Ancestral Character Reconstruction Research Articles

Phylogeny and biogeography support ancient vicariance and subsequent dispersal out of Africa in Palpimanidae spiders (Araneae)

Abstract The Palpimanidae are one of five extant Palpimanoidea families that occur mainly in South America and Africa, although there are lineages in Madagascar, islands of the Indian Ocean, and parts of Asia. Here we examine the role of plate tectonics in shaping the distribution of Palpimanidae. We perform molecular sequencing via target enrichment, which makes use of fragmented DNA, because most specimens are stored in natural history museum collections and were not properly preserved for molecular sequencing. We perform phylogenetic analysis, divergence dating, and ancestral range reconstructions to assess whether continental vicariance shaped the evolution of Palpimanidae. We also examine evolution of eye loss via ancestral character reconstruction. We report the first Palpimanidae phylogeny based on genomic data that samples the majority of Chediminae genera. Results suggest that Palpimanidae originated in the Triassic, with diversification spanning the Jurassic to the Cretaceous. Vicarance played a role in early diversification, with later range expansion out of Africa. Based on morphology and phylogenetic analysis, Sceliraptor is synonymized with Sarascelis, Sceliscelis is synonymized with Scelidocteus, and the new genus and species Sitamacho tao gen. nov., sp. nov. is described, with three species transferred from Hybosida.

Reversed oxidative TCA (roTCA) for carbon fixation by an Acidimicrobiia strain from a saline lake.

Acidimicrobiia are widely distributed in nature and suggested to be autotrophic via the Calvin-Benson-Bassham (CBB) cycle. However, direct evidence of chemolithoautotrophy in Acidimicrobiia is lacking. Here, we report a chemolithoautotrophic enrichment from a saline lake, and the subsequent isolation and characterization of a chemolithoautotroph, Salinilacustristhrix flava EGI L10123T, which belongs to a new Acidimicrobiia family. Although strain EGI L10123T is autotrophic, neither its genome nor Acidimicrobiia metagenome-assembled genomes (MAGs) from the enrichment culture encode genes necessary for the CBB cycle. Instead, genomic, transcriptomic, enzymatic, and stable-isotope probing data hinted at the activity of the reversed oxidative TCA (roTCA) coupled with the oxidation of sulfide as the electron donor. Phylogenetic analysis and ancestral character reconstructions of Acidimicrobiia suggested that the essential CBB gene rbcL was acquired through multiple horizontal gene transfer events from diverse microbial taxa. In contrast, genes responsible for sulfide- or hydrogen-dependent roTCA carbon fixation were already present in the last common ancestor of extant Acidimicrobiia. These findings imply the possibility of roTCA carbon fixation in Acidimicrobiia and the ecological importance of Acidimicrobiia. Further research in the future is necessary to confirm whether these characteristics are truly widespread across the clade.

Nuclear phylogenomics of Asteraceae with increased sampling provides new insights into convergent morphological and molecular evolution

Convergent morphological evolution is widespread in flowering plants, and understanding this phenomenon relies on well-resolved phylogenies. Nuclear phylogenetic reconstruction using transcriptome datasets has been successful in various angiosperm groups, but it is limited to taxa with available fresh materials. Asteraceae, which are one of the two largest angiosperm families and are important for both ecosystems and human livelihood, show multiple examples of convergent evolution. Nuclear Asteraceae phylogenies have resolved relationships among most subfamilies and many tribes, but many phylogenetic and evolutionary questions regarding subtribes and genera remain, owing to limited sampling. Here, we increased the sampling for Asteraceae phylogenetic reconstruction using transcriptomes and genome-skimming datasets and produced nuclear phylogenetic trees with 706 species representing two-thirds of recognized subtribes. Ancestral character reconstruction supports multiple convergent evolutionary events in Asteraceae, with gains and losses of bilateral floral symmetry correlated with diversification of some subfamilies and smaller groups, respectively. Presence of the calyx-related pappus may have been especially important for the success of some subtribes and genera. Molecular evolutionary analyses support the likely contribution of duplications of MADS-box and TCP floral regulatory genes to innovations in floral morphology, including capitulum inflorescences and bilaterally symmetric flowers, potentially promoting the diversification of Asteraceae. Subsequent divergences and reductions in CYC2 gene expression are related to the gain and loss of zygomorphic flowers. This phylogenomic work with greater taxon sampling through inclusion of genome-skimming datasets reveals the feasibility of expanded evolutionary analyses using DNA samples for understanding convergent evolution.

Integrated phylogenetic analyses reveal the evolutionary, biogeographic, and diversification history of Asian warty treefrog genus Theloderma (Anura, Rhacophoridae).

Asian warty treefrogs, genus Theloderma, are morphologically variable arboreal frogs endemic to Southeast Asia and Southern China. However, integrated systematic studies are lacking, and knowledge of the genus in terms of diversity, origin, and historical diversification remains limited. To address these knowledge gaps, we used three mitochondrial and five nuclear gene fragments to reconstruct the Theloderma phylogeny, estimate divergence times, and examine the biogeography of the genus. Phylogenetic and species delimitation analyses suggest that the genus Theloderma comprises three major clades corresponding to two subgenera and seven species groups, and mPTP identified at least 12 putative cryptic species, suggesting that species diversity has been underestimated. Biogeographic analyses indicated that most recent common ancestor of Theloderma originated in the Indochina Peninsula during the Middle Oligocene (ca. 27.77 Ma) and the splitting of Clade A to C occurred in the Late Oligocene (ca. 23.55-25.57 Ma). Current biogeographic patterns result from two distinct processes: insitu diversification in the Indochina Peninsula and dispersal in multiple areas, namely southward dispersal to the Malay Peninsula and Borneo, northeastward dispersal to Southern China, northward dispersal to the Himalayas, and dispersal from Southern China to the Indochina Peninsula. Ancestral character reconstruction suggests that the ancestor of Theloderma may have possessed a small body size, rough dorsal skin, and absence of vomerine teeth and hand webbing, and that these four characters have undergone multiple evolutions. Principal component analysis based on eight bioclimatic variables did not clearly distinguish the three major clades of Theloderma, suggesting that species in these clades may occupy similar climatic ecological niches. Our research highlights the importance of orogeny and paleoclimatic changes, in shaping amphibian biodiversity in mountain ecosystems.

The ten-million-year explosion: Paleocognitive reconstructions of domain-general cognitive ability (G) in extinct primates

The correlation between primate “Big G” scores and brain volume in 68 extant species was employed to estimate probable G values for an additional 68 extinct and 1 extant species with endocranial volume data, employing phylogenetic bracketing. Three different methods were used to generate bracketed estimates, which all showed high convergence. The average of these G estimates (for the extinct primates) coupled with the values from the extant species were found to correlate strongly with neurocognitive measures of both extant and extinct primate taxa, specifically Transfer Index scores (an indicator of cognitive flexibility) and the neuroanatomical covariance ratio (a measure of neural integration). Ancestral character reconstruction incorporating G values was made possible with a phylogenetic tree containing data on the relationships among extant and extinct primates. Negative correlations were found between G and branch length, indicating that higher-G species do not persist as long as lower-G ones, consistent with the presence of the grey-ceiling effect (brain mass negatively predicts maximum population growth rate, and therefore a heightened vulnerability to extinction). Cladogenesis rates were also positively associated with G. Both associations were robust to models that controlled for false positive rates. Comparative models revealed that G evolved in extinct and extant primates in a punctuated pattern. The biggest increase in G occurred after the split between the members of the tribes Hominini and Gorillini 10 million years ago. Hence at the macroevolutionary scale, there can be said to have been a “ten-million-year explosion” in primate G leading up to modern humans.

Evolutionary features of microspore and pollen grain development in Cyperaceae.

Asymmetric meiosis leading to the release of pollen grains as pseudomonads is a synapomorphy in Cyperaceae, but differences in microspore development are relevant in the family's evolutionary history. Cyperaceae members present atypical microsporogenesis, in which one meiotic product is functional while the other three degenerate, culminating in pseudomonad pollen formation. Differences during development, such as pseudomonad shape and degenerative microspore positioning, are seen throughout the family, but no phylogenetic interpretation has been made regarding these variances thus far. In this study, we analyzed the early- and late-diverging sedge genera Hypolytrum and Eleocharis, respectively, while comparing them to data available in the literature and conducting an ancestral character reconstruction for pseudomonad traits. Light microscopy results show that pseudomonad development in Hypolytrum is homologous to several other sedge genera, presenting apical degenerative microspores. However, pseudomonads are round and centrally arranged in the anther locule in this case, which consists of a pleisiomorphic trait for the family. The basal positioning of degenerative microspores is restricted to Rhynchospora, consisting of an apomorphic trait for this genus. Despite these differences, ultrastructural analysis of Eleocharis pseudomonad revealed shared features with other genera studied, which include variations in chromatin condensation and cytoplasmic turnover in functional cells. These common features seem related to the different cellular fates seen during microspore development and further corroborate the synapomorphic status of pseudomonads in sedges.

An updated classification for the hyper-diverse genus Corydalis (Papaveraceae: Fumarioideae) based on phylogenomic and morphological evidence.

The genus Corydalis, with ca. 530 species, has long been considered taxonomically challenging because of its great variability. Previous molecular analyses, based on a few molecular markers and incomplete taxonomic sampling, were clearly inadequate to delimit sections and subgenera. We have performed phylogenetic analyses of Corydalis and related taxa, using 65 shared protein-coding plastid genes from 313 accessions (including 280 samples of ca. 226 species of Corydalis) and 152 universal low-copy nuclear genes from 296 accessions (including 271 samples of Corydalis) covering all 42 previously recognized sections and five independent "series". Phylogenetic trees were inferred using Bayesian Inference and Maximum Likelihood. Eight selected morphological characters were estimated using ancestral state reconstructions. Results include: (i) of the three subgenera of Corydalis, two are fully supported by both the plastid and nuclear data; the third, subg. Cremnocapnos, is weakly supported by plastid DNA only, whereas in the nuclear data the two included sections form successive outgroups to the rest of the genus; (ii) among all 42 sections and five "series", 25 sections and one "series" are resolved as monophyletic in both data sets; (iii) the common ancestor of Corydalis is likely to be a perennial plant with a taproot, yellow flowers with a short saccate spur, linear fruits with recurved fruiting pedicels, and seeds with elaiosomes; (iv) we provide a new classification of Corydalis with four subgenera (of which subg. Bipapillatae is here newly described), 39 sections, 16 of which are consistent with the previous classification, 16 sections have been recircumscribed, one section has been reinstated and six new sections are established. Characters associated with lifespan, underground structures, floral spur, fruit and elaiosomes are important for the recognition of subgenera and sections. These new phylogenetic analyses combined with ancestral character reconstructions uncovered previously unrecognized relationships, and greatly improved our understanding of the evolution of the genus.

Revisiting the evolutionary trend toward the mammalian lower jaw in non-mammalian synapsids in a phylogenetic context.

The mammalian lower jaw comprises a single bone, the dentary, which is a unique feature among vertebrates. The lower jaws of extinct non-mammalian synapsids were composed of the dentary and several postdentary bones. Synapsid fossils exhibit variation in the dentary size relative to the overall lower jaw. An evolutionary trend toward dentary enlargement and postdentary reduction in non-mammalian synapsids has long been documented but has not been established using modern phylogenetic comparative methods. In this study, we examine the evolutionary pattern of dentary size relative to the lower jaw through phylogenetic analyses of measurements in a broad range of non-mammalian synapsid taxa. Our analyses revealed an evolutionary trend toward dentary area enlargement relative to the overall lower jaw in the lateral view across all non-mammalian synapsids. This trend is likely due to vertical expansion of the dentary given that the same trend is not evident when looking at anterior to posterior measurements of the dentary relative to the lower jaw as a whole in lateral view. Ancestral character reconstructions revealed that the evolution of the measurements was not unidirectional in non-mammalian synapsids. Our results provide no evidence of an evolutionary trend toward the dentary enlargement at the expense of postdentary bones across non-mammalian synapsids. This implies that the evolutionary origin of the mammalian lower jaw is not adequately explained by the evolutionary trend of dentary enlargement throughout non-mammalian synapsids. Instead, selection that occurred during the transition from non-mammalian cynodonts to early mammals may have produced the mammalian lower jaw.

Modeling Drug Resistance Emergence and Transmission in HIV-1 in the UK.

A deeper understanding of HIV-1 transmission and drug resistance mechanisms can lead to improvements in current treatment policies. However, the rates at which HIV-1 drug resistance mutations (DRMs) are acquired and which transmitted DRMs persist are multi-factorial and vary considerably between different mutations. We develop a method for the estimation of drug resistance acquisition and transmission patterns. The method uses maximum likelihood ancestral character reconstruction informed by treatment roll-out dates and allows for the analysis of very large datasets. We apply our method to transmission trees reconstructed on the data obtained from the UK HIV Drug Resistance Database to make predictions for known DRMs. Our results show important differences between DRMs, in particular between polymorphic and non-polymorphic DRMs and between the B and C subtypes. Our estimates of reversion times, based on a very large number of sequences, are compatible but more accurate than those already available in the literature, with narrower confidence intervals. We consistently find that large resistance clusters are associated with polymorphic DRMs and DRMs with long loss times, which require special surveillance. As in other high-income countries (e.g., Switzerland), the prevalence of sequences with DRMs is decreasing, but among these, the fraction of transmitted resistance is clearly increasing compared to the fraction of acquired resistance mutations. All this indicates that efforts to monitor these mutations and the emergence of resistance clusters in the population must be maintained in the long term.

Molecular phylogeny of Onobrychis sect. Onobrychis (Fabaceae-Hedysareae) with insights into its taxonomy and character evolution

Onobrychis sect. Onobrychis, as the most species-rich section of the genus Onobrychis, ranges from Central Asia to Europe with a south extension into North Africa. In this study, we concentrated on the systematics of previously poorly represented O.sect. Onobrychis using a substantial taxon sampling. To test the monophyly of the section as well as to clarify interspecific relationships, the nuclear ribosomal DNA internal transcribed spacer(nrDNA ITS) and three chloroplast sequences (ndhF-rlp32, rpl32-trnL(UAG), and trnG(UCC)-trnS(GCU)) were employed. Phylogenetic analyses were performed by maximum likelihood and Bayesian inferences. The results showed that O.sect. Onobrychis is monophyletic and composed of two main well-supported clades which are redefined as two subsections (O.subsect. Onobrychis, O. subsect. Macropterae).Ancestral character reconstruction of morphological traits: life form, wing to standard ratio, chromosome number, and ploidy level, indicated that short-winged petal is a synapomorphy for a group of species in the section while other traits have undergone parallel evolution. Possessing the basic chromosome number x= 7 has arisen twice, independently. It seems that perenniality and polyploidy are probably the key innovations in the diversification of O. sect. Onobrychis.

Karyotype evolution in Ronderosia grasshoppers (Orthoptera: Acrididae)

Abstract Grasshoppers in the genus Ronderosia (divergence time < 5 Mya) provide a special opportunity to examine the impact of chromosome rearrangements (CRs) in evolution and speciation, because they exhibit extensively rearranged karyotypes involving autosome-autosome and autosome-X chromosomes, the latter leading to the formation of a neo-sex chromosome system (neo-SCS). Despite the potential role of CRs in speciation, the general patterns of karyotype changes resulting from CRs in Ronderosia are still unclear. Here we investigated karyotype evolution in eight Ronderosia species using cytogenetic and phylogenetic models of ancestral character reconstruction. The overall findings suggest a central role for CRs in generating variability in chromosome number and neo-SCS across Ronderosia grasshoppers. It also suggests that incipient species with little morphological change, such as those observed in Ronderosia species, could accumulate marked differences in their karyotypes contributing to post-zygotic reproductive isolation. Besides the evolution of chromosome number in the phylogeny of the genus, chromosome change due to centric fusion is coupled with the divergence pattern observed in the group. While a cladogenetic event could represent a case where karyotypic differences result in speciation, further in-depth genomic studies are needed to better understand how these dramatic restructurings of karyotypes may have occurred.

Insights into the evolutionary history and taxonomic status of Sinopteris (Pteridaceae)

As an endemic Chinese genus, Sinopteris C. Chr. & Ching was once considered an early diverged taxon of cheilanthoid ferns, and its taxonomic status has long been controversial. In this study, eight datasets spanning the complete chloroplast genomes and three nuclear genes were used to reconstruct the phylogeny of Sinopteris and its relatives. In addition, combining morphological analyses, divergence time estimation, and ancestral trait reconstruction, the origin and evolutionary history of Sinopteris were comprehensively discussed. Based on the complete chloroplast genome dataset, our analyses yielded a phylogram with all clades strongly supported (ML-BS = 100, BI-PP = 1.0), and the topology was almost identical to that based on the concatenated sequences of nrDNA, CRY2, and IBR3. Two species of Sinopteris were united and sister to Aleuritopteris niphobola (C. Chr.) Ching. They constituted a stable monophyletic group embedded in Aleuritopteris Fée. This was also consistent with the results of morphological analyses. Divergence time estimation indicated that the clade of Aleuritopteris and Sinopteris originated in the early Miocene (ca. 16.80 Ma) and experienced two rapid diversifications, which could coincide with environmental heterogeneity caused by the progressive uplift of the Himalayas and the intense uplift of the Hengduan Mountains. Sinopteris originated in the late Miocene (ca. 6.96 Ma), accompanied by the sharp intensifications of Asian Monsoon, and began to diversify at 2.34 Ma, following the intense uplift of the Hengduan Mountains. Ancestral character reconstruction showed that monangial sori and subsessile sporangia were clearly late derived states rather than early diverged states. Both the molecular phylogenetic and morphological analyses support the inclusion of Sinopteris in Aleuritopteris.

The anatomy of the seed-coat includes diagnostic characters in the subtribe Eugeniinae (Myrteae, Myrtaceae).

The subtribe Eugeniinae comprises of two genera, Eugenia (ca. 1,100 species) and Myrcianthes (ca. 40 species). Eugenia is the largest genus of neotropical Myrtaceae and its latest classification proposes 11 sections. This study describes the seed anatomy of forty-one species of Eugeniinae in order to provide possible diagnostic characteristics. Following standard anatomical techniques, flower buds, flowers, and fruits were processed and analyzed using microtome sections and light microscopy. The phylogeny used the regions ITS, rpl16, psbA-trnH, trnL-rpl32, and trnQ-rps16, following recent studies in the group. Ancestral character reconstruction uncovered that: (1) the ancestral ovule in Eugeniinae was campylotropous (98.9% probability), bitegmic (98.5% probability), and unitegmic ovules arose on more than one lineage independently within Eugenia; (2) the pachychalazal seed-coat appeared with a 92% probability of being the ancestral type; (3) non-lignified seed-coat (24,5% probability) and aerenchymatous mesotesta (45.8% probability) are diagnostic characters in Myrcianthes pungens (aerenchymatous mesotesta present in the developing seed-coat) and in the species of E. sect. Pseudeugenia until the species of E. sect. Schizocalomyrtus and it is the type of seed-coat that predominates in most basal sections on the tree; (4) the partial sclerification (only in the exotesta—exotestal seed-coat) is mainly observed in species of E. sect. Excelsae, E. sect. Jossinia (group X), and E. sect. Racemosae (22.2% probability); (5) and in the species of the recent lineages of Eugenia, with a probability of 27.2%, predominate the exomesotestal or testal construction of the seed-coat [character observed in almost all species analyzed of E. sect. Jossinia (group Y) and E. sect. Umbellatae]. A dehiscent fruit is considered as a plesiomorphic state in Myrtaceae; the ancestor of this family had seeds with a completely sclerified testa, and the other testa types described for the current species with dehiscent and indehiscent fruits are simplified versions of this ancestral type. Perhaps, this means that the sclerified layers in the seed-coat have remained in whole or in part as a plesiomorphic condition for taxa with a capsule and bacca. Maintaining the plesiomorphic condition may have represented a selective advantage at some point in the evolutionary history of the family and its groups.

Characterization of the Complete Mitochondrial Genome of Eight Diurnal Hawkmoths (Lepidoptera: Sphingidae): New Insights into the Origin and Evolution of Diurnalism in Sphingids.

Simple SummaryWe newly sequenced the mitochondrial genomes of 22 species from three subfamilies in the family Sphingidae. The mitogenomes of six diurnal species were newly sequenced, assembled, and annotated in this study (Hemaris radians, Macroglossum bombylans, Macroglossum fritzei, Macroglossum pyrrhosticta, Neogurelca himachala, and Sataspes xylocoparis) and, together with those of two previously reported diurnal hawkmoths (Cephonodes hylas and Macroglossum stellatarum), were analyzed in terms of sequence length, nucleotide composition, relative synonymous codon usage, non-synonymous/synonymous substitution ratio, gene spacing, and repeat sequences. The phylogenetic relationships among the subfamilies of Sphingidae were analyzed to explore the origin, divergence time, and evolutionary status of diurnal sphingids. In addition, we performed divergence time estimation and reconstruction of ancestral characters. Diurnal behavior of hawkmoths first originated 29.19 million years ago (Mya) and may have been influenced by the combination of herbaceous flourishing, which occurred 26–28 Mya, the uplift of the Tibetan Plateau, and the large-scale evolution of bats in the Oligocene to Pre-Miocene. Moreover, diurnalism in hawkmoths had multiple independent origins.In this study, the mitochondrial genomes of 22 species from three subfamilies in the Sphingidae were sequenced, assembled, and annotated. Eight diurnal hawkmoths were included, of which six were newly sequenced (Hemaris radians, Macroglossum bombylans, M. fritzei, M. pyrrhosticta, Neogurelca himachala, and Sataspes xylocoparis) and two were previously published (Cephonodes hylas and Macroglossum stellatarum). The mitochondrial genomes of these eight diurnal hawkmoths were comparatively analyzed in terms of sequence length, nucleotide composition, relative synonymous codon usage, non-synonymous/synonymous substitution ratio, gene spacing, and repeat sequences. The mitogenomes of the eight species, ranging in length from 15,201 to 15,461 bp, encode the complete set of 37 genes usually found in animal mitogenomes. The base composition of the mitochondrial genomes showed A+T bias. The most commonly used codons were UUA (Leu), AUU (Ile), UUU (Phe), AUA (Met), and AAU (Asn), whereas GCG (Ala) and CCG (Pro) were rarely used. A phylogenetic tree of Sphingidae was constructed based on both maximum likelihood and Bayesian methods. We verified the monophyly of the four current subfamilies of Sphingidae, all of which had high support. In addition, we performed divergence time estimation and ancestral character reconstruction analyses. Diurnal behavior in hawkmoths originated 29.19 million years ago (Mya). It may have been influenced by the combination of herbaceous flourishing, which occurred 26–28 Mya, the uplift of the Tibetan Plateau, and the large-scale evolution of bats in the Oligocene to Pre-Miocene. Moreover, diurnalism in hawkmoths had multiple independent origins in Sphingidae.

Paepalanthus (Eriocaulaceae) without scapes and spathes, a survey with the description of a new species

Eriocaulaceae are mainly characterized by their unisexual flowers arranged in capituliform inflorescences at the apex of scapes surrounded by closed spathes. The lack of the scapes and spathes is a rare condition in the family, restricted to a small number of species in Eriocaulon, Paepalanthus, and Syngonanthus. In this manuscript we explore the species of Paepalanthus lacking spathes and scapes based on morphological and molecular data (ITS, trnL-trnF, and psbA-trnH). The specimens with dimerous flowers form a monophyletic group represented by the only two species of P. subg. Thelxinoë. The accessions with trimerous flowers cluster with species of the polyphyletic P. ser. Leptocephali. As expected, the ancestral character reconstruction indicates that the loss of scapes and spathes occurred at least twice in Paepalanthus, but probably occurred four to five times in the family. Based on molecular data, SEM images, and morphological analysis, most of the specimens lacking scapes were assigned to previously described species with the exception of the trimerous-flowered specimens treated here as a new species, P. paganuccii. Besides the full treatment for this new species, we provide an annotated checklist for all Paepalanthus species without scapes, with nomenclatural novelties, a distribution map, illustrations, photos, and a comparative table.

Absence of some cytochrome P450 (CYP) and hydroxysteroid dehydrogenase (HSD) enzymes in hagfishes

Corticosteroids are synthesized from cholesterol by steroidogenic enzyme catalysts belonging to two main families: the cytochrome p450s (CYPs) and hydroxysteroid dehydrogenases (HSDs). The action of these steroidogenic enzymes allows the genesis of the terminal active corticosteroids 11-deoxycortisol (S), 1ɑ-hydroxycorticosterone (1α-OH-B), or cortisol in different fish species. However, for Cyclostomes like hagfishes, the terminal corticosteroid is still undefined. In this study, we examined the presence or absence of CYPs and HSDs as traits in fishes to gain insight about the primary corticosteroid synthesis pathways of the hagfishes. We used published cytochrome c oxidase I (COXI) amino acid sequences to construct a phylogeny of fishes and then mapped the CYPs and HSDs as morphological traits onto the tree to predict the ancestral character states through ancestral character reconstruction (ACR). There is a clear phylogenetic signal for CYP (i.e., CYP11a1, 17, 21, and 11b) and HSD (i.e., 11-βHSD and 3β-HSD) derivatives of interest throughout the more derived fishes. Using trait-based ACR, we also found that hagfishes possess genes for 3β-HSD, CYP11a1, CYP17, and CYP21. Importantly, the presence of CYP21 implies that hagfish can synthesize 11-deoxycorticosterone (11-DOC) and S. Previous research demonstrated that despite hagfish having CYP21, neither 11-DOC nor S could be detected in hagfish. This discrepancy between the presence of steroidogenic enzymes and products brings into question the expression and/or function of CYP21 in hagfishes.

Recurrent evolutionary changes in pollen shape and ornamentation of the vataireoid clade (Papilionoideae, Fabaceae), an early-branching lineage of florally divergent genera

Abstract Pollen traits have clarified the classification and understanding of the evolutionary history of different groups of angiosperms, due to their wide morphological diversity. To increase our knowledge on the morphological evolution of the vataireoid clade (Fabaceae, Papilionoideae), a lineage comprising 30 species of Neotropical trees in the genera Luetzelburgia, Sweetia, Vatairea and Vataireopsis, pollen materials of 26 species were acetolysed and analysed using light and scanning electron microscopy. Characters were defined to reconstruct the pollen evolution in each of the ancestral nodes of the phylogenetic tree of the clade using Bayesian stochastic character mapping. All vataireoid genera have relatively variable pollen morphology, ranging from small to medium and subtriangular to circular amb, and their aperture margo may be present or absent. The exine ornamentation varies greatly across the clade: perforate (L. praecox and S. fruticosa); rugulate (L. andradelimae); rugulate-foveolate-perforate (Vatairea guianensis and Vatairea heteroptera); reticulate (Vataireopsis araroba) and microreticulate in most species. Analyses of ancestral character reconstruction revealed multiple events of independent evolution in all pollen traits during the diversification of the vataireoid clade in the last 16 Myr, since the Miocene. Just as evolutionary transitions in floral architecture were so recurrent in the vataireoid clade, pollen evolution was relatively labile and exhibited little phylogenetic conservatism in shape or ornamentation.

Untargeted Metabolomics Sheds Light on the Diversity of Major Classes of Secondary Metabolites in the Malpighiaceae Botanical Family.

Natural products produced by plants are one of the most investigated natural sources, which substantially contributed to the development of the natural products field. Even though these compounds are widely explored, the literature still lacks comprehensive investigations aiming to explore the evolution of secondary metabolites produced by plants, especially if classical methodologies are employed. The development of sensitive hyphenated techniques and computational tools for data processing has enabled the study of large datasets, being valuable assets for chemosystematic studies. Here, we describe a strategy for chemotaxonomic investigations using the Malpighiaceae botanical family as a model. Our workflow was based on MS/MS untargeted metabolomics, spectral searches, and recently described in silico classification tools, which were mapped into the latest molecular phylogeny accepted for this family. The metabolomic analysis revealed that different ionization modes and extraction protocols significantly impacted the chemical profiles, influencing the chemotaxonomic results. Spectral searches within public databases revealed several clades or genera-specific molecular families, being potential chemical markers for these taxa, while the in silico classification tools were able to expand the Malpighiaceae chemical space. The classes putatively annotated were used for ancestral character reconstructions, which recovered several classes of metabolites as homoplasies (i.e., non-exclusive) or synapomorphies (i.e., exclusive) for all sampled clades and genera. Our workflow combines several approaches to perform a comprehensive evolutionary chemical study. We expect it to be used on further chemotaxonomic investigations to expand chemical knowledge and reveal biological insights for compounds classes in different biological groups.

A first phylogenetic hypothesis for the diverse genus Conophytum (Ruschieae, Ruschioideae, Aizoaceae) suggests convergent evolution of floral syndromes

Pollinator specialisation is one of the major drivers of angiosperm diversification in the Greater Cape Floristic Region (GCFR) of South Africa. Conophytum (Aizoaceae) is a flagship genus for the underexplored arid regions of the GCFR (ca. 108 spp.) with 83.9% of its species endemic to this region and has a floral structure that is unique within the rapidly diversified Ruschieae (Aizoaceae). Floral traits, together with leaf characters divide the genus into 16 sections. We present here the first phylogenetic hypothesis for Conophytum, based on molecular data. The combined phylogenetic data for six plastid regions (matK, rpl16, rps16, trnL-F, trnQ-rps16 and trnS-trnG) were analysed using Bayesian inference, maximum likelihood and maximum parsimony to test the evolution of this floral diversity and current sectional classification. Conophytum was recovered as monophyletic including the three small genera Berrisfordia L.Bolus, Herreanthus Schwantes and Ophthalmophyllum Dinter & Schwantes. Six strongly supported clades were recovered within Conophytum: while several of the sections were broadly retrieved within these clades, the more morphologically variable sections such as Minuscula and Wettsteinia were not supported by our data. The distributions of the six clades largely overlap and are generally confined to the arid parts of the GCFR within South Africa, with only one clade extending into the southern wetter parts. Ancestral character reconstructions showed that floral traits evolved multiple times with low phylogenetic signal recovered for autumn- and winter-flowering and flower type, while the other traits indicated no phylogenetic signal. This lack of phylogenetic signal suggests that drivers of diversity in Conophytum are possibly linked to adaptation to pollinators, with a high rate of inferred pollination shifts observed, i.e. 1.7 shifts per species, mirroring that of Lapeirousia (Iridacaeae) in the GCFR. The evolution of unique tubular flowers and shift to autumn-flowering were recovered as traits indicating strong phylogenetic signal and may have enabled Conophytum to exploit a range of pollinators through diversification in floral morphologies.

Morphology, molecular phylogeny and systematics of the diatom genus Fallacia (Sellaphoraceae), with descriptions of three new species1.

Fallacia is distinguished morpho-anatomically from Navicula sensu lato based on the possession of an H-shaped chloroplast, lateral sterna and a finely porous conopeum, but whether this genus is monophyletic is still in question. Three new Fallacia species are described based on morphology and SSU rRNA and rbcL gene sequences: Fallacia tateyamensis sp. nov., Fallacia bosoensis sp. nov. and Fallacia laevis sp. nov. We performed the first comprehensive molecular and morphological phylogenetic analyses of 31 Fallacia species based on 11 new sequences from six species and 23 morphological characters. We also documented the detailed morphogenesis of Fallacia for the first time. Fallacia is not monophyletic. Both morphological and DNA sequence data supported the separation of Rossia from Fallacia, while the phylogenetic position of Pseudofallacia is uncertain. We recognized four morphogroups in Fallacia by morphological and molecular phylogenetic analyses. Ancestral character reconstruction indicated that diatoms in Sellaphoraceae evolved from the possession of two lateral narrow parallel depressions covered by narrow nonporous conopea, to lyre-shaped canals covered by wide porous conopea. Lanceolate canals and the presence of areolae in canals evolved multiple times independently.