Site fidelity by bees drives pollination facilitation in sequentially blooming plant species.

Plant species can influence the pollination and reproductive success of coflowering neighbors that share pollinators. Because some individual pollinators habitually forage in particular areas, it is also possible that plant species could influence the pollination of neighbors that bloom later. When flowers of a preferred forage plant decline in an area, site-fidelity may cause individual flower feeders to stay in an area and switch plant species rather than search for preferred plants in a new location. A newly blooming plant species may quickly inherit a set of visitors from a prior plant species, and therefore experience higher pollination success than it would in an area where the first species never bloomed. To test this, we manipulated the placement and timing of two plant species, Delphinium barbeyi and later-blooming Gentiana parryi. We recorded the responses of individually marked bumble bee pollinators. About 63% of marked individuals returned repeatedly to the same areas to forage on Delphinium. When Delphinium was experimentally taken out of bloom, most of those site-faithful individuals (78%) stayed and switched to Gentiana. Consequently, Gentiana flowers received more visits in areas where Delphinium had previously flowered, compared to areas where Delphinium was still flowering or never occurred. Gentiana stigmas received more pollen in areas where Delphinium disappeared than where it never bloomed, indicating that Delphinium increases the pollination of Gentiana when they are separated in time. Overall, we show that individual bumble bees are often site-faithful, causing one plant species to increase the pollination of another even when separated in time, which is a novel mechanism of pollination facilitation.

Flower orientation influences the consistency of bumblebee movement within inflorescences.

BACKGROUND AND AIMS: Plant species differ greatly in the three-dimensional arrangements of their flowers (inflorescence architecture). However, the nature of selection responsible for this diversity is poorly understood. Studies that examine among-species variation suggest that inflorescence architecture affects pollinator behaviour, and so should influence plant mating. However, few studies consider the consequences of within-population architectural variation for pollinator behaviour. METHODS: We manipulated inflorescence architecture of Delphinium glaucum to contrast bumblebee responses to normal and one-sided (secund) inflorescences. KEY RESULTS: The 'dimensionality' of manipulated inflorescences did not affect the number of flowers that bees visited; however, bees moved upward proportionally more on secund inflorescences. CONCLUSIONS: This study shows that realistic within-population variation in inflorescence architecture can manipulate pollinator behaviour. These results bear important consequences for plant mating success and the coordinated evolution of inflorescence architecture and floral specialization within inflorescences. These results also question why secund inflorescences are rare, for which we propose four testable explanations.

Heterospecific pollen deposition in Delphinium barbeyi: linking stigmatic pollen loads to reproductive output in the field.

BACKGROUND AND AIMS: Most pollinators are generalists and therefore are likely to transfer heterospecific pollen among co-flowering plants. Most work on the impacts of heterospecific pollen deposition on plant fecundity has utilized hand-pollination experiments in greenhouse settings, and we continue to know very little about the reproductive effects of heterospecific pollen in field settings. METHODS: We explored how patterns of naturally deposited heterospecific pollen relate to the reproductive output of Delphinium barbeyi, a common subalpine perennial herb in the Rocky Mountains (USA). We assessed a wide range of naturally occurring heterospecific pollen proportions and pollen load sizes, and linked stigmatic pollen deposition directly to seed set in individual carpels in the field. KEY RESULTS: We found that heterospecific pollen deposition in D. barbeyi is common, but typically found at low levels across stigmas collected in our sites. Neither conspecific nor heterospecific pollen deposition was related to carpel abortion. By contrast, we saw a significant positive relationship between conspecific pollen amount and viable seed production, as well as a significant negative interaction between the effects of conspecific pollen and heterospecific pollen amount, whereby the effect of conspecific pollen on viable seed production became weaker with greater heterospecific deposition on stigmas. CONCLUSIONS: To our knowledge, this is the first demonstration of a relationship between heterospecific pollen and seed production in a field setting. In addition, it is the first report of an interaction between conspecific and heterospecific pollen quantities on seed production. These findings, taken with the results from other studies, suggest that greenhouse hand-pollination studies and field studies should be more tightly integrated in future work to better understand how heterospecific pollen transfer can be detrimental for plant reproduction.

Yeasts in nectar enhance male fitness in a montane perennial herb.

Floral nectar of many plant species is prone to colonization by microbial organisms such as yeasts. Their presence and metabolism of nectar chemical components have the potential to modify a suite of floral traits important for pollinator attraction, including nectar quality and scent. However, studies on the direct and indirect effects of nectar-inhabiting microorganisms on pollinator behavior and plant reproductive success remain rare. To determine their potential to affect pollinator behavior and plant fitness, we experimentally manipulated the common nectar-inhabiting yeast Metschnikowia reukaufii in the nectar of Delphinium nuttallianum, a short-lived montane perennial herb. We detected positive, indirect, pollinator-mediated effects of yeasts on male plant fitness measured as pollen donation using powdered fluorescent dyes. However, we detected no direct or indirect effects on components of female fitness. Matching effects on male plant fitness, pollinators responded positively to the presence of yeasts, removing more nectar from flowers treated with M. reukaufii. Our results provide evidence of effects of nectar-inhabiting yeasts on male plant fitness and highlight the importance of microorganisms in mediating plant-pollinator interactions and subsequent plant fitness.

Single pollinator species losses reduce floral fidelity and plant reproductive function.

Understanding the functional impacts of pollinator species losses on plant populations is critical given ongoing pollinator declines. Simulation models of pollination networks suggest that plant communities will be resilient to losing many or even most of the pollinator species in an ecosystem. These predictions, however, have not been tested empirically and implicitly assume that pollination efficacy is unaffected by interactions with interspecific competitors. By contrast, ecological theory and data from a wide range of ecosystems show that interspecific competition can drive variation in ecological specialization over short timescales via behavioral or morphological plasticity, although the potential implications of such changes in specialization for ecosystem functioning remain unexplored. We conducted manipulative field experiments in which we temporarily removed single pollinator species from study plots in subalpine meadows, to test the hypothesis that interactions between pollinator species can shape individual species' functional roles via changes in foraging specialization. We show that loss of a single pollinator species reduces floral fidelity (short-term specialization) in the remaining pollinators, with significant implications for ecosystem functioning in terms of reduced plant reproduction, even when potentially effective pollinators remained in the system. Our results suggest that ongoing pollinator declines may have more serious negative implications for plant communities than is currently assumed. More broadly, we show that the individual functional contributions of species can be dynamic and shaped by the community of interspecific competitors, thereby documenting a distinct mechanism for how biodiversity can drive ecosystem functioning, with potential relevance to a wide range of taxa and systems.

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.

Reproductive and physiological responses to simulated climate warming for four subalpine species.

* The carbon costs of reproduction were examined in four subalpine herbaceous plant species for which number and size of flowers respond differently under a long-term infrared warming experiment. * Instantaneous measurements of gas exchange and an integrative model were used to calculate whole-plant carbon budgets and reproductive effort (RE). * Of the two species for which flowering was reduced, only one (Delphinium nuttallianum) exhibited higher RE under warming. The other species (Erythronium grandiflorum) flowers earlier when freezing events under warming treatment could have damaged floral buds. Of the two species for which flowering rates were not reduced, one (Helianthella quinquenervis) had higher RE, while RE was unaffected for the other (Erigeron speciosus). Each of these different responses was the result of a different combination of changes in organ size and physiological rates in each of the species. * Results show that the magnitude and direction of responses to warming differ greatly among species. Such results demonstrate the importance of examining multiple species to understand the complex interactions among physiological and reproductive responses to climate change.

Facilitation drives local abundance and regional distribution of a rare plant in a harsh environment.

The importance of facilitation to local community dynamics is becoming increasingly recognized. However, the predictability of positive interactions in stressful environments, the balance of competition and facilitation along environmental gradients, and the scaling of local positive interactions to regional distributions are aspects of facilitation that remain unresolved. I explored these questions in a habitat specialist, Delphinium uliginosum, and a moss, Didymodon tophaceus, both found in small serpentine wetlands. I tested three hypotheses: (1) moss facilitates germination, growth, and/or fecundity of D. uliginosum; (2) facilitation is stronger at the harsher ends of gradients in soil moisture, toxicity, and/or biomass; and (3) facilitation is reflected in positive associations at the levels of local abundance and regional occurrence. Although considerable competitive interactions occurred in later life stages, moss strongly facilitated D. uliginosum seedling emergence. There was no evidence that this facilitative effect weakened, or switched to competition, in benign environments. D. uliginosum was more locally abundant and more frequently present, across a large portion of its range, with than without moss, indicating a net facilitative effect in the face of competitive influences. Facilitated recruitment, possibly by seed retention, was found to be an important control on abundance and distribution in this rare species.

Expression of ethylene receptors Dl-ERS1-3 and Dl-ERS2, and ethylene response during flower senescence in Delphinium.

To clarify the relationships of flower senescence, especially sepal abscission, and ethylene receptor gene expression in different flower parts, we isolated two cDNAs encoding ethylene receptors Dl-ERS1-3 and Dl-ERS2 from Delphinium flowers. Deduced polypeptides possessed no response regulator domain, indicating that they belong to a family of ethylene response sensor (ERS) ethylene receptors. Dl-ERS1-3 and Dl-ERS2 exhibited constitutive levels during flower senescence. Exogenous ethylene increased transcript levels in sepals, which are influenced by ethylene but not in gynoecia and receptacles, which produce ethylene. It was suggested that expression of ethylene receptor genes under ethylene exposure was differentially regulated in each organ of the flower.

Co-expression of an ethylene receptor gene, ERS1, and ethylene signaling regulator gene, CTR1, in Delphinium during abscission of florets.

We are trying to determine the mechanisms responsible for ethylene-induced floret abscission in cut flowers of Delphinium and recently identified an ethylene receptor gene, ERS1, and studied its response to ethylene treatment. In order to identify additional components of the ethylene response network in Delphinium, we performed 3' and 5' rapid amplification of cDNA ends (RACE) using the consensus sequence of the serine/threonine kinase domain of the ethylene signaling regulator gene (CTR1) involved in the constitutive triple response (CTR) to ethylene. The full-length cDNA (2754 nt) encoded a protein of 800 amino acids, which contained the expected serine/threonine kinase domain, the consensus ATP-binding site, and the serine/threonine kinase catalytic site. The protein had quite high (>50%) overall identity to CTR1 from Arabidopsis and tomato, and 70-75% identity in the catalytic site. The amount of mRNA encoding both CTR1 and ERS1 more than doubled within 6 h in cut florets incubated in the presence of exogenous ethylene. Similarly, the amount of ERS1 transcript doubled in florets within 6 d of harvesting, presumably in response to endogenous ethylene, while CTR1 mRNA increased to about 40% over the same period. However, in the presence of silver thiosulfate (STS), an ethylene inhibitor, the level of both transcripts remained essentially unchanged for the first 8 d before declining to very low levels. Florets on the control plants had almost completely abscised by 6 d, but the florets on STS-treated plants had not abscised by 20 d, by which time the flowers were almost dead. The data are consistent with the hypothesis that endogenous ethylene evokes the accumulation of both these transcripts (and their encoded proteins), thereby speeding up abscission and reducing the useful shelf life of the cut flowers.