Considering decoupled phenotypic diversification between ontogenetic phases in macroevolution: An example using Triggerfishes (Balistidae)

Across the Tree of Life, most studies of phenotypic disparity and diversification have been restricted to adult organisms. However, many lineages have distinct ontogenetic phases that do not reflect the same traits as their adult forms. Non-adult disparity patterns are particularly important to consider for coastal ray-finned fishes, which often have juvenile phases with distinct phenotypes. These juvenile forms are often associated with sheltered nursery environments, with phenotypic shifts between adults and juvenile stages that are readily apparent in locomotor morphology. However, whether this ontogenetic variation in locomotor morphology reflects a decoupling of diversification dynamics between life stages remains unknown. Here we investigate the evolutionary dynamics of locomotor morphology between adult and juvenile triggerfishes. Integrating a time-calibrated phylogenetic framework with geometric morphometric approaches and measurement data of fin aspect ratio and incidence, we reveal a mismatch between morphospace occupancy, the evolution of morphological disparity, and the tempo of trait evolution between life stages. Collectively, our results illuminate how the heterogeneity of morpho-functional adaptations can decouple the mode and tempo of morphological diversification between ontogenetic stages.

Skull morphological variation in a British stranded population of false killer whale (Pseudorca crassidens): a three-dimensional geometric morphometric approach

The false killer whale (Pseudorca crassidens (Owen, 1846)) is a globally distributed delphinid that shows geographical differentiation in its skull morphology. We explored cranial morphological variation in a sample of 85 skulls belonging to a mixed sex population stranded in the Moray Firth, Scotland, in 1927. A three-dimensional digitizer (Microscribe 2GX) was used to record 37 anatomical landmarks on the cranium and 25 on the mandible to investigate size and shape variation and to explore sexual dimorphism using geometric morphometric. Males showed greater overall skull size than females, whereas no sexual dimorphism could be identified in cranial and mandibular shape. Allometric skull changes occurred in parallel for both males and females, supporting the lack of sexual shape dimorphism for this particular sample. Also, fluctuating asymmetry did not differ between crania of males and females. This study confirms the absence of sexual shape dimorphism and the presence of a sexual size dimorphism in this false killer whale population.

The first epiphytic macrolichen and its implication to the interacting with Mesozoic forest ecosystem

Lichens are well known as pioneer organisms or stress-tolerant extremophiles playing a core role in the early formation of terrestrial ecosystems, of which epiphytic lichens make a distinct contribution to the water-cycle and nutrient cycling in forest ecosystem. But due to the scarcity of relevant fossil records, the evolutionary history of epiphytic lichens is poorly documented. Herein, based on the new material of Daohugouthallus ciliiferus, we demonstrated that the hitherto oldest macrolichen inhabited a gymnosperm branch, representing the first unambiguous Jurassic epiphytic lichen. Combing the fossil and extant macrolichen representatives, we performed the geometric morphometric analysis and comprehensive comparison to infer the systematic status of this rare Jurassic macrolichen. The results declared that D. ciliiferus cannot be assigned to any known macrolichen lineages for its elder age and particular habits, and therefore a new family, Daohugouthallaceae was proposed. This work updated the current knowledge to the historical evolution of epiphytic lichens, implying the macrolichens may have diversified much earlier than the generally accepted K–Pg boundary. In addition, our new finding also provided direct evidence for tracing the continuing joint development of epiphytic lichens and forest ecosystem since the Jurassic of 165 Mya.

Recovery of Haemal Lordosis in European Seabass Dicentrarchus labrax (Linnaeus 1758)

Lordosis of the haemal part of the vertebral column is a frequent abnormality in reared fish. Haemal lordosis develops during the late larval and early juvenile life stages of fish, mainly due to the high swimming activity of the fish in the rearing tanks. In the present study, we have examined whether haemal lordosis recovers during the growth of European seabass. Furthermore, we aimed to develop simple morphometric indices (PrAn1 and PrAn2) that might link the severity of lordosis at the juvenile stage with fish morphological quality at harvesting. At 111 days post-hatching (dph, 53 ± 4 mm standard length, SL), 600 seabass juveniles with lordotic (L, 200 fish) or normal (N, 400 fish) external morphology were selected and introduced in a common tank. At 150 dph (75 ± 7 mm SL), 350 fish were randomly selected, pit-tagged and transferred to a sea cage for on-growing up to 502 dph (234 ± 16 mm SL). The morphological examination of the fish at 150 and 502 dph revealed that 60% (46 out of 77) of L juveniles turned into normal phenotype by the end of on-growing period. Interestingly, 56% of the fish with recovered external morphology (N-Rec) presented either a completely normal vertebral column (31%) or minor abnormalities of individual vertebrae (25%). Following the results of geometric morphometric analysis, the differences in the body shape between N-Rec and N fish were not statistically significant (p > 0.05, canonical variate analysis). The examined morphometric indices were effective in discriminating the normal fish from 58% (PrAn1) to 65% (PrAn2) of lordotic juveniles. Results are discussed with respect to the mechanism of lordosis recovery, and spotlights on their application for quality control and cull out of the abnormal fish in commercial hatcheries.

Testing the accuracy of 3D automatic landmarking via genome-wide association studies

Various advances in 3D automatic phenotyping and landmark-based geometric morphometric methods have been made. While it is generally accepted that automatic landmarking compromises the capture of the biological variation, no studies have directly tested the actual impact of such landmarking approaches in analyses requiring a large number of specimens and for which the precision of phenotyping is crucial to extract an actual biological signal adequately. Here, we use a recently developed 3D atlas-based automatic landmarking method to test its accuracy in detecting QTLs associated with craniofacial development of the house mouse skull and lower jaws for a large number of specimens (circa 700) that were previously phenotyped via a semi-automatic landmarking method complemented with manual adjustment. We compare both landmarking methods with univariate and multivariate mapping of the skull and the lower jaws. We find that most significant SNPs and QTLs are not recovered based on the data derived from the automatic landmarking method. Our results thus confirm the notion that information is lost in the automated landmarking procedure although somewhat dependent on the analyzed structure. The automatic method seems to capture certain types of structures slightly better, such as lower jaws whose shape is almost entirely summarized by its outline and could be assimilated as a 2D flat object. By contrast, the more apparent 3D features exhibited by a structure such as the skull are not adequately captured by the automatic method. We conclude that using 3D atlas-based automatic landmarking methods requires careful consideration of the experimental question.

Geometric Morphometrics of Bilateral Asymmetry in Eunotia bilunaris (Eunotiales, Bacillariophyceae) as a Tool for the Quantitative Assessment of Teratogenic Deviations in Frustule Shapes

A number of pennate diatom genera typically have teratogenic deformations of their siliceous frustules due to the effects of environmental stress, such as high concentrations of heavy metals and low pH. However, the quantitative assessment of these deformations has rarely been applied. One species in which aberrations have frequently been reported is Eunotia bilunaris, which typically has bilaterally symmetric frustules with dorso-ventral differentiation. In this study, we aimed to illustrate the geometric morphometric analysis of symmetry as a tool for assessing the severity of teratogenic deformations. These were quantified by Procrustes superimposition of equidistant points placed along the valvar outlines in pairs of configurations based on their bilateral reflection symmetry. The shape deformations were mostly confined to central parts of the ventral outlines and were captured both by the symmetric and asymmetric subspaces of the variation. The amount of bilateral asymmetry in individual cells was negatively related to frustule size via the allometric power law relationship, illustrating that asymmetry increased in the asexual diminution series. The presented analysis provides a framework for the quantitative assessment of frustule deformations in eunotioid diatoms that can be used for the comparative scoring of teratogenic deviations among cells, populations, or species.

The oscillating Mucin-type protein DPY-6 has a conserved role in nematode mouth and cuticle formation

Nematodes show an extraordinary diversity of mouth structures and strikingly different feeding strategies, which has enabled an invasion of all ecosystems. However, nearly nothing is known about the structural and molecular architecture of the nematode mouth (stoma). Pristionchus pacificus is an intensively studied nematode that exhibits unique life history traits, including predation, teeth-like denticle formation, and mouth-form plasticity. Here, we used a large-scale genetic screen to identify genes involved in mouth formation. We identified Ppa-dpy-6 to encode a Mucin-type hydrogel-forming protein that is macroscopically involved in the specification of the cheilostom, the anterior part of the mouth. We used a recently developed protocol for geometric morphometrics of miniature animals to characterize these defects further and found additional defects that affect mouth form, shape, and size resulting in an overall malformation of the mouth. Additionally, Ppa-dpy-6 is shorter than wild-type with a typical Dumpy phenotype, indicating a role in the formation of the external cuticle. This concomitant phenotype of the cheilostom and cuticle provides the first molecular support for the continuity of these structures and for the separation of the cheilostom from the rest of the stoma. In C. elegans, dpy-6 was an early mapping mutant but its molecular identity was only determined during genome-wide RNAi screens and not further investigated. Strikingly, geometric morphometric analysis revealed previously unrecognized cheilostom and gymnostom defects in Cel-dpy-6 mutants. Thus, the Mucin-type protein DPY-6 represents to the best of our knowledge, the first protein involved in nematode mouth formation with a conserved role in cuticle deposition. This study opens new research avenues to characterize the molecular composition of the nematode mouth, which is associated with extreme ecological diversification.

Three-Dimensional Dental Analysis for Sex Estimation in the Italian Population: A Pilot Study Based on a Geometric Morphometric and Artificial Neural Network Approach

Dental dimorphism can be used for discriminating sex in forensic contexts. Geometric morphometric analysis (GMA) allows the evaluation of the shape and size, separately, of uneven 3D objects. This study presents experiments using a novel combination of GMA and an artificial neural network (ANN) for sex classification, applied to premolars of Caucasian Italian adults (50 females and 50 males). General Procrustes superimposition (GPS) and the partial least square (PLS) method were performed, respectively, to study the shape variance between sexes and to eliminate landmark variations. The “set-aside” approach was used to assess the accuracy of the proposed neural networks. As the main findings of the pilot study, the proposed method applied to the first upper premolar correctly classified 90% of females and 73% of males of the test sample. The accuracy was 0.84 and 0.80 for the training and test samples, respectively. The sexual dimorphism resulting from GMA was low, although statistically significant. GMA combined with the ANN demonstrated better sex classification ability than previous odontometric or dental morphometric methods. Future research could overcome some limitations by considering a larger sample of subjects and other kinds of teeth and experimenting with the use of computer vision for automatic landmark positioning.