Multilocus Dataset Research Articles

Molecular taxonomic identification and species-level phylogeny of the narrow-mouthed frogs of the genus Rhombophryne (Anura: Microhylidae: Cophylinae) from Madagascar

The study of diamond frogs (genus Rhombophryne, endemic to Madagascar) has been historically hampered by the paucity of available specimens, because of their low detectability in the field. Over the last 10 years, 13 new taxa have been described, and 20 named species are currently recognized. Nevertheless, undescribed diversity within the genus is probably large, calling for a revision of the taxonomic identification of published records and an update of the known distribution of each lineage. Here we generate DNA sequences of the mitochondrial 16S rRNA gene of all specimens available to us, revise the genetic data from public databases, and report all deeply divergent mitochondrial lineages of Rhombophryne identifiable from these data. We also generate a multi-locus dataset (including five mitochondrial and eight nuclear markers; 9844 bp) to infer a species-level phylogenetic hypothesis for the diversification of this genus and revise the distribution of each lineage. We recognize a total of 10 candidate species, two of which are identified here for the first time. The genus Rhombophryne is here proposed to be divided into six main species groups, and phylogenetic relationships among some of them are not fully resolved. These frogs are primarily distributed in northern Madagascar, and most species are known from only few localities. A previous record of this genus from the Tsingy de Bemaraha (western Madagascar) is interpreted as probably due to a mislabelling and should not be considered further unless confirmed by new data. By generating this phylogenetic hypothesis and providing an updated distribution of each lineage, our findings will facilitate future species descriptions, pave the way for evolutionary studies, and provide valuable information for the urgent conservation of diamond frogs.

Metahyphopichia suwanaadthiae sp. nov., an anamorphic yeast species in the order Saccharomycetales and reassignment of Candida silvanorum to the genus Metahyphopichia.

Seven yeast strains, representing a single novel anamorphic species, were isolated in Thailand. They consisted of five strains (DMKU-MRY16T, DMKU-SK18, DMKU-SK25, DMKU-SK30 and DMKU-SK32) obtained from five different mushrooms, and two strains (ST-224 and 11-14.2) derived from insect frass and soil, respectively. The pairwise sequence analysis indicated that all seven strains had identical sequences in the D1/D2 domains of the large subunit (LSU) rRNA gene and the internal transcribed spacer (ITS) region. Metahyphopichia silvanorum was the most closely related species, but with 11.9-12.4% nucleotide substitutions in the D1/D2 domains of the LSU rRNA gene and 13.1-13.3% nucleotide substitutions in the ITS region. The phylogenetic analyses based on the concatenated sequences of the ITS region and the D1/D2 domains of the LSU rRNA gene showed that the seven strains form a well-separated subclade in a clade containing M. silvanorum and Metahyphopichia laotica with high bootstrap support. A phylogenetic analysis of a multilocus dataset including the small subunit (SSU) rRNA gene, the ITS region, the D1/D2 domains of the LSU rRNA gene, translation elongation factor 1-alpha gene, actin gene and the RNA polymerase II subunit 2 gene, confirmed the presence of the monophyletic clade that also includes M. silvanorum and M. laotica, and strongly supported the phylogenetic isolation of the seven strains from its neighbouring species. Therefore, the seven strains were assigned as a single novel species of the genus Metahyphopichia, according to their phylogenetic relationships. The name Metahyphopichia suwanaadthiae sp. nov. is proposed to accommodate the seven strains. The holotype is DMKU-MRY16T (TBRC 11775T=NBRC 114386T=PYCC 8655T). The MycoBank number of the novel species is MB 841280. In addition, Candida silvanorum is reassigned to the genus Metahyphopichia. The MycoBank number of M. silvanorum comb. nov. is MB 841279.

Cryptic Species Diversity and Phylogenetic Relationship in the Rust Genus Chrysomyxa from China.

Chrysomyxa rusts are fungal pathogens widely distributed in the Northern hemisphere, causing spruce needle and cone rust diseases, and they are responsible for significant economic losses in China. Taxonomic delimitation and precise species identification are difficult within this genus because some characters often overlap in several species. Adequate species delimitation, enhanced by the use of DNA-based methodologies, will help to establish well-supported species boundaries and enable the identification of cryptic species. Here, we explore the cryptic species diversity in the rust genus Chrysomyxa from China. Species delimitation analyses are conducted using a distance-based method (ABGD) and three tree-based methods (GMYC, bPTP, and mPTP) based on combined LSU and ITS sequences of over 60 specimens. Although there is some incongruence among species delimitation methods, two new species and three putative cryptic species are identified. The key to 20 Chrysomyxa species distributed in China is presented. These results suggest that a significant level of undiscovered cryptic diversity is likely to be found in Chrysomyxa from China. Future studies should consider multiple analytical methods when dealing with multi-locus datasets.

World Travelers: Parthenogenesis and Ecological Tolerance Enable Multiple Colonization Events by the Widespread Short-Tailed Whipscorpion, Stenochrus portoricensis (Schizomida: Hubbardiidae)

Abstract Whereas morphology remains a powerful tool for the diagnosis and description of short-tailed whip scorpions, or schizomids (Order Schizomida Petrunkevitch, 1945), especially when adults of both sexes are available, the systematics of some schizomid taxa is difficult to resolve due to a lack of characters in these morphologically conserved arachnids. Stenochrus portoricensis Chamberlin, 1922, defined on a single character of the female spermathecae, is the most widespread schizomid in the New World. Numerous records in the Neotropics, from the southern United States to Brazil, throughout the Caribbean, and further afield, including the Galapagos Islands and Europe, raise the question as to whether S. portoricensis is indeed a single widespread species or a complex of multiple species with conserved morphology? The present study uses a multilocus dataset and the broadest geographical sample currently available to address the phylogeography of S. portoricensis with molecular divergence dating and ancestral area reconstruction of all currently known species of Stenochrus Chamberlin, 1922. Analyses recovered S. portoricensis as paraphyletic. Two species previously synonymized are revalidated and transferred to Stenochrus. Population structure analyses recovered the remaining samples of S. portoricensis as a single monophyletic species with low genetic divergence and comprising two subclades. Ancestral area reconstruction suggests a Mesoamerican origin for Stenochrus, which contains a widespread species, recently introduced to multiple localities. Introductions to Europe and the Caribbean occurred from a single clade in the Yucatán Peninsula, Mexico, within which genetic divergence is minimal, confirming the hypothesis of multiple independent introductions with successful colonization facilitated by parthenogenetic reproduction.

Ophiocordycepsaphrophoridarum sp. nov., a new entomopathogenic species from Guizhou, China.

BackgroundOphiocordyceps is the largest genus in the family Ophiocordicipitaceae, including many entomopathogenic species. In recent years, many species have been described in this genus, with a wide range of host insects. Entomopathogenic fungi include ecologically, economically and medicinally important species, but a large portion of their diversity remains to be discovered and described.New informationIn this study, a new species, Ophiocordycepsaphrophoridarum sp. nov, parasitising Aphrophoridae sp. (Hemiptera) is proposed from China, based on evidence from morphology and molecular phylogenetic analyses. This species is characterised by fibrous, pigmented stromata, cylindrical asci and filiform ascospores. Compared to its closest relative, O.tricentri, the new species has wider perithecia and longer asci. Molecular phylogenetic analyses of a multilocus dataset (consisting of SSU, ITS, LSU, TEF1, RPB1 and RPB2) confirm its placement in Ophiocordyceps. Ophiocordycepsaphrophoridarum is morphologically described and illustrated with colour photographs. Morphological comparisons with closely-related species are also presented in tabulated format.

Phylogeography and morphometric variation in the Cinnamon Hummingbird complex: Amazilia rutila (Aves: Trochilidae)

BackgroundThe Mesoamerican dominion is a biogeographic area of great interest due to its complex topography and distinctive climatic history. This area has a large diversity of habitats, including tropical deciduous forests, which house a large number of endemic species. Here, we assess phylogeographic pattern, genetic and morphometric variation in the Cinnamon Hummingbird complex Amazilia rutila, which prefers habitats in this region. This resident species is distributed along the Pacific coast from Sinaloa—including the Tres Marías Islands in Mexico to Costa Rica, and from the coastal plain of the Yucatán Peninsula of Mexico south to Belize.MethodsWe obtained genetic data from 85 samples of A. rutila, using 4 different molecular markers (mtDNA: ND2, COI; nDNA: ODC, MUSK) on which we performed analyses of population structure (median-joining network, STRUCTURE, FST, AMOVA), Bayesian and Maximum Likelihood phylogenetic analyses, and divergence time estimates. In order to evaluate the historic suitability of environmental conditions, we constructed projection models using past scenarios (Pleistocene periods), and conducted Bayesian Skyline Plots (BSP) to visualize changes in population sizes over time. To analyze morphometric variation, we took measurements of 5 morphological traits from 210 study skins. We tested for differences between sexes, differences among geographic groups (defined based on genetic results), and used PCA to examine the variation in multivariate space.ResultsUsing mtDNA, we recovered four main geographic groups: the Pacific coast, the Tres Marías Islands, the Chiapas region, and the Yucatán Peninsula together with Central America. These same groups were recovered by the phylogenetic results based on the multilocus dataset. Demography based on BSP results showed constant population size over time throughout the A. rutila complex and within each geographic group. Ecological niche model projections onto past scenarios revealed no drastic changes in suitable conditions, but revealed some possible refuges. Morphometric results showed minor sexual dimorphism in this species and statistically significant differences between geographic groups. The Tres Marías Islands population was the most differentiated, having larger body size than the remaining groups.ConclusionsThe best supported evolutionary hypothesis of diversification within this group corresponds to geographic isolation (limited gene flow), differences in current environmental conditions, and historical habitat fragmentation promoted by past events (Pleistocene refugia). Four well-defined clades comprise the A. rutila complex, and we assess the importance of a taxonomic reevaluation. Our data suggest that both of A. r. graysoni (Tres Marías Islands) and A. r. rutila (Pacific coast) should be considered full species. The other two strongly supported clades are: (a) the Chiapas group (southern Mexico), and (b) the populations from Yucatán Peninsula and Central America. These clades belong to the corallirostris taxon, which needs to be split and properly named.

Revisiting Linnaean and Wallacean Shortfalls in Mindanao Fanged Frogs: The Limnonectes magnus Complex Consists of Only Two Species

We revisit the question of species diversity among Mindanao Fanged Frogs of the Limnonectes magnus complex consisting of L. magnus, L. diuatus, L. ferneri, and a previously hypothesized putative new species, inferred in the first molecular phylogenetic studies of the genus almost 2 decades ago. Using a multilocus molecular deoxyribonucleic acid sequence data set and comprehensive sampling of 161 individuals from throughout the Mindanao Pleistocene aggregate island complex landmasses (a distinct faunal region of the southern Philippines) we characterize geographically structured genetic diversity, focusing on the phylogenetic placement of individuals from each species' type locality. We also present new morphometric data from large samples of freshly collected material from the type localities of each included species; together with examination of the name-bearing original type specimens, we conclude that an overestimation of species diversity has occurred and has been exacerbated by the indiscriminate acceptance of the hypothesis of the existence of widespread cryptic species in this group. We place L. ferneri in synonymy with L. diuatus, clarify the identification of the latter taxon with respect to L. magnus, and apply this name to the widespread, generalist, highly variable giant Fanged Frog distributed throughout the Mindanao faunal region of the southern Philippines.

Taxonomic Revision of Scaly-toed Geckos (Reptilia: Gekkonidae: Lepidodactylus) in the Northern Philippines, with Descriptions of Four New Species

Recent higher level phylogenetic analyses of gekkonid lizards of the genus Lepidodactylus uncovered an array of unrecognized species diversity, particularly within the Philippine archipelago. Novel phylogenetic analyses of multilocus data sets suggest that as many as five, previously undescribed, species-level lineages of Scaly-toed Geckos occur in just the northern portions of the archipelago. Here, we evaluate Lepidodactylus species diversity in the Lepidodactylus yami–balioburius clade and describe four new forest species from Luzon Island and surrounding minor island groups. Interestingly, these species are the first endemic Lepidodactylus taxa described from Luzon proper and peripheral islands. In this first review of Philippine Scaly-toed Gecko diversity in nearly half a century, we use a suite of morphological characters along with molecular data to delimit evolutionary lineages. All species described in this paper can be distinguished from congeners by an array of discrete external traits; all are also monophyletic groups, separated in our phylogenetic analyses of the mitochondrial ND2 gene. This study increases significantly the number of known Scaly-toed Geckos in the Philippines from 7 to 11, which is likely still an underestimate of the species diversity in this understudied clade.

Phylogeny and biogeography of the pantropical whip spider family Charinidae (Arachnida: Amblypygi)

AbstractThe present contribution addresses the phylogeny and biogeography of the pantropical whip spider family Charinidae Quintero, 1986, the most species-rich in the arachnid order Amblypygi Thorell, 1883, based on morphology and multilocus DNA sequences, analysed simultaneously using parsimony, maximum likelihood and Bayesian inference. The morphological matrix comprises 138 characters, scored for four outgroup taxa and 103 ingroup terminals representing all genera and 64% of the species of Charinidae. The multilocus dataset comprises sequences from two nuclear and three mitochondrial gene loci for four outgroup taxa and 48 ingroup representing 30 (23%) taxa of Charinidae. Charinidae are monophyletic, with Weygoldtia Miranda et al., 2018 sister to a monophyletic group comprising Charinus Simon, 1892 and Sarax Simon, 1892, neither of which are reciprocally monophyletic. Charinidae diverged from other amblypygid families in the Late Carboniferous, c. 318 Mya, on the supercontinent Pangaea. Weygoldtia diverged from the common ancestor of Charinus and Sarax during the Late Permian, c. 257 Mya, when changes in climate reduced tropical forests. The divergence of Charinus and Sarax coincides with the fragmentation of Pangaea, c. 216 Mya. Sarax colonized South-East Asia via Australia. The charinid fauna of New Caledonia originated before the Oligocene, when the island separated from Australia, c. 80 Mya.

Molecular phylogeny and evolution of Calaphidinae (Hemiptera: Aphididae).

Calaphidinae is the second-largest subfamily in the family Aphididae. Despite their species diversity and some taxonomic controversy, no phylogenetic studies have been conducted on them thus far. Herein, we report the first molecular phylogeny of Calaphidinae and two related lineages, Phyllaphidinae and Saltusaphidinae, based on five genes (3418 bp) for 126 taxa. Maximum parsimony, maximum-likelihood and Bayesian inference phylogenetic analyses were performed on the multilocus dataset. Divergence time estimation, biogeographical reconstruction, ancestral host plant reconstruction and PhyloType analyses were performed to identify evolutionary trends in Calaphidinae. Our phylogenetic results lead to several conclusions: Phyllaphidinae is a sister group to Calaphidinae s.l.; Calaphidinae is paraphyletic with respect to the former "Saltusaphidinae"; the ingroup clade was subdivided into nine newly recognized lineages; and three subtribes of Calaphidinae (Monaphidina, Calaphdina and Panaphidina) and many genera were not recovered as monophyletic. A new classification is proposed with eight tribal divisions that reflect our phylogenetic results, including three new tribes (Pterocallidini trib.n., Pseudochromaphidini trib.n. and Shivaphidini trib.n.) and three new statuses (Saltusaphidini stat.n., Therioaphidini stat.n. and Myzocallidini stat.n.). The ancestral reconstruction results imply that the ingroup taxa's common ancestor originated in the Eastern Palaearctic and might have fed on Fagaceae in the Late Cretaceous. Later, multiple host shifts and an expanding geographical distribution led to the current species diversity of Calaphidinae. Our reconstructions suggest that species diversification cannot solely be explained by speciation via host shifts and that geographical isolation probably also played a key role. Our results provide new insight into the natural classification and history of the host plant associations and biogeography of Calaphidinae s.l.

DrosoPhyla: Resources for Drosophilid Phylogeny and Systematics.

The vinegar fly Drosophila melanogaster is a pivotal model for invertebrate development, genetics, physiology, neuroscience, and disease. The whole family Drosophilidae, which contains over 4,400 species, offers a plethora of cases for comparative and evolutionary studies. Despite a long history of phylogenetic inference, many relationships remain unresolved among the genera, subgenera, and species groups in the Drosophilidae. To clarify these relationships, we first developed a set of new genomic markers and assembled a multilocus data set of 17 genes from 704 species of Drosophilidae. We then inferred a species tree with highly supported groups for this family. Additionally, we were able to determine the phylogenetic position of some previously unplaced species. These results establish a new framework for investigating the evolution of traits in fruit flies, as well as valuable resources for systematics.

Species delimitation and phylogenetic analyses reveal cryptic diversity within Cerebratulus marginatus (Nemertea: Pilidiophora)

The heteronemertean Cerebratulus marginatus Renier, 1804 is a well-known representative of the family Lineidae (Nemertea) inhabiting sandy or muddy sediments throughout the world. Due to its wide geographic distribution and the lack of distinctive external morphological features among different populations, the species has been considered cosmopolitan. Increasing evidence indicates that nemertean species, including Cerebratulus marginatus, described based on morphology are often not supported by genetic data and represent complexes of cryptic species, rather than cosmopolitan species. Here, we use species delimitation and phylogenetic analyses to evaluate the speciation patterns within the putative C. marginatus species complex. We examined 25 specimens morphologically ascribed to C. marginatus and closely related species using a multilocus dataset including two mitochondrial markers (16S rRNA, COI) and three nuclear markers (18S rRNA, 28S rRNA, H3). Our analyses identified at least four supported independent lineages among the populations analysed, indicating that samples identified as C. marginatus might represent indeed a complex of cryptic species. Therefore, our results do not support the cosmopolitanism of C. marginatus and provide additional evidence suggesting that species identifications in nemerteans should not be based exclusively on morphological or anatomical features which do not accurately reflect evolutionary patterns. Our study highlights the importance of including molecular data in addition to morphological information to accurately delimit species boundaries, thereby improving nemertean taxonomy by providing a more accurate picture of the true species diversity and geographic distribution of this typically neglected phylum.

Multilocus phylogenies reveal three new truffle-like taxa and the traces of interspecific hybridization in Octaviania (Boletaceae, Boletales)

ABSTRACTAmong many convergently evolved sequestrate fungal genera in Boletaceae (Boletales, Basidiomycota), the genus Octaviania is the most diverse. We recently collected many specimens of Octaviania subg. Octaviania, including several undescribed taxa, from Japan and the Americas. Here we describe two new species in subgenus Octaviania, O. tenuipes and O. tomentosa, from temperate to subtropical evergreen Fagaceae forests in Japan based on morphological observation and robust multilocus phylogenetic analyses (nrDNA ITS and partial large subunit [LSU], translation elongation factor 1-α gene [TEF1] and the largest subunit of RNA polymerase II gene [RPB1]). Based on specimens from the Americas as well as studies of the holotype, we also taxonomically re-evaluate O. asterosperma var. potteri. Our analysis suggests that O. asterosperma var. potteri is a distinct taxon within the subgenus Octaviania so we recognize this as O. potteri stat. nov. We unexpectedly collected O. potteri specimens from geographically widespread sites in the USA, Japan and Colombia. This is the first verified report of Octaviania from the South American continent. Our molecular analyses also revealed that the RPB1 sequence of one O. tenuipes specimen was identical to that of a closely related species, O. japonimontana, and that one O. potteri specimen from Minnesota had an RPB1 sequence of an unknown species of O. subg. Octaviania. Additionally, one O. japonimontana specimen had an unusually divergent TEF1 sequence. Gene-tree comparison and phylogenetic network analysis of the multilocus dataset suggest that these heterogenous sequences are most likely the result of previous inter- and intra-specific hybridization. We hypothesize that frequent hybridization events in Octaviania may have promoted the high genetic and species diversity found within the genus.

Disjoint Tree Mergers for Large-Scale Maximum Likelihood Tree Estimation

The estimation of phylogenetic trees for individual genes or multi-locus datasets is a basic part of considerable biological research. In order to enable large trees to be computed, Disjoint Tree Mergers (DTMs) have been developed; these methods operate by dividing the input sequence dataset into disjoint sets, constructing trees on each subset, and then combining the subset trees (using auxiliary information) into a tree on the full dataset. DTMs have been used to advantage for multi-locus species tree estimation, enabling highly accurate species trees at reduced computational effort, compared to leading species tree estimation methods. Here, we evaluate the feasibility of using DTMs to improve the scalability of maximum likelihood (ML) gene tree estimation to large numbers of input sequences. Our study shows distinct differences between the three selected ML codes—RAxML-NG, IQ-TREE 2, and FastTree 2—and shows that good DTM pipeline design can provide advantages over these ML codes on large datasets.

Geography is more important than life history in the recent diversification of the tiger salamander complex

The North American tiger salamander species complex, including its best-known species, the Mexican axolotl, has long been a source of biological fascination. The complex exhibits a wide range of variation in developmental life history strategies, including populations and individuals that undergo metamorphosis; those able to forego metamorphosis and retain a larval, aquatic lifestyle (i.e., paedomorphosis); and those that do both. The evolution of a paedomorphic life history state is thought to lead to increased population genetic differentiation and ultimately reproductive isolation and speciation, but the degree to which it has shaped population- and species-level divergence is poorly understood. Using a large multilocus dataset from hundreds of samples across North America, we identified genetic clusters across the geographic range of the tiger salamander complex. These clusters often contain a mixture of paedomorphic and metamorphic taxa, indicating that geographic isolation has played a larger role in lineage divergence than paedomorphosis in this system. This conclusion is bolstered by geography-informed analyses indicating no effect of life history strategy on population genetic differentiation and by model-based population genetic analyses demonstrating gene flow between adjacent metamorphic and paedomorphic populations. This fine-scale genetic perspective on life history variation establishes a framework for understanding how plasticity, local adaptation, and gene flow contribute to lineage divergence. Many members of the tiger salamander complex are endangered, and the Mexican axolotl is an important model system in regenerative and biomedical research. Our results chart a course for more informed use of these taxa in experimental, ecological, and conservation research.

Large‐scale diversity reassessment, evolutionary history, and taxonomic revision of the green macroalgae family Udoteaceae (Bryopsidales, Chlorophyta)

AbstractUdoteaceae is a morphologically diverse family of the order Bryopsidales. Despite being very widespread geographically, this family is little known as compared with the closely related Halimedaceae or Caulerpaceae. Using the most extensive Udoteaceae collection to date and a multilocus genetic data set (tufA, rbcL, and 18S rDNA), we reassessed the species diversity of the family, as well as the phylogenetic relationships, the diagnostic morphoanatomical characters, and evolutionary history of its genera, toward a proposed taxonomic revision. Our approach included a combination of molecular and morphological criteria, including species delimitation methods, phylogenetic reconstruction, and mapping of trait evolution. We successfully delimited 62 species hypotheses, of which 29 were assigned (existing) species names and 13 represent putative new species. Our results also led us to revise the genera Udotea s.s., Rhipidosiphon s.s., and Chlorodesmis s.s., to validate the genus Rhipidodesmis, and to propose three new genera: Glaukea gen. nov., Ventalia gen. nov., and Udoteopsis gen. nov. We also identified two large species complexes, which we refer to as the “Penicillus–Rhipidosiphon–Rhipocephalus–Udotea complex” and the “Poropsis–Penicillus–Rhipidodesmis complex”. Using a time‐calibrated phylogeny, we estimated the origin of the family Udoteaceae at Late Triassic (ca. 216 Ma), whereas most of the genera originated during Paleogene. Our morphological inference results indicated that the thallus of the Udoteaceae ancestor was likely entirely corticated and calcified, composed of a creeping axis with a multisiphonous stipe and a pluristromatic flabellate frond. The frond shape, cortication, and calcification are still symplesiomorphies for most extant Udoteaceae genera and represent useful diagnostic characters.

Species Tree Estimation from Gene Trees by Minimizing Deep Coalescence and Maximizing Quartet Consistency: A Comparative Study and the Presence of Pseudo Species Tree Terraces.

Species tree estimation from multilocus data sets is extremely challenging, especially in the presence of gene tree heterogeneity across the genome due to incomplete lineage sorting (ILS). Summary methods have been developed which estimate gene trees and then combine the gene trees to estimate a species tree by optimizing various optimization scores. In this study, we have extended and adapted the concept of phylogenetic terraces to species tree estimation by "summarizing" a set of gene trees, where multiple species trees with distinct topologies may have exactly the same optimality score (i.e., quartet score, extra lineage score, etc.). We particularly investigated the presence and impacts of equally optimal trees in species tree estimation from multilocus data using summary methods by taking ILS into account. We analyzed two of the most popular ILS-aware optimization criteria: maximize quartet consistency (MQC) and minimize deep coalescence (MDC). Methods based on MQC are provably statistically consistent, whereas MDC is not a consistent criterion for species tree estimation. We present a comprehensive comparative study of these two optimality criteria. Our experiments, on a collection of data sets simulated under ILS, indicate that MDC may result in competitive or identical quartet consistency score as MQC, but could be significantly worse than MQC in terms of tree accuracy-demonstrating the presence and impacts of equally optimal species trees. This is the first known study that provides the conditions for the data sets to have equally optimal trees in the context of phylogenomic inference using summary methods. [Gene tree; incomplete lineage sorting; phylogenomic analysis, species tree; summary method.].

Diversity of Diaporthe species associated with melanose disease on citrus trees in Jiangxi Province, China

Citrus melanose is an important disease that seriously damages citrus production. It is caused by Diaporthe species, which are known as important pathogens, endophytes or saprobes with broad host ranges. Over 170 species of Diaporthe were reported around the world and the complexity of species structures makes it hard to be identified. Phylogenetic analysis based on translation elongation factor 1-ɑ (TEF1) gene, s-tubulin (TUB) gene, calmodulin (CaM) gene, histone (HIS) gene and ITS locus was a reliable method to identify Diaporthe species In this study, the species structure of Diaporthe species from different tissues within individual trees was investigated. Altogether, 494 single spore isolates were obtained, and 92 of them were selected to sequence the five loci (ITS, TEF1, HIS, TUB, CaM) for phylogenetic analysis. Based on the combined multi-locus dataset, phylogenetic trees constructed by using two methods (MP and ML) showed that D. citri was the dominant species no matter on fruits, leaves or twigs. Diaporthe species showed the greatest diversity on fruits and the least diversity on twigs. D. eres was only isolated from twigs. The results of this study may enhance our understanding for the species diversity of Diaporthe species on different tissues from same trees, such information would be useful for the control of Diaporthe diseases in the future.

Sinuicella denisonii, a new genus and species in thePeltigeraceaefrom western North America

AbstractThe new genusSinuicella, an early successional lichen, was found on bare soil in Oregon, USA. The thallus is minute fruticose, grey to nearly black, branching isotomic dichotomous, branches round, 20–90 μm wide in water mount. The cortex is composed of interlocking cells shaped like jigsaw puzzle pieces. Spores are hyaline, 1-septate, 25–40(–50) × 6.5–9(–11) μm. Maximum likelihood phylogenetic analyses on multilocus data sets, first spanning the entire orderPeltigeralesand then restricted toPeltigeraceaewith extended sampling fromSolorinaandPeltigera, revealed the placement ofSinuicellaoutside of currently recognized genera, sister toPeltigera, with high support. Based on the phylogenetic, morphological and ecological distinctness ofSinuicella, we formally introduce a new genus represented by the single speciesS. denisonii. The cyanobiont ofS. denisoniiisNostocfrom phylogroup XL, Clade 2, Subclade 3 based on therbcLXmarker.

Investigating species boundaries in Colletotrichum

Colletotrichum is one of the most important plant pathogenic genera that is responsible for numerous diseases which can have a profound impact on the agricultural sector. Species delineation is difficult due to a lack of distinctive phenotypic variation. Therefore, in this study three different genomic approaches based on phylogenetic, evolutionary and coalescent-based methods are applied to establish robust species boundaries. The reliability of five different DNA barcodes was also assessed to provide further insights into species delineation. The ITS region can resolve the placement of taxa up to the species complex level. The GAPDH and TUB2 markers are determined to be the most informative for most complexes. However, no single marker could discriminate between species in all complexes, therefore different molecular approaches based on multi-locus datasets are recommended. This is the first study to provide an estimated divergence time for all species complexes in Colletotrichum. The estimated divergence time for species complexes ranged between 4.8 to 32.2 MYA. Based on the high level of congruent results obtained from the different molecular approaches, a new species complex, the Colletotrichum agaves complex is introduced. This complex consists of five taxa which are characterised by the presence of straight or slightly curved conidia with obtuse apices. This study shows that coalescent approaches and multi-locus phylogeny are crucial to establish species boundaries in Colletotrichum. The taxonomic placement of three singleton taxa Colletotrichum axonopodi, C. cariniferi and C. parallelophorum is revised. We accept 248 species and provide recommendations regarding species boundaries in the graminicola–caudatum complex.