Neural crest cells as a source of microevolutionary variation.

Vertebrates have some of the most complex and diverse features in animals, from varied craniofacial morphologies to colorful pigmentation patterns and elaborate social behaviors. All of these traits have their developmental origins in a multipotent embryonic lineage of neural crest cells. This "fourth germ layer" is a vertebrate innovation and the source of a wide range of adult cell types. While others have discussed the role of neural crest cells in human disease and animal domestication, less is known about their role in contributing to adaptive changes in wild populations. Here, we review how variation in the development of neural crest cells and their derivatives generates considerable phenotypic diversity in nature. We focus on the broad span of traits under natural and sexual selection whose variation may originate in the neural crest, with emphasis on behavioral factors such as intraspecies communication that are often overlooked. In all, we encourage the integration of evolutionary ecology with developmental biology and molecular genetics to gain a more complete understanding of the role of this single cell type in trait covariation, evolutionary trajectories, and vertebrate diversity.

Quantitative morphological analysis framework of infant cranial sutures and fontanelles based on CT images.

Characterizing the suture morphological variation is a crucial step to investigate the influence of sutures on infant head biomechanics. This study aimed to establish a comprehensive quantitative framework for accurately capturing the cranial suture and fontanelle morphologies in infants. A total of 69 CT scans of 2-4 month-old infant heads were segmented to identify semilandmarks at the borders of cranial sutures and fontanelles. Morphological characteristics, including length, width, sinuosity index (SI), and surface area, were measured. For this, an automatic method was developed to determine the junction points between sutures and fontanelles, and thin-plate-spline (TPS) was utilized for area calculation. Different dimensionality reduction methods were compared, including nonlinear and linear principal component analysis (PCA), as well as deep-learning-based variational autoencoder (VAE). Finally, the significance of various covariates was analyzed, and regression analysis was performed to establish a statistical model relating morphological parameters with global parameters. This study successfully developed a quantitative morphological framework and demonstrate its application in quantifying morphologies of infant sutures and fontanelles, which were shown to significantly relate to global parameters of cranial size, suture SI, and surface area for infants aged 2-4 months. The developed framework proved to be reliable and applicable in extracting infant suture morphology features from CT scans. The demonstrated application highlighted its potential to provide valuable insights into the morphologies of infant cranial sutures and fontanelles, aiding in the diagnosis of suture-related skull fractures. Infant suture, Infant fontanelle, Morphological variation, Morphology analysis framework, Statistical model.

Morphological and molecular characterization of Stomachicola muraenesocis Yamaguti, 1934 (Digenea: Hemiuridae) from the daggertooth pike conger Muraenesox cinereus (Forsskål).

Hemiurid digeneans conspecific with Stomachicola muraenesocis Yamaguti, 1934 (the type species of the genus Stomachicola Yamaguti, 1934) were collected from the stomach of the daggertooth pike conger Muraenesox cinereus (Forsskål) off the Persian Gulf of Iran. This study aimed to provide a detailed characterization of Stom. muraenesocis, including measurements, illustrations and scanning electron microscopy (s.e.m.) representations. Comparisons with the original and previous descriptions revealed morphological and metrical variations in several features (i.e. body size and shape, arrangement of reproductive organs, soma to ecsoma length ratio, position of genital opening, number of vitelline tubules and extension of uterine coils) between Stom. muraenesocis from different hosts and localities. This study presents the first molecular sequence data associated with the small (18S) and large (28S) subunit nuclear ribosomal RNA genes (rDNA) for Stom. muraenesocis. Phylogenetic analyses of the 18S dataset placed Stom. muraenesocis as sister lineage to a clade formed of a group of species of Lecithaster Lühe, 1901 (Lecithasteridae Odhner, 1905). In contrast, phylogenetic analyses based on the 28S consistently recovered a sister relationship between Stom. muraenesocis and representatives of the Hemiuridae Looss, 1899. Further comprehensive phylogenetically based classification in light of morphology and taxonomic history of the Hemiuridae and Lecithasteridae is required to infer phylogenetic affinities and historical biogeography of Stomachicola. A comprehensive list of previously reported species of Stomachicola together with their associated hosts, localities and morphometric data is provided.

Taxonomical use of scale ornamentation: Challenges by intraspecific and intra-individual variations in four adult specimens of Polypterus bichir.

Many actinopterygian fish groups, including fossil and extant polypteriforms and lepisosteiforms, fossil halecomorphs, and some basal teleosts, have stout bony scales covered by layers of ganoin-an enamel layer ornamented with minute tubercles. Ganoid scales preserve well as disarticulated remains and notably constitute most of the fossil record for polypteriform in both South America and Africa. Based on two variables (tubercle size and distance between tubercles), some authors reported that the ganoin tubercle ornamentation in these scales is constant within a species and differs between species and allows distinguishing species or at least groups of species. However, despite its promising potential for assessing polypteriform paleodiversity, this tool has remained unused, probably because the variables are not well defined, and intraspecific variation does not seem to have been considered. To address this gap, we aimed to test the intraspecific and intra-individual variation in the ornamentation of ganoid scales in the type species Polypterus bichir. We propose three different parameters to describe the tubercle ornamentation: the distance between contiguous tubercles centers, their density, and their relative spatial organization. With these parameters, we investigate the variation in ganoin ornamentation among four specimens and across different regions of the body. Our results show that the distribution of the tubercles is highly variable within a same species, regardless of the body region, and sometimes even between different sectors of a same scale. Moreover, the variation observed in P. bichir overlaps with the distribution described in the literature for several extant and fossil species. Thus far, the ornamentation of ganoid scales is not a reliable diagnostical feature for polypterids.

Morphological variability of wild-growing crown imperial (Fritillaria imperialis L.) germplasm in central region of Iran-implications for in-situ conservation initiatives.

BACKGROUND: Crown imperial (Fritillaria imperialis L.) is a threatened bulbous plant which has great ornamental and medicinal values and importance. In the present study, a total of 100 specimens of wild-growing F. imperialis from 10 natural areas of Markazi province, Iran, representing one of the main centers of genetic diversity of this species, were evaluated using 37 phenotypic attributes during April 2021. RESULTS: High level of genetic variation within populations (75%) and low levels of genetic variation among populations (25%) was revealed. The highest coefficient of variation (CV) was found in leaf trichome (82.00%) and then margin of crown leaves (80.44%). In addition, flower color (CV = 50.86%), flower number (CV = 44.61%), peduncle diameter (CV = 33.44%), and plant length (CV = 32.55%)-all important from an ornamental point of view- showed relatively high CV values. The CV was the lowest for flower shape, filament color, bulb shape, bulblet number, and floral scent. Ward cluster analysis identified two main clusters, containing 14 and 86 specimens, respectively. The first group consisted mainly of specimens from the adjacent Shahbaz and Rasvand populations. According to the principal component analysis (PCA), the first six components of data accounted for 88.36% of total variance. The Shahbaz-1, Shahbaz-2, Shahbaz-6, Shahbaz-7, Shahbaz-9, and Bolagh-8 specimens showed the highest variation and were separated from others, which they can be used further in breeding programs, while Sarchal-2, Bolagh-3, and Chepeqli-4 specimens showed the lowest variability. Moreover, the studied populations were clustered into four distinct groups, each including populations that were geographically close to one another. CONCLUSIONS: Although the examined specimens revealed high genetic diversity herein, the results indicated that wild-growing populations of F. imperialis are still at risk suffering from overcollection in the most of studied areas, especially in Deh-Sad and Tureh.

Making meaning from fragmentary fossils: Early Homo in the Early to early Middle Pleistocene.

In celebration of the 50th anniversary of the Journal of Human Evolution, we re-evaluate the fossil record for early Homo (principally Homo erectus, Homo habilis, and Homo rudolfensis) from early diversification and dispersal in the Early Pleistocene to the ultimate demise of H. erectus in the early Middle Pleistocene. The mid-1990s marked an important historical turning point in our understanding of early Homo with the redating of key H. erectus localities, the discovery of small H. erectus in Asia, and the recovery of an even earlier presence of early Homo in Africa. As such, we compare our understanding of early Homo before and after this time and discuss how the order of fossil discovery and a focus on anchor specimens has shaped, and in many ways biased, our interpretations of early Homo species and the fossils allocated to them. Fragmentary specimens may counter conventional wisdom but are often overlooked in broad narratives. We recognize at least three different cranial and two or three pelvic morphotypes of early Homo. Just one postcranial morph aligns with any certainty to a cranial species, highlighting the importance of explicitly identifying how we link specimens together and to species; we offer two ways of visualizing these connections. Chronologically and morphologically H. erectus is a member of early Homo, not a temporally more recent species necessarily evolved from either H. habilis or H. rudolfensis. Nonetheless, an ancestral-descendant notion of their evolution influences expectations around the anatomy of missing elements, especially the foot. Weak support for long-held notions of postcranial modernity in H. erectus raises the possibility of alternative drivers of dispersal. New observations suggest that the dearth of faces in later H. erectus may mask taxonomic diversity in Asia and suggest various later mid-Pleistocene populations could derive from either Asia or Africa. Future advances will rest on the development of nuanced ways to affiliate fossils, greater transparency of implicit assumptions, and attention to detailed life history information for comparative collections; all critical pursuits for future research given the great potential they have to enrich our evolutionary reconstructions for the next fifty years and beyond.

Morphological modification of silver nanoparticles against multi-drug resistant gram-negative bacteria and cytotoxicity effect in A549 lung cancer cells through in vitro approaches.

In the present study, the individual cultures of Proteus mirabilis (P. mirabilis) and Klebsiella pneumoniae (K. pneumoniae) were treated with morphologically modified silver nanoparticles (Ag NPs) and were found to display zones of inhibition of ~ 8 mm, 16 mm, 20 mm, and 22 mm (P. mirabilis) and 6 mm, 14 mm, 20 mm, and 24 mm (K. pneumoniae) at concentrations of 25 µg/ml, 50 µg/mL, 75 µg/mL, and 100 µg/mL, respectively. In addition, turbidity tests were performed based on O. D. values, which exhibited 92% and 90% growth inhibitions at 100 µg/mL concentration for P. mirabilis and K. pneumoniae, respectively. Furthermore, the IC50 concentration of Ag NPs was established for A549 lung cancer cells and found to be at 500 µg/mL. Evidently, the morphological variation of Ag NPs treated A549 lung cancer cells was exhibited with differential morphology studied by phase-contrast microscopy. The results demonstrated that the synthesized Ag NPs was not only efficient against gram-positive bacteria but also against gram-negative bacteria and A549 cancer cells, suggesting that the potential of these biosynthesized Ag NPs is a future drug discovery source for inhibiting bacteria and cancer cells.

The Australopithecus assemblage from Sterkfontein Member 4 (South Africa) and the concept of variation in palaeontology.

Interpreting morphological variation within the early hominin fossil record is particularly challenging. Apart from the fact that there is no absolute threshold for defining species boundaries in palaeontology, the degree of variation related to sexual dimorphism, temporal depth, geographic variation or ontogeny is difficult to appreciate in a fossil taxon mainly represented by fragmentary specimens, and such variation could easily be conflated with taxonomic diversity. One of the most emblematic examples in paleoanthropology is the Australopithecus assemblage from the Sterkfontein Caves in South Africa. Whereas some studies support the presence of multiple Australopithecus species at Sterkfontein, others explore alternative hypotheses to explain the morphological variation within the hominin assemblage. In this review, I briefly summarize the ongoing debates surrounding the interpretation of morphological variation at Sterkfontein Member 4 before exploring two promising avenues that would deserve specific attention in the future, that is, temporal depth and nonhuman primate diversity.

Investigating the effects of compound paralogous EPHB receptor mutations on mouse facial development.

BACKGROUND: Variation in facial shape may arise from the combinatorial or overlapping actions of paralogous genes. Given its many members, and overlapping expression and functions, the EPH receptor family is a compelling candidate source of craniofacial morphological variation. We performed a detailed morphometric analysis of an allelic series of E14.5 Ephb1-3 receptor mutants to determine the effect of each paralogous receptor gene on craniofacial morphology. RESULTS: We found that Ephb1, Ephb2, and Ephb3 genotypes significantly influenced facial shape, but Ephb1 effects were weaker than Ephb2 and Ephb3 effects. Ephb2-/- and Ephb3-/- mutations affected similar aspects of facial morphology, but Ephb3-/- mutants had additional facial shape effects. Craniofacial differences across the allelic series were largely consistent with predicted additive genetic effects. However, we identified a potentially important nonadditive effect where Ephb1 mutants displayed different morphologies depending on the combination of other Ephb paralogs present, where Ephb1+/- , Ephb1-/- , and Ephb1-/- ; Ephb3-/- mutants exhibited a consistent deviation from their predicted facial shapes. CONCLUSIONS: This study provides a detailed assessment of the effects of Ephb receptor gene paralogs on E14.5 mouse facial morphology and demonstrates how the loss of specific receptors contributes to facial dysmorphology.

The Role of Morphological Variation in Evolutionary Robotics: Maximizing Performance and Robustness.

Exposing an Evolutionary Algorithm that is used to evolve robot controllers to variable conditions is necessary to obtain solutions which are robust and can cross the reality gap. However, we do not yet have methods for analyzing and understanding the impact of the varying morphological conditions which impact the evolutionary process, and therefore for choosing suitable variation ranges. By morphological conditions, we refer to the starting state of the robot, and to variations in its sensor readings during operation due to noise. In this article, we introduce a method that permits us to measure the impact of these morphological variations and we analyze the relation between the amplitude of variations, the modality with which they are introduced, and the performance and robustness of evolving agents. Our results demonstrate that (i) the evolutionary algorithm can tolerate morphological variations which have a very high impact, (ii) variations affecting the actions of the agent are tolerated much better than variations affecting the initial state of the agent or of the environment, and (iii) improving the accuracy of the fitness measure through multiple evaluations is not always useful. Moreover, our results show that morphological variations permit generating solutions which perform better both in varying and non-varying conditions.

Assessment of genetic diversity of ninety-one bottle gourd [Lagenaria siceraria (Mol.) Standl.] genotypes from fourteen different agro-climatic zones of India using agro-morphological traits and SSR markers.

BACKGROUND: The evaluation of the magnitude of genetic diversity present in the germplasm collection is prerequisite for bottle gourd improvement programme. The characterization of the diversity pattern of Indian bottle gourd accessions will facilitate the optimal use of genetic resources for breeding improved cultivars. METHODS: In the present study, the magnitude of genetic diversity was evaluated in ninety-one genotypes of bottle gourd collected across fourteen different agro-climatic zones of India. RESULTS: Significant variations were observed for all the studied ten quantitative and nine qualitative traits. The ninety-one genotypes were grouped into nine clusters based on cluster analysis of morphological characteristics. Eigen value from principal component analysis depicted first seven quantitative traits accounted for more than 97.5 cumulative percent of the total variations. The first two components accounted for 50 cumulative percent of the total variation, which signifies a high degree of correlation between the analyzed traits. Molecular diversity with the 40 SSR markers screened revealed 11 polymorphic markers in the genotypes studied. Population structure analysis divulged five populations, conforming to the Principal Coordinate Analysis. Molecular analysis revealed genetically diverse genotypes along with the morphologically divergent genotypes from the quantitative traits and highest inter-cluster distance would be the most appropriate parents for exploiting heterosis. CONCLUSIONS: The results of this study will facilitate the optimal use of genetic resources for breeding improved cultivars of bottle gourd and the adoption of the identified superior genotypes directly by the breeders.

A new variety of chondrocoracoideus muscle, or an additional head of pectoralis major muscle.

The pectoralis major and pectoralis minor muscles are located in the anterior chest wall. This region is characterized by high morphological variability. During dissection an additional muscle was found, originating from the lateral border of the pectoralis major muscle. After fusion it passed into the tendinous part coursing under the insertion of the pectoralis major muscle, then formed a common junction with the short head of the biceps brachii muscle, the distal attachment of which is on the coracoid process. Such an accessory structure could lead to neurovascular compression and cause thoracic outlet syndrome, of which pain is usually the first symptom. This muscle has not been described in the literature so far and for that reason we can name the present case as an unique structure.

Coexistence of accessory band with the second type of extensor hallucis longus.

The main muscles responsible for extension of the toes are the extensor digitorum longus and extensor hallucis longus. Morphological variants of both of these muscles are mostly related to the numbers of tendons and their unusual points of insertion. The case presented shows an accessory band deriving from the extensor digitorum longus and fusing with the extensor hallucis longus, a concomitant additional tendon of the latter inserting to the proximal phalanx of the hallux. Knowledge of these anatomical patterns is important not only from the anatomical but also the clinical point of view and can be useful in lower limb surgery.

Evolvability of the vertebrate craniofacial skeleton.

The skull is a vertebrate novelty. Morphological adaptations of the skull are associated with major evolutionary transitions, including the shift to a predatory lifestyle and the ability to masticate while breathing. These adaptations include the chondrocranium, dermatocranium, articulated jaws, primary and secondary palates, internal choanae, the middle ear, and temporomandibular joint. The incredible adaptive diversity of the vertebrate skull indicates an underlying bauplan that promotes evolvability. Comparative studies in craniofacial development suggest that the craniofacial bauplan includes three secondary organizers, two that are bilaterally placed at the Hinge of the developing jaw, and one situated in the midline of the developing face (the FEZ). These organizers regulate tissue interactions between the cranial neural crest, the neuroepithelium, and facial and pharyngeal epithelia that regulate the development and evolvability of the craniofacial skeleton.

Human burials at the Kisese II rockshelter, Tanzania.

OBJECTIVES: The Late Pleistocene and early Holocene in eastern Africa are associated with complex evolutionary and demographic processes that contributed to the population variability observed in the region today. However, there are relatively few human skeletal remains from this time period. Here we describe six individuals from the Kisese II rockshelter in Tanzania that were excavated in 1956, present a radiocarbon date for one of the individuals, and compare craniodental morphological diversity among eastern African populations. MATERIALS AND METHODS: This study used standard biometric analyses to assess the age, sex, and stature of the Kisese II individuals. Eastern African craniodental morphological variation was assessed using measures of dental size and a subset of Howells' cranial measurements for the Kisese II individuals as well as early Holocene, early pastoralist, Pastoral Neolithic, and modern African individuals. RESULTS: Our results suggest a minimum of six individuals from the Kisese II collections with two adults and four juveniles. While the dating for most of the burials is uncertain, one individual is directly radiocarbon dated to ~7.1 ka indicating that at least one burial is early Holocene in age. Craniodental metric comparisons indicate that the Kisese II individuals extend the amount of human morphological diversity among Holocene eastern Africans. CONCLUSIONS: Our findings contribute to a growing body of evidence that Late Pleistocene and early Holocene eastern Africans exhibited relatively high amounts of morphological diversity. However, the Kisese II individuals suggest morphological similarity at localized sites potentially supporting increased regionalization during the early Holocene.

Secular change in morphological cranial and mandibular trait frequencies in European Americans born 1824-1987.

OBJECTIVES: Secular change in cranial and postcranial morphometrics and morphological traits has been documented in several studies. However, to date, few studies have addressed temporal changes occurring in the expression of cranial morphological traits commonly used in ancestry estimation. This study examines secular change in the expression of 23 cranial and mandibular morphological traits; accounting for age-at-death, sex, and year-of-birth. MATERIALS & METHODS: Data were collected on 23 morphological cranial and mandibular traits for European American individuals (19-97 years of age) from the Hamann-Todd Skeletal Collection (n = 518) and the William M. Bass Donated Skeletal Collection (n = 602). Individuals were divided into six birth-year cohorts: 1824-1849 (Cohort 1), 1850-1874 (Cohort 2), 1875-1899 (Cohort 3), 1900-1924 (Cohort 4), 1925-1949 (Cohort 5), and 1950-1987 (Cohort 6). RESULTS: Statistical analyses, including Pearson's chi-square, correspondence analysis, and ordinal regression, demonstrate that secular changes have occurred in 11 traits, including: anterior nasal spine (ANS); malar tubercle (MT); nasal bone contour (NBC); postbregmatic depression (PBD); supranasal suture (SPS); transverse palatine suture (TPS); zygomaticomaxillary suture (ZS); ascending ramus shape (ARS); gonial angle flare (GAF); mandibular tori (MDT); and posterior ramus edge inversion (PREI), with changes occurring in both sexes for ANS, MT, TPS, ZS, GAF, MDT, and PREI. Significant changes in trait expression were found predominately between Cohorts 3 and 4, and Cohorts 4 and 5. While the sex of an individual affected the expression of ANS, MT, NBC, PBD, SPS, ZS, ARS, GAF, and PREI, age-at-death only affected MT and PREI. DISCUSSION: This study demonstrates that secular change in morphological cranial and mandibular traits has occurred over the last two centuries in European Americans, with the most considerable change appearing at the turn of the twentieth century. Changes in morphological trait expression over a relatively short period of time correspond with changes seen in craniometric analyses and correlate with the industrialization of society and environmental and cultural changes, such as medical advancements, nutrition, and population health/stress.

Estimating mobility using sparse data: Application to human genetic variation.

Mobility is one of the most important processes shaping spatiotemporal patterns of variation in genetic, morphological, and cultural traits. However, current approaches for inferring past migration episodes in the fields of archaeology and population genetics lack either temporal resolution or formal quantification of the underlying mobility, are poorly suited to spatially and temporally sparsely sampled data, and permit only limited systematic comparison between different time periods or geographic regions. Here we present an estimator of past mobility that addresses these issues by explicitly linking trait differentiation in space and time. We demonstrate the efficacy of this estimator using spatiotemporally explicit simulations and apply it to a large set of ancient genomic data from Western Eurasia. We identify a sequence of changes in human mobility from the Late Pleistocene to the Iron Age. We find that mobility among European Holocene farmers was significantly higher than among European hunter-gatherers both pre- and postdating the Last Glacial Maximum. We also infer that this Holocene rise in mobility occurred in at least three distinct stages: the first centering on the well-known population expansion at the beginning of the Neolithic, and the second and third centering on the beginning of the Bronze Age and the late Iron Age, respectively. These findings suggest a strong link between technological change and human mobility in Holocene Western Eurasia and demonstrate the utility of this framework for exploring changes in mobility through space and time.

Developmental processes regulate craniofacial variation in disease and evolution.

Variation in development mediates phenotypic differences observed in evolution and disease. Although the mechanisms underlying phenotypic variation are still largely unknown, recent research suggests that variation in developmental processes may play a key role. Developmental processes mediate genotype-phenotype relationships and consequently play an important role regulating phenotypes. In this review, we provide an example of how shared and interacting developmental processes may explain convergence of phenotypes in spliceosomopathies and ribosomopathies. These data also suggest a shared pathway to disease treatment. We then discuss three major mechanisms that contribute to variation in developmental processes: genetic background (gene-gene interactions), gene-environment interactions, and developmental stochasticity. Finally, we comment on evolutionary alterations to developmental processes, and the evolution of disease buffering mechanisms.

Genetic and morphological divergence at a biogeographic break in the beach-dwelling brooder Excirolana hirsuticauda Menzies (Crustacea, Peracarida).

BACKGROUND: There is a biogeographic break located at 30°S in the southeast Pacific, in a coastal area of strong environmental discontinuities. Several marine benthic taxa with restricted dispersal have a coincident phylogeographic break at 30°S, indicating that genetic structure is moulded by life history traits that limit gene flow and thereby promote divergence and speciation. In order to evaluate intraspecific divergence at this biogeographic break, we investigated the genetic and morphological variation of the directly developing beach isopod Excirolana hirsuticauda along 1900 km of the southeast Pacific coast, across 30°S. RESULTS: The COI sequences and microsatellite data both identified a strong discontinuity between populations of E. hirsuticauda to the north and south of 30°S, and a second weaker phylogeographic break at approximately 35°S. The three genetic groups were evidenced by different past demographic and genetic diversity signatures, and were also clearly distinguished with microsatellite data clustering. The COI sequences established that the genetic divergence of E. hirsuticauda at 30°S started earlier than divergence at 35°. Additionally, the three groups have different past demographic signatures, with probable demographic expansion occurring earlier in the southern group (south of 35°S), associated with Pleistocene interglacial periods. Interestingly, body length, multivariate morphometric analyses, and the morphology of a fertilization-related morphological character in males, the appendix masculina, reinforced the three genetic groups detected with genetic data. CONCLUSIONS: The degree of divergence of COI sequences, microsatellite data, and morphology was concordant and showed two geographic areas in which divergence was promoted at differing historical periods. Variation in the appendix masculina of males has probably promoted reproductive isolation. This variation together with gene flow restrictions promoted by life history traits, small body size, oceanographic discontinuities and sandy-beach habitat continuity, likely influenced species divergence at 30°S in the southeast Pacific coast. The degree of genetic and morphological differentiation of populations to the north and south of 30°S suggests that E. hirsuticauda harbours intraspecific divergence consistent with reproductive isolation and an advanced stage of speciation. The speciation process within E. hirsuticauda has been shaped by both restrictions to gene flow and a prezygotic reproductive barrier.

Evolution of Ceratozamia cycads: A proximate-ultimate approach.

Evidence suggests that past climatic fluctuations affected speciation of extant cycads. However, empirical genetic and morphological evidence explaining patterns and processes of species diversification are scarce. There are some explanations for the origin and evolution of the genus Ceratozamia, but with inconclusive results. To elucidate the evolution of Ceratozamia, we used genetic and phenotypic sources as empirical data, which were applied in a 'proximate-ultimate' framework (ecological and evolutionary scale, respectively). Our results suggested that the evolutionary mechanisms of speciation were shaped by deterministic (natural selection-adaptation) driven by climatic conditions associated to water stress, and probably enhanced by stochastic processes (gene drift and inbreeding). In general terms, punctuated evolution models were those that best explained the patterns of speciation throughout the phylogenetic history of the lineages encompassed in the genus Ceratozamia. Finally, we provide empirical evidence on the tempo and mode of the evolution of a 'living plant fossil'.