and/or difficult establishment under colder conditions. Visitation rates on the lower part of the elevational range greatly exceeded community rates recorded several decades ago when the planet was cooler. Current pollen limitation for some species in some habitats might underestimate historical levels.

Plastic Responses Contribute to Explaining Altitudinal and Temporal Variation in Potential Flower Longevity in High Andean Rhodolirion montanum.

The tendency for flower longevity to increase with altitude is believed by many alpine ecologists to play an important role in compensating for low pollination rates at high altitudes due to cold and variable weather conditions. However, current studies documenting an altitudinal increase in flower longevity in the alpine habitat derive principally from studies on open-pollinated flowers where lower pollinator visitation rates at higher altitudes will tend to lead to flower senescence later in the life-span of a flower in comparison with lower altitudes, and thus could confound the real altitudinal pattern in a species´ potential flower longevity. In a two-year study we tested the hypothesis that a plastic effect of temperature on flower longevity could contribute to an altitudinal increase in potential flower longevity measured in pollinator-excluded flowers in high Andean Rhodolirium montanum Phil. (Amaryllidaceae). Using supplemental warming we investigated whether temperature around flowers plastically affects potential flower longevity. We determined tightly temperature-controlled potential flower longevity and flower height for natural populations on three alpine sites spread over an altitudinal transect from 2350 and 3075 m a.s.l. An experimental increase of 3.1°C around flowers significantly decreased flower longevity indicating a plastic response of flowers to temperature. Flower height in natural populations decreased significantly with altitude. Although temperature negatively affects flower longevity under experimental conditions, we found no evidence that temperature around flowers explains site variation in flower longevity over the altitudinal gradient. In a wetter year, despite a 3.5°C temperature difference around flowers at the extremes of the altitudinal range, flower longevity showed no increase with altitude. However, in a drier year, flower longevity increased significantly with altitude. The emerging picture suggests an increase in flower longevity along the altitudinal gradient is less common for potential flower longevity than for open-pollination flower longevity. Independently of any selection that may occur on potential longevity, plastic responses of flowers to environmental conditions are likely to contribute to altitudinal variation in flower longevity, especially in dry alpine areas. Such plastic responses could push flowers of alpine species towards shorter life-lengths under climate change, with uncertain consequences for successful pollination and plant fitness in a warming world.

Stage-dependent patterns of drought tolerance and gas exchange vary between sexes in the alpine willow, Salix glauca.

Females and males of sexually dimorphic species have distinct resource demands due to differential allocation to reproduction. Sexual allocation theory predicts that functional traits will diverge between sexes to support these demands. However, such dimorphism may be masked by the impact of current reproduction on source-sink interactions between vegetative and reproductive organs. We ask whether natural selection has led to genetic dimorphism in homologous physiological traits between sexes of the dioecious willow shrub, Salix glauca. In a common garden experiment we compared physiological responses to drought stress by male and female ramets in the absence of confounding demands from reproductive structures. Ramets experienced similar pre-dawn leaf water status (Psi(l)) as parental genets in flower within the natural population, indicating that experimental dry-down mirrored environmental conditions in nature. Male and female ramets achieved similar instantaneous water use efficiency, based on the ratio of carbon gain to water loss, under wet and dry conditions. However, female ramets experienced greater water stress (i.e., more negative Psi(l)) than males under dry conditions. Lower Psi(l) for female ramets may partly reflect the maintenance of conductance under drought; males, in contrast, maintain Psi(l) under drought by reducing conductance. Differences between sexes in terms of conductance and leaf water status of the vegetative ramets were absent in a concomitant comparison of parental flowering plants. Our results show (1) genetic divergence in physiology between sexes of S. glauca occurs in the absence of gender-specific reproductive sinks, (2) males are the more physiologically plastic sex with respect to water use, and (3) paradoxically, divergence in water relations between sexes is not detectable at sexual maturity under natural conditions.

Ecological correlates of secondary sexual dimorphism in Salix glauca (Salicaceae).

The ecological causation hypothesis for secondary sexual dimorphism was tested in Salix glauca, a dioecious willow shrub. Plants growing in a Colorado Rocky Mountain (USA) krummholz mosaic of mesic and xeric patches were monitored for four consecutive years. Ecological causation is predicated on unique resource demands associated with sexual function. In S. glauca, seeds have twofold higher N and P concentrations compared to pollen. P, but not N, allocation costs differed between sexes at plant and flower scales. Ecological causation also predicts spatial segregation of sexes along underlying habitat gradients. In five populations of S. glauca, sexes displayed significant spatial segregation. The theory also predicts that sexes differ in performance across gradients of environmental stress or resource availability. Consistent with this hypothesis, females had lower drought tolerance than males under years of extreme aridity. Furthermore, over 10 years, annual shoot growth for females was greatest in mesic habitat patches, while males grew at a consistent rate regardless of habitat aridity. Because current shoot growth is correlated with future catkin production, habitat specialization likely provides a fitness payoff in females. Overall, this long-term study provides some of the strongest evidence to date for ecological causation of secondary sexual dimorphism in plants.