Scleractinian corals owe their ecological success to long-enigmatic genomic innovations. By reconstructing ancestral linkage groups (ALGs) across 222 high-quality chromosome-level metazoan genomes at key evolutionary nodes spanning the bilaterian–cnidarian–sponge lineage, we propose the concepts of Precambrian “proto-cell type” and post-Cambrian “derived cell type” based on ALG–chromosome correspondence, showing that scleractinian corals represent a typical proto-cell type by preserving the most conserved Precambrian chromosomal core. From the ancestral 28-chromosome karyotype, they evolved a stable 14-chromosome architecture through progressive fusions while maintaining intact synteny for over 450 million years—the greatest such conservation in animals. This stability enabled coupled evolution of endosymbiosis and biomineralization, as revealed by pan-genomic single-cell transcriptomic analyses. These findings illuminate metazoan karyotype evolution and inform coral reef conservation.

Coral reef ecosystems face escalating threats from anthropogenic global climate challenges, leading to frequent bleaching events. A key issue in coral transplantation is the inability of fragments to rapidly grow to sizes that can resist environmental pressures. The observation of accelerated growth during the early stages of coral regeneration provides new insights for addressing this challenge. To investigate the underlying molecular mechanisms, we study the fast-growing stony coral Acropora muricata. Using single-cell RNA sequencing, bulk RNA sequencing, and high-resolution micro-computed tomography, we identify a critical regeneration phase around 2-4 weeks post-injury. Single-cell transcriptome analysis reveals 11 function-specific cell clusters. Pseudotime analysis indicates epidermal cell differentiation into calicoblasts. Bulk RNA-seq results highlight a temporal limitation in coral's rapid regeneration. Through integrated multi-omics analysis, this study emphasizes the importance of a comprehensive understanding of coral regeneration, providing insights beyond fundamental knowledge and offering potential protective strategies to promote coral growth.

Acroporidae and Pocilloporidae are the predominant reef-building corals of Indo-Pacific coral reefs. Coral colony is the basic geometric construction of coral reef, while sustained formation of their calcium carbonate skeletons is the heart of healthy reefs. However, the reef forming strategies in coral growth is still obscure. In this study, we reconstructed 32 representative samples using high-resolution computed tomography and created canal networks inside skeletal data to investigate coral growth regulation and related parameters. In Acropora and Montipora colonies, the polyps are connected as an integral network by complex canals that perform and regulate physiological processes including budding, branching, and mineralising. Furthermore, we visualised coral growth axes and rings, revealing colony growth pattern regulation. We also drew a skeleton grey-gradient heat map and calculated coral skeleton ratios to reveal skeletal diversity, devising a method to quantitatively analyse coral growth. On the basis of the canal network reconstructions, we hypothesised coral growth strategies and compared the similarities and differences among the four genera. This work extends the knowledge of how corals grow their skeleton, what the major controls are and how skeletal growth varies amongst species, ensuring further protections of tropical coral reefs.

The coral reef ecosystem faces increasing threats under global climate challenges. One of the core issues is the inability of fragments to quickly grow into a size that can resist environmental pressures in coral transplantation. The observation of accelerated growth during the early stages of coral regeneration provides new insights for addressing this challenge. To investigate related molecular mechanisms, our study focused on the fast-growing stony coral Acropora muricata (with chromosome-scale reference genome). Employing diverse techniques, including single-cell RNA sequencing (scRNA-seq), we unveiled related intricate cellular dynamics. Single-cell analysis revealed notable shifts in calicoblasts and epidermal cells around 2-4 weeks post-injury. Gene expression analysis revealed enrichment in immune response and biomineralization pathways. Pseudotime analysis explained the differentiation of epidermal cells into calicoblasts, while time-course analysis identified key genes associated with dynamic biomineralization changes. This study enhances our understanding of coral regeneration, offering insights for protective strategies to foster coral growth. ### Competing Interest Statement The authors have declared no competing interest.

Ocean acidification is increasing in frequency and is considered one of the most important causes of severe damage to global coral reefs. Therefore, there is an urgent need to study the impact of acid stress on the growth patterns of major reef-building corals. Here, we studied the skeleton forming strategies of four widely distributed coral species in a simulated acidified habitat with a pH of 7.6–7.8. We reconstructed and visualized the skeleton building process, quantified elemental calcium loss, and determined gene expression changes. The results suggest that different reef-building corals have diverse growing strategies in acidified seawater. A unique ‘cavity-like’ forming process starts from the inside of the skeletons of Acropora muricata , which sacrifices skeleton density to protect its polyp-canal system. The forming patterns in Pocillopora damicornis , Montipora capricornis , and M. foliosa were characterized by ‘osteoporosis’, exhibiting disordered skeletal structures, insufficient synthesis of adhesion proteins, and low bone mass, correspondingly. In addition, we found that skeletal areas near coral polyps suffered less and had later acidified damage than other skeletal areas in the colony. These results help to understand the skeleton-forming strategies of several major coral species under acid stress, thereby laying a foundation for coral reef protection and restoration under increasing ocean acidification.### Competing Interest StatementThe authors have declared no competing interest.

The mechanisms underlying the toxicity of environmental stress are unclear for marine macrobenthos. Copper/Cu has posed the most serious threats to amphioxus, an ancient and model benthic cephalochordate. Herein, a dynamic change in the physiological parameters (GR, SOD, ATP, and MDA) was detected with ROS accumulation in Branchiostoma belcheri exposed to 0.3 mg·L-1 Cu. Transcriptomes and microRNAomes of B. belcheri were generated to investigate the molecular mechanisms by which this amphioxus copes with Cu exposure. Time-specific genes identified at different time points after exposure were involved in the stimulus and immune response, detoxification and ionic homeostasis, aging and the nervous system, sequentially, with prolongation of exposure time, forming a dynamic process of molecular response to Cu stress. In total, 57 differentially expressed miRNAs were identified under Cu stress. Transcriptomics-miRNAomics analyses indicate that these miRNAs targeted genes associated with many key biological processes such as xenobiotics degradation, oxidative stress, and energy metabolism. The constructed miRNA-mRNA-pathway network uncovered a broad post-transcriptional regulatory mechanism in B. belcheri to cope with Cu stress. Overall, this integrated analyses show that enhanced defense response, accelerated ROS elimination, and repressed ATP production constitute a comprehensive strategy to cope with Cu toxicity in the ancient macrobenthos.

Introduction: Coral reefs, among the most invaluable ecosystems in the world, face escalating threats from climate change and anthropogenic activities. To decipher the genetic underpinnings of coral adaptation and resilience, we undertook comprehensive transcriptome profiling of two emblematic coral species, Montipora foliosa and Montipora capricornis, leveraging PacBio Iso-Seq technology. These species were strategically selected for their ecological significance and their taxonomic proximity within the Anthozoa class.Methods: Our study encompassed the generation of pristine transcriptomes, followed by thorough functional annotation via diverse databases. Subsequently, we quantified transcript abundance and scrutinized gene expression patterns, revealing notable distinctions between the two species.Results: Intriguingly, shared orthologous genes were identified across a spectrum of coral species, highlighting a substantial genetic conservation within scleractinian corals. Importantly, a subset of genes, integral to biomineralization processes, emerged as exclusive to scleractinian corals, shedding light on their intricate evolutionary history. Furthermore, we discerned pronounced upregulation of genes linked to immunity, stress response, and oxidative-reduction processes in M. foliosa relative to M. capricornis. These findings hint at the presence of more robust mechanisms in M. foliosa for maintaining internal equilibrium and effectively navigating external challenges, underpinning its potential ecological advantage. Beyond elucidating genetic adaptation in corals, our research underscores the urgency of preserving genetic diversity within coral populations.Discussion: These insights hold promise for informed conservation strategies aimed at safeguarding these imperiled ecosystems, bearing ecological and economic significance. In synthesis, our study seamlessly integrates genomic inquiry with ecological relevance, bridging the gap between molecular insights and the imperative to conserve coral reefs in the face of mounting threats.

Coral reefs are facing unprecedented threats due to global climate change, particularly elevated sea surface temperatures causing coral bleaching. Understanding coral responses at the molecular level is crucial for predicting their resilience and developing effective conservation strategies. In this study, we conducted a comprehensive gene expression analysis of four coral species to investigate their long-term molecular response to heat stress. We identified distinct gene expression patterns among the coral species, with laminar corals exhibiting a stronger response compared to branching corals. Heat shock proteins (HSPs) showed an overall decreasing expression trend, indicating the high energy cost associated with sustaining elevated HSP levels during prolonged heat stress. Peroxidases and oxidoreductases involved in oxidative stress response demonstrated significant upregulation, highlighting their role in maintaining cellular redox balance. Differential expression of genes related to calcium homeostasis and bioluminescence suggested distinct mechanisms for coping with heat stress among the coral species. Furthermore, the impact of heat stress on coral biomineralization varied, with downregulation of carbonic anhydrase and skeletal organic matrix proteins indicating reduced capacity for biomineralization in the later stages of heat stress. Our findings provide insights into the molecular mechanisms underlying coral responses to heat stress and highlight the importance of considering species-specific responses in assessing coral resilience. The identified biomarkers may serve as indicators of heat stress and contribute to early detection of coral bleaching events. These findings contribute to our understanding of coral resilience and provide a basis for future research aimed at enhancing coral survival in the face of climate change.

Modern scleractinian corals are classified into robust, complex, and basal clades through comparative molecular studies. However, only few morphological or biological criteria can systematically determine the evolutionary trajectories of these major scleractinian coral clades. Here, we obtained the structural information of 21 scleractinian coral species representing robust and complex clades: High-resolution micro-computed tomography was used to reconstruct the polyp-canal systems in their colonies and to visualize the dynamic polyp growth processes. We found that the emergence of mesh-like canals may distinguish representatives of complex and robust clades. The differences in polyp-canal connections suggest distinct evolutionary trajectories among coral species: The formation of the canal network promoted the development of more complex coral structures, and coral polyps within this network formed calices of very similar volume, following precise axial growth directions. The influence of individual polyps on the coral colony becomes less significant as coral structures become more complex, and coral species with more complicated polyp-canal systems occupied niches more efficiently. This work supplements current evolutionary studies on reef-building corals, providing insight for further studies on coral growth patterns.

Coral transcriptomic data largely rely on short-read sequencing, which severely limits the understanding of coral molecular mechanisms and leaves many important biological questions unresolved. Here, we sequence the full-length transcriptomes of four common and frequently dominant reef-building corals using the PacBio Sequel II platform. We obtain information on reported gene functions, structures, and expression profiles. Among them, a comparative analysis of biomineralization-related genes provides insights into the molecular basis of coral skeletal density. The gene expression profiles of the symbiont Symbiodiniaceae are also isolated and annotated from the holobiont sequence data. Finally, a phylogenetic analysis of key circadian clock genes among 40 evolutionarily representative species indicates that there are four key members in early metazoans, including cry genes; Clock or Npas2; cyc or Arntl; and tim, while per, as the fifth member, occurs in Bilateria. In summary, this work provides a foundation for further work on the manipulation of skeleton production or symbiosis to promote the survival of these important organisms.

Corals should make excellent models for cross-kingdom research because of their natural animal-photobiont holobiont composition, yet a lack of studies and experimental data restricts their use. Here we integrate new full-length transcriptomes and small RNAs of four common reef-building corals with the published Cladocopium genomes to gain deeper insight into gene regulation in coral-Symbiodiniaceae holobionts. Eleven novel Symbiodiniaceae miRNAs get identified, and enrichment results of their target genes show that they might play a role in downregulating rejection from host coral cells, protecting symbiont from autophagy and apoptosis in parallel. This work provides evidence for the early origin of cross-kingdom regulation as a mechanism of self-defense autotrophs can use against heterotrophs, sheds more light on coral-Symbiodiniaceae holobionts, and contributes valuable data for further coral research.

Amphioxus, as the best living proxy to the chordate ancestor, is considered an irreplaceable model organism for evolutionary studies of chordates and deuterostomes. In this study, a high-quality genome of the Beihai amphioxus, Branchiostoma belcheri beihai, was de novo assembled and annotated. Within four amphioxus genomes, a wide range of gene novelties were identified, revealing new genes that share unexpectedly high similarities with those from non-metazoan species. These gene innovation events have played roles in a range of amphioxus adaptations, including innate immunity responses, adaptation to anaerobic environments, and regulation of calcium balance. The gene novelties related to innate immunity, such as a group of lipoxygenases and a DEAD-box helicase, boosted amphioxus immune responses. The novel genes for alcohol dehydrogenase and ferredoxin could aid in the anaerobic tolerance of amphioxus. A proximally arrayed cluster of EF-hand calcium-binding protein genes were identified to resemble those of bacteria. The copy number of this gene cluster was linearly correlated to the sea salinity of the collection region, suggesting that it may enhance their survival at different calcium concentrations. Collectively, this comprehensive study on gene novelties of amphioxus reveals insights into the early genome evolution of chordates and deuterostomes and provides valuable resources for future research.

Background Amphioxus is a model organism for vertebrate evolutionary research. The significant contrast between morphological phenotypic similarity and high-level genetic polymorphism among amphioxus populations has aroused scientists' attention. Here we resequenced 21 amphioxus genomes to over 100X depth and mapped them to a haploid reference. Results More than 11.5 million common SNPs were detected in the amphioxus population, which mainly affect genes enriched in ion transport, signal transduction and cell adhesion, while protein structure analysis via AlphaFold2 revealed that these SNPs fail to bring effective structural variants. Conclusions Our work provides explanation for “amphioxus polymorphism paradox” in a micro view, and generates an enhanced genomic dataset for amphioxus research.

Reef-building corals play an important role in the marine ecosystem, and analyzing their proteomes from a structural perspective will exert positive effects on exploring their biology. Here we integrated mass spectrometry with newly published ColabFold to obtain digital structural proteomes of dominant reef-building corals. 8,382 proteins co-expressed in A. muricata, M. foliosa and P. verrucosa were identified, then 8,166 of them got predicted structures after around 4,060 GPU hours of computation. The resulting dataset covers 83.6% of residues with a confident prediction, while 25.9% have very high confidence. Our work provides insight-worthy predictions for coral research, confirms the reliability of ColabFold in practice, and is expected to be a reference case in the impending high-throughput era of structural proteomics.

The origin and early evolution of animal development remain among the many deep, unresolved problems in evolutionary biology. As a compelling case for the existence of pre-Cambrian animals, the Ediacaran embryo-like fossils (EELFs) from the Weng'an Biota (approx. 609 Myr old, Doushantuo Formation, South China) have great potential to cast light on the origin and early evolution of animal development. However, their biological implications can be fully realized only when their phylogenetic positions are correctly established, and unfortunately, this is the key problem under debate. As a significant feature of developmental biology, the cell division pattern (CDP) characterized by the dynamic spatial arrangement of cells and associated developmental mechanisms is critical to reassess these hypotheses and evaluate the diversity of the EELFs; however, their phylogenetic implications have not been fully realized. Additionally, the scarcity of fossil specimens representing late developmental stages with cell differentiation accounts for much of this debate too. Here, we reconstructed a large number of EELFs using submicron resolution X-ray tomographic microscopy and focused on the CDPs and associated developmental mechanisms as well as features of cell differentiation. Four types of CDPs and specimens with cell differentiation were identified. Contrary to the prevailing view, our results together with recent studies suggest that the diversity and complexity of developmental mechanisms documented by the EELFs are much higher than is often claimed. The diverse CDPs and associated development features including palintomic cleavage, maternal nutrition, asymmetric cell divisions, symmetry breaking, establishment of polarity or axis, spatial cell migration and differentiation constrain some, if not all, EELFs as total-group metazoans. This article is part of the theme issue ‘The impact of Chinese palaeontology on evolutionary research’.

Reef-building corals play an important role in marine ecosystems. However, owing to climate change, ocean acidification, and predation by invasive crown-of-thorns starfish, these corals are declining. As marine animals comprise polyps, reproduction by asexual budding is pivotal in scleractinian coral growth. The fibroblast growth factor (FGF) signaling pathway is essential in coral budding morphogenesis. Here, we sequenced the full-length transcriptomes of four common and frequently dominant reef-building corals and screened out the budding-related FGF and FGFR genes. Thereafter, three-dimensional (3D) models of FGF and FGFR proteins as well as FGF-FGFR binding models were reconstructed. Based on our findings, the FGF8-FGFR3 binding models in Pocillopora damicornis, Montipora capricornis, and Acropora muricata are typical receptor tyrosine kinase-signaling pathways that are similar to the Kringelchen (FGFR) in hydra. However, in P. verrucosa, FGF8 is not the FGFR3 ligand, which is found in other hydrozoan animals, and its FGFR3 must be activated by other tyrosine kinase-type ligands. Overall, this study provides background on the potentially budding propagation signaling pathway activated by the applications of biological agents in reef-building coral culture that could aid in the future restoration of coral reefs.

Distinct expression of the miRNAs has rarely been explored in basal cell carcinoma (BCC) of skin, and the regulatory role of miRNAs in BCC development remains quite opaque. Here, we collected control tissues from adjacent noncancerous skin ( n = 15 ; control group) and tissues at tumor centers from patients with cheek BCC ( n = 15 ; BCC group) using punch biopsies. After six small RNA sequencing- (sRNA-seq-) based miRNA expression profiles were generated for both BCC and controls, including three biological replicates, we conducted comparative analysis on the sRNA-seq dataset, discovering 181 differentially expressed miRNAs (DEMs) out of the 1,873 miRNAs in BCCs. In order to validate the sRNA-seq data, expression of 15 randomly selected DEMs was measured using the TaqMan probe-based quantitative real-time PCR. Functional analysis of predicted target genes of DEMs in BCCs shows that these miRNAs are primarily involved in various types of cancers, immune response, epithelial growth, and morphogenesis, as well as energy production and metabolism, indicating that BCC development is caused, at least in part, by changes in miRNA regulation for biological and disease processes. In particular, the “basal cell carcinoma pathways” were found to be enriched by predicted DEM targets, and regulatory relationships between DEMs and their targeted genes in this pathway were further uncovered. These results revealed the association between BCCs and abundant miRNA molecules that regulate target genes, functional modules, and signaling pathways in carcinogenesis.

Abstract Colonies are the basic geometric building blocks of coral reefs. However, the forming regulations of both colonies and reefs are still not understood adequately. Therefore, in this study, we reconstructed 25 samples using high‐resolution micro‐computed tomography to investigate coral growth patterns and parameters. Our skeleton and canal reconstructions revealed the characteristics of different coral species, and we further visualized the growth axes and growth rings to understand the coral growth directions. We drew a skeleton grayscale map and calculated the coral skeleton void ratios to ascertain the skeletal diversity, devising a method to quantify coral growth. On the basis of the three‐dimensional (3D) reconstructions and growth parameters, we investigated the growth strategies of different coral species. This research increases the breadth of knowledge on how reef‐building corals grow their colonies, providing information on reef‐forming regulations. The data in this paper contain a large amount of coral growth information, which can be used in further research on reef‐forming patterns under different conditions. The method used in this study can also be applied to animals with porous skeletons.

Coral reefs are cornerstone of global marine ecosystems, providing shelter for over one third of marine organisms. Currently, along with global warming and increased human activities, large-scale decline of coral reefs has become a severe ecosystem problem, and now quantitative detection of heat shock protein (HSP) gene by nanotechnology has become a research hotspot in this field. However, Acropora muricata is one of the most important dominant reef-building corals in Indo- Pacific region, encounter an urgent obstacle on the HSP detection research by nanoscience and nanotechnology for lack of sequence background. Here, we combined PacBio single molecular real-time (SMRT) and HiSeq X Ten sequencing technologies to perform full-length transcriptome sequencing of heat shock proteins in Acropora muricata, a reef-building coral dominant in many Indo-Pacific reefs, to annotate them. Thirteen functional heat shock proteins (HSPs) were identified using phylogenetic analysis, classified into three subgroups as HSP60, HSP70 and HSP90. HSPs are widely distributed in all animal phyla, having evolved from the last prokaryotic common ancestor. Additionally, phylogenetic and tertiary nanostructure analyses suggested that HSP70 is the most diverse HSP in A. muricata, with extensive sequence and structure differences indicating adaptations to warming water and suggesting its utility in studies of El Niño and other warming events. A greater understanding of the HSP gene family is likely to also be of value in studies of coral nanotechnological detection that can be used to protect reef ecosystems.

寒武纪大爆发是生命演化史上重要的创新事件.我国澄江动物群(距今约5.18亿年)代表了寒武纪大爆发的高潮,而产自寒武系最底部,时代上早于澄江动物群的宽川铺生物群(距今约5.35亿年)则为研究寒武纪大爆发的第一幕提供了重要的埋藏窗口.宽川铺生物群产自陕南宁强、西乡等地寒武系底部宽川铺组钙质磷块岩和磷质碎屑灰岩中,以保存了大量动物胚胎和软躯体微型动物而闻名.其中四方塔型壳(Quadrapyrgites)是宽川铺生物群中常见的软躯体动物化石,因与同层位的橄榄蛋(Olivooides)在发育过程和形态上具有相似性而被认为是后者的姐妹群.但它们在动物树上的位置仍然存在争议.为检验前人提出的各种亲缘关系假说,本文综合使用电子扫描显微镜成像和超高分辨率X射线显微断层成像技术,重建了四方塔型壳胚胎标本的三维结构,首次揭示了其内陷式原肠作用类型.新的数据支持了四方塔型壳属于刺细胞动物而非环神经动物的观点.