Hydnora (Hydnoraceae) comprises a few parasitic species exceptional among the autotrophic members of the perianth-bearing Piperales. Flowers in the genus are thick, fleshy, sapromyophilous, and develop into massive, polyspermous fruits. They are formed directly along underground rhizomes that parasitize species of Euphorbiaceae and Fabaceae. Due to its peculiar floral morphology and lack of leaves, Hydnora is often dubbed ‘the strangest plant in the world’. Here we generated the first transcriptomes of Hydnora visseri from the dissected rhizome, perianth, osmophore, stamen, carpel, and fruit. Our results suggest that Hydnora possesses one of the simplest developmental genetic toolkits for flowering and floral organ identity among angiosperms, further emphasizing its uniqueness. We detected that most of the photoperiodic flowering integrators are expressed. In contrast, regulators of the autonomous pathway and circadian clock were notably absent from the transcriptomes. Conversely, we identified an intact genetic toolkit linked to floral organ identity and fruit development in Hydnora. Through positional homology and gene expression data, we inferred that the perianth of Hydnora corresponds to the calyx, and that the osmophores are late sepal elaborations. Additionally, the expression patterns of genes responsible for stamen, carpel, and ovule identity align with the canonical ABCDE model. Finally, we recorded large-scale duplications in putative perianth identity genes prior to the diversification of all perianth-bearing Piperales. This study serves as an additional comparative point for assessing the evolutionary onset of holoparasitic plants, as Hydnora and its sister genus Prosopanche are likely the earliest branching representatives of this lifestyle across angiosperms.Supplementary InformationThe online version contains supplementary material available at 10.1186/s13227-025-00252-8.

Lineage-specific adult structures form through modifications of pre-existing juvenile body parts during postembryonic development in insects. It remains unclear how these novel traits originate from ancestral structures within the constrained body plan. In the coffin-headed cricket Loxoblemmus equestris, an ancestral rounded head shape directly transforms into a flattened derived form in a sex-specific manner. To understand the origin of novel traits, we investigated the development of the adult head in L. equestris as a model of lineage-specific novelty. We found that head morphologies remained sexually monomorphic until the final molt, and the male-specific head shape emerged in the frons region during the transition to adulthood in L. equestris. Two- and three-dimensional morphological analyses revealed that the sexual dimorphism in the frons epithelial folding patterns appeared in the late final nymphal instar. These results suggest that the male-specific novel head development is linked to the final molt in L. equestris. We tested this hypothesis by knocking down the metamorphic gene network (MGN) comprised of Krüppel-homolog 1 (Kr-h1), broad (br), and Ecdysone induced protein 93F (E93). Despite the timing shifts of the nymph-to-adult transition caused by knockdown of the MGN, male-specific head structures are formed only after the final molt. These results demonstrate that the novel male head structures are formed during the final molt through the formation of sex-specific epithelial patterns in L. equestris. This highlights the unique metamorphic lifecycle with the final molt as a driver that has created lineage- and sex-specific adult forms in insects.

BackgroundPhagocytosis is a universal physiological process in eukaryotes with many important biological functions. In nudibranch gastropods, a novel form of phagocytosis called nematocyst sequestration is specialized for the uptake of venomous stinging organelles stolen from their cnidarian prey. This process is highly selective. Here we use the emerging model nudibranch species Berghia stephanieae and Hermissenda opalescens to identify genes enriched within the body regions where nematocyst sequestration occurs, and investigate how the expression profile of phagocytosis, immune, and digestive genes differs between nematocyst-sequestering regions relative to those where other phagocytic functions occur.ResultsWe identified 166 genes with significantly higher expression in sequestering regions in B. stephanieae, including genes associated with development, membrane transport, and metabolism. Of these, at least 31 overlap with transcripts upregulated in H. opalescens sequestering tissues. Using hybridization chain reaction in situs, we show that at least two of these genes were localized to sequestering cells in B. stephanieae, including a putative C-type lectin receptor and a collagen. Genes annotated with phagocytosis, digestion, or immunity GO terms were often expressed in both sequestering and non-sequestering tissues, suggesting that they may also play a role in sequestration processes.ConclusionOur results suggest that phagocytosis genes likely play a role in the sequestration phenotype, and that a small subset of genes (e.g., collagen) may play unique functions yet to be uncovered. We also show that genes categorized as functioning in endocytosis, immunity, and digestion have lower overall expression in sequestering tissues, supporting the hypothesis that sequestering tissues show a narrowing of function compared to digestive tissues. This study lays the foundation for further inquiry into mechanisms of organelle sequestration in nudibranchs and other organisms.

The Notch signalling pathway is a conserved regulator of cellular processes, including differentiation, proliferation, and apoptosis, across Metazoa. While its roles in bilaterians are well-characterized, the evolutionary history and functional diversification of Notch signalling in early branching metazoans, including cnidarians and parasitic cnidarians, myxozoans, remain underexplored. This study examines the presence and structural diversity of Notch pathway components across early diverging metazoan lineages, with a particular focus on Myxozoa—parasitic cnidarians characterized by extreme morphological and genomic reduction. Comparative analyses of 58 metazoan species revealed broad conservation of core Notch components, alongside losses in ctenophores, placozoans, poriferans, and cnidarians. Myxozoa retain only 14 of the 28 canonical pathway components, lacking key elements, such as MAML, Hes/Hey, and DVL. Phylogenetic analyses of Notch receptors highlight structural divergence, particularly in extracellular domains, reflecting lineage-specific variations. The phylogeny of Delta and Jagged ligands reveals an evolutionary trajectory, with Delta ligands showing early diversification within metazoans, and their structural variability has been explored, though their functional roles remain unknown. The Notch receptor was detected at the subcellular level in proliferative stages of Sphaerospora molnari through immunolocalization studies, suggesting it is active in these cells. Our findings contribute to understanding the evolution of the Notch signalling pathway, highlighting its conserved role in developmental regulation across early branching Metazoa.

A challenge for evolutionary developmental (evo-devo) biology is to expand the breadth of research organisms used to investigate how animal diversity has evolved through changes in embryonic development. New experimental systems should couple a relevant phylogenetic position with available molecular tools and genomic resources. As a phylum of the sister group to chordates, echinoderms extensively contributed to our knowledge of embryonic patterning, organ development and cell-type evolution. Echinoderms display a variety of larval forms with diverse shapes, making them a suitable group to compare the evolution of embryonic developmental strategies. However, because of the laboratory accessibility and the already available techniques, most studies focus on sea urchins and sea stars mainly. As a comparative approach, the field would benefit from including information on other members of this group, like the sea cucumbers (holothuroids), for which little is known on the molecular basis of their development. Here, we review the spawning and culture methods, the available morphological and molecular information, and the current state of genomic and transcriptomic resources on sea cucumbers. With the goal of making this system accessible to the broader community, we discuss how sea cucumber embryos and larvae can be a powerful system to address the open questions in evo-devo, including understanding the origins of bilaterian structures.

BackgroundIn social insects, interactions among colony members trigger caste differentiation with morphological modifications. In termite caste differentiation, caste-specific morphologies (such as mandibles in soldiers, genital organs in reproductives or wings in alates) are well developed during post-embryonic development under endocrine controls (e.g., juvenile hormone and ecdysone). Since body part-specific morphogenesis in caste differentiation is hormonally regulated by global factors circulated throughout the body, positional information should be required for the caste-specific and also body part-specific morphogenesis. To identify factors providing the positional information, expression and functional analyses of eight Hox genes were carried out during the three types of caste differentiation (i.e., soldier, neotenic and alate differentiation) in a termite, Hodotermopsis sjostedti.ResultsSpatio-temporal patterns of Hox gene expression during caste differentiation were elucidated by real-time qPCR, showing the caste-specific upregulations of Hox genes during the differentiation processes. Among eight Hox genes, Deformed (Dfd) was upregulated specifically in mandibles in soldier differentiation, abdominal-A (abd-A) and Abdominal-B (Abd-B) were upregulated in the abdomen in neotenic differentiation, while Sex-comb reduced (Scr) and Antennapedia (Antp) were upregulated during alate differentiation. Furthermore, RNAi knockdown of Dfd in soldier differentiation and of abd-A and Abd-B in neotenic differentiation distorted the modifications of caste-specific morphologies.ConclusionsGene expression and functional analyses in this study revealed that, in the caste differentiation in termites, upregulation of Hox genes provide positional identities of body segments, resulting in the caste-specific morphogenesis. The acquisition of such developmental modifications would have enabled the evolution of sophisticated caste systems in termites.

The vertebrate head skeleton has evolved a myriad of forms since their divergence from invertebrate chordates. The connection between novel gene expression and cell types is therefore of importance in this process. The transformation of the jawed vertebrate (gnathostome) head skeleton from oral cirri to jointed jaw elements required a diversity of cartilages as well as changes in the patterning of these tissues. Although lampreys are a sister clade to gnathostomes, they display skeletal diversity with distinct gene expression and histologies, a useful model for addressing joint evolution. Specifically, the lamprey tissue known as mucocartilage has noted similarities with the jointed elements of the mandibular arch in jawed vertebrates. We thus asked whether the cells in lamprey mucocartilage and gnathostome joint tissue could be considered homologous. To do this, we characterized new genes that are involved in gnathostome joint formation and characterized the histochemical properties of lamprey skeletal types. We find that most of these genes are minimally found in mucocartilage and are likely later innovations, but we do identify new activity for gdf5/6/7b in both hyaline and mucocartilage, supporting its role as a chondrogenic regulator. Contrary to previous works, our histological assays do not find any perichondrial fibroblasts surrounding mucocartilage, suggesting that mucocartilage is non-skeletogenic tissue that is partially chondrified. Interestingly, we also identify new histochemical features of the lamprey otic capsule that diverge from normal hyaline. Paired with our new insights into lamprey mucocartilage, we propose a broader framework for skeletal evolution in which an ancestral soxD/E and gdf5/6/7 network directs mesenchyme along a spectrum of cartilage-like features.

BackgroundThe polyplacophoran mollusks (chitons) possess serially arranged shell plates. This feature is unique among mollusks and believed to be essential to explore the evolution of mollusks as well as their shells. Previous studies revealed several cell populations in the dorsal epithelium (shell field) of polyplacophoran larvae and their roles in the formation of shell plates. Nevertheless, they provide limited molecular information, and shell field morphogenesis remains largely uninvestigated.ResultsIn the present study, we investigated shell field development in the chiton Acanthochitona rubrolineata based on morphological characteristics and molecular patterns. A total of four types of tissue could be recognized from the shell field of A. rubrolineata. The shell field comprised not only the centrally located, alternatively arranged plate fields and ridges, but also the tissues surrounding them, which were the precursors of the girdle and we termed as the girdle field. The girdle field exhibited a concentric organization composed of two circularly arranged tissues, and spicules were only developed in the outer circle. Dynamic engrailed expression and F-actin (filamentous actin) distributions revealed relatively complicated morphogenesis of the shell field. The repeated units (plate fields and ridges) were gradually established in the shell field, seemingly different from the manners used in the segmentation of Drosophila or vertebrates. The seven repeated ridges also experienced different modes of ontogenesis from each other. In the girdle field, the presumptive spicule-formation cells exhibited different patterns of F-actin aggregations as they differentiate.ConclusionsThese results reveal the details concerning the structure of polyplacophoran shell field as well as its morphogenesis. They would contribute to exploring the mechanisms of polyplacophoran shell development and molluscan shell evolution.

BackgroundBrachiopods and molluscs are lophotrochozoans with hard external shells which are often believed to have evolved convergently. While palaeontological data indicate that both groups are descended from biomineralising Cambrian ancestors, the closest relatives of brachiopods, phoronids and bryozoans, are mineralised to a much lower extent and are comparatively poorly represented in the Palaeozoic fossil record. Although brachiopod and mollusc shells are structurally analogous, genomic and proteomic evidence indicates that their formation involves a complement of conserved, orthologous genes. Here, we study a set of genes comprised of 3 homeodomain transcription factors, one signalling molecule and 6 structural proteins which are implicated in mollusc and brachiopod shell formation, search for their orthologs in transcriptomes or genomes of brachiopods, phoronids and bryozoans, and present expression patterns of 8 of the genes in postmetamorphic juveniles of the rhynchonelliform brachiopod T. transversa.ResultsTranscriptome and genome searches for the 10 target genes in the brachiopods Terebratalia transversa, Lingula anatina, Novocrania anomala, the bryozoans Bugula neritina and Membranipora membranacea, and the phoronids Phoronis australis and Phoronopsis harmeri resulted in the recovery of orthologs of the majority of the genes in all taxa. While the full complement of genes was present in all brachiopods with a single exception in L. anatina, a bloc of four genes could consistently not be retrieved from bryozoans and phoronids. The genes engrailed, distal-less, ferritin, perlucin, sp1 and sp2 were shown to be expressed in the biomineralising mantle margin of T. transversa juveniles.ConclusionsThe gene expression patterns we recovered indicate that while mineralised shells in brachiopods and molluscs are structurally analogous, their formation builds on a homologous process that involves a conserved complement of orthologous genes. Losses of some of the genes related to biomineralisation in bryozoans and phoronids indicate that loss of the capacity to form mineralised structures occurred already in the phoronid–bryozoan stem group and supports the idea that mineralised skeletons evolved secondarily in some of the bryozoan subclades.

Background Alternative patterns of secondary growth in stems of Nyctaginaceae is present in all growth habits of the family and have been known for a long time. However, the interpretation of types of cambial variants have been controversial, given that different authors have given them different developmental interpretations. The different growth habits coupled with an enormous stem anatomical diversity offers the unique opportunity to investigate the evolution of complex developments, to address how these anatomies shifted within habits, and how the acquisition of novel cambial variants and habit transitions impacted the diversification of the family.MethodsWe integrated developmental data with a phylogenetic framework to investigate the diversity and evolution of stem anatomy in Nyctaginaceae using phylogenetic comparative methods, reconstructing ancestral states, and examining whether anatomical shifts correspond to species diversification rate shifts in the family.ResultsTwo types of cambial variants, interxylary phloem and successive cambia, were recorded in Nyctaginaceae, which result from four different ontogenies. These ontogenetic trajectories depart from two distinct primary vascular structures (regular or polycyclic eustele) yet, they contain shared developmental stages which generate stem morphologies with deconstructed boundaries of morphological categories (continuum morphology). Unlike our a priori hypotheses, interxylary phloem is reconstructed as the ancestral character for the family, with three ontogenies characterized as successive cambia evolving in few taxa. Cambial variants are not contingent on habits, and their transitions are independent from species diversification.ConclusionsOur findings suggest that multiple developmental mechanisms, such as heterochrony and heterotopy, generate the transitions between interxylary phloem and successive cambia. Intermediate between these two extremes are present in Nyctaginaceae, suggesting a continuum morphology across the family as a generator of anatomical diversity.