High toxin concentration in pollen may deter collection by bees in butterfly-pollinated Rhododendron molle.

BACKGROUNDS AND AIMS: The hypothesis that plants evolve features that protect accessible pollen from consumption by flower visitors remains poorly understood. METHODS: To explore potential chemical defenses against pollen consumption, we examined the pollinator assemblage, foraging behaviour, visitation frequency and pollen transfer efficiency in Rhododendron molle, a highly toxic shrub containing Rhodojaponin III. Nutrient (protein and lipid) and toxic components in pollen and other tissues were measured. KEY RESULTS: Overall in the five populations, floral visits by butterflies and bumblebees were relatively more frequent than visits by honeybees. All foraged for nectar but not pollen. Butterflies did not differ from bumblebees in the amount of pollen removed per visit, but deposited more pollen per visit. Pollination experiments indicated that R. molle was self-compatible, but both fruit and seed production were pollen limited. Our analysis indicated that the pollen was not protein-poor and had a higher concentration of the toxic compound Rhodojaponin III than petals and leaves, which compound was undetectable in nectar. CONCLUSION: Pollen toxicity in Rhododendron flowers may discourage pollen robbers (bees) from taking the freely accessible pollen grains, while the toxin-free nectar rewards effective pollinators, promoting pollen transfer. This preliminary study supports the hypothesis that chemical defense in pollen would be likely to evolve in species without physical protection from pollinivores.

Testing the effect of individual scent compounds on pollinator attraction in nature using quasi-isogenic Capsella lines.

PREMISE: Floral scent, usually consisting of multiple compounds, is a complex trait, and its role in pollinator attraction has received increasing attention. However, disentangling the effect of individual floral scent compounds is difficult due to the complexity of isolating the effect of single compounds by traditional methods. METHODS: Using available quasi-isogenic lines (qILs) that were generated as part of the original mapping of the floral scent volatile-related loci CNL1 (benzaldehyde) and TPS2 (ß-ocimene) in Capsella, we generated four genotypes that should only differ in these two compounds. Plants of the four genotypes were introduced into a common garden outside the natural range of C. rubella or C. grandiflora, with individuals of a self-compatible C. grandiflora line as pollen donors, whose different genetic background facilitates the detection of outcrossing events. Visitors to flowers of all five genotypes were compared, and the seeds set during the common-garden period were collected for high-throughput amplicon-based sequencing to estimate their outcrossing rates. RESULTS: Benzaldehyde and ß-ocimene emissions were detected in the floral scent of corresponding genotypes. While some pollinator groups showed specific visitation preferences depending on scent compounds, the outcrossing rates in seeds did not vary among the four scent-manipulated genotypes. CONCLUSIONS: The scent-manipulated Capsella materials constructed using qILs provide a powerful system to study the ecological effects of individual floral scent compounds under largely natural environments. In Capsella, individual benzaldehyde and ß-ocimene emission may act as attractants for different types of pollinators.

The evolution of flower-pollinator trait matching, and why do some alpine gingers appear to be mismatched?

BACKGROUND AND AIMS: Morphological matching between flower and pollinator traits has been documented in diverse plant lineages. Indeed, the matching of corolla tube length and pollinator tongue length has been cited repeatedly as a classic case of coevolution. However, there are many possible evolutionary routes to trait matching. Our aim here is both to review the evolutionary mechanisms of plant-pollinator trait matching and to investigate a specific case of trait matching/mismatching in a genus of alpine gingers. METHODS: Roscoea gingers with long corolla tubes in the western Himalayas have pollinators with correspondingly long tongues, but the match between corolla tube and pollinator tongue lengths is not seen in the eastern Himalayas. Six floral traits were measured, including corolla tube depth, an internal trait controlling pollinator access to nectar. We calculated coefficients of variation and phylogenetically controlled correlation patterns of these traits in six Roscoea species in order to gain possible insights into stabilizing selection and modularization of these traits. KEY RESULTS: The distal (nectar-containing) portion of the corolla tube exhibited lower coefficients of variations than did the basal portion. This is consistent with the hypothesis that pollinators mediate stabilizing selection on the distal, but not basal, portion of the corolla tube. This result, combined with phylogenetic data, suggests that the elevated liquid level of nectar in the distal tube evolved subsequent to dispersal into the eastern Himalayan region and loss of long-tongue pollinators. After accounting for phylogeny, corolla tube length, anther length, style length and labellum width were all intercorrelated. Corolla-tube depth was not part of this covariational module, however, suggesting separate adaptation to short-tongued pollinators. CONCLUSIONS: The reduction in functional corolla tube depth in the Roscoea appears to be related to the loss of long-tongued pollinators associated with dispersal to the eastern Himalayas and pollination by short-tongued pollinators. The apparent mismatch between floral tubes and pollinator tongues is a case of cryptic trait matching between flowers and pollinators, underscoring the importance of combining floral anatomy with pollination ecology in assessing plant-pollinator trait matching.

Do Silene species with exposed stigmas tolerate interference by heterospecific pollen?

PREMISE: Co-flowering species that have not evolved an avoidance mechanism may have tolerance to heterospecific pollen (HP) deposition as an adaptive strategy to minimize any deleterious effects of HP transfer, but empirical evidence for the tolerance hypothesis remains scarce. METHODS: To estimate the potential effects of heterospecific pollen deposition (HPD) on female reproductive success, we counted conspecific (CP) and HP pollen grains deposited on stigmas and assessed subsequent seed set of both open- and hand-pollinated flowers in three co-flowering Silene species with exposed stigmas that usually received numerous HP grains on the elongated receptive area. RESULTS: The percentage of HP grains per flower (HP%) varied from 16.6% to 43.0% among three species. Silene chungtienensis had lower HP%, and the CP-HP relationship was neutral; S. gracilicaulis and S. yunnanensis had a relatively higher HP% with a positive CP-HP relationship. The effects of CP and HP number on natural seed set were positive for all three species, but HP% had stronger negative effects in S. chungtienensis and S. gracilicaulis. In hand-pollinated flowers of the three Silene species, seed set did not decrease with HP whether CP was in excess or insufficient, indicating no negative effects of HPD on seed production. CONCLUSIONS: Consistent with the tolerance hypothesis, our results indicated that species with higher HP interference are likely to be tolerant to an increase in HP%. These species with generalist-pollinated flowers and exposed large stigmas may benefit from an increase of conspecific pollen deposition, despite the associated increase in heterospecific pollen deposition.

Pollen-feeding behavior of diverse insects on Geranium delavayi, a flower with large, accessible pollen grains.

PREMISE: Why have pollen grains evolved to be exceptionally large in some species? Pollen-feeding hypothesis suggests that if the proportion of pollen amounts for feeding is reduced in a flower, the low allocation to pollen number would allow pollen grains to be larger. METHODS: To examine whether species with large pollen grains experience low pollen consumption, the behavior of insects feeding on nectar and pollen was observed and pollen transfer efficiency was estimated for four visitor types in Geranium delavayi. To see whether bees actively collected pollen, the numbers of grains in pollen baskets and on the body were compared. Both nutritional value (total protein and lipid) and chemical defense (phenolic metabolites) in pollen against pollen feeders were measured. RESULTS: Bumblebees and honeybees foraged for nectar, rarely groomed pollen into corbiculae, and had >5× higher pollen transfer efficiency than smaller solitary bees and flies, which were pollen eaters that removed more pollen but deposited less. Pollen grains were characterized by low protein and high lipid content with a low protein-lipid ratio, an unfavorable combination for bumblebees. Three secondary metabolites were significantly higher in pollen grains (7.77 mg/g) than in petals (1.08 mg/g) or in nectar (0.44 mg/g), suggesting stronger chemical defense in pollen. CONCLUSIONS: Our results indicated that large bees took nectar but little of the nutritionally poor and highly toxic pollen. These data support one prediction of the pollen-feeding hypothesis, that species with few and large pollen grains would also have low pollen-consumption rates.

Degree of style coiling is associated with corolla-tube length in the nectarless flowers of Roscoea schneideriana.

The plant-pollinator 'arms race' model posits that a major driver of the evolution of elongated corollas in flowers is reciprocal selection for 'morphological fit' between pollinator-tongue length and access distance to nectar (usually corolla-tube length). Evidence for the pollinator-mediated selection on tube length and evolution of multiple, correlated floral traits remains inconclusive. To gain possible insights into the strength of stabilizing selection by assessing standing phenotypic variation, we measured a series of functionally important floral traits, including corolla tube length and 'effective' tube depth and degree of style coiling. We then calculated coefficients of variation (CV) for these traits in three field populations of R. schneideriana. Unlike in most long-tubed flowers, the bottom part of the corolla tube is completely occupied by the style, with no room for nectar. The length of this portion of the corolla tube was more variable (higher CV) than the upper part of the corolla tube, suggesting that functional tube depth was under stronger stabilizing selection. The degree of style coiling was negatively related to the corolla-tube length in all three populations of R. schneideriana, suggesting that there may be conflicting selection acting on style length and corolla-tube length, which are otherwise usually tightly correlated. Given the lack of nectar in the flowers of this species, the long corolla tubes and long styles may represent morphological holdovers from ancestors that were pollinated by long-tongued pollinators, as is still seen in related species in the western Himalayas.

Touch-sensitive stamens enhance pollen dispersal by scaring away visitors.

Animal-pollinated plants have to get pollen to a conspecific stigma while protecting it from getting eaten. Touch-sensitive stamens, which are found in hundreds of flowering plants, are thought to function in enhancing pollen export and reducing its loss, but experimental tests are scarce. Stamens of Berberis and Mahonia are inserted between paired nectar glands and when touched by an insect's tongue rapidly snap forward so that their valvate anthers press pollen on the insect's tongue or face. We immobilized the stamens in otherwise unmodified flowers and studied pollen transfer in the field and under enclosed conditions. On flowers with immobilized stamens, the most common bee visitor stayed up to 3.6× longer, yet removed 1.3× fewer pollen grains and deposited 2.1× fewer grains on stigmas per visit. Self-pollen from a single stamen hitting the stigma amounted to 6% of the grains received from single bee visits. Bees discarded pollen passively placed on their bodies, likely because of its berberine content; nectar has no berberine. Syrphid flies fed on both nectar and pollen, taking more when stamens were immobilized. Pollen-tracking experiments in two Berberis species showed that mobile-stamen-flowers donate pollen to many more recipients. These results demonstrate another mechanism by which plants simultaneously meter out their pollen and reduce pollen theft.

Compassion fatigue, burnout, compassion satisfaction and depression among emergency department physicians and nurses: a cross-sectional study.

OBJECTIVES: Emergency department physicians and nurses are at high risk of compassion fatigue, burnout and depression. The purpose of this study was to examine the inter-relationship between compassion fatigue, burnout, compassion satisfaction and depression in emergency department physicians and nurses. DESIGN: A cross-sectional study. SETTING: This study was conducted in five tertiary hospitals in five different cities across the province of Sichuan, China, in 2021. PARTICIPANTS: A total of 342 emergency department physicians and nurses participated in the study. MAIN OUTCOME MEASURES: Compassion fatigue, burnout, compassion satisfaction and depression scores. RESULTS: Among the study participants, 100% were found to have depressive symptoms, 27.8% had low compassion satisfaction, 2.3% had high burnout and 3.8% had compassion fatigue. In the final multiple linear regression model, marital status (p=0.008; 95% CI -5.205 to -0.789), history of chronic disease (p=0.003; 95% CI -6.461 to -1.386), compassion satisfaction (p<0.001; 95% CI 0.593 to 1.274), burnout (p=0.019; 95% CI 0.084 to 0.930) and compassion fatigue (p<0.001; 95% CI -1.527 to -1.053) among emergency department physicians and nurses were considered to be significant predictors of depression. CONCLUSIONS: The prevalence of depression among emergency department physicians and nurses is high in the province of Sichuan, China. Compassion fatigue, burnout and compassion satisfaction were significantly associated with depression in emergency department physicians and nurses. Hospital administrations should consider these findings to develop appropriate psychological interventions and strategies, to prevent, alleviate or treat severe depression among emergency department physicians and nurses in the province of Sichuan.

Foraging behavior and pollination efficiency of generalist insects in an understory dioecious shrub Helwingia japonica.

PREMISE: It has been hypothesized that pollination success in animal-pollinated dioecious plants relies on opportunistic pollinators with no discrimination against female flowers. However, empirical studies of pollinator foraging behavior and pollination effectiveness in dioecious species are few. METHODS: To investigate potential pollinators in Helwingia japonica, a dioecious shrub with small, inconspicuous flowers, we compared floral visitors and visit frequency to female and male plants for three flowering seasons in two field populations in subtropical forests in southwest China. Pollen placement on the insect bodies of four groups (solitary bees and other bees, fungus gnats, and other flies) was compared, and insect foraging behavior was observed. Pollen removal and conspecific and heterospecific pollen deposition per visit were measured to compare pollination effectiveness among the four groups. RESULTS: Floral visitors usually did not discriminate between male and female flowers and did not gather pollen into collections. Our measurements of pollen transfer efficiency showed that solitary bees were the most effective pollinators with the highest conspecific pollen deposition. These insects seemed to be opportunistic visitors because pollen grains of H. japonica were distributed evenly over different regions of the visitor's body, and heterospecific pollen accounted for over 50% of total pollen loads on stigmas in the two populations. CONCLUSIONS: Our investigations indicated that potential pollinators were generalists and did not discriminate against female flowers, as predicted for dioecious species pollinated by insects. A perspective of pollen removal by floral visitors offers insights into the evolution of plant sexual systems.

Pollen grain size associated with pollinator feeding strategy.

Angiosperm pollen grain diameter varies greatly from a few microns to over 100, but the selective forces driving the interspecific variation in pollen size remain unclear. Although both pre- and post-pollination hypotheses have been proposed, empirical evidence remains scarce. Here we propose that visits by pollen-foraging pollinators have selected against large pollen grains. An association between pollinator behaviour and pollen grain size was confirmed by field studies of 80 flowering species in natural communities, showing that pollinators positively collected pollen in those species with relatively smaller pollen grains but rarely did so in species with larger ones. Allowing for the confounding effects of pollinator type, flower size or style length and pollen grain number, we found a significant effect of pollen-foraging behaviour on variation in pollen grain size, particularly in bee-pollinated plants. While these results suggest that many plant species whose pollen is collected or consumed by pollinators produce small pollen grains, it remains unclear whether pollen grain size is directly affected by pollinator foraging habit or indirectly mediated by pollen number trade-offs.

Both small and large plants are likely to produce staminate (male) flowers in a hermaphrodite lily.

Resource allocation to female and male function may vary among individual plants in species with variable sex expression. Size-dependent sex allocation has been proposed in hermaphrodites, in which female-biased allocation may increase with plant size. In many hermaphrodites with large floral displays, however, later-produced flowers tend to be functionally male. This paradoxical relationship between female and male function and plant size remains poorly understood. The subalpine lily Lilium lankongense has individuals of three sexual types: males with only staminate flowers, hermaphrodites with only perfect flowers, and andromonoecious plants with both perfect and staminate flowers. Here we tested theoretical predictions of size-dependent sex allocation in L. lankongense by measuring plant height and flower number of individuals of each sex at five field sites in the mountainous region of Shangri-La, southwestern China. To investigate variation in phenotypic gender, we identified sex expression of 457 individuals one year later. Our investigation showed that male plants, which usually produced one flower, were significantly smaller than andromonoecious and hermaphrodite plants. In addition, the total flower numbers of andromonoecious and hermaphrodite plants increased significantly with plant size. Large individuals were more likely to produce terminal staminate flowers, as there were more flowers in andromonoecious than in hermaphrodite individuals. Non-flowered plants were significantly smaller than flowering ones. Perfect flowers had significantly larger petals and pistils than staminate flowers, but they did not differ in dry weight of stamens. Our findings indicate that when plants are small, the less costly sex is favored, consistent with the 'size-advantage hypothesis'. When plants are large, both female and male investments change isometrically, as later-produced flowers tend to be functionally male.

Transitions between the Terrestrial and Epiphytic Habit Drove the Evolution of Seed-Aerodynamic Traits in Orchids.

Orchids are globally distributed, a feature often attributed to their tiny dustlike seeds. They were ancestrally terrestrial but in the Eocene expanded into tree canopies, with some lineages later returning to the ground, providing an evolutionarily replicated system. Because seeds are released closer to the ground in terrestrial species than in epiphytic ones, seed traits in terrestrials may have been under selective pressure to increase seed dispersal efficiency. In this study, we test the expectations that seed airspace-a trait known to increase seed flotation time in the air-is (i) larger in terrestrial lineages and (ii) has increased following secondary returns to a terrestrial habit. We quantified and scored 20 seed traits in 121 species and carried out phylogenetically informed analyses. Results strongly support both expectations, suggesting that aerodynamic traits even in dust seeds are under selection to increase dispersal ability, following shifts in average release heights correlated with changes in habit.

Pollinator effectiveness and importance between female and male mining bee (Andrena).

Bees are often considered to be effective pollinators in both agricultural and natural ecosystems but could be ineffective pollinators in that they collect large quantities of pollen for food provision but deliver little to stigmas. Male bees do not collect pollen to feed larvae, and their pollination role has been underappreciated. Here we compare pollination effectiveness, visit frequency and pollen foraging behaviour between female and male individuals of a mining bee, Andrena emeishanica, visiting a nectariferous spring flower (Epimedium pubescens). Female bees were observed to forage for both pollen and nectar, but male bees foraged only for nectar. Female bees had large hairy hind tibiae with conspicuous scopae, and nearly 90% of the pollen grains they collected went onto the hind legs. Male bees removed less pollen from anthers than female bees but deposited more pollen on stigmas per visit. The higher pollen transfer efficiency of male bees was due to 48.4% of pollen grains remaining ungroomed on the thorax and abdomen, available for stigma contact, but their visitation rate to flowers was much lower. Our results indicate that male solitary bees could transfer more pollen on the stigma per visit but were less important (transferred less pollen in total, because they made fewer visits per unit time) than females.

Color-matching between pollen and corolla: hiding pollen via visual crypsis?

Visual signals attractive to friends may also attract enemies.  The bright colors of anthers and pollen have generally been thought to attract pollinators. We hypothesize that visual crypsis of anthers, and particularly pollen, should be favored in flowering plants because protection from pollen collectors reduces the loss of male gametes. To understand adaptive strategies relating to the color of pollen, we measured the color of pollen, undehisced anther sacs, and their background, the corolla, with a spectrometer for 104 insect-pollinated flowering species from a natural community in Hengduan Mountains, southwest China. The colors of anthers, pollen and corollas were diverse in these species. The color diversity of exposed pollen was significantly higher than that of concealed pollen (i.e. where anthers are enclosed or shielded by corollas). The color contrast between pollen and corolla was significantly smaller in species with exposed pollen than in those with concealed pollen. Unlike anther color, exposed pollen color tended to match its background corolla color. Our phylogenetic comparative analysis showed contrasting effects of pollen color patterns between flowers with exposed pollen and those with concealed pollen, revealing a strategy of hiding pollen from pollen thieves via visual crypsis.

Bumblebee Rejection of Toxic Pollen Facilitates Pollen Transfer.

Many bees are effective pollen collectors; however, pollen grains collected by bees for larval food are lost for plant sexual reproduction. Recognition of these conflicting interests between bees and flowers is essential for understanding of reproduction for both bees and flowers [1-3]. Plant defense compounds in pollen may function to reduce pollen waste by deterring ineffective pollinators [4-6], but this hypothesis remains unexamined. Here, we provide evidence that secondary metabolites in pollen function as chemical defense by deterring some bees from gathering pollen. In two Dipsacus species, a defense compound, dipsacus saponin [7], occurs in pollen but not in nectar. We observed that bumblebees disliked grooming bitter-tasting pollen with a high saponin content. Manipulation of saponin concentrations in nectar and measurements of corbicular pollen showed that the bumblebee species differed in their tolerance to saponin. Those species susceptible to saponin groomed little Dipsacus pollen into their pollen loads, and their ungroomed pollen was observed to be effectively delivered to stigmas. By rewarding bees with edible nectar, but not pollen, plants solve the conflict of pollen partitioning between sexual and reward functions. Ungroomed toxic pollen on the bee body promotes pollen transfer efficiency, facilitating pollination.

Nectar supplementation changes pollinator behaviour and pollination mode in Pedicularis dichotoma: implications for evolutionary transitions.

Backgrounds and Aims: Gain or loss of floral nectar, an innovation in floral traits, has occurred in diverse lineages of flowering plants, but the causes of reverse transitions (gain of nectar) remain unclear. Phylogenetic studies show multiple gains and losses of floral nectar in the species-rich genus Pedicularis. Here we explore how experimental addition of nectar to a supposedly nectarless species, P. dichotoma, influences pollinator foraging behaviour. Methods: The liquid (nectar) at the base of the corolla tube in P. dichotoma was investigated during anthesis. Sugar components were measured by high-performance liquid chromatography. To understand evolutionary transitions of nectar, artificial nectar was added to corolla tubes and the reactions of bumble-bee pollinators to extra nectar were examined. Key Results: A quarter of unmanipulated P. dichotoma plants contained measurable nectar, with 0.01-0.49 µL per flower and sugar concentrations ranging from 4 to 39 %. The liquid surrounding the ovaries in the corolla tubes was sucrose-dominant nectar, as in two sympatric nectariferous Pedicularis species. Bumble-bees collected only pollen from control (unmanipulated) flowers of P. dichotoma, adopting a sternotribic pollination mode, but switched to foraging for nectar in manipulated (nectar-supplemented) flowers, adopting a nototribic pollination mode as in nectariferous species. This altered foraging behaviour did not place pollen on the ventral side of the bees, and sternotribic pollination also decreased. Conclusion: Our study is the first to quantify variation in nectar production in a supposedly 'nectarless' Pedicularis species. Flower manipulations by adding nectar suggested that gain (or loss) of nectar would quickly result in an adaptive behavioural shift in the pollinator, producing a new location for pollen deposition and stigma contact without a shift to other pollinators. Frequent gains of nectar in Pedicularis species would be beneficial by enhancing pollinator attraction in unpredictable pollination environments.

Pollen aggregation by viscin threads in Rhododendron varies with pollinator.

Pollen grains can be dispersed singly or variously aggregated in groups. Whether the evolution of pollen aggregation is driven by the pollinator remains unexplored. We hypothesize that an extensive pollen aggregation is favored under a scarcity of pollinators. Variation in pollen aggregation by viscin threads in 13 Rhododendron species was measured as it is related to pollen removal in a visit. Visitation rates of functional pollinator groups that vary in their grooming behavior were investigated in each species. Pollen deposited on stigmas in the field was also sampled. Seven Rhododendron species were infrequently pollinated by low-intensity grooming animals, including birds, butterflies and moths. The other six species were more frequently pollinated by bees with a high intensity of pollen grooming. Bird- and Lepidoptera-pollinated species produced longer pollen-connecting threads that connected more pollen grains. Phylogenetically independent contrast analysis of the 13 species showed that pollinator visitation frequency was negatively related to amounts of pollen removal per visit but not to stigmatic pollen loads. The finding of interspecific patterns in pollen removal related to pollinator visitation frequency suggests pollinator-mediated selection on pollen packaging strategies, supporting the hypothesis of floral evolution via pollen export.