Nitrogen-fixing bacteria boost floral attractiveness in a tropical legume species during nutrient limitation.

PREMISE: Legumes establish mutualistic interactions with pollinators and nitrogen (N)-fixing bacteria that are critical for plant reproduction and ecosystem functioning. However, we know little about how N-fixing bacteria and soil nutrient availability affect plant attractiveness to pollinators. METHODS: In a two-factorial greenhouse experiment to assess the impact of N-fixing bacteria and soil types on floral traits and attractiveness to pollinators in Chamaecrista latistipula (Fabaceae), plants were inoculated with N-fixing bacteria (NF+) or not (NF-) and grown in N-rich organic soil (+N organic soil) or N-poor sand soil (-N sand soil). We counted buds and flowers and measured plant size during the experiment. We also measured leaf, petal, and anther reflectance with a spectrophotometer and analyzed reflectance curves. Using the bee hexagon model, we estimated chromatic contrasts, a crucial visual cues for attracting bees that are nearby and more distant. RESULTS: NF+ plants in -N sand soil had a high floral display and color contrasts. On the other hand, NF- plants and/or plants in +N organic soil had severely reduced floral display and color contrasts, decreasing floral attractiveness to bee pollinators. CONCLUSIONS: Our findings indicate that the N-fixing bacteria positively impact pollination, particularly when nutrients are limited. This study provides insights into the dynamics of plant-pollinator interactions and underscores the significant influence of root symbionts on key floral traits within tropical ecosystems. These results contribute to understanding the mechanisms governing mutualisms and their consequences for plant fitness and ecological dynamics.

An evolutionary disruption of the buzz pollination syndrome in neotropical montane plants.

PREMISE: Under pollinator limitations, specialized pollination syndromes may evolve toward contrasting responses: a generalized syndrome with increased pollinator attraction, pollinator reward, and pollen transfer capacity; or the selfing syndrome with increased self-pollen deposition, but reduced pollinator attraction and pollen transfer capacity. The buzz-pollination syndrome is specialized to explore female vibrating bees as pollinators. However, vibrating bees become less-active pollinators at montane areas of the Atlantic Forest (AF) domain. This study investigated whether the specialized buzz-pollination syndrome would evolve toward an alternative floral syndrome in montane areas of the AF domain, considering a generalized and the selfing syndromes as alternative responses. METHODS: We utilized a lineage within the buzz-pollinated Miconia as study system, contrasting floral traits between montane AF-endemic and non-endemic species. We measured and validated floral traits that were proxies for pollinator attraction, reward access, pollen transfer capacity, and self-pollen deposition. We inferred the evolution of floral trait via phylogenetic comparative methods. RESULTS: AF-endemic species have selectively evolved greater reward access and more frequently had generalist pollination. Nonetheless, AF-endemic species also have selectively evolved toward lower pollen transfer capacity and greater self pollination. These patterns indicated a complex evolutionary process that has jointly favored a generalized and the selfing syndromes. CONCLUSIONS: The buzz pollination syndrome can undergo an evolutionary disruption in montane areas of the AF domain. This floral syndrome is likely more labile than often assumed, allowing buzz-pollinated plants to reproduce in environments where vibrating bees are less-reliable pollinators.

An assessment of methods to combine evolutionary history and conservation: A case study in the Brazilian campo rupestre.

Premise: Conservation policies typically focus on biodiversity hotspots. An alternative approach involves analyzing the evolutionary history of lineages in geographic areas along with their threat levels to guide conservation efforts. Mountains exhibit high levels of plant species richness and micro-endemism, and biogeographic studies commonly point to recent and rapid evolutionary radiations in these areas. Using a nearly endemic clade of legumes, our study evaluates conservation prioritization approaches in the campo rupestre, a Neotropical ecosystem associated with mountaintops that is located between two biodiversity hotspots. Methods: We compared the EDGE and EDGE2 metrics, which combine the evolutionary distinctiveness and the extinction risk of a species in a single value. These metrics are compared with traditional metrics used to assess conservation priority, such as phylogenetic diversity. Results: The EDGE values reported are lower than those of other studies using this metric, mostly due to the prevalence of threatened species with short phylogenetic branch lengths (low values of evolutionary distinctiveness). Certain areas of campo rupestre with relatively high phylogenetic diversity and EDGE values do not correspond to areas with high species richness, agreeing with previous studies on biodiversity hotspots. Discussion: Our study highlights the necessity of conservation of the campo rupestres as well as advantages and disadvantages of using EDGE, EDGE2, and phylogenetic diversity for appropriate selection of conservation areas with rapid evolutionary radiations. The selection of the metrics will depend primarily on the life history of the focus group and the data availability, as well as the conservation approach.

Lianas in tropical dry seasonal forests have a high hydraulic efficiency but not always a higher embolism resistance than lianas in rainforests.

BACKGROUND AND AIMS: Lianas have higher relative abundance and biomass in drier seasonal forests than in rainforests, but whether this difference is associated with their hydraulic strategies is unclear. Here, we investigate whether lianas of seasonally dry forests are safer and more efficient in water transport than rainforest lianas, explaining patterns of liana abundance. METHODS: We measured hydraulic traits on five pairs of congeneric lianas of the tribe Bignonieae in two contrasting forest sites: the wet 'Dense Ombrophilous Forest' in Central Amazonia (~2 dry months) and the drier 'Semideciduous Seasonal Forest' in the inland Atlantic Forest (~6 dry months). We also gathered a broader database, including 197 trees and 58 liana species from different tropical forests, to compare hydraulic safety between habits and forest types. KEY RESULTS: Bignonieae lianas from both forests had high and similar hydraulic efficiency but exhibited variability in resistance to embolism across forest types when phylogenetic relationships were taken into account. Three genera had higher hydraulic safety in the seasonal forest than in the rainforest, but species across both forests had similar positive hydraulic safety margins despite lower predawn water potential values of seasonal forest lianas. We did not find the safety-efficiency trade-off. Merging our results with previously published data revealed a high variability of resistance to embolism in both trees and lianas, independent of forest types. CONCLUSIONS: The high hydraulic efficiency of lianas detected here probably favours their rapid growth across tropical forests, but differences in hydraulic safety highlight that some species are highly vulnerable and may rely on other mechanisms to cope with drought. Future research on the lethal dehydration threshold and the connection between hydraulic resistance strategies and liana abundance could offer further insights into tropical forest dynamics under climatic threats.

Resource availability and disturbance frequency shape evolution of plant life forms in Neotropical habitats.

Organisms use diverse strategies to thrive in varying habitats. While life history theory partly explains these relationships, the combined impact of resource availability and disturbance frequency on life form strategy evolution has received limited attention. We use Chamaecrista species, a legume plant lineage with a high diversity of plant life forms in the Neotropics, and employ ecological niche modeling and comparative phylogenetic methods to examine the correlated evolution of plant life forms and environmental niches. Chamaephytes and phanerophytes have optima in environments characterized by moderate water and nutrient availability coupled with infrequent fire disturbances. By contrast, annual plants thrive in environments with scarce water and nutrients, alongside frequent fire disturbances. Similarly, geophyte species also show increased resistance to frequent fire disturbances, although they thrive in resource-rich environments. Our findings shed light on the evolution of plant strategies along environmental gradients, highlighting that annuals and geophytes respond differently to high incidences of fire disturbances, with one enduring it as seeds in a resource-limited habitat and the other relying on reserves and root resprouting systems in resource-abundant habitats. Furthermore, it deepens our understanding of how organisms evolve associated with their habitats, emphasizing a constraint posed by low-resource and high-disturbance environments.

The role of intraspecific variation in bumblebee body size and behavior on buzz pollination of a tropical legume species.

PREMISE: The outcomes of generalized mutualisms rely on partner trait variation. In pollination mutualisms, although less explored, intraspecific variation in pollinator traits can be pivotal for successful pollination. We investigated the role of intraspecific body size and behavioral trait variations of bumblebee Bombus morio on the pollination of a buzz-pollinated legume species, Chamaecrista latistipula. METHODS: To explore the impact of body size and behavior of B. morio on the pollination of C. latistipula, we observed visits to virgin flowers and quantified the pollen removal and deposition (pollination performance) and fruit and seed production (reproductive fitness). By analyzing video and sound recordings, we measured B. morio body size and behavior on each flower, including bee vibration descriptors. RESULTS: We observed intraspecific behavioral differences among B. morio bumblebees associated with different body sizes. Larger bumblebees had half the handling time and vibrational pulses, less angular displacement within flowers, and larger relative peak amplitudes during vibrations than smaller bumblebees did. Regardless of their large variation in size and behavior, bumblebees were equally effective in removing pollen and pollinating flowers. The high female reproductive fitness was independent of bumblebee body size and behavior, likely due to the interaction between both. On the other hand, larger bumblebees visited flowers for shorter periods, probably promoting higher male reproductive fitness. CONCLUSIONS: This study is the first to highlight the large intraspecific variation of bumblebee body size and behavior in buzz-pollinated flowers in the field. Together, body size and behavior effects unexpectedly cancel each other, generating a high buzz pollination efficiency.

Plant species with larger extrafloral nectaries produce better quality nectar when needed and interact with the best ant partners.

Few studies have explored the phenotypic plasticity of nectar production on plant attractiveness to ants. Here, we investigate the role of extrafloral nectary (EFN) size on the productivity of extrafloral nectar in three sympatric legume species. We hypothesized that plant species with larger EFNs (i) have higher induced nectar secretion after herbivory events, and (ii) are more likely to interact with more protective (i.e. dominant) ant partners. We target 90 plants of three Chamaecrista species in the field. We estimated EFN size and conducted field experiments to evaluate any differences in nectar traits before and after leaf damage to investigate the phenotypic plasticity of nectar production across species. We conducted multiple censuses of ant species feeding on EFNs over time. Plant species increased nectar descriptors after leaf damage, but in different ways. Supporting our hypothesis, C. duckeana, with the largest EFN size, increased all nectar descriptors, with most intense post-herbivory-induced response, taking its place as the most attractive to ants, including dominant species. EFN size variation was an excellent indicator of nectar productivity across species. The higher control over reward production in plants with larger sized EFNs reflects an induction mechanism under damage that reduces costs and increases the potential benefits of indirect biotic defences.

From species descriptions to diversity patterns: the validation of taxonomic data as a keystone for ant diversity studies reproducibility and accuracy.

Research findings in natural sciences need to be comparable and reproducible to effectively improve our understanding of ecological and behavioural patterns. In this sense, knowledge frontiers in biodiversity studies are directly tied to taxonomic research, especially in species-rich tropical regions. Here we analysed the taxonomic information available in 470 studies on Brazilian ant diversity published in the last 50 years. We aimed to quantify the proportion of studies that provide enough data to validate taxonomic identification, explore the frequency of studies that properly acknowledge their taxonomic background, and investigate the primary resources for ant identification in Brazil. We found that most studies on Brazilian ant diversity (73.6%) explicitly stated the methods used to identify their specimens. However, the proportion of papers that provide complete data for the repository institutions and vouchered specimens is vanishingly small (5.8%). Additionally, only 40.0% of the studies consistently presented taxon authorities and years of description, rarely referencing taxonomic publications correctly. In turn, the number of specialists and institutions consulted for ant identification in Brazil has increased in the last years, along with the number of studies that explicitly provide their taxonomic procedures for ant identification. Our findings highlight a shift between generations regarding the recognition of taxonomy as fundamental science, deepening our understanding of biodiversity.

The evolution of ecological specialization underlies plant endemism in the Atlantic Forest.

BACKGROUND AND AIMS: The evolution of ecological specialization is favoured under divergent selection imposed by increased environmental heterogeneity, although specialization can limit the geographical range of organisms, thus promoting endemism. The Atlantic Forest (AF) is an ancient montane domain with high plant endemism, containing different environments for plant specialization. Miconia is the most diverse genus of woody flowering plant within the AF domain, including AF-endemic and non-endemic lineages. We hypothesized that Miconia species have faced increased environmental heterogeneity and consequently have been selected towards increased specialization in the AF domain, and this increased specialization has greatly reduced species geographical ranges, ultimately promoting endemism. Hence, we made the following predictions: (1) AF-endemic species should face greater environmental heterogeneity than non-endemic species; (2) AF-endemic species should be more specialized than non-endemic species; (3) specialization should lead to smaller geographical ranges; (4) specialization and small geographical ranges among AF-endemic species should conform to a selection-driven evolutionary scenario rather than to a neutral evolutionary scenario; and (5) small geographical ranges among AF-endemic species should date back to the occupation of the AF domain rather than to more recent time periods. METHODS: We used geographical, environmental and phylogenetic data on a major Miconia clade including AF-endemic and non-endemic species. We calculated Rao's Q to estimate the environmental heterogeneity faced by species. We used georeferenced occurrences to estimate the geographical ranges of species. We applied environmental niche modelling to infer species niche breadth. We inferred the most likely evolutionary scenario for species geographical range and niche breadth via a model-fitting approach. We used ancestral reconstructions to evaluate species geographical range throughout time. KEY RESULTS: Atlantic Forest-endemic species faced 33-60 % more environmental heterogeneity, with the increase being associated with montane landscapes in the AF. The AF-endemic species were 60 % more specialized overall, specifically over highly variable environmental gradients in AF montane landscapes. Specialization strongly predicted small geographical ranges among AF-endemic species and was a major range-limiting factor among endemic lineages. The AF-endemic species have evolved towards specialization and small geographical ranges under a selection-driven regime, probably imposed by the great environmental heterogeneity in AF montane landscapes. The AF-endemic species underwent a major reduction of geographical range immediately after their evolution, indicating a long-standing effect of selective pressures in the AF domain. CONCLUSION: Environmental heterogeneity imposes selective pressures favouring ecological specialization and small geographical ranges among plant lineages in the AF domain. This selection-driven process has probably promoted plant endemism in the AF domain throughout its history.

Data standardization of plant-pollinator interactions.

BACKGROUND: Animal pollination is an important ecosystem function and service, ensuring both the integrity of natural systems and human well-being. Although many knowledge shortfalls remain, some high-quality data sets on biological interactions are now available. The development and adoption of standards for biodiversity data and metadata has promoted great advances in biological data sharing and aggregation, supporting large-scale studies and science-based public policies. However, these standards are currently not suitable to fully support interaction data sharing. RESULTS: Here we present a vocabulary of terms and a data model for sharing plant-pollinator interactions data based on the Darwin Core standard. The vocabulary introduces 48 new terms targeting several aspects of plant-pollinator interactions and can be used to capture information from different approaches and scales. Additionally, we provide solutions for data serialization using RDF, XML, and DwC-Archives and recommendations of existing controlled vocabularies for some of the terms. Our contribution supports open access to standardized data on plant-pollinator interactions. CONCLUSIONS: The adoption of the vocabulary would facilitate data sharing to support studies ranging from the spatial and temporal distribution of interactions to the taxonomic, phenological, functional, and phylogenetic aspects of plant-pollinator interactions. We expect to fill data and knowledge gaps, thus further enabling scientific research on the ecology and evolution of plant-pollinator communities, biodiversity conservation, ecosystem services, and the development of public policies. The proposed data model is flexible and can be adapted for sharing other types of interactions data by developing discipline-specific vocabularies of terms.

A sophisticated case of division of labour in the trimorphic stamens of the Cassia fistula (Leguminosae) flower.

Buzz-pollinated pollen flowers have pollen as the primary resource for pollinators and must deal with a conflict between the exploitation of pollen grains by bees and pollination success. It has been hypothesized that heterostemony allows division of labour between stamens as a solution to the pollen dilemma. To test the division of labour hypothesis, we chose Cassia fistula, which has a trimorphic androecium and analysed androecium development, pollen grain release mechanisms and visitor behaviour. We explored the reflectance of floral organs and carried out an exclusion experiment to test the attractiveness of each stamen morph to the bee species. Finally, we explored the structural, ultrastructural and functional variation between the pollen grains, including pollen viability across stamen morphs. The differences among the three stamen morphs, which is developed from two whorls of the stamen, are the first evidence of the division of labour in our study system. Large Bombus and Xylocopa bees actively and exclusively exploited the pollen grains from the central poricidal anthers generating pollen deposition on their bodies. The reflectance pattern of floral organs indicated a targeting of these large bees to the central anthers, corroborated by the anther manipulative experiment where only the exclusion of the anthers positioned in the flower centre, especially the intermediate stamens, reduced bee visits. Both results revealed a division of labour, in which the intermediate stamen morph was responsible for both floral attractiveness and pollen resources. Only the largest stamen morph produced germinable pollen grains, highlighting their role as pollinating stamens. The smallest stamen morph has a less clear function, likely representing an economy in pollen production for feeding function. Our findings suggest that the evolution of the trimorphic androecium is associated with division of labour in large pollen flowers and can represent a strong strategy for circumventing the pollen dilemma, optimizing the feeding function by reducing pollen grain investment from central anthers.

Interspecific anatomical differences result in similar highly flexible stems in Bignoniaceae lianas.

PREMISE: Lianas are intriguing forest components in the tropics worldwide. They are characterized by thin and flexible stems, which have been related to a unique stem anatomy. Here, we hypothesized that the anatomical diversity of lianas, varying in shapes, proportions, and dimensions of tissues and cell types, would result in different stem bending stiffnesses across species. To test this hypothesis, we chose four abundant liana species of central Amazonia belonging to the monophyletic tribe Bignonieae (Bignoniaceae) and compared their basal stems for their anatomical architectures and bending properties. METHODS: Measurements of anatomical architecture and bending stiffness (structural Young's modulus) included light microscopy observations and three-point bending tests, which were performed on basal stems of eight individuals from four Bignonieae species. All analyses, including comparisons among species and relationships between stem stiffness and anatomical architecture, were performed using linear models. RESULTS: Although the anatomical architecture of each species consists of different qualitative and quantitative combinations of both tissues and cell types in basal stems, all species analyzed showed similarly lower bending stiffnesses. This similarity was shown to be directly related to high bark contribution to the second moment of area, vessel area and ray width. CONCLUSIONS: Similar values of stem bending stiffness were encountered in four liana species analyzed despite their variable anatomical architectures. This pattern provides new evidence of how different quantitative combinations of tissue and cell types in the basal stems of lianas can generate similarly low levels of stiffness in a group of closely related species.

Morphology, secretion composition, and ecological aspects of stipular colleters in Rubiaceae species from tropical forest and savanna.

Colleters are secretory structures that produce and release mucilage or a mucilage-resin mixture protecting meristems and young structures against desiccation, herbivores, and pathogens. The secretions may vary in colleters of same or different types, indicating that the functionality of colleters may be more specific than previously thought. In this study, we compared 17 Rubiaceae species from savanna and forest environment focusing on colleter secretions and its ecological role. First, we evaluated the morphology, distribution, and histochemistry of stipular colleters using light and scanning electron microscopy. Additionally, we investigated the phenology, microclimate, and the proportion of damaged apices in the savanna and forest species. We recorded standard-type colleters, variable in distribution and size, in 14 of the 17 studied species. The secretion varied from predominantly hydrophilic, mixed to predominantly lipophilic. During the budding period, secretion covered the vegetative apices. Savanna species had a prevalence of lipid secretion in habitats with higher luminosity, which had a lower proportion of damaged apices. In contrast, forest species occurred in habitats with lower luminosity and had a higher proportion of damaged apices, in general with the absence of lipids in the colleters. These results highlight that colleters with similar morphology clearly differed in secretions among species, especially between species from savanna and forest, in which the colleters appear potentially associated with protection against irradiation in savanna, but not in the forest environment.

Geographic mosaic of plant evolution: extrafloral nectary variation mediated by ant and herbivore assemblages.

Herbivory is an ecological process that is known to generate different patterns of selection on defensive plant traits across populations. Studies on this topic could greatly benefit from the general framework of the Geographic Mosaic Theory of Coevolution (GMT). Here, we hypothesize that herbivory represents a strong pressure for extrafloral nectary (EFN) bearing plants, with differences in herbivore and ant visitor assemblages leading to different evolutionary pressures among localities and ultimately to differences in EFN abundance and function. In this study, we investigate this hypothesis by analyzing 10 populations of Anemopaegma album (30 individuals per population) distributed through ca. 600 km of Neotropical savanna and covering most of the geographic range of this plant species. A common garden experiment revealed a phenotypic differentiation in EFN abundance, in which field and experimental plants showed a similar pattern of EFN variation among populations. We also did not find significant correlations between EFN traits and ant abundance, herbivory and plant performance across localities. Instead, a more complex pattern of ant-EFN variation, a geographic mosaic, emerged throughout the geographical range of A. album. We modeled the functional relationship between EFNs and ant traits across ant species and extended this phenotypic interface to characterize local situations of phenotypic matching and mismatching at the population level. Two distinct types of phenotypic matching emerged throughout populations: (1) a population with smaller ants (Crematogaster crinosa) matched with low abundance of EFNs; and (2) seven populations with bigger ants (Camponotus species) matched with higher EFN abundances. Three matched populations showed the highest plant performance and narrower variance of EFN abundance, representing potential plant evolutionary hotspots. Cases of mismatched and matched populations with the lowest performance were associated with abundant and highly detrimental herbivores. Our findings provide insights on the ecology and evolution of plant-ant guarding systems, and suggest new directions to research on facultative mutualistic interactions at wide geographic scales.

Trichome structure and evolution in Neotropical lianas.

BACKGROUND AND AIMS: Trichomes are epidermal outgrowths generally associated with protection against herbivores and/or desiccation that are widely distributed from ferns to angiosperms. Patterns of topological variation and morphological evolution of trichomes are still scarce in the literature, preventing valid comparisons across taxa. This study integrates detailed morphoanatomical data and the evolutionary history of the tribe Bignonieae (Bignoniaceae) in order to gain a better understanding of current diversity and evolution of trichome types. METHODS: Two sampling schemes were used to characterize trichome types: (1) macromorphological characterization of all 105 species currently included in Bignonieae; and (2) micromorphological characterization of 16 selected species. Individual trichome morphotypes were coded as binary in each vegetative plant part, and trichome density and size were coded as multistate. Ancestral character state reconstructions were conducted using maximum likelihood (ML) assumptions. KEY RESULTS: Two main functional trichome categories were found: non-glandular and glandular. In glandular trichomes, three morphotypes were recognized: peltate (Pg), stipitate (Sg) and patelliform/cupular (P/Cg) trichomes. Non-glandular trichomes were uniseriate, uni- or multicellular and simple or branched. Pg and P/Cg trichomes were multicellular and non-vascularized with three clearly distinct cell layers. Sg trichomes were multicellular, uniseriate and long-stalked. ML ancestral character state reconstructions suggested that the most recent common ancestor (MRCA) of Bignonieae probably had non-glandular, Pg and P/Cg trichomes, with each trichome type presenting alternative histories of appearance on the different plant parts. For example, the MRCA of Bignonieae probably had non-glandular trichomes on the stems, prophylls, petiole, petiolule and leaflet veins while P/Cg trichomes were restricted to leaflet blades. Sg trichomes were not present in the MRCA of Bignonieae independently of the position of these trichomes. These trichomes had at least eight independent origins in tribe. CONCLUSIONS: The patterns of trichome evolution indicate that most morphotypes are probably homologous in Bignonieae and could be treated under the same name based on its morphological similarity and common evolutionary history, in spite of the plethora of names that have been previously designated in the literature. The trichome descriptions presented here will facilitate comparisons across taxa, allowing inferences on the relationsthips between trichome variants and future studies about their functional properties.

Evolution of extrafloral nectaries: adaptive process and selective regime changes from forest to savanna.

Much effort has been devoted to understanding the function of extrafloral nectaries (EFNs) for ant-plant-herbivore interactions. However, the pattern of evolution of such structures throughout the history of plant lineages remains unexplored. In this study, we used empirical knowledge on plant defences mediated by ants as a theoretical framework to test specific hypotheses about the adaptive role of EFNs during plant evolution. Emphasis was given to different processes (neutral or adaptive) and factors (habitat change and trade-offs with new trichomes) that may have affected the evolution of ant-plant associations. We measured seven EFN quantitative traits in all 105 species included in a well-supported phylogeny of the tribe Bignonieae (Bignoniaceae) and collected field data on ant-EFN interactions in 32 species. We identified a positive association between ant visitation (a surrogate of ant guarding) and the abundance of EFNs in vegetative plant parts and rejected the hypothesis of phylogenetic conservatism of EFNs, with most traits presenting K-values < 1. Modelling the evolution of EFN traits using maximum likelihood approaches further suggested adaptive evolution, with static-optimum models showing a better fit than purely drift models. In addition, the abundance of EFNs was associated with habitat shifts (with a decrease in the abundance of EFNs from forest to savannas), and a potential trade-off was detected between the abundance of EFNs and estipitate glandular trichomes (i.e. trichomes with sticky secretion). These evolutionary associations suggest divergent selection between species as well as explains K-values < 1. Experimental studies with multiple lineages of forest and savanna taxa may improve our understanding of the role of nectaries in plants. Overall, our results suggest that the evolution of EFNs was likely associated with the adaptive process which probably played an important role in the diversification of this plant group.