Morphology,phylogeny and taxonomy of South American bothropoid pitvipers (Serpentes,Viperidae)

Carrasco, P.A., Mattoni, C.I., Leynaud, G.C. & Scrocchi, G.J. (2012). Morphology, phylogeny and taxonomy of South American bothropoid pitvipers (Serpentes, Viperidae). —Zoologica Scripta, 41, 109–124. South American bothropoids comprise a monophyletic and greatly diverse group of pitvipers that were initially included in the genus Bothrops and later assigned to five genera. Until recently, most phylogenetic analyses of bothropoids used exclusively mitochondrial DNA sequences, whereas few of them have included morphological traits. Moreover, the systematic affinities of some species remain unclear. In this study, we performed a parsimony analysis of morphological data obtained from the examination of 111 characters related to lepidosis, colour pattern, osteology, and hemipenial morphology of 35 of the 48 species that compose the bothropoid group. The morphological data analysed contain novel information about several species, including the incertae sedis. Morphology was analysed separately and combined with 2393 molecular characters obtained from published sequences of four mitochondrial genes. Five characters of the ecology were also included. A sensitivity analysis was performed using different weighting criteria for the characters. The congruence among different sources of evidence was evaluated through partitioned and total evidence analyses, the analyses of reduced datasets and the use of incongruence length difference test. With few exceptions, results showed groups of species similar to those obtained in previous studies; however, incongruences between morphological and molecular characters, and within the molecular partition, were revealed. This conflict affects the relationship between particular groups of species, leading to alternative phylogenetic hypotheses for bothropoids: hierarchical radiation or two major lineages within the group. The results also showed that Bothrops sensu stricto is paraphyletic. We discuss previous taxonomic approaches and, considering both phylogenetic hypotheses, we propose an arrangement that rectifies the paraphyly of Bothrops: maintaining Bothrocophias, assigning Bothrops andianus to this genus; and recognising the sister clade as Bothrops, synonymising Bothriopsis, Bothropoides and Rhinocerophis.     

Morphological and molecular evidence for phylogeny and classification of South American pitvipers,genera Bothrops,Bothriopsis, and Bothrocophias (Serpentes: Viperidae)

Species in the genus Bothrops s. l. are extraordinarily variable in ecology and geography, compared with other genera in the subfamily Crotalinae. In contrast to the trend of splitting large and variable groups into smaller, more ecologically and phenotypically cohesive genera, the genus Bothrops has remained speciose. In addition, previous phylogenetic analyses have found Bothrops to be paraphyletic with respect to the genus Bothriopsis. Taxonomic arguments exist for synonymizing Bothriopsis with Bothrops, and for splitting Bothrops into smaller genera, but the greatest hindrance to taxonomic revision has been incomplete phylogenetic information. We present a phylogeny of Bothrops, Bothriopsis, and Bothrocophias based on 85 characters of morphology and 2343 bp of four mitochondrial gene regions, and with significantly greater taxonomic coverage than previous studies. The combined data provide improved support over independent datasets, and support the existence of discrete species groups within Bothrops. The monophyly and distinctness of these groups warrant recognition at the generic level, and we propose a new taxonomic arrangement to reflect these findings. © 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 156 , 617–640.     

Profilings of subproteomes of lectin-binding proteins of nine Bothrops venoms reveal variability driven by different glycan types

Snake venom proteomes have long been investigated to explore a multitude of biologically active components that are used for prey capture and defense, and are involved in the pathological effects observed upon mammalian envenomation. Glycosylation is a major protein post-translational modification in venoms and contributes to the diversification of proteomes. We have shown that Bothrops venoms are markedly defined by their content of glycoproteins, and that most N-glycan structures of eight Bothrops venoms contain sialic acid, while bisected N-acetylglucosamine was identified in Bothrops cotiara venom. To further investigate the mechanisms involved in the generation of different venoms by related snakes, here the glycoproteomes of nine Bothrops venoms (Bothrops atrox, B. cotiara, Bothrops erythromelas, Bothrops fonsecai, B. insularis, Bothrops jararaca, Bothrops jararacussu, Bothrops moojeni and Bothrops neuwiedi) were comparatively analyzed by enrichment with three lectins of different specificities, recognizing bisecting N-acetylglucosamine- and sialic acid-containing glycoproteins, and mass spectrometry. The lectin capture strategy generated venom fractions enriched with several glycoproteins, including metalloprotease, serine protease, and L- amino acid oxidase, in addition to various types of low abundant enzymes. The different contents of lectin-enriched proteins underscore novel aspects of the variability of the glycoprotein subproteomes of Bothrops venoms and point to the role of distinct types of glycan chains in generating different venoms by closely related snake species.     

Systématique moléculaire des phlebotominae : étude pilote. paraphylie du genre phlebotomus

Phylogenetic relationships among Phlebotominae were inferred through a pilot study using parsimony analysis of the D2 domain of ribosomal DNA sequences: 455 pairs of bases were sequenced in nine species of Phlebotomine sandflies which belong to the genera Lutzomyia, Phlebotomus and Sergentomyia. Two taxa are used as outgroups: Psychoda sp. and Nemapalpus flavus which is the sister group of the Phlebotominae. The South American genus Lutzomyia appears to be monophyletic. The Mediterranean species Sergentomyia dentata is its sister group and is not clustered with the Old World genus Phlebotomus. The latter is a paraphyletic genus with an early individualisation of the branch including the closely related subgenera Phlebotomus and Paraphlebotomus, and a late individualisation of the subgenus Larroussius. These results have some consequences on the biogeography of the leishmaniasis in the Old World.     

Using 18S rDNA to resolve diaptomid copepod (Copepoda: Calanoida: Diaptomidae) phylogeny: an example with the North American genera

The phylogenetic relationships among the numerous genera of diaptomid copepods remain elusive due to difficulties in obtaining sufficient numbers of phylogenetically informative morphological characters for cladistic analysis. Molecular phylogenetic techniques offer high potential to resolve phylogenetic relationships in the absence of sufficient morphological characters because of the ease in which many characters can be unambiguously coded. I present the first molecular phylogeny for diaptomid copepod genera using 18S rDNA. Specifically, I test Light’s (1939) hypothesis regarding the interrelationships among the North American diaptomid genera. The 18S phylogeny is remarkably consistent with Light’s hypothesis. The endemic North American genera represent a monophyletic group exclusive of the non-endemic genera. Moreover, his hypothesized basal genus for the North America genera, Hesperodiaptomus, is the basal genus in this analysis. However, his Leptodiaptomus group is not reciprocally monophyletic with his Hesperodiaptomus group, but is rather a derived member of the latter group. Finally, the genus Mastigodiaptomus is found to be more closely allied with the non-endemic genera, as Light suggested. This phylogeny contributes heavily to the understanding of phylogenetic relationships among North American diaptomids and has large implications for the systematics of diaptomids in general. The use of 18S rDNA sequences in phylogenetic analyses of diaptomid copepods can be used to confirm the monophyly of recognized genera, the interrelationships among genera, and subsequent biogeographic interpretation of the family’s diversification. The use of molecular data, such as 18S rDNA sequences, to test phylogenetic hypotheses based on a very limited number of morphological characters will be a particularly useful approach to phylogenetic analysis in this system.     

Ontogenetic Variation in Biological Activities of Venoms from Hybrids between Bothrops erythromelas and Bothrops neuwiedi Snakes

Lance-headed snakes are found in Central and South America, and they account for most snakebites in Brazil. The phylogeny of South American pitvipers has been reviewed, and the presence of natural and non-natural hybrids between different species of Bothrops snakes demonstrates that reproductive isolation of several species is still incomplete. The present study aimed to analyze the biological features, particularly the thrombin-like activity, of venoms from hybrids born in captivity, from the mating of a female Bothrops erythromelas and a male Bothrops neuwiedi, two species whose venoms are known to display ontogenetic variation. Proteolytic activity on azocoll and amidolytic activity on N-benzoyl-DL-arginine-p-nitroanilide hydrochloride (BAPNA) were lowest when hybrids were 3 months old, and increased over body growth, reaching values similar to those of the father when hybrids were 12 months old. The clotting activity on plasma diminished as hybrids grew; venoms from 3- and 6-months old hybrids showed low clotting activity on fibrinogen (i.e., thrombin-like activity), like the mother venom, and such activity was detected only when hybrids were older than 1 year of age. Altogether, these results point out that venom features in hybrid snakes are genetically controlled during the ontogenetic development. Despite the presence of the thrombin-like enzyme gene(s) in hybrid snakes, they are silenced during the first six months of life.     

A key to the Mexican and Central America Genera of Anthonomini (Curculionidae,Curculioninae)

Presently the only keys available for identification of genera of Anthonomini are limited to those of the United States of America and Canada. A dichotomous key is presented to identify all genera of Mexican and Central American Anthonomini. Previous keys do not include the genera Achia, Botanebius, Loncophorus, Loncophorellus and Melexerus. A brief synopsis is given for each genus and photographs of representative species are included.     

Weighting and Congruence: A Case Study Based on Three Mitochondrial Genes in Pitvipers

Weighting strategies in a total evidence approach are often conducted in order to remove homoplasy, with the implicit hope to increase congruence between data partitions. Incongruence was measured using the test of Farriset al.(J. S. Farris, M. Källersjö, A. G. Kluge, and C. Bult, 1995,Cladistics10, 315–319) within and between three mitochondrial genes from pitvipers (Crotalinae) in partitioning each codon position for the coding genes. Incongruence between codon positions of a gene can be worse than incongruence between genes at analogous positions. Third positions of codons behave very differently in terms of incongruence from one gene to another while showing similar patterns in saturation tests. Instead of removing characters in order to discard homoplasy, which is hopeless and does not increase general congruence, we advocate for the removal of those substitutions that are incongruent with the rest. The genusCalloselasmaand its sister group the genusHypnaleare the most basal Crotalinae. Asiatic pitvipers are paraphyletic, while American pitvipers are monophyletic.     

Revision of the bulbous Iridaceae of North America

The native bulbous Iridaceae of North America excluding Mexico comprise seven species in four genera. The type species ofAlophia Herb. (1836) is shown to belong toEustylis Engelm. & Gray (1836), hence the latter genus is reduced to synonymy. The well known genusAlophia (sensu auct. non Herb.) (Herbertia Sweet) now takes the nameTrifurcia. As a result of the new interpretation ofAlophia, new combinations are made for the North American and Latin American species ofTrifurcia andAlophia. New chromosome counts are given forNemastylis floridana (2n = 56) andTrifurcia lahue (2n = 56).     

Considering gene flow when using coalescent methods to delimit lineages of North American pitvipers of the genus Agkistrodon

Examining species diversity and mechanisms of speciation using coalescent models provides a framework for how regional diversity is accrued, even in well‐studied areas such as the Nearctic. It is likely, that gene flow among closely‐related species with adjacent distributions may be common. However, the absence of gene flow is a primary assumption of many phylogeographical methods that produce species trees and delimit species using Bayesian or likelihood functions in a coalescent framework. In the present study, we examine delimitation when gene flow between species is present using empirical datasets from two species of North American pitvipers of the genus Agkistrodon. We also use niche modelling to determine whether these young lineages occur in distinct environmental niches. To manage the problem of gene flow between species, we first identify admixed individuals, demonstrate that gene flow has occurred, and then identify the impact of alternative population assignments of admixed individuals on delimitation posterior probabilities. In addition, we examine the influence of mitochondrial genes relative to other loci combined in coalescent analyses that delimit species. Here, we find that the copperheads (Agkistrodon contortrix) and the cottonmouths (Agkistrodon piscivorus) are each composed of two distinct species, with each occupying different niches. Importantly, we find that species can be delimited when the amount of gene flow between lineages is low, although the methods are acutely sensitive to population assignment of individuals. © 2014 The Linnean Society of London     

An investigation of the cranial evolution of Asian pitvipers (Serpentes: Crotalinae), with comments on the phylogenetic position of Peltopelor macrolepis

Guo, P., Jadin, R.C., Malhotra, A. and Li, C. 2009. An investigation of the cranial evolution of Asian pitvipers (Serpentes: Crotalinae), with comments on the phylogenetic position of Peltopelor macrolepis—Acta Zoologica (Stockholm) 91 : 402–407. We investigated the evolution of 12 cranial characters of 31 species of Asian pitvipers by examining the character state changes on a consensus tree modified from broadly consistent molecular results. We found that these characters appear stable with only one intraspecific polymorphism. Nine of the 12 characters form useful synapomorphies, whereas three are ambiguous and evolutionarily plastic. Clades that are supported with numerous apomorphies are the Trimeresurus group [consisting of the recently defined genera Trimeresurus sensu Malhotra and Thorpe (2004) , Parias, Popeia, Viridovipera, Himalayophis, and Cryptelytrops] and the genera Protobothrops, Parias, and Viridovipera. Two species previously considered as congeners but now known to be distantly related, Ovophis monticola and ‘Ovophis’okinavensis, have nearly identical character states, demonstrating substantial convergence in cranial characters. Finally, we attempt to infer the phylogenetic position of Peltopelor macrolepis by comparing its cranial features with that of other pitvipers.     

PEP car☐ykinase containing species in the Brachiaria group of the subfamily panicoideae

In the Gramineae, a survey of species among the Brachiaria group of the subfamily Panicoideae, tribe Paniceae revealed that they are PEP car?ykinase containing species. This group includes the genera Brachiaria, Eriochloa and Urochloa. With the exception of the genus Panicum, these are the only genera within the Panicoideae found to contain PEP car?ykinase species. It is suggested that the PEP car?ykinase species of the genus Panicum, P. fasciculatum, P. maximum, P. molle and P. texanum, might be best placed in the Brachiaria group.     

Pericarp structure and phylogeny of tribe Mentheae (Lamiaceae)

Pericarp structure is investigated in 143 specimens belonging to 59 out of ca. 65 genera and 134 out of ca. 2,000 species of Lamiaceae subfamily Nepetoideae tribe Mentheae. Some of the data presented by earlier authors are believed to be incorrect. The variation in pericarp structure has been found to be strongly correlated to the variations in DNA and gross morphological characters. The groups of two-staminate genera (Salvia, etc.) and four-staminate genera of Salviinae (Lepechinia and Chaunostoma) can be distinguished from each other and from other labiates by differences in pericarp structure. The other two subtribes, Menthinae and Nepetinae, differ considerably, but their variations strongly overlap. The differences in pericarp structure also suggest that Menthinae can be divided into three main monophyletic groups based on other data: (1) the main bulk of the tribe, (2) a group of Prunella and Cleonia and perhaps also Horminum, and (3) the characteristic genus Lycopus. Pericarp data disagree with a suggestion that the genus Melissa should be included in the subtribe Salviinae. Modifications of the subtribal classification of Mentheae are discussed.     

Ascaridoid nematodes of amphibians and reptiles: Ophidascaris Baylis, 1920

The genus Ophidascaris is revised and divided into five groups of species. A key for the species groups is provided. Group 1 (‘filaria’ group) occurs in pythons and a key is provided for differentiating eight species based on fresh and preserved specimens and on developmental patterns. O. papillifera (Linstow, 1898) is redescribed from the type-specimens and is considered to be close or identical to O. niuginiensis, for which Candoia carinatus is recorded as a new host. A key for the differentiation of species in Groups 2 to 5 is based on preserved specimens only. Group 2 (‘obconica’ group) contains (i) O. obconica and O. trichuriformis [= caballeroi] in South American colubrids (further investigation will probably show that O. trichuriformis is a synonym of O. obconica); new host records are Xenodon severus, X. neuwiedii, X. colubrinus, Leptodeira annulata, Thamnodynastes pallidus, Leimadophis poecilogyrus and Boa constrictor; (ii) O. ashi n. sp. (new species name for ‘O. labiatopapillosa’) in North American colubrids, new host records are Nerodia valida, Heterodon nasicus and Storeria occipitomaculata; (iii) O. mombasica in African colubrids; new host records are Psammophis phillipsii and P. sibilans; (iv) O. solenopoion in Madagascan colubrids; (v) O. pyrrhus in Australian elapids; new host records are Cacophis squamulosus, Cryptophis nigrescens, Demansia atra, D. olivacea, Hemiaspis signata, Hoplocephalus bitorquatus, H. stephensi, H. bungaroides and Tropedechis carinatus, it is also recorded in Australia in the colubrid Styporhyncus mairii (new host record) and in Demansia papuensis papuensis and D. olivacea papuensis in Papua New Guinea; (vi) O. piscatori in Asian colubrids; (vii) O. excavata [? = schikhobalovi] in Agkistrodon spp. and possibly other aquatic snakes in Asia; new host record is Agkistrodon halys blomhoffi. Group 3 (‘radiosa’ group) in African viperids contains O. radiosa [ = intorta] in Bitis spp. Specimens from Atheris nitschei, Causus rhombeatus and the colubrid Boaedon lineatus were similar, but showed differences indicating possibility of other species in this group. Group 4 (‘najae’ group) in African elapids and Asian elapids and colubrids contains O. najae [ = daubaylisi]; new host records are Ophiophagus hannah, Boiga cyanea, Elaphe carinata. Group 5 (‘arndti’ group) in South American crotalids and colubrids contains O. arndti [ = travassosi and sprenti] in Crotalus spp. and Bothrops spp.; new host record is B. atrox in Panama. No morphological differentiation except size could be detected between O. arndti in crotalines and O. sicki in colubrids, but in view of difference in the feeding habits of their host, both species names were tentatively sustained; new host records for O. sicki are Xenodon neuwiedii, Leimadophis poecilogyrus, Pseudoboa cloelia, Philodryas patagoniensis and Micrurus frontalis; O. ochoteranai was regarded as a species inquirenda. The following species previously placed in Amplicaecum are placed in Ophidascaris; excavata Hsu & Hoeppli, 1931; schikhobalovi Mozgovoy, 1950; robertsi Sprent & Mines, 1960; longispiculum Oshmarin & Demshin, 1972; orientalis Wang, 1965. The position of Ophidascaris in relation to other ascaridoids is discussed: it is placed within the subfamily Ascaridinae sensu Sprent (1983) containing all other ascaridoids of terrestrial animals. It is concluded that Ophidascaris is in a relatively recent stage of evolution. The most likely centre of dispersal for the genus appears to have been Central Africa with spread in one direction to Asia and thence to the New World and in another direction to Madagascar and Australia.     

Structural bases for a complete myotoxic mechanism: Crystal structures of two non-catalytic phospholipases A2-like from Bothrops brazili venom

Bothrops brazili is a snake found in the forests of the Amazonian region whose commercial therapeutic anti-bothropic serum has low efficacy for local myotoxic effects, resulting in an important public health problem in this area. Catalytically inactive phospholipases A₂-like (Lys49-PLA₂s) are among the main components from Bothrops genus venoms and are capable of causing drastic myonecrosis. Several studies have shown that the C-terminal region of these toxins, which includes a variable combination of positively charged and hydrophobic residues, is responsible for their activity. In this work we describe the crystal structures of two Lys49-PLA₂s (BbTX-II and MTX-II) from B. brazili venom and a comprehensive structural comparison with several Lys49-PLA₂s. Based on these results, two independent sites of interaction were identified between protein and membrane which leads to the proposition of a new myotoxic mechanism for bothropic Lys49-PLA₂s composed of five different steps. This proposition is able to fully explain the action of these toxins and may be useful to develop efficient inhibitors to complement the conventional antivenom administration.     

Floral microcharacters in Lessingianthus (Vernonieae,Asteraceae) and their taxonomic implications

Micromorphological characters of 113 species of the South American genus Lessingianthus H.Rob. (Vernonieae, Asteraceae) were analyzed to evaluate their reliability as taxonomic markers. The corolla pubescence was studied in detail for the first time in the genus. Glandular and non-glandular trichomes were studied and described. The trichome types allow differentiation among related species, but do not reflect the relationships among species groups. On the other hand, the basal stylar node only occurs in a distinctive group of species within the genus. The apical appendage of the anthers is non-glandular in all species of the genus. Several character states analyzed in Lessingianthus are often widespread in other related genera of Vernonieae. Therefore, Lessingianthus can be circumscribed only by a combination of micro- and macromorphological features.     

Comparison of Phylogeny,Venom Composition and Neutralization by Antivenom in Diverse Species of Bothrops Complex

In Latin America, Bothrops snakes account for most snake bites in humans, and the recommended treatment is administration of multispecific Bothrops antivenom (SAB – soro antibotrópico). However, Bothrops snakes are very diverse with regard to their venom composition, which raises the issue of which venoms should be used as immunizing antigens for the production of pan-specific Bothrops antivenoms. In this study, we simultaneously compared the composition and reactivity with SAB of venoms collected from six species of snakes, distributed in pairs from three distinct phylogenetic clades: Bothrops, Bothropoides and Rhinocerophis. We also evaluated the neutralization of Bothrops atrox venom, which is the species responsible for most snake bites in the Amazon region, but not included in the immunization antigen mixture used to produce SAB. Using mass spectrometric and chromatographic approaches, we observed a lack of similarity in protein composition between the venoms from closely related snakes and a high similarity between the venoms of phylogenetically more distant snakes, suggesting little connection between taxonomic position and venom composition. P-III snake venom metalloproteinases (SVMPs) are the most antigenic toxins in the venoms of snakes from the Bothrops complex, whereas class P-I SVMPs, snake venom serine proteinases and phospholipases A₂ reacted with antibodies in lower levels. Low molecular size toxins, such as disintegrins and bradykinin-potentiating peptides, were poorly antigenic. Toxins from the same protein family showed antigenic cross-reactivity among venoms from different species; SAB was efficient in neutralizing the B. atrox venom major toxins. Thus, we suggest that it is possible to obtain pan-specific effective antivenoms for Bothrops envenomations through immunization with venoms from only a few species of snakes, if these venoms contain protein classes that are representative of all species to which the antivenom is targeted.     

Molecular phylogenetics of tribe Synandreae, a North American lineage of lamioid mints (Lamiaceae)

The five mint genera Brazoria, Macbridea, Physostegia, Synandra and Warnockia (Lamioideae: Lamiaceae) are all North American endemics. Together with the monotypic European genus Melittis and the Asian genus Chelonopsis, these taxa have been classified as subtribe Melittidinae. Previous morphological studies have failed to uncover synapomorphic characters for this group. We sequenced the plastid trnL‐trnF region and trnS‐trnG spacer and the nuclear ribosomal 5S non‐transcribed spacer (5S‐NTS) to assess phylogenetic relationships within Melittidinae. Standard parsimony and direct optimization (POY) analyses show Melittis, the type genus of the subtribe, as sister to Stachys. Thus, the monophyly of subtribe Melittidinae is not supported either by molecular or morphological data. However, the North American endemics form a monophyletic group that can be recognized as the recircumscribed tribe Synandreae. The molecular relationships among these genera are corroborated by both morphological and cytological data. The expected close relationship between the south‐central endemics Warnockia and Brazoria and their sister relationship to the widespread genus Physostegia is confirmed. Nevertheless, most of the North American endemics are restricted to the south‐east of the continent. Dispersal westwards and northwards is correlated with an increase in chromosome numbers. No specific Eurasian origin (i.e., transatlantic or transpacific) can be determined, but Synandreae are clearly distinct from the large Stachys clade, and therefore represent a separate migration into North America. © The Willi Hennig Society 2007.     

Molecular-signature analyses support the establishment of the actinobacterial genus Sphaerimonospora (Mingma et al. 2016)

The genera Microbispora and Sphaerimonospora were examined for GyrB and RecA amino-acid signatures to determine whether molecular-signature analyses support the recent establishment of the genus Sphaerimonospora. The creation of Sphaerimonospora was based mainly upon morphological differences between Microbispora and Sphaerimonospora and the clustering of the type strains of the two genera in phylogenetic trees based on a multilocus sequence analysis. The molecular-signature analyses showed that all members of Sphaerimonospora can be distinguished from all members of Microbispora at 14 amino acid positions in the GyrB protein and at four positions in the shorter RecA protein. These amino acid differences can be used as signatures to differentiate the members of these genera from each other and thus provide support for the establishment of the genus Sphaerimonospora. This is the first demonstration of the use of molecular signatures to support the establishment of a new genus in the family Streptosporangiaceae. Following the transfer of Microbispora mesophila and Microbispora thailandensis from Microbispora to Sphaerimonospora, all species in the genus Microbispora are characterised by the insertion of a small, hydrophobic amino acid after position 208 in the GyrB protein. This insertion is absent from the GyrB protein of members of the genus Sphaerimonospora.     

Taxonomic update of Clitocybula sensu lato with a new generic classification

The taxonomy and phylogeny of the hydropoid clade (genera Clitocybula s.l., Megacollybia, Leucoinocybe gen. nov., Hydropus, Trogia, Gerronema, Porotheleum and Lignomphalia gen. nov.) in Europe is studied using morphological and molecular approaches; the first three genera in detail including all known European species. Only two European species remain in Clitocybula s.str., Clitocybula lacerata and Clitocybula familia. The European C. lacerata is a species complex which should be treated as C. lacerata agg. at the current state of knowledge. A neotype originating from type area was designated to fix the application of the name. The presence of American species Clitocybula abundans in Europe is insufficiently proved. “Clitocybula dryadicola ? belongs to the genus Hydropus, and Clitocybula tilieti has an unclear systematic position. The results showed that Megacollybia and Leucoinocybe represent independent genera separated from Clitocybula. The genus Leucoinocybe is validly published with two European species, Leucoinocybe lenta and Leucoinocybe taniae. “Clitocybula flavoaurantia” proved to be conspecific with the latter species. The genus Lignomphalia is published for “Pseudoomphalina lignicola”, a lignicolous omphalinoid species. The Indian “Clitocybula sulcata” is transferred to Leucoinocybe and “Clitocybula atrialba” to Gerronema. The first European records of Megacollybia marginata are published.