Rocky Mountain Biological Laboratory Research Articles

METHODS: Validating an immunoassay to measure fecal glucocorticoid metabolites in yellow-bellied marmots

Quantifying physiological stress in wild animals is essential for understanding their health, reproductive success, and survival in a variable environment. The yellow-bellied marmot (Marmota flaviventer) study at the Rocky Mountain Biological Laboratory near Crested Butte, Colorado, USA is the world's second longest study of free-living mammals. Historically, we used a validated corticosterone radioimmunoassay (RIA) to measure fecal glucocorticoid metabolites (FGMs) as a proxy for physiological stress. However, the costs and risks associated with working with radioisotopes drove us to consider a more sustainable method. Here we evaluate the suitability of two competitive corticosterone enzyme assays (EIA), one from Cayman Chemical Company (CCC) and one from Arbor Assays (AA), to measure marmot FGMs via their cross-reaction. The findings revealed that the AA EIA better matched the RIA in terms of accuracy across high and low FGM concentrations, had superior assay parameters, showed the highest correlations with RIA results and effectively captured the annual variations in FGM concentrations, thus demonstrating its reliability for use in longitudinal studies. We further analytically validated the AA EIA for FGMs and confirmed its efficacy and lack of matrix effects, thus establishing its suitability for ongoing and future studies of FGMs in marmots. The transition to the AA EIA from the RIA ensures continued data integrity while enhancing safety and environmental sustainability.

The timing of reproduction is responding plastically, not genetically, to climate change in yellow-bellied marmots (Marmota flaviventer).

With global climates changing rapidly, animals must adapt to new environmental conditions with altered weather and phenology. The key to adapting to these new conditions is adjusting the timing of reproduction to maximize fitness. Using a long-term dataset on a wild population of yellow-bellied marmots (Marmota flaviventer) at the Rocky Mountain Biological Laboratory (RMBL), we investigated how the timing of reproduction changed with changing spring conditions over the past 50 years. Marmots are hibernators with a 4-month active season. It is thus crucial to reproduce early enough in the season to have time to prepare for hibernation, but not too early, as snow cover prevents access to food. Importantly, climate change in this area has, on average, increased spring temperatures by 5°C and decreased spring snowpack by 50 cm over the past 50 years. We evaluated how female marmots adjust the timing of their reproduction in response to changing conditions and estimated the importance of both microevolution and plasticity in the variation in this timing. We showed that, within a year, the timing of reproduction is not as tightly linked to the date a female emerges from hibernation as previously thought. We reported a positive effect of spring snowpack but not of spring temperature on the timing of reproduction. We found inter-individual variation in the timing of reproduction, including low heritability, but not in its response to changing spring conditions. There was directional selection for earlier reproduction since it increased the number and proportion of pups surviving their first winter. Taken together, the timing of marmot reproduction might evolve via natural selection; however, plastic changes will also be extremely important. Further, future studies on marmots should not operate under the assumption that females reproduce immediately following their emergence.

Climate data from the Rocky Mountain Biological Laboratory (1975-2022).

The Rocky Mountain Biological Laboratory (RMBL) (Colorado, USA) is the site for many research projects spanning decades, taxa, and research fields from ecology to evolutionary biology to hydrology and beyond. Climate is a focus of much of this work and provides important context for the rest. There are five major sources of data on climate in the RMBL vicinity, each with unique variables, formats, and temporal coverage. These data sources include 1) RMBL resident billy barr, 2) the National Oceanic and Atmospheric Administration (NOAA), 3) the United States Geological Survey (USGS), 4) the United States Department of Agriculture (USDA), and 5) Oregon State University's PRISM Climate Group. Both the NOAA and the USGS have automated meteorological stations in Crested Butte, CO, approximately 10 km from the RMBL while the USDA has an automated meteorological station on Snodgrass Mountain, approximately 2.5 km from the RMBL. Each of these data sets has unique spatial and temporal coverage and formats. Despite the wealth of work on climate-related questions using data from the RMBL, previous researchers have each had to access and format their own climate records, make decisions about how to handle missing data, and recreate data summaries. Here we provide a single curated climate data set of daily observations covering the years 1975 to 2022 that blends information from all five sources and includes annotated scripts documenting decisions for handling data. This synthesized climate data will facilitate future research, reduce duplication of effort, and increase our ability to compare results across studies. The data set includes information on precipitation (water and snow), snowmelt date, temperature, wind speed, soil moisture and temperature, and stream flows, all of which are publicly available from a combination of sources. In addition to the formatted raw data, we provide several new variables that are commonly used in ecological analyses, including growing degree days, growing season length, a cold severity index, hard frost days, an index of El Niño-Southern Oscillation, and aridity (standardized precipitation evapotranspiration index). These new variables are calculated from the daily weather records. As appropriate, data are also presented as minima, maxima, means, residuals, and cumulative measures, for a variety of time scales including days, months, seasons, and years. The Rocky Mountain Biological Laboratory is a global research hub. Scientists on-site at the RMBL come from many countries and produce about 50 peer-reviewed publications each year, researchers from around the world also routinely make use of data from the RMBL for synthetic work, and educators around the United States use data from RMBL for teaching modules. This curated and combined data set will be of use to a wide audience. Along with the synthesized combined data set we include the raw data and the R code for cleaning the raw data and creating the monthly and yearly data sets, which facilitate adding additional years or data using the same standardized protocols. No copyright or proprietary restrictions are associated with the use of this data set; please cite this data paper when the data are used in publications or scientific events.

Biogeochemical characteristics and hydroperiod affect carbon dioxide flux rates from exposed high‐elevation pond sediments

AbstractWhile inundated, small ponds (< 1000 m2 area) account for disproportionately large contributions of CO2 efflux to the global carbon budget and also store carbon in anoxic sediments. However, pond hydrology is shifting toward increasingly dry conditions in alpine and temperate zones, which might lead to increased exposure of shallow pond sediments. We analyzed sediment CO2 efflux rates in dried sediments of multiple ponds of varying hydrology and sediment characteristics at montane and subalpine elevations near the Rocky Mountain Biological Laboratory in Colorado. Average CO2 efflux rates from exposed sediments, 331.5 ± 11.5 mmol m−2 d−1 at the montane sites and 142.8 ± 45.1 mmol m−2 d−1 at the subalpine sites, were 10 times higher than average CO2 efflux rates from pond water. Principal components analysis to reduce dimensionality of sediment characteristics revealed that random inter‐pond differences rather than exposure timing or hydroperiod drove variation among sediments. In linear mixed effects models of CO2 flux rates, significant predictors included sediment moisture and temperature, pH, total organic carbon, and organic matter content at all pond hydroperiod classifications and sites. However, the sediment characteristics explaining the most variance differed among sites and hydroperiods and included nitrate concentrations, pH, bulk density, and temperature. We conclude that pond sediments are heterogeneous both within and among ponds in close proximity, and drivers of relatively high CO2 efflux rates differ among pond hydroperiods and elevations. This work emphasizes that local differences can impact predictions of CO2 flux from lentic sediments which are becoming increasingly exposed.

Effects of climate change on phenologies and distributions of bumble bees and the plants they visit

AbstractSurveys of bumble bees and the plants they visit, carried out in 1974 near the Rocky Mountain Biological Laboratory in Colorado, were repeated in 2007, thus permitting the testing of hypotheses arising from observed climate change over the intervening 33‐yr period. As expected, given an increase in average air temperature with climate warming and a declining temperature with increasing elevation, there have been significant shifts toward higher elevation for queens or workers or both, for most bumble bee species, for bumble bee queens when species are combined, and for two focal plant species, with no significant downward shifts. However, contrary to our hypotheses, we failed to observe significant altitudinal changes for some bumble bee species and most plant species, and observed changes in elevation were often less than the upward shift of 317 m required to maintain average temperature. As expected, community flowering phenology shifted toward earlier in the season throughout our study area, but bumble bee phenology generally did not change, resulting in decreased synchrony between bees and plants. However, we were unable to confirm the narrower expectation that phenologies of bumble bee workers and community flowering coincided in 1974 but not in 2007. As expected, because of reduced synchrony between bumble bees and community flowering, bumble bee abundance was reduced in 2007 compared with 1974. Hence, climate change in our study area has apparently resulted primarily in reduced abundance and upward shift in distribution for bumble bees and shift toward earlier seasonality for plant flowering. Quantitative disagreements between climate change expectations and our observations warrant further investigation.

“Men to match the mountains” The story behind the TriBeta Johnson award

B eta Beta Beta (TriBeta) is a National Biological Honor Society, dedicated to the recognition and support of outstanding undergraduate students in biology and the life sciences. The driving force behind the society is men and women who have unselfishly given their time, talents and personal resources. This is a brief history and background of the prestigious John C. Johnson Award, which is presented to students for outstanding posters at District, Regional and National Conventions sponsored by TriBeta. The John C. Johnson legacy has a rich and colorful history involving Dr. John C. Johnson, Sr. and his son, Dr. John C. ‘‘Chris’’ Johnson, Jr. Both Dr. Johnsons made significant contributions for the advancement of TriBeta and the biological sciences. The Johnson legacy continues with Chris’s daughter, Carol M. Johnson, who is the Vice President of the Rocky Mountain Biological Laboratory Board of Trustees. The Johnsons and their families have spanned a century making memorable and indelible impressions on teaching and research regarding nature and the biological science they loved. Theirs is a story of benevolent sharing of knowledge, fortified with insight and foresight into the environment and the natural world. Their unselfish contributions have been memorialized with the coveted TriBeta Johnson Award. The storied history behind the Johnson Award began a century ago in the state of Colorado with Dr. John C. Johnson, Sr., followed by his son, Dr. John C. ‘‘Chris’’ Johnson, Jr.

W. S. Cooper As I Knew Him: Teacher, Mentor and Friend

W. S. Cooper As I Knew Him: Teacher, Mentor and Friend

The Effect of the Foresummer Drought on Carbon Exchange in Subalpine Meadows

Climate in subalpine meadows of the Rocky Mountains can be characterized by an early (foresummer) drought that occurs after snowmelt (May) and lasts until the start of the summer monsoon season (July). Climate change models predict an increase in the length and severity of this dry period due to earlier snowmelt dates, rising air temperatures, and shifts in the start and/or intensity of the North American monsoon. However, it is unknown how changes in the severity of this early season dry period will affect ecosystem carbon exchange. To address the importance of early season drought, we combined a watering manipulation with 11 years of ecosystem carbon exchange data across an elevational gradient at the Rocky Mountain Biological Laboratory in Gothic, Colorado. Long-term trends reveal that earlier snowmelt dates lead to a decrease in net ecosystem productivity (NEP), in part because of the positive effect on early growing season drought conditions. Manipulating the strength of the foresummer drought by watering revealed that the timing of growing season precipitation is more important than the total amount for determining cumulative NEP. The strength of the foresummer drought did not significantly impact ecosystem respiration rates, but plants that experienced a strong foresummer drought exhibited more water stress, and lower instantaneous rates of NEP, even during the rainy season. Our results highlight the central role of the foresummer drought in determining rates of carbon exchange throughout the growing season, and the potential for an increasingly negative balance of carbon in subalpine meadows under future climate change.

Using Rich Context and Data Exploration to Improve Engagement with Climate Data and Data Literacy: Bringing a Field Station into the College Classroom

ABSTRACT Authentic scientific data, when richly contextualized, can provide the basis for compelling learning experiences. Many undergraduate students either do not have access to primary data, or if they do, the data are so abstract that student engagement is limited. Here, we describe contextual information and data-rich, student-centered activities we developed to give life to data sets from an intensely studied place, the Rocky Mountain Biological Laboratory (RMBL). The project Web site, Digital RMBL, highlights charismatic organisms, scientists, and long-term data sets as a tool for engaging students who are unable to physically visit a field station. The Biology of Climate Change module, the focus of this paper, has been tested in college-level classrooms by 10 collaborating faculty and 243 undergraduate students at a variety of colleges and universities across the nation. Authentic long-term data sets, primary literature, data visualizations, and a flexible format suitable for laboratory sections have led to very high usability ratings by collaborating faculty. In student surveys, a surprising number of undergraduate science majors (20%–30%) report that they have never worked with authentic scientific data—even at major research universities. The percentage of students who have not worked with data is much higher at collaborating 2 y institutions (60%–80%). The majority of students report that they appreciate the opportunity to explore long-term climate science data sets despite the frustrations they experience with the “messiness” of authentic scientific data. The impact of this climate change activity is achieved through engagement with people, place, and research subjects, followed by student-centered data exploration that builds personal interest and scientific discovery skills. This paper outlines one model in which scientists can meet funding agency requirements to share data publicly while providing excellent opportunities for improving climate and data literacy at the college level.

Yellow-bellied marmots do not compensate for a late start: the role of maternal allocation in shaping life-history trajectories

Yellow-bellied marmots, Marmota flaviventris, are obligate hibernators with a relatively short active season. Animals born earlier in the season have higher chances of fulfilling the energetic requirements to survive the long winter. Therefore, the onset of breeding should have a profound impact on juvenile survival. However, there are different ways to compensate for a late start. Late breeders might allocate more resources to late born offspring, making up for the bad start, or juveniles might show compensatory growth. They are not exclusive hypotheses and both can lead to juveniles entering hibernation with a similar body condition. We used data from a long term study in and around the Rocky Mountain Biological Laboratory in Colorado to test these two hypotheses. Animals are individually marked and trapped regularly. We compared mass at weaning, mass at the end of the season, growth rates and survival between animals born earlier and later in the season. We found no evidence of increased maternal input: late breeders had lighter offspring at weaning than early breeders, and late born juveniles did not increase their growth rates. Therefore, late born individuals ended the season with lower body mass, and were thus, less likely to survive the winter. In conclusion, life-history trajectories of juvenile yellow-bellied marmots were determined by maternal allocation, whereas post- weaning decisions did not modify their fate.

Do stream mayflies exhibit trade-offs between food acquisition and predator avoidance behaviors?

Organisms often experience trade-offs between behaviors necessary for food acquisition and predator avoidance. In mesocosms and microcosms, we manipulated the risk and resource environments of grazing mayflies (Baetis bicaudatus) common in streams near the Rocky Mountain Biological Laboratory (RMBL) in western Colorado, USA. We exposed mayflies to treatments with or without chemical cues from predatory brook trout (Salvelinus fontinalis) and high or low levels of benthic algal resources to test the nonconsumptive effects of predators and food availability on grazer foraging behavior, predator-avoidance behavior, growth rates, and the effect of grazers on accrual of algal food resources. Total drift (daytime + nighttime) of Baetis was significantly reduced under conditions of fish cues and high food levels. However, daytime drift, a risky food-search behavior, was negligible in all cases except the no-fish-cue/low-food treatment. Therefore, predator-induced nocturnal periodicity of drift behavior was influenced by variation in food levels. At low-food levels, Baetis growth rates and grazer effect on algal accrual were reduced, but neither measure of grazer performance (growth rates nor grazer impacts) was affected by fish cues or their interaction with food levels. We conclude that mayflies faced with potential trade-offs between food acquisition and predator-avoidance behaviors exhibit risky behavior (high daytime drift) only when predator-risk cues are absent and resources are low. Moreover, we observed no short-term costs of predator-induced suppression of foraging movements under low-resource conditions. These results imply that previously observed negative effects of trout on size and fecundity of mature Baetis are not explained by slower growth rates.

Yellow-bellied marmots: insights from an emergent view of sociality.

Ecological factors explain variation in sociality both within and between species of marmots-large alpine ground squirrels. Fifty years of study, by me and my colleagues, of the yellow-bellied marmots (Marmota flaviventris) at the Rocky Mountain Biological Laboratory, near Crested Butte, CO, USA, has created opportunities to see how sociality changes with population and group size. Over the past decades, we have witnessed a natural experiment whereby the population tripled in size. If we view sociality as an emergent process, then demography acts as a constraint on interactions between individuals, and the threefold increase in population size should have consequences for group structure. We have used social network statistics to study the causes and consequences of social interactions by capitalizing on this demographic variation. Such an emergent view is ideally studied in an integrative Tinbergian way that focuses on both mechanism and function. We have determined that some social attributes are heritable, that social cohesion is established through age and kin structure, that well-embedded females (but not males) are less likely to disperse, and that there are fitness consequences of social attributes. Together, this integrative relationship-centred view expands on the traditional ecological model of sociality and offers a framework that can be applied to other systems.

Consequences of early snowmelt in Rocky Mountains

Snow melted significantly earlier in the Rocky Mountains in 2012 than in previous years, with serious consequences for plants and animals, scientists reported at the AGU Fall Meeting. David Inouye of the University of Maryland, College Park, and the Rocky Mountain Biological Laboratory said that “the timing of winter's end is changing.” He has been observing snowmelt dates and flowering of plants at a site at 2900 meters altitude. This year's snowmelt occurred 23 April, whereas the previous year, snow melted 19 June, he reported.

Fecal glucocorticoid metabolites in wild yellow-bellied marmots: Experimental validation, individual differences and ecological correlates

Natural selection is expected to shape phenotypic traits that permit organisms to respond appropriately to the environments in which they live. One important mechanism by which animals cope with changes in their environment is through physiological responses to stressors mediated by glucocorticoid hormones. Here we perform biological and physiological validations of a minimally-invasive technique for assessing fecal corticosterone metabolites (FCMs) in captive and wild groups of yellow-bellied marmots (Marmota flaviventris). Then we draw from ten years of data on these obligate hibernators at the Rocky Mountain Biological Laboratory in Colorado, USA to assess the extent to which seasonal and daily changes explain naturalistic variation in baseline levels of FCMs. Interestingly, beyond important population-level variation with respect to year, season, time of day, sex, age and reproductive state, we found repeatable inter-individual differences in FCMs, suggesting this hormonal trait might be a meaningful target of selection. FCM levels were 68% lower in captive than wild marmots, suggesting that the natural environment in which these animals occur is generally more challenging or less predictable than life in captivity. Most live-trapping events failed to represent stressors for wild marmots such that repeated measurements of traits were possible with minimal “stress” to subjects. We also document the natural ranges of annual and seasonal variation necessary for understanding the extent to which anthropogenic assaults represent stressors for wild mammals. Taken together, this study provides a foundation for understanding the evolution of hormonal traits and has important welfare and conservation implications for field biologists.

Ecology on the Web

Ecology on the Web

Ecosystem engineering by beavers affects mayfly life histories

1. The North American beaver has been studied as a model ecosystem engineer for many decades. Previous studies have documented physical, chemical and biological impacts attributed to beaver engineering in both aquatic and terrestrial environments. This study focused on the effects of ecosystem engineering by beavers on life histories of a common mayfly and on the potential consequences for mayfly populations. 2. We studied 18 montane beaver ponds of varying size and shape in western Colorado near the Rocky Mountain Biological Laboratory. Our goal was to test whether variation in beaver pond morphology (pond size and shape) explains downstream changes in stream temperature, mayfly size and timing of emergence. 3. Downstream water temperatures varied predictably with pond morphology, being colder downstream of high-head dams and warmer downstream of low-head dams. Pond morphology was also a significant predictor of variation in the size of mature female Baetis bicaudatus (the most abundant mayfly), with larger females emerging downstream of high-head dams and smaller females downstream of low-head dams. The size of male B. bicaudatus was not significantly related to pond morphology or stream temperature. There was no relationship between pond morphology and variation in the timing of emergence of Baetis (males or females) between upstream and downstream reaches. 4. Our results have implications for the effects of beaver ponds on Baetis individual fitness because large Baetis females are more fecund. Therefore, predictable female size variation associated with beaver pond morphology makes it possible to model the effects of beaver activity on local contributions of Baetis to the regional pool of reproductive adults at the catchment scale. Additionally, predictable changes in the size of emerging mayflies may have important consequences for the magnitude of aquatic to terrestrial resource subsidies in beaver-modified systems.

Changes in snowmelt date and summer precipitation affect the flowering phenology of Erythronium grandiflorum (glacier lily; Liliaceae)

Climate change has affected species worldwide, including alterations in phenology, migration patterns, distribution, and survival. Because Erythronium grandiflorum is an early-season bloomer, alterations in its phenology may have serious implications for many North American Rocky Mountain communities, including changes in resource availability for pollinators and herbivores. • We investigated whether changes in the snowmelt date, summer temperature, and summer precipitation have altered the timing and abundance of flowering in E. grandiflorum by collecting long-term data on floral abundance from 1975-2008 in a series of 2 × 2 m plots at the Rocky Mountain Biological Laboratory (RMBL) in Gothic, Colorado in the United States. • Snowmelt date and mean summer temperature were negatively correlated. Over the 30-yr study, the snowmelt date advanced by 4.14 d/decade, and mean summer temperature increased by 0.38°C/decade. Summer precipitation was variable, showing no change. The first, peak, and last flowering dates of E. grandiflorum advanced an average of 3.2 d/decade. Furthermore, earlier snowmelt and greater summer precipitation in the previous year led to earlier flowering in E. grandiflorum. There was no change in flowering abundance in this species, indicating it may be controlled by a complex set of abiotic and biotic variables. • Our study indicates that snowmelt is arriving earlier at the RMBL, which has caused earlier flowering in E. grandiflorum. Because alterations in phenology can disrupt important ecological interactions, information on potential phenological shifts in species that interact with E. grandiflorum is essential in determining the net effect of climate-driven alterations in phenology.

Variation in the impact of climate change on flowering phenology and abundance: An examination of two pairs of closely related wildflower species

Variability in plant phenological responses to climate change is likely to lead to changes in many ecological relationships as the climate continues to change. We used a 34-yr record of flowering times and flower abundance for four species (two Delphinium [Ranunculaceae] species and two Mertensia [Boraginaceae] species) from a subalpine plant community near the Rocky Mountain Biological Laboratory to test the hypothesis that the phenologies of early-flowering species change more rapidly in response to climatological and other abiotic cues than do late-flowering species, a pattern previously found in plant communities in North America and Europe. We also explored a related hypothesis, that flower abundance of late-flowering species is more responsive to changes in climate than that of early-flowering species. The Delphinium species did not support these hypotheses, but the Mertensia species did. The difference between the peak flowering times of the early and late Mertensia species is expanding, leading to a period of diminished resources for pollinators that specialize on this genus. Mertensia ciliata populations are already severely declining in our study area, possibly as a result of earlier snowmelt. Together, these results show that the reported differences between early- and late-flowering species may be widespread, but they are not ubiquitous.

The Effects of Climate Change on the Phenological Interactions of Plants and Pollinators

AbstractSymposium title: Interdisciplinary Canary: Linkages between Ecology and Sustainable Decision Making in a Dynamic Environment*1) Background/Question/Methods* The responses of pollinators to climate change could include changes in phenology of migratory pollinators and in the routes or destinations for their migration, changes in the phenology and distribution of non-migratory species, and changes in the host plants they visit for nectar and pollen. Plants face similar challenges with regard to changes in their distributions, their reproductive phenology, and interactions with both co-flowering species and pollinators (competition, facilitation, etc.). Unless pollinators and their host plants are responding similarly to changing environmental cues that affect their phenology, their historical patterns of interaction, both mutualistic and competitive, are likely to change. Long-term data are essential to investigating which if any of these potential outcomes are occurring. A 36-year record of abundance and phenology of flowering of 90+ wildflower species, surveys of the altitudinal distribution of bumble bees in the 1970s and the past few years, and data from a long-term Malaise trap sampling program, all near the Rocky Mountain Biological Laboratory (West Elk mountains, Colorado) are used for this investigation. Results/Conclusions* Although the flowering phenology of all species examined to date is affected by a single environmental event, disappearance of the winter snowpack (range 22 April -19 June since 1975), either their responses to that single cue are not uniform, or different species respond to additional cues in addition to snowmelt (e.g., growing degree days). Thus the community of co-flowering species varies temporally and quantitatively among years; differential sensitivity to frost damage is an example of an environmental variable that generates the quantitative variation among years, and is in turn affected by date of snowmelt. Arrival dates of migratory Broad-tailed Hummingbirds are significantly correlated with the amount of snow remaining on 30 April, and with the day of first flowering of Erythronium grandiflorum (glacier lily), the first flower that they visit at this site in the spring. Altitudinal distributions of at least some bumble bee species, and of the flowers they feed on, are also changing, with one bee species occurring about 600m higher than it did 30 years ago and one wildflower (Mertensia cilata) disappearing from lower altitudes where it was historically common. As these communities of plants and pollinators respond to environmental changes with changes in phenology and distribution, new interactions will be created and old ones will be lost.

EFFECTS OF CLIMATE CHANGE ON PHENOLOGY, FROST DAMAGE, AND FLORAL ABUNDANCE OF MONTANE WILDFLOWERS

The timing of life history traits is central to lifetime fitness and nowhere is this more evident or well studied as in the phenology of flowering in governing plant reproductive success. Recent changes in the timing of environmental events attributable to climate change, such as the date of snowmelt at high altitudes, which initiates the growing season, have had important repercussions for some common perennial herbaceous wildflower species. The phenology of flowering at the Rocky Mountain Biological Laboratory (Colorado, USA) is strongly influenced by date of snowmelt, which makes this site ideal for examining phenological responses to climate change. Flower buds of Delphinium barbeyi, Erigeron speciosus, and Helianthella quinquenervis are sensitive to frost, and the earlier beginning of the growing season in recent years has exposed them to more frequent mid-June frost kills. From 1992 to 1998, on average 36.1% of Helianthella buds were frosted, but for 1999-2006 the mean is 73.9%; in only one year since 1998 have plants escaped all frost damage. For all three of these perennial species, there is a significant relationship between the date of snowmelt and the abundance of flowering that summer. Greater snowpack results in later snowmelt, later beginning of the growing season, and less frost mortality of buds. Microhabitat differences in snow accumulation, snowmelt patterns, and cold air drainage during frost events can be significant; an elevation difference of only 12 m between two plots resulted in a temperature difference of almost 2 degrees C in 2006 and a difference of 37% in frost damage to buds. The loss of flowers and therefore seeds can reduce recruitment in these plant populations, and affect pollinators, herbivores, and seed predators that previously relied on them. Other plant species in this environment are similarly susceptible to frost damage so the negative effects for recruitment and for consumers dependent on flowers and seeds could be widespread. These findings point out the paradox of increased frost damage in the face of global warming, provide important insights into the adaptive significance of phenology, and have general implications for flowering plants throughout the region and anywhere climate change is having similar impacts.