Recent genetic, phenetic and ecological divergence across the Mesoamerican highlands: a study case with Diglossa baritula (Aves: Thraupidae).

The topographical, geological, climatic and biodiversity complexity of Mesoamerica has made it a primary research focus. The Mesoamerican highlands is a region with particularly high species richness and within-species variation. The Cinnamon-bellied Flowerpiercer, Diglossa baritula (Wagler, 1832), is a species endemic to the Mesoamerican highlands, with three allopatric subspecies currently recognized. To characterize divergence within this species, we integrated genomics, morphology, coloration and ecological niche modeling approaches, obtained from sampling individuals across the entire geographic distribution of the species. Our results revealed a clear genomic divergence between the populations to the east versus the west of the Isthmus of Tehuantepec. In contrast to the genomic results, morphology and coloration analyses showed intermediate levels of differentiation, indicating that population groups within D. baritula have probably been under similar selective pressures. Our morphology results indicated that the only sexually dimorphic morphological variable is the wing chord, with males having a longer wing chord than females. Finally, ecological data indicated that there are differences in ecological niche within D. baritula. Our data suggest that D. baritula could contain two or more incipient species at the intermediate phase of the speciation continuum. These results highlight the importance of the geographical barrier of the Isthmus of Tehuantepec and Pleistocene climatic events in driving isolation and population divergence in D. baritula. The present investigation illustrates the speciation potential of the D. baritula complex and the capacity of Mesoamerican highlands to create cryptic biodiversity and endemism.

Implications of dominance hierarchy on hummingbird-plant interactions in a temperate forest in Northwestern Mexico.

The structuring of plant-hummingbird networks can be explained by multiple factors, including species abundance (i.e., the neutrality hypothesis), matching of bill and flower morphology, phenological overlap, phylogenetic constraints, and feeding behavior. The importance of complementary morphology and phenological overlap on the hummingbird-plant network has been extensively studied, while the importance of hummingbird behavior has received less attention. In this work, we evaluated the relative importance of species abundance, morphological matching, and floral energy content in predicting the frequency of hummingbird-plant interactions. Then, we determined whether the hummingbird species' dominance hierarchy is associated with modules within the network. Moreover, we evaluated whether hummingbird specialization (d') is related to bill morphology (bill length and curvature) and dominance hierarchy. Finally, we determined whether generalist core hummingbird species are lees dominant in the community. We recorded plant-hummingbird interactions and behavioral dominance of hummingbird species in a temperate forest in Northwestern Mexico (El Palmito, Mexico). We measured flowers' corolla length and nectar traits and hummingbirds' weight and bill traits. We recorded 2,272 interactions among 13 hummingbird and 10 plant species. The main driver of plant-hummingbird interactions was species abundance, consistent with the neutrality interaction theory. Hummingbird specialization was related to dominance and bill length, but not to bill curvature of hummingbird species. However, generalist core hummingbird species (species that interact with many plant species) were less dominant. The frequency of interactions between hummingbirds and plants was determined by the abundance of hummingbirds and their flowers, and the dominance of hummingbird species determined the separation of the different modules and specialization. Our study suggests that abundance and feeding behavior may play an important role in North America's hummingbird-plant networks.

Isolation by resistance explains genetic diversity in the Arremon brushfinches of northern Mesoamerica.

Genetic differentiation between and within natural populations is the result of the joint effects of neutral and adaptative processes. In addition, the spatial arrangement of the landscape promotes connectivity or creates barriers to gene flow, directly affecting speciation processes. In this study, we carried out a landscape genomics analysis using NextRAD data from a montane forest specialist bird complex, the Mesoamerican Chestnut-capped/Green-striped Brushfinch of the genus Arremon. Specifically, we examined population genomic structure using different assignment methods and genomic differentiation and diversity, and we tested alternative genetic isolation hypotheses at the individual level (e.g., isolation by barrier, IBB; isolation by environment, IBE; isolation by resistance, IBR). We found well-delimited genomic structuring (K = 5) across Mesoamerican montane forests in the studied group. Individual-level genetic distances among major montane ranges were mainly explained by IBR hypotheses in this sedentary Neotropical taxon. Our results uncover genetic distances/differentiation and patterns of gene flow in allopatric species that support the role of tropical mountains as spatial landscape drivers of biodiversity. IBR clearly supports a pattern of conserved niche-tracking of suitable habitat conditions and topographic complexity throughout glacial-interglacial dynamics.

Quantifying phenological diversity: a framework based on Hill numbers theory.

Background: Despite the great concern triggered by the environmental crisis worldwide, the loss of temporal key functions and processes involved in biodiversity maintenance has received little attention. Species are restricted in their life cycles by environmental variables because of their physiological and behavioral properties; thus, the timing and duration of species' presence and their activities vary greatly between species within a community. Despite the ecological relevance of such variation, there is currently no measure that summarizes the key temporal aspects of biological diversity and allows comparisons of community phenological patterns. Here, we propose a measure that synthesizes variability of phenological patterns using the Hill numbers-based attribute diversity framework. Methods: We constructed a new phenological diversity measure based on the aforementioned framework through pairwise overlapping distances, which was supplemented with wavelet analysis. The Hill numbers approach was chosen as an adequate way to define a set of diversity values of different order q, a parameter that determines the sensitivity of the diversity measure to abundance. Wavelet transform analysis was used to model continuous variables from incomplete data sets for different phenophases. The new measure, which we call Phenological Hill numbers (PD), considers the decouplings of phenophases through an overlapping area value between pairs of species within the community. PD was first tested through simulations with varying overlap in phenophase magnitude and intensity and varying number of species, and then by using one real data set. Results: PD maintains the diversity patterns of order q as in any other diversity measure encompassed by the Hill numbers framework. Minimum PD values in the simulated data sets reflect a lack of differentiation in the phenological curves of the community over time; by contrast, the maximum PD values reflected the most diverse simulations in which phenological curves were equally distributed over time. PD values were consistent with the homogeneous distribution of the intensity and concurrence of phenophases over time, both in the simulated and the real data set. Discussion: PD provides an efficient, readily interpretable and comparable measure that summarizes the variety of phenological patterns observed in ecological communities. PD retains the diversity patterns of order q characteristic of all diversity measures encompassed by the distance-based Hill numbers framework. In addition, wavelet transform analysis proved useful for constructing a continuous phenological curve. This methodological approach to quantify phenological diversity produces simple and intuitive values for the examination of phenological diversity and can be widely applied to any taxon or community's phenological traits.

Forbidden links, trait matching and modularity in plant-hummingbird networks: Are specialized modules characterized by higher phenotypic floral integration?

BACKGROUND: Plant-pollinator mutualistic networks show non-random structural properties that promote species coexistence. However, these networks show high variability in the interacting species and their connections. Mismatch between plant and pollinator attributes can prevent interactions, while trait matching can enable exclusive access, promoting pollinators' niche partitioning and, ultimately, modularity. Thus, plants belonging to specialized modules should integrate their floral traits to optimize the pollination function. Herein, we aimed to analyze the biological processes involved in the structuring of plant-hummingbird networks by linking network morphological constraints, specialization, modularity and phenotypic floral integration. METHODS: We investigated the understory plant-hummingbird network of two adjacent habitats in the Lacandona rainforest of Mexico, one characterized by lowland rainforest and the other by savanna-like vegetation. We performed monthly censuses to record plant-hummingbird interactions for 2 years (2018-2020). We also took hummingbird bill measurements and floral and nectar measurements. We summarized the interactions in a bipartite matrix and estimated three network descriptors: connectance, complementary specialization (H2'), and nestedness. We also analyzed the modularity and average phenotypic floral integration index of each module. RESULTS: Both habitats showed strong differences in the plant assemblage and network dynamics but were interconnected by the same four hummingbird species, two Hermits and two Emeralds, forming a single network of interaction. The whole network showed low levels of connectance (0.35) and high specialization (H2' = 0.87). Flower morphologies ranged from generalized to specialized, but trait matching was an important network structurer. Modularity was associated with morphological specialization. The Hermits Phaethornis longirostris and P. striigularis each formed a module by themselves, and a third module was formed by the less-specialized Emeralds: Chlorestes candida and Amazilia tzacatl. The floral integration values were higher in specialized modules but not significantly higher than that formed by generalist species. CONCLUSIONS: Our findings suggest that biological processes derived from both trait matching and "forbidden" links, or nonmatched morphological attributes, might be important network drivers in tropical plant-hummingbird systems while morphological specialization plays a minor role in the phenotypic floral integration. The broad variety of corolla and bill shapes promoted niche partitioning, resulting in the modular organization of the assemblage according to morphological specialization. However, more research adding larger datasets of both the number of modules and pollination networks for a wider region is needed to conclude whether phenotypic floral integration increases with morphological specialization in plant-hummingbird systems.

Temporal dynamics of the hummingbird-plant interaction network of a dry forest in Chamela, Mexico: a 30-year follow-up after two hurricanes.

BACKGROUND: Interactions among species are a driving force of community structure. The species composition of animal-plant interaction networks can be highly dynamic on a temporal scale, even though the general network structure is usually not altered. However, few studies have examined how interaction networks change over long periods of time, particularly after extreme natural events. We analyzed herein the structure of the hummingbird-plant interaction network in a dry forest of Chamela, Mexico, comparing the structure in 1985-1986 with that in 2016-2017 following the passage of two hurricanes (category 2 Jova in 2011 and category 4 Patricia in 2015). METHODS: The fieldwork was carried out in the Chamela-Cuixmala Biosphere Reserve in Jalisco, Mexico. In the last 30 years, three severe drought events and two hurricanes have affected this region. Previously, from 1985-1986, hummingbird-plant interactions were recorded monthly for one year in the study area. Then, from 2016-2017, we replicated the sampling in the same localities. We compared the network parameters describing the plant-hummingbird interactions of each period using adjacency matrices. RESULTS: We found differences in the number and identity of interacting species, especially plants. The plant species missing in 2016-2017 were either the least connected in the original network (1985-1986) or belonged to groups such as cacti, epiphytes, or trees. The new plant species incorporated in the 2016-2017 network were herbs, vines, and shrubs, or other species barely connected. These changes in the composition are consistent with reports on vegetation damage after strong hurricanes at other study sites. Conversely, all hummingbird species remained in the network, with the exception of Heliomaster constantii, which was primarily connected to a plant species absent in the 2016-2017 network. Migratory and habitat generalist species (i.e., Archilochus spp.) showed higher abundances following the disturbance events. CONCLUSIONS: Most of the parameters describing the hummingbird-plant network structure remained unchanged after 30 years, with the exception of an increase in plant robustness and hummingbird niche overlap. However, the network's generalist core was affected by the loss of some species. Also, core plant species such as Ipomoea bracteata, Combretum farinosum, and Justicia candicans were found to be important for maintaining the hummingbird-plant interaction network. The temporal perspective of this study provides unique insights into the conservation of plant-hummingbird networks across time and extreme natural events.

Are hummingbirds generalists or specialists? Using network analysis to explore the mechanisms influencing their interaction with nectar resources.

Mutualistic interactions are powerful drivers of biodiversity on Earth that can be represented as complex interaction networks that vary in connection pattern and intensity. One of the most fascinating mutualisms is the interaction between hummingbirds and the plants they visit. We conducted an exhaustive search for articles, theses, reports, and personal communications with researchers (unpublished data) documenting hummingbird visits to flowers of nectar-rewarding plants. Based on information gathered from 4532 interactions between 292 hummingbird species and 1287 plant species, we built an interaction network between nine hummingbird clades and 100 plant families used by hummingbirds as nectar resources at a continental scale. We explored the network architecture, including phylogenetic, morphological, biogeographical, and distributional information. As expected, the network between hummingbirds and their nectar plants was heterogeneous and nested, but not modular. When we incorporated ecological and historical information in the network nodes, we found a generalization gradient in hummingbird morphology and interaction patterns. The hummingbird clades that most recently diversified in North America acted as generalist nodes and visited flowers with ornithophilous, intermediate and non-ornithophilous morphologies, connecting a high diversity of plant families. This pattern was favored by intermediate morphologies (bill, wing, and body size) and by the low niche conservatism in these clades compared to the oldest clades that diversified in South America. Our work is the first effort exploring the hummingbird-plant mutualistic network at a continental scale using hummingbird clades and plant families as nodes, offering an alternative approach to exploring the ecological and evolutionary factors that explain plant-animal interactions at a large scale.

Hummingbird migration and flowering synchrony in the temperate forests of northwestern Mexico.

BACKGROUND: Many species of birds are morphologically and physiologically adapted for migration. Migratory movements of birds can range from thousands of kilometers, such as when birds migrate from wintering to breeding sites in summer, to several kilometers, such as when birds migrate among habitats in a single mountain system. The main factor that influences bird migration is the seasonal fluctuation of food resources; climate, predation, competition for resources and endogenous programming are also important factors. Hummingbirds are highly dependent on nectar, so their migration is likely correlated with the blooming of plant species. The ecological implications of altitudinal migration in the mountains of North America as well as the latitudinal migration of Selasphorus rufus through Mexico are still poorly understood. To explore these issues, over three non-consecutive years, we evaluated interannual variation in the phenologies of a latitudinal migrant (S. rufus) and an altitudinal migrant (Amazilia beryllina) and their visited plants. METHODS: We assessed the relationship between two migratory hummingbirds and flower abundance in 20 fixed-radius plots (25 m radius). All available flowers were counted along transects (40 × 5 m) inside each fixed-radius plot. Sampling was performed every 10 days from November 12 through February 20 of 2010-2011, 2013-2014 and 2015-2016, resulting in a total of 11 samples of each plot per period. Phenological variation and the relationships among hummingbird abundance, flower abundance and vegetation type were evaluated using a generalized additive mixed model. RESULTS: S. rufus abundance was related to sampling time in the first and third periods; this relationship was not significant in the second period. A. beryllina abundance was related with the sampling time over all three periods. The abundance of S. rufus hummingbirds was significantly related to the number of Salvia iodantha flowers. The abundance of A. beryllina hummingbirds was related to the number of S. iodantha and Cestrum thyrsoideum flowers and the total number of flowers. We found a non-significant correlation between S. rufus and A. beryllina abundance and vegetation types. CONCLUSION: Contrary to expectations, the long-distance migration of S. rufus was not consistent over the sampling periods. The migration of S. rufus through the study region may be altered by changes in climate, as has occurred with other species of migratory birds. In the present study, the migration of S. rufus was correlated with the blooming of S. iodantha. In comparison, the altitudinal migrant A. beryllina responded to the availability of floral resources but was not associated with a particular plant. The migration of this latter species in the area probably depends on multiple factors, including climatic and demographic factors, but is particularly dependent on the supply of floral resources and competition for these resources.

Reproductive ecology and isolation of Psittacanthus calyculatus and P. auriculatus mistletoes (Loranthaceae).

BACKGROUND: Relationships between floral biology and pollinator behavior are important to understanding species diversity of hemiparasitic Psittacanthus mistletoes (c. 120 species). We aimed to investigate trait divergence linked to pollinator attraction and reproductive isolation (RI) in two hummingbird-pollinated and bird-dispersed Psittacanthus species with range overlap. METHODS: We investigated the phylogenetic relationships, floral biology, pollinator assemblages, seed dispersers and host usage, and the breeding system and female reproductive success of two sympatric populations of P. calyculatus and P. auriculatus, and one allopatric population of P. calyculatus. Flowers in sympatry were also reciprocally pollinated to assess a post-mating component of RI. RESULTS: Hummingbird assemblages differed between calyculatus populations, while allopatric plants of calyculatus opened more but smaller flowers with longer lifespans and produced less nectar than those in sympatry. Bayesian-based phylogenetic analysis indicated monophyly for calyculatus populations (i.e. both populations belong to the same species). In sympatry, calyculatus plants opened more and larger flowers with longer lifespans and produced same nectar volume than those of auriculatus; populations shared pollinators but seed dispersers and host usage differed between species. Nectar standing crops differed between sympatric populations, with lower visitation in calyculatus. Hand pollination experiments indicated a predominant outcrossing breeding system, with fruit set after interspecific pollination two times higher from calyculatus to auriculatus than in the opposite direction. CONCLUSIONS: Given the low genetic differentiation between calyculatus populations, observed trait divergence could have resulted from changes regarding the local communities of pollinators and, therefore, expected divergence for peripheral, allopatric populations. Using RI estimates, there were fewer heterospecific matings than expected by chance in P. calyculatus (RI4A = 0.629) as compared to P. auriculatus (RI4A = 0.20). When considering other factors of ecological isolation that affect co-occurrence, the RI4C values indicate that isolation by hummingbird pollinators was less effective (0.20) than isolation by host tree species and seed dispersers (0.80 and 0.60, respectively), suggesting that host usage is the most important ecological isolation factor between the two species. Accordingly, the absolute and relative cumulative strength values indicated that the host tree species' barrier is currently contributing the most to maintaining these species in sympatry.

Myiarchus flycatchers are the primary seed dispersers of Bursera longipes in a Mexican dry forest.

We evaluated the seed dispersal of Bursera longipes by birds along a successional gradient of tropical dry forest (TDF) in southwestern Mexico. B. longipes is an endemic tree to the TDF in the Balsas basin. The relative abundance of frugivorous birds, their frequency of visits to B. longipes and the number of removed fruits were recorded at three study sites with different stages of forest succession (early, intermediate and mature) characterized by distinct floristic and structural elements. Flycatchers of the Myiarchus and Tyrannus genera removed the majority of fruits at each site. Overall, visits to B. longipes were less frequent at the early successional site. Birds that function as legitimate dispersers by consuming whole seeds and regurgitating or defecating intact seeds in the process also remove the pseudoaril from seeds, thereby facilitating the germination process. The highest germination percentages were recorded for seeds that passed through the digestive system of two migratory flycatchers: M. cinerascens and M. nutingii. Perch plants, mainly composed of legumes (e.g., Eysenhardtia polystachya, Acacia cochliacantha, Calliandra eryophylla, Mimosa polyantha), serve also as nurse plants since the number of young individuals recruited from B. longipes was higher under these than expected by chance. This study shows that Myiarchus flycatchers are the most efficient seed dispersers of B. longipes across all successional stages. This suggests a close mutualistic relationship derived from adaptive processes and local specializations throughout the distribution of both taxa, as supported by the geographic mosaic theory of coevolution.

Geographic differentiation in the pollination system of the columnar cactus Pachycereus pecten -aboriginum.

The pollination biology of the cactus Pachycereus pecten-aboriginum was studied in a tropical location in western Mexico (ca. 18° N latitude) to compare with data from a northern population (ca. 28° N latitude). Throughout this range, the nectar-feeding bat Leptonycteris curasoae is resident within the tropics but migratory in its northern range. The hypothesis was tested that if a predictable bat presence has been an important force in the evolution of pollination systems in columnar cacti, P. pecten-aboriginum will have a specialized pollination system within the tropics and a generalized pollination system in northern populations. In both areas, pollination experiments showed that P. pecten-aboriginum has a self-incompatible, hermaphroditic breeding system. In the tropical area, flowers open at night and close early in the morning. Nectar is secreted only during the night, and flowers are exclusively pollinated by three species of nectar-feeding bats, indicating a specialized pollination system. In contrast, anthesis and nectar secretion in northern populations occur during the night and day, allowing visitation and effective pollination by both nocturnal and diurnal pollinators. This study provides evidence of divergence mediated by pollinator predictability (resident vs. migrant), through shifts from short to long anthesis and nectar production periods from southern to northern populations.

Carbohydrate analysis of floral nectar using medium infrared.

An instrumental method based on a chemometric model of the medium region of the infrared (MIR) was developed to analyse total sugar content and the proportions of glucose, fructose and sucrose. In order to construct the model, a set of 127 standard aqueous solutions of different sugars in the concentration range 0-20% (w/v) were prepared and analysed in the interval 4900-700 cm(-1). The MIR was transformed by normalisation, correction of baseline using the second derivative, and suppression of the signals of water and carbon dioxide. The region between 1150 and 950 cm(-1) showed the highest correlation between signal and concentration. The correlation coefficient for total sugar content was 0.956, whilst those for glucose, fructose and sucrose were 0.982, 0.972 and 0.992, respectively. The method was validated using a set of 28 samples of nectar which had been assayed by chromatographic and refractometric methods. The method shows potential utility for the prediction of nectar sugar components.