Fossil Calibrations Research Articles

Bayesian Selection of Relaxed-clock Models: Distinguishing Between Independent and Autocorrelated Rates.

In Bayesian molecular-clock dating of species divergences, rate models are used to construct the prior on the molecular evolutionary rates for branches in the phylogeny, with independent and autocorrelated rate models being commonly used. The two classes of models, however, can result in markedly different divergence time estimates for the same dataset, and thus selecting the best rate model appears important for obtaining reliable in- ferences of divergence times. However, the properties of Bayesian rate model selection are not well understood, in particular when the number of sequence partitions analysed increases and when age calibrations (such as fossil calibrations) are misspecified. Further- more, Bayesian rate model selection is computationally expensive as it requires calculation of marginal likelihoods by MCMC sampling, and therefore methods that can speed up the model selection procedure without compromising its accuracy are desirable. In this study, we use a combination of computer simulations and real data analysis to investigate the sta- tistical behaviour of Bayesian rate model selection and we also explore approximations of the likelihood to improve computational efficiency in large phylogenomic datasets. Our simulations demonstrate that the posterior probability for the correct rate model converges to one as more molecular sequence partitions are analysed and when no calibrations are used, as expected due to asymptotic Bayesian model selection theory. Furthermore, we also show the model selection procedure is robust to slight misspecification of calibrations, and reliable inference of the correct rate model is possible in this case. However, we show that when calibrations are seriously misspecified, calculated model probabilities are com- pletely wrong and may converge to one for the wrong rate model. Finally, we demonstrate that approximating the phylogenetic likelihood under an arcsine branch-length transform can dramatically reduce the computational cost of rate model selection without compro- mising accuracy. We test the approximate procedure on two large phylogenies of primates (372 species) and flowering plants (644 species), replicating results obtained on smaller datasets using exact likelihood. Our findings and methodology can assist users in selecting the optimal rate model for estimating times and rates along the Tree of Life.

Ediacaran origin and Ediacaran-Cambrian diversification of Metazoa.

The timescale of animal diversification has been a focus of debate over how evolutionary history should be calibrated to geologic time. Molecular clock analyses have invariably estimated a Cryogenian or Tonian origin of animals while unequivocal animal fossils first occur in the Ediacaran. However, redating of key Ediacaran biotas and the discovery of several Ediacaran crown-Metazoa prompt recalibration of molecular clock analyses. We present revised fossil calibrations and use them in molecular clock analyses estimating the timescale of metazoan evolutionary history. Integrating across uncertainties including phylogenetic relationships, clock model, and calibration strategy, we estimate Metazoa to have originated in the early Ediacaran, Eumetazoa in the middle Ediacaran, and Bilateria in the upper Ediacaran, with many crown-phyla originating across the Ediacaran-Cambrian interval or elsewise fully within the Cambrian. These results are in much closer accord with the fossil record, coinciding with marine oxygenation, but they reject a literal reading of the fossil record.

Phylogenetic analysis of Syrphidae (Insecta, Diptera) based on low-coverage whole genomes

Abstract The hoverflies are one of the most abundant families in Diptera and an important category of resource insects. They can provide important ecosystem services such as pollination and beneficial biological control agents. In the four-subfamily classification system of the Syrphidae, the monophyly of the Eristalinae within Syrphidae has not been recovered. In this study, we performed new sequencing and analysis of low-coverage whole genomes for 25 species of Syrphidae. Phylogenetic reconstructions were conducted based on different datasets and analytical strategies. Across all our analyses, the monophyly of the Syrphinae is supported but we do not recover the monophyly of the Eristalinae, consistent with previous research findings. Additionally, employing fossil calibrations for divergence time estimation, our results indicate that the origin of the Syrphidae occurred between the Albian and Aptian stages, approximately between 120.5 and 109.3 million years ago, the origin of the subfamily Syrphinae was dated in the Early Eocene, about 50.8–47.8 million years ago, in accordance with the diversification of their prey. This study will help us understand the higher-level phylogenetic and evolutionary relationships within Syrphidae.

A Molecular Temporal Evolutionary Framework of Land Plants and the Age of Angiosperms

A series of Bayesian molecular clock analyses with varying model combinations, parameters, and tree topologies were conducted to establish a temporal evolutionary framework of land plants and to estimate the age of angiosperms. The data consisted of five organellar and nuclear genes, 633 taxa, and 23 fossil calibration points (FCPs). The analyses were evaluated using an internal search thoroughness measure, effective sample sizes, and two external criteria: the consensus land plant phylogeny and a newly developed method that used the Hennigian reciprocal illumination principle to compare molecular age estimates of 21 nodes representing major land plant clades and the root split in green plants with those of their fossils in an a posteriori fashion. The results show that an analysis using a general time reversible (GTR) DNA substitution model, log normal branch-rate distribution, and Yule tree prior represented the best model combination, with the Birth-Death tree prior being equally optimal. This analysis estimated the age of the land plant crown group as 496.95 million years (MY), with a 95% highest posterior density (HPD) of 465.66–531.43 MY, and the age of the angiosperm crown group as 219.93 MY, with a 95% HPD of 184.09–256.82 MY. The former agrees with the fossil evidence, but the latter is significantly older than the fossil ages. A thorough examination of fossil evidence of angiosperms, as well as their evolutionary history in light of recent knowledge of morphology, physiology, and atmospheric CO2 concentration in the Mesozoic era, suggests that angiosperm radiation might have been delayed after their origin and that the current fossil record probably reflects only the products of that radiation, not the origin of angiosperms. Overall, the study shows that both molecular clock analyses and fossil studies, while agreeing on some aspects but disagreeing on others, can help develop a complete understanding of the temporal evolutionary history of land plants.

Cicada minimum age tree: Cryptic speciation and exponentially increasing base substitution rates in recent geologic time.

We developed a new time-calibrated tree incorporating primarily endemic along with some cryptic Ryukyu islands cicada data, following the recent publication of global cicada data by Marshall et al. (2018), Łukasik et al. (2018), Simon et al. (2019), Price et al. (2019), and Hill et al. (2021). A total of 352 specimens were analyzed using BEAST v1. X software with a relaxed clock model. Fossil calibrations as old as Triassic were adopted largely following Johnson et al. (2018) and Moulds (2018), and a Quaternary geological event calibration was adopted following Osozawa et al. (2012, 2021b) and input into BEAST v1. X. Our timetree suggests that Tettigarctidae had a cicada basal lineage as old as 200.63 Ma, with Derotettiginae the next oldest lineage at 99.2 Ma. Tibicininae is a sister of the remaining subfamilies of Tettigomyiinae, Cicadettinae, and Cicadidae, and their species level differentiation and radiation began at 40.57 Ma. The Cicadinae clade consists of specific tribes with paraphyletic relationship, and the vicariance and adaptive radiation generated many cryptic species in each tribe. We estimated base substitution rate as a function of age, and the result strongly indicates an exponential increase of base substitution rate in recent geologic time. The consequent increase in cicada biodiversity, including generation of cryptic species in the Ryukyu Islands and surroundings, may have been driven by the generation and spreading of C4 grasses and coeval Quaternary climate change.

Historical biogeography of North American killifishes (Cyprinodontiformes) recapitulates geographical history in the Gulf of México watershed

Abstract We analysed phylogenetic relationships within a major clade of Cyprinodontiformes (Teleostei) that includes five families of North American killifishes. We used DNA sequences from five genes for 130 species, with four fossil calibrations and three secondary calibrations, to generate a time-calibrated phylogeny. We estimated diversification rates, ancestral areas, and ancestral habitats for each node. Findings were interpreted within a detailed biogeographical synthesis. The results indicate that the clade arose in the Eocene along the Gulf of México coast. The speciation rate was uniform through time, except for acceleration in Cyprinodontidae after ~10.9 Mya. In other families, neither viviparity nor marine-to-freshwater transition was associated with accelerated speciation. Sea-level fluctuations might have created a speciation pump by stimulating cycles of dispersal and vicariance along the coast. Diversification also included many cases of inland immigration from coastal ancestors. For upland lineages, ancient river drainages accord with lineage distributions, including enigmatic disjunctions in Goodeidae and Fundulus. Diversification in uplands occurred via barrier displacement within alluvial or tectonically active landscapes. Killifishes also display high environmental tolerance and persist within harsh, peripheral environments unsuitable for most other fishes. Hence, a combination of clade antiquity, adaptability, dynamic geography, and persistence can explain the living diversity of New World killifishes.

Mitogenome-Based Phylogeny with Divergence Time Estimates Revealed the Presence of Cryptic Species within Heptageniidae (Insecta, Ephemeroptera).

Heptageniidae are known for their flat heads and bodies and are divided into three subfamilies. Despite the extensive diversity within this group and considerable efforts made to understand their evolutionary history, the internal classifications and origin time of Heptageniidae remains controversial. In this study, we newly sequenced 17 complete mitogenomes of Heptageniidae to reconstruct their phylogenetic positions within this family. Because of the ambiguous time of origin, our study also estimated the divergence time within Heptageniidae based on five fossil calibration points. The results of BI and ML trees all highly supported the monophyly of Heptageniidae and three subfamilies. The phylogenetic relationship of Rhithrogeninae + (Ecdyonurinae + Heptageniinae) was also recovered. The divergence time showed that Heptageniidae originated from 164.38 Mya (95% HPD, 150.23-181.53 Mya) in the mid-Jurassic, and Rhithrogeninae originated from 95.54 Mya (95% HPD, 73.86-120.19 Mya) in the mid-Cretaceous. Ecdyonurinae and Heptageniinae began to diverge at 90.08 Mya (95% HPD, 68.81-113.16 Mya) in the middle Cretaceous. After morphological identification, analysis of the mitogenome's composition, genetic distance calculation, phylogenetic analysis, and divergence time calculation, we suggest that two different populations of Epeorus montanus collected from Aksu, Xinjiang Uygur Autonomous Region (40°16' N, 80°26' E) and Xinyuan, Xinjiang Uygur Autonomous Region (43°20' N, 83°43' E) in China are cryptic species of E. montanus, but further detailed information on their morphological characteristics is needed to fully identify them.

Tangled webs and spider-flowers: Phylogenomics, biogeography, and seed morphology inform the evolutionary history of Cleomaceae.

Cleomaceae is an important model clade for studies of evolutionary processes including genome evolution, floral form diversification, and photosynthetic pathway evolution. Diversification and divergence patterns in Cleomaceae remain tangled as research has been restricted by its worldwide distribution, limited genetic sampling and species coverage, and a lack of definitive fossil calibration points. We used target sequence capture and the Angiosperms353 probe set to perform a phylogenetic study of Cleomaceae. We estimated divergence times and biogeographic analyses to explore the origin and diversification of the family. Seed morphology across extant taxa was documented with multifocal image-stacking techniques and morphological characters were extracted, analyzed, and compared to fossil records. We recovered a well-supported and resolved phylogenetic tree of Cleomaceae generic relationships that includes 236 (~86%) species. We identified 11 principal clades and confidently placed Cleomella as sister to the rest of the family. Our analyses suggested that Cleomaceae and Brassicaceae diverged ~56 mya, and Cleomaceae began to diversify ~53 mya in the Palearctic and Africa. Multiple transatlantic disjunct distributions were identified. Seeds were imaged from 218 (~80%) species in the family and compared to all known fossil species. Our results represent the most comprehensive phylogenetic study of Cleomaceae to date. We identified transatlantic disjunctions and proposed explanations for these patterns, most likely either long-distance dispersals or contractions in latitudinal distributions caused by climate change over geological timescales. We found that seed morphology varied considerably but mostly mirrored generic relationships.

The role of deep hybridization in fern speciation: Examples from the Thelypteridaceae.

Hybridization is recognized as an important mechanism in fern speciation, with many allopolyploids known among congeners, as well as evidence of ancient genome duplications. Several contemporary instances of deep (intergeneric) hybridization have been noted, invariably resulting in sterile progeny. We chose the christelloid lineage of the family Thelypteridaceae, recognized for its high frequency of both intra- and intergeneric hybrids, to investigate recent hybrid speciation between deeply diverged lineages. We also seek to understand the ecological and evolutionary outcomes of resulting lineages across the landscape. By phasing captured reads within a phylogenomic data set of GoFlag 408 nuclear loci using HybPhaser, we investigated candidate hybrids to identify parental lineages. We estimated divergence ages by inferring a dated phylogeny using fossil calibrations with treePL. We investigated ecological niche conservatism between one confirmed intergeneric allotetraploid and its diploid progenitors using the centroid, overlap, unfilling, and expansion (COUE) framework. We provide evidence for at least six instances of intergeneric hybrid speciation within the christelloid clade and estimate up to 45 million years of divergence between progenitors. The niche quantification analysis showed moderate niche overlap between an allopolyploid species and its progenitors, with significant divergence from the niche of one progenitor and conservatism to the other. The examples provided here highlight the overlooked role that allopolyploidization following intergeneric hybridization may play in fern diversification and range and niche expansions. Applying this approach to other fern taxa may reveal a similar pattern of deep hybridization resulting in highly successful novel lineages.

A targeted gene phylogenetic framework to investigate diversification in the highly diverse yet geographically restricted red devil spiders (Araneae, Dysderidae).

The family Dysderidae is a highly diverse group of nocturnal ground-dwelling and active-hunter spiders. Dysderids are mostly restricted to the Western Palearctic, and particularly rich and abundant around the Mediterranean region. Interestingly, the distribution of species richness among its 24 genera and three subfamilies is highly biased-80% of its 644 documented species belong to just two genera, Dysdera (326) and Harpactea (211). Dysderidae provides an excellent study case for evolutionary and ecological research. It includes cases of trophic specialization, which are uncommon among spiders, and exhibit other remarkable biological (e.g. holocentric chromosomes), behavioural (e.g. cryptic female choice), evolutionary (e.g. adaptive radiation) and ecological features (e.g. recurrent colonization of the subterranean environment). The lack of a quantitative hypothesis on its phylogenetic structure has hampered its potential as a testing ground for evolutionary, biogeographical and ecological hypotheses. Here, we present the results of a target, multi-locus phylogenetic analysis, using mitochondrial (cox1, 16s and 12s) and nuclear genes (h3, 28s and 18s), of the most exhaustive taxonomic sample within Dysderidae (104 spp.) to date and across related families (Synspermiata) (83 spp.). We estimate divergence times using a combination of fossil and biogeographic node calibrations and use this timeline to identify shifts in diversification rates. Our results support the monophyly of the Dysderidae subfamilies Rhodinae and Dysderinae but reject Harpacteinae as currently defined. Moreover, the clades recovered within Harpacteinae do not support its current taxonomy. The origin of the family most likely post-dated the break-up of Pangea, and cave colonization may be older than previously considered. After correcting for the taxonomic artefacts, we identified a significant shift in diversification rates at the base of the genus Dysdera. Although the unique coexistence of specialist and generalist diets within the lineage could be suggested as the potential driver for the rate acceleration, further quantitative analyses would be necessary to test this hypothesis.

Cretaceous amber inclusions illuminate the evolutionary origin of tardigrades

Tardigrades are a diverse phylum of microscopic invertebrates widely known for their extreme survival capabilities. Molecular clocks suggest that tardigrades diverged from other panarthropods before the Cambrian, but their fossil record is extremely sparse. Only the fossil tardigrades Milnesium swolenskyi (Late Cretaceous) and Paradoryphoribius chronocaribbeus (Miocene) have resolved taxonomic positions, restricting the availability of calibration points for estimating for the origin of this phylum. Here, we revise two crown-group tardigrades from Canadian Cretaceous-aged amber using confocal fluorescence microscopy, revealing critical morphological characters that resolve their taxonomic positions. Formal morphological redescription of Beorn leggi reveals that it features Hypsibius-type claws. We also describe Aerobius dactylus gen. et sp. nov. based on its unique combination of claw characters. Phylogenetic analyses indicate that Beo. leggi and Aer. dactylus belong to the eutardigrade superfamily Hypsibioidea, adding a critical fossil calibration point to investigate tardigrade origins. Our molecular clock estimates suggest an early Paleozoic diversification of crown-group Tardigrada and highlight the importance of Beo. leggi as a calibration point that directly impacts estimates of shallow nodes. Our results suggest that independent terrestrialization of eutardigrades and heterotardigrades occurred around the end-Carboniferous and Lower Jurassic, respectively. These estimates also provide minimum ages for convergent acquisition of cryptobiosis.

Whole genome phylogenomics helps to resolve the phylogenetic position of the Zygothrica genus group (Diptera, Drosophilidae) and the causes of previous incongruences

Incomplete Lineage Sorting (ILS) and introgression are among the two main factors causing incongruence between gene and species trees. Advances in phylogenomic studies have allowed us to overcome most of these issues, providing reliable phylogenetic hypotheses while revealing the underlying evolutionary scenario. Across the last century, many incongruent phylogenetic reconstructions were recovered for Drosophilidae, employing a limited sampling of genetic markers or species. In these studies, the monophyly and the phylogenetic positioning of the Zygothrica genus group stood out as one of the most controversial questions. Thus, here, we addressed these issues using a phylogenomic approach, while accessing the influence of ILS and introgressions on the diversification of these species and addressing the spatio-temporal scenario associated with their evolution. For this task, the genomes of nine specimens from six Neotropical species belonging to the Zygothrica genus group were sequenced and evaluated in a phylogenetic framework encompassing other 39 species of Drosophilidae. Nucleotide and amino acid sequences recovered for a set of 2,534 single-copy genes by BUSCO were employed to reconstruct maximum likelihood (ML) concatenated and multi-species coalescent (MSC) trees. Likelihood mapping, quartet sampling, and reticulation tests were employed to infer the level and causes of incongruence. Lastly, a penalized-likelihood molecular clock strategy with fossil calibrations was performed to infer divergence times. Taken together, our results recovered the subdivision of Drosophila into six different lineages, one of which clusters species of the Zygothrica genus group (except for H. duncani). The divergence of this lineage was dated to Oligocene ∼ 31 Mya and seems to have occurred in the same timeframe as other key diversification within Drosophila. According to the concatenated and MSC strategies, this lineage is sister to the clade joining Drosophila (Siphlodora) with the Hawaiian Drosophila and Scaptomyza. Likelihood mapping, quartet sampling, reticulation reconstructions as well as introgression tests revealed that this lineage was the target of several hybridization events involving the ancestors of different Drosophila lineages. Thus, our results generally show introgression as a major source of previous incongruence. Nevertheless, the similar diversification times recovered for several of the Neotropical Drosophila lineages also support the scenario of multiple and simultaneous diversifications taking place at the base of Drosophilidae phylogeny, at least in the Neotropics.

Molecular phylogenetics of the superfamily Stromboidea (Caenogastropoda): New insights from increased taxon sampling

AbstractThe superfamily Stromboidea is a clade of morphologically distinctive gastropods which include the iconic Strombidae, or ‘true conchs’. In this study, we present the most taxonomically extensive phylogeny of the superfamily to date, using fossil calibrations to produce a chronogram and extant geographical distributions to reconstruct ancestral ranges. From these results, we confirm the monophyly of all stromboidean families; however, six genera are not monophyletic using current generic assignments (Strombidae: Lentigo, Canarium, Dolomena, Doxander; Xenophoridae: Onustus, Xenophora). Within Strombidae, analyses resolve an Indo‐West Pacific (IWP) clade sister to an East Pacific/Atlantic clade, together sister to a second, larger IWP clade. Our results also indicate two pulses of strombid diversification within the Miocene, and a Tethyan/IWP origin for Strombidae—both supported by the fossil record. However, conflicts between divergence time estimates and the fossil record warrant further exploration. Species delimitation analyses using the COI barcoding gene support several taxonomic changes. We synonymise Euprotomus aurora with Euprotomus bulla, Strombus alatus with Strombus pugilis, Dolomena abbotti with Dolomena labiosa, and Dolomena operosa with Dolomena vittata. We identified cryptic species complexes within Terebellum terebellum, Lambis lambis, “Canarium” wilsonorum, Dolomena turturella and Maculastrombus mutabilis. We reinstate Rimellopsis laurenti as a species (previously synonymised with R. powisii) and recognise Harpago chiragra rugosus and Lambis truncata sowerbyi valid at the rank of species. Finally, we establish several new combinations to render Lentigo, Dolomena, and Canarium monophyletic: Lentigo thersites, Dolomena robusta, Dolomena epidromis, Dolomena turturella, Dolomena taeniata, Dolomena vanikorensis, D. vittata, “Canarium” wilsonorum, Hawaiistrombus scalariformis, Maculastrombus mutabilis, Maculastrombus microurceus.

Phylogeny and diversification analyses of extant planthopper families (Hemiptera: Fulgoromorpha) based on a mitochondrial dataset reveal ancient lineages originating during the Jurassic and originally feeding on gymnosperms

Abstract Phylogenetic analyses of planthoppers were conducted using a mitogenome dataset to examine the evolutionary relationships of current families based on both maximum likelihood (ML) and Bayesian inference methods. Delphacoidea, including Cixiidae and Delphacidae, and Fulgoroidea, including all other families, were consistently identified as monophyletic groups. However, Nogodinidae displayed a polyphyletic pattern, with various lineages emerging sister to several issidoid families. Variations were observed across different analyses in the positioning of (Dictyopharidae + Fulgoridae) and the relationships of Ricaniidae sister either to Issidae or to (Flatidae + Acanaloniidae). Several statistical tests indicate that the ML topology has the highest level of statistical confidence. The (Dictyopharidae + Fulgoridae) clade is best positioned as sister to (Derbidae + (Achilidae + (Tropiduchidae + other “higher” planthoppers families))) and the Ricaniidae as sister to (Flatidae + Acanaloniidae). With 13 fossil calibration points, origin times for Delphacoidea and Fulgoroidea were dated back to the Late Permian, approximately 257.46 million years ago. Delphacidae and Cixiidae split during the Late Triassic, while Fulgoroidea diversified earlier during the Middle Triassic. Divergence and diversification times are also provided for all other extant planthopper families. These results suggest that Delphacoidea and Fulgoroidea likely diverged from the older planthopper fossil families prior to the emergence of angiosperms and may have initially fed on gymnosperms and ancient ferns. The diversification of extant planthopper families occurred during the Jurassic and Cretaceous periods, influenced by evolving climatic conditions, the decline in gymnosperms, and the increasing diversity of angiosperm plants.

A comprehensive molecular phylogeny of Cephalotrichum and Microascus provides novel insights into their systematics and evolutionary history.

The genera Cephalotrichum and Microascus contain ecologically, morphologically and lifestyle diverse fungi in Microascaceae (Microascales, Sordariomycetes) with a world-wide distribution. Despite previous studies having elucidated that Cephalotrichum and Microascus are highly polyphyletic, the DNA phylogeny of many traditionally morphology-defined species is still poorly resolved, and a comprehensive taxonomic overview of the two genera is lacking. To resolve this issue, we integrate broad taxon sampling strategies and the most comprehensive multi-gene (ITS, LSU, tef1 and tub2) datasets to date, with fossil calibrations to address the phylogenetic relationships and divergence times among major lineages of Microascaceae. Two previously recognised main clades, Cephalotrichum (24 species) and Microascus (49 species), were re-affirmed based on our phylogenetic analyses, as well as the phylogenetic position of 15 genera within Microascaceae. In this study, we provide an up-to-date overview on the taxonomy and phylogeny of species belonging to Cephalotrichum and Microascus, as well as detailed descriptions and illustrations of 21 species of which eight are newly described. Furthermore, the divergence time estimates indicate that the crown age of Microascaceae was around 210.37 Mya (95 % HPD: 177.18-246.96 Mya) in the Late Triassic, and that Cephalotrichum and Microascus began to diversify approximately 27.07 Mya (95 % HPD: 20.47-34.37 Mya) and 70.46 Mya (95 % HPD: 56.96-86.24 Mya), respectively. Our results also demonstrate that multigene sequence data coupled with broad taxon sampling can help elucidate previously unresolved clade relationships. Citation: Wei TP, Wu YM, Zhang X, et al. 2024. A comprehensive molecular phylogeny of Cephalotrichum and Microascus provides novel insights into their systematics and evolutionary history. Persoonia 52: 119-160. https://doi.org/10.3767/persoonia.2024.52.05 .

Diversity, morphology, and phylogeny of freshwater mussels of the genus Nodularia (Bivalvia: Unionidae) from China, with descriptions of four new species

AbstractFreshwater bivalves (Bivalvia, Unionida) are one of the most threatened groups of animals in the world. Defining species boundaries and understanding the phylogeny and genetic diversity of these species is key to guiding their conservation and management. However, the presence of significant phenotypic plasticity and convergence within this group complicates species delimitation. This includes the freshwater mussel genus Nodularia, endemic to East Asia, for which a comprehensive understanding of species diversity and phylogenetic relationships remains elusive due to inadequate sampling in previous studies, particularly in China, a widely recognized biodiversity hotspot for freshwater mussels. Here, we conduct comprehensive taxonomic and phylogenetic analyses of Nodularia species based on extensive sampling across 23 provinces in China and multiple data sources, including shell morphology, soft body anatomy, six‐gene (COI + ND1 + 16S + 18S + 28S + histone H3) and mitogenome datasets. The integrative systematics approach used here reveals 10 distinct species in this genus, four of which are new to science, i.e. Nodularia hanensis sp. nov., Nodularia huana sp. nov., Nodularia fusiformans sp. nov., Nodularia dualobtusus sp. nov. and two of which are new records for China, i.e. Nodularia dorri (Wattebled [Journal de Conchyliologie, 34, 1886, 54]) and Nodularia micheloti (Morlet [Journal de Conchyliologie, 34, 1886, 75]). We also propose that the nominal species Nodularia jourdyi (Morlet [Journal de Conchyliologie, 34, 1886, 75]) syn. nov. is a new synonym for Nodularia douglasiae (Griffith & Pidgeon, 1833) based on molecular data. BI, ML, and BEAST analyses based on the six‐gene dataset and mitochondrial phylogenomics consistently support the following phylogenetic relationships: (N. dorri + (N. hanensis sp. nov. + N. micheloti)) + (N. breviconcha + (N. huana sp. nov. + (N. fusiformans sp. nov. + ((N. nuxpersicae + N. nipponensis) + (N. dualobtusus sp. nov. + N. douglasiae))))). The molecular clock with fossil calibration indicates that Nodularia originated in the Late Cretaceous period (ca. 73.78 Mya). It then diverged into two independent clades during the Middle Paleogene (ca. 45.01 Mya), followed by a rapid burst of extant speciation during the Neogene (mean age 28.28 to 4.79 Mya). Nodularia breviconcha is the earliest differentiated taxon among the 10 Nodularia taxa, appearing during the Paleogene‐Neogene transition (28.28 Mya; 95% HPD = 14.35–48.44 Mya). Taken together, we provide a robust systematic framework for Nodularia species, addressing phylogenetic relationships, taxonomy, and evolutionary history of this group.

Comparative Mitogenome of Phylogenetic Relationships and Divergence Time Analysis within Potamanthidae (Insecta: Ephemeroptera).

Potamanthidae belongs to the superfamily Ephemeroidea but has no complete mt genome released in the NCBI (except for two unchecked and one partial mt genome). Since the sister clade to Potamanthidae has always been controversial, we sequenced seven mt genomes of Potamanthidae (two species from Rhoenanthus and five species from Potamanthus) in order to rebuild the phylogenetic relationships of Potamanthidae in this study. The divergence time of Potamanthidae was also investigated by utilizing five fossil calibration points because of the indeterminate origin time. In addition, because Rhoenanthus coreanus and Potamanthus luteus are always in low-temperature environments, we aimed to explore whether these two species were under positive selection at the mt genome level. Amongst the 13 PCGs, CGA was used as the start codon in COX1, whereas other genes conformed to initiating with an ATN start codon. From this analysis, UUA (L), AUU (I), and UUU (F) had the highest usage. Furthermore, the DHU arm was absent in the secondary structure of S1 in all species. By combining the 13 PCGs and 2 rRNAs, we reconstructed the phylogenetic relationship of Potamanthidae within Ephemeroptera. The monophyly of Potamanthidae and the monophyly of Rhoenanthus and Potamanthus were supported in the results. The phylogenetic relationship of Potamanthidae + (Ephemeridae + Polymitarcyidae) was also recovered with a high prior probability. The divergence times of Potamanthidae were traced to be 90.44 Mya (95% HPD, 62.80-121.74 Mya), and the divergence times of Rhoenanthus and Potamanthus originated at approximately 64.77 Mya (95% HPD, 43.82-88.68 Mya), thus belonging to the late Pliocene Epoch or early Miocene Epoch. In addition, the data indicated that R. coreanus was under negative selection and that ATP8 and ND2 in Potamanthidae had a high evolutionary rate.

A multi-locus phylogeny for the Diamesinae (Chironomidae: Diptera) provides new insights into evolution of an amphitropical clade

Abstract Diamesinae is a subfamily of Chironomidae, whose species live in cold lotic or oligotrophic lentic habitats with global distribution excepting Antarctica. The cool stenothermic ecology of nearly all diamesines produces a typical amphitropical pattern of absence at tropical latitudes, except at high elevation. Recent attention has focused on the species discovery, while evolutionary relationships at the generic and tribal level have remained inadequately understood. Current classification implies analogous evolutionary diversification in each hemisphere (boreal and austral). To test this concept, we used six genetic markers (18S, 28S, CAD1, COI-5p, COI-3p and COII) and fossil calibrations to produce a well-supported and resolved time-calibrated phylogeny of the subfamily. Austral and boreal diamesines indeed are reciprocally monophyletic lineages, with estimated Jurassic divergence (130-196, 164 Ma). The boreal Protanypodini, previously understood to be a tribe within Diamesinae, is excluded and elevated here to subfamily rank as Protanypodinae stat. nov. Ancestral austral diamesines probably originated in South America and successively reached New Zealand, Australia and South Africa during the Cretaceous-Paleogene. The Holarctic tribes Diamesini and Boreoheptagyiini probably originated in the Eastern Palaearctic with further dispersal/vicariance into the Western Palaearctic, Nearctic, East, and very likely dispersed southwards to montane East Africa and Borneo.

Cicada minimum age tree: Cryptic speciation and exponentially increasing base substitution rates in recent geologic time

We developed a new time-calibrated tree incorporating primarily endemic along with some cryptic Ryukyu islands cicada data, following the recent publication of global cicada data by Marshall et al. (2018), Łukasik et al. (2018), Simon et al. (2019), Price et al. (2019), and Hill et al. (2021). A total of 352 specimens were analyzed using BEAST v1. X software with a relaxed clock model. Fossil calibrations as old as Triassic were adopted largely following Johnson et al. (2018) and Moulds (2018), and a Quaternary geological event calibration was adopted following Osozawa et al. (2012, 2021b) and input into BEAST v1. X. Our timetree suggests that Tettigarctidae had a cicada basal lineage as old as 200.63 Ma, with Derotettiginae the next oldest lineage at 99.2 Ma. Tibicininae is a sister of the remaining subfamilies of Tettigomyiinae, Cicadettinae, and Cicadidae, and their species level differentiation and radiation began at 40.57 Ma. The Cicadinae clade consists of specific tribes with paraphyletic relationship, and the vicariance and adaptive radiation generated many cryptic species in each tribe. We estimated base substitution rate as a function of age, and the result strongly indicates an exponential increase of base substitution rate in recent geologic time. The consequent increase in cicada biodiversity, including generation of cryptic species in the Ryukyu Islands and surroundings, may have been driven by the generation and spreading of C4 grasses and coeval Quaternary climate change.

Out of the ocean: the timescale of molluscan evolution based on phylogenomics revealed the ages of mollusks’ evolutionary transitions into the novel environment

Being the phylum with the second largest biodiversity, mollusks are widely distributed in marine, freshwater, and land, conquered almost all habitats on the earth. Throughout geological time, several molluscan lineages independently colonized freshwater and land, evolving independently in new habitats. Resolving the timing of their colonization for novel environments is the basis of understanding the complex evolution of Mollusca. Herein, we obtained an elaborate single–copy orthologs set including 107 genes with a total length of 3,447,675 amino acid sites across 134 mollusks to reconstruct the phylogeny of Mollusca. The phylogenetic analysis recovered the “Aculifera+ Conchifera” topology with well–supported. By the divergence times estimation with fossil calibration, we revealed Cambrian rapid diversification of all molluscan classes. What’s more, our results provided the times of three major independent colonization of novel environments and consistent with previous studies. The freshwater mussels (Bivalvia: Unionida) colonized freshwater about 233 Mya, ancient origin and late diversification make this order to become the main represent of the freshwater bivalves. The orders Stylommatophora and Basommatophora (Gastropoda: Pulmonata) respectively colonized land and freshwater about 201 Mya, the Triassic–Jurassic mass extinction may have provided the opportunities for their colonization. Ampullariidae (Gastropoda) colonized freshwater about 156 Mya, their lungs may have contributed to the adaption of tropical freshwater environment and make them survive. Our results probably offer the most comprehensive time–scale of molluscan evolution, could provide some insights into the habitat transition and evolution of Mollusca.