The first of 1000 equally most parsimonious trees (TL = 367; CI = 0.638; RI = 0.928; RC = 0.591) resulting from a parsimony analysis of the LSU (28S) sequence alignment. The bootstrap support values are indicated at the nodes (parsimony bootstrap / distance with HKY85 model bootstrap; only values >74%) and the scale bar represents the number of changes. Thickened branches reflect those branches present in the strict consensus tree. Orders are indicated in darker blue and orange blocks and family names in light blue and light brown blocks. Species names of interest to this study are shown in bold text. The tree was rooted to Dothidea sambuci (GenBank AY544691). 

The first of 1000 equally most parsimonious trees (TL = 367; CI = 0.638; RI = 0.928; RC = 0.591) resulting from a parsimony analysis of the LSU (28S) sequence alignment. The bootstrap support values are indicated at the nodes (parsimony bootstrap / distance with HKY85 model bootstrap; only values >74%) and the scale bar represents the number of changes. Thickened branches reflect those branches present in the strict consensus tree. Orders are indicated in darker blue and orange blocks and family names in light blue and light brown blocks. Species names of interest to this study are shown in bold text. The tree was rooted to Dothidea sambuci (GenBank AY544691). 

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The type species of the genus Tiarosporella, T. paludosa, is epitypified and confirmed as a member of the Botryosphaeriaceae. Based on morphology and DNA sequence data of the large subunit nuclear ribosomal RNA gene (LSU, 28S) and the internal transcribed spacers (ITS) and 5.8S rRNA gene of the nrDNA operon, the genus Tiarosporella is shown to be p...

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... 2012) and Macrophomina (Sarr et al. 2014). Phylogenetically however, it proved to be allied to genera in the complex in the Botryosphaeriaceae (Fig. 1), which was quite unexpected. Subram. & K. Ramakr., Journal of the Indian Botanical Society 36: 83 ...
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... phylogenies were generated; the first was based on 56 LSU sequences (including the outgroup Dothidea sambuci GenBank AY544681) and was used to determine the familial and ordinal relationships of the studied species (Fig. 1), the second was based on a combined ITS and LSU alignment of 44 isolates (including the outgroup Saccharata proteae strain CBS 115206) and was used to determine the genus relationships and species identification within the Botryosphaeriaceae (Fig. 2), and the third was based on a combined ITS, LSU, SSU, TEF and TUB alignment of 18 Darkera isolates and was used for species identification (Fig. 3). The first analysis (LSU) (including the outgroup sequence) and the resulting dataset of 773 characters, including alignment gaps which were treated as fifth base, consisted of 604 constant characters, 36 variable parsimony- uninformative characters and 133 parsimony-informative characters. The maximum of 1000 equally most parsimonious trees were retained (TL = 367; CI = 0.638; RI = 0.928; RC = 0.591), the first of which is presented in Fig. 1. The overall topology was identical between the distance tree (data not shown) and the presented parsimony tree (Fig. 1) with some minor rearrangements of terminal clades in the different families. Overall, the parsimony analysis yielded less well-supported nodes compared to the distance analysis. The Dermateaceae was well-supported in both analyses, whereas the Phacidiaceae was only supported in the distance analysis. The Darkera clade itself is well-supported in both analyses, although the deeper structure of the sub-clades of the Phacidiaceae collapses into a basal polytomy in the parsimony analysis (see strict consensus branches in Fig. 1). The Phyllostictaceae is well-supported in both analyses, whereas the Botryosphaeriaceae is strongly supported in the distance analysis (98 % bootstrap support) but less so in the parsimony analysis (76 % bootstrap support). The LSU phylogeny based on the current dataset alone does not provide a well-supported topology for the Botryosphaeriaceae and therefore the data was combined with ITS for the second analysis. The second analysis (combined ITS and LSU alignment) (including the outgroup sequence) and the resulting dataset of 1243 characters, including alignment gaps which were treated as fifth base, consisted of 976 constant characters, 92 variable parsimony-uninformative characters and 175 parsimony-informative characters. Twenty-two equally most parsimonious trees were obtained (TL = 619; CI = 0.577; RI = 0.809; RC = 0.467), the first of which is presented in Fig. 2. In this phylogeny, all genera that are presented by more than one strain or species are supported with a parsimony bootstrap support value of at least 80 %; the only exception is Diplodia which is split into two lineages without support for the connecting node. The tiarosporella-like strains are polyphyletic in the tree and therefore novel genera are introduced below to accommodate those not clustering in the Tiarosporella clade. Except for Tiarosporella tritici (= Eutiarosporella tritici , see below) and T. africana (= Eut. africana , see below), all species in the ITS-LSU phylogeny could be resolved. In the case of this exception, the two species can easily be distinguished based on their TEF or TUB sequences (data not shown). The third analysis (combined ITS, LSU, SSU, TEF and TUB alignment) was based on the resulting dataset of 2879 characters, including alignment gaps which were treated as fifth base, consisted of 2857 constant characters, 3 variable parsimony-uninformative characters and 19 parsimony-informative characters (TL = 22; CI = 1.0; RI = 1.0; RC = 1.0). Only a single most parsimonious tree was obtained, presented in Fig. 3, which clearly separated the strains belonging to Darkera picea from those belonging to D. parca ...
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... phylogenies were generated; the first was based on 56 LSU sequences (including the outgroup Dothidea sambuci GenBank AY544681) and was used to determine the familial and ordinal relationships of the studied species (Fig. 1), the second was based on a combined ITS and LSU alignment of 44 isolates (including the outgroup Saccharata proteae strain CBS 115206) and was used to determine the genus relationships and species identification within the Botryosphaeriaceae (Fig. 2), and the third was based on a combined ITS, LSU, SSU, TEF and TUB alignment of 18 Darkera isolates and was used for species identification (Fig. 3). The first analysis (LSU) (including the outgroup sequence) and the resulting dataset of 773 characters, including alignment gaps which were treated as fifth base, consisted of 604 constant characters, 36 variable parsimony- uninformative characters and 133 parsimony-informative characters. The maximum of 1000 equally most parsimonious trees were retained (TL = 367; CI = 0.638; RI = 0.928; RC = 0.591), the first of which is presented in Fig. 1. The overall topology was identical between the distance tree (data not shown) and the presented parsimony tree (Fig. 1) with some minor rearrangements of terminal clades in the different families. Overall, the parsimony analysis yielded less well-supported nodes compared to the distance analysis. The Dermateaceae was well-supported in both analyses, whereas the Phacidiaceae was only supported in the distance analysis. The Darkera clade itself is well-supported in both analyses, although the deeper structure of the sub-clades of the Phacidiaceae collapses into a basal polytomy in the parsimony analysis (see strict consensus branches in Fig. 1). The Phyllostictaceae is well-supported in both analyses, whereas the Botryosphaeriaceae is strongly supported in the distance analysis (98 % bootstrap support) but less so in the parsimony analysis (76 % bootstrap support). The LSU phylogeny based on the current dataset alone does not provide a well-supported topology for the Botryosphaeriaceae and therefore the data was combined with ITS for the second analysis. The second analysis (combined ITS and LSU alignment) (including the outgroup sequence) and the resulting dataset of 1243 characters, including alignment gaps which were treated as fifth base, consisted of 976 constant characters, 92 variable parsimony-uninformative characters and 175 parsimony-informative characters. Twenty-two equally most parsimonious trees were obtained (TL = 619; CI = 0.577; RI = 0.809; RC = 0.467), the first of which is presented in Fig. 2. In this phylogeny, all genera that are presented by more than one strain or species are supported with a parsimony bootstrap support value of at least 80 %; the only exception is Diplodia which is split into two lineages without support for the connecting node. The tiarosporella-like strains are polyphyletic in the tree and therefore novel genera are introduced below to accommodate those not clustering in the Tiarosporella clade. Except for Tiarosporella tritici (= Eutiarosporella tritici , see below) and T. africana (= Eut. africana , see below), all species in the ITS-LSU phylogeny could be resolved. In the case of this exception, the two species can easily be distinguished based on their TEF or TUB sequences (data not shown). The third analysis (combined ITS, LSU, SSU, TEF and TUB alignment) was based on the resulting dataset of 2879 characters, including alignment gaps which were treated as fifth base, consisted of 2857 constant characters, 3 variable parsimony-uninformative characters and 19 parsimony-informative characters (TL = 22; CI = 1.0; RI = 1.0; RC = 1.0). Only a single most parsimonious tree was obtained, presented in Fig. 3, which clearly separated the strains belonging to Darkera picea from those belonging to D. parca ...
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... phylogenies were generated; the first was based on 56 LSU sequences (including the outgroup Dothidea sambuci GenBank AY544681) and was used to determine the familial and ordinal relationships of the studied species (Fig. 1), the second was based on a combined ITS and LSU alignment of 44 isolates (including the outgroup Saccharata proteae strain CBS 115206) and was used to determine the genus relationships and species identification within the Botryosphaeriaceae (Fig. 2), and the third was based on a combined ITS, LSU, SSU, TEF and TUB alignment of 18 Darkera isolates and was used for species identification (Fig. 3). The first analysis (LSU) (including the outgroup sequence) and the resulting dataset of 773 characters, including alignment gaps which were treated as fifth base, consisted of 604 constant characters, 36 variable parsimony- uninformative characters and 133 parsimony-informative characters. The maximum of 1000 equally most parsimonious trees were retained (TL = 367; CI = 0.638; RI = 0.928; RC = 0.591), the first of which is presented in Fig. 1. The overall topology was identical between the distance tree (data not shown) and the presented parsimony tree (Fig. 1) with some minor rearrangements of terminal clades in the different families. Overall, the parsimony analysis yielded less well-supported nodes compared to the distance analysis. The Dermateaceae was well-supported in both analyses, whereas the Phacidiaceae was only supported in the distance analysis. The Darkera clade itself is well-supported in both analyses, although the deeper structure of the sub-clades of the Phacidiaceae collapses into a basal polytomy in the parsimony analysis (see strict consensus branches in Fig. 1). The Phyllostictaceae is well-supported in both analyses, whereas the Botryosphaeriaceae is strongly supported in the distance analysis (98 % bootstrap support) but less so in the parsimony analysis (76 % bootstrap support). The LSU phylogeny based on the current dataset alone does not provide a well-supported topology for the Botryosphaeriaceae and therefore the data was combined with ITS for the second analysis. The second analysis (combined ITS and LSU alignment) (including the outgroup sequence) and the resulting dataset of 1243 characters, including alignment gaps which were treated as fifth base, consisted of 976 constant characters, 92 variable parsimony-uninformative characters and 175 parsimony-informative characters. Twenty-two equally most parsimonious trees were obtained (TL = 619; CI = 0.577; RI = 0.809; RC = 0.467), the first of which is presented in Fig. 2. In this phylogeny, all genera that are presented by more than one strain or species are supported with a parsimony bootstrap support value of at least 80 %; the only exception is Diplodia which is split into two lineages without support for the connecting node. The tiarosporella-like strains are polyphyletic in the tree and therefore novel genera are introduced below to accommodate those not clustering in the Tiarosporella clade. Except for Tiarosporella tritici (= Eutiarosporella tritici , see below) and T. africana (= Eut. africana , see below), all species in the ITS-LSU phylogeny could be resolved. In the case of this exception, the two species can easily be distinguished based on their TEF or TUB sequences (data not shown). The third analysis (combined ITS, LSU, SSU, TEF and TUB alignment) was based on the resulting dataset of 2879 characters, including alignment gaps which were treated as fifth base, consisted of 2857 constant characters, 3 variable parsimony-uninformative characters and 19 parsimony-informative characters (TL = 22; CI = 1.0; RI = 1.0; RC = 1.0). Only a single most parsimonious tree was obtained, presented in Fig. 3, which clearly separated the strains belonging to Darkera picea from those belonging to D. parca ...
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... phylogenies were generated; the first was based on 56 LSU sequences (including the outgroup Dothidea sambuci GenBank AY544681) and was used to determine the familial and ordinal relationships of the studied species (Fig. 1), the second was based on a combined ITS and LSU alignment of 44 isolates (including the outgroup Saccharata proteae strain CBS 115206) and was used to determine the genus relationships and species identification within the Botryosphaeriaceae (Fig. 2), and the third was based on a combined ITS, LSU, SSU, TEF and TUB alignment of 18 Darkera isolates and was used for species identification (Fig. 3). The first analysis (LSU) (including the outgroup sequence) and the resulting dataset of 773 characters, including alignment gaps which were treated as fifth base, consisted of 604 constant characters, 36 variable parsimony- uninformative characters and 133 parsimony-informative characters. The maximum of 1000 equally most parsimonious trees were retained (TL = 367; CI = 0.638; RI = 0.928; RC = 0.591), the first of which is presented in Fig. 1. The overall topology was identical between the distance tree (data not shown) and the presented parsimony tree (Fig. 1) with some minor rearrangements of terminal clades in the different families. Overall, the parsimony analysis yielded less well-supported nodes compared to the distance analysis. The Dermateaceae was well-supported in both analyses, whereas the Phacidiaceae was only supported in the distance analysis. The Darkera clade itself is well-supported in both analyses, although the deeper structure of the sub-clades of the Phacidiaceae collapses into a basal polytomy in the parsimony analysis (see strict consensus branches in Fig. 1). The Phyllostictaceae is well-supported in both analyses, whereas the Botryosphaeriaceae is strongly supported in the distance analysis (98 % bootstrap support) but less so in the parsimony analysis (76 % bootstrap support). The LSU phylogeny based on the current dataset alone does not provide a well-supported topology for the Botryosphaeriaceae and therefore the data was combined with ITS for the second analysis. The second analysis (combined ITS and LSU alignment) (including the outgroup sequence) and the resulting dataset of 1243 characters, including alignment gaps which were treated as fifth base, consisted of 976 constant characters, 92 variable parsimony-uninformative characters and 175 parsimony-informative characters. Twenty-two equally most parsimonious trees were obtained (TL = 619; CI = 0.577; RI = 0.809; RC = 0.467), the first of which is presented in Fig. 2. In this phylogeny, all genera that are presented by more than one strain or species are supported with a parsimony bootstrap support value of at least 80 %; the only exception is Diplodia which is split into two lineages without support for the connecting node. The tiarosporella-like strains are polyphyletic in the tree and therefore novel genera are introduced below to accommodate those not clustering in the Tiarosporella clade. Except for Tiarosporella tritici (= Eutiarosporella tritici , see below) and T. africana (= Eut. africana , see below), all species in the ITS-LSU phylogeny could be resolved. In the case of this exception, the two species can easily be distinguished based on their TEF or TUB sequences (data not shown). The third analysis (combined ITS, LSU, SSU, TEF and TUB alignment) was based on the resulting dataset of 2879 characters, including alignment gaps which were treated as fifth base, consisted of 2857 constant characters, 3 variable parsimony-uninformative characters and 19 parsimony-informative characters (TL = 22; CI = 1.0; RI = 1.0; RC = 1.0). Only a single most parsimonious tree was obtained, presented in Fig. 3, which clearly separated the strains belonging to Darkera picea from those belonging to D. parca ...
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... three phylogenies were generated; the first was based on 56 lSu sequences (including the outgroup Dothidea sambuci GenBank aY544681) and was used to determine the familial and ordinal relationships of the studied species ( Fig. 1), the second was based on a combined ItS and lSu alignment of 44 isolates (including the outgroup Saccharata proteae strain CBS 115206) and was used to determine the genus relationships and species identification within the Botryosphaeriaceae (Fig. 2), and the third was based on a combined ItS, lSu, SSu, teF and tuB alignment of 18 ...
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... characters, including alignment gaps which were treated as fifth base, consisted of 604 constant characters, 36 variable parsimonyuninformative characters and 133 parsimony-informative characters. the maximum of 1000 equally most parsimonious trees were retained (tl = 367; CI = 0.638; rI = 0.928; rC = 0.591), the first of which is presented in Fig. 1. the overall topology was identical between the distance tree (data not shown) and the presented parsimony tree ( Fig. 1) with some minor rearrangements of terminal clades in the different families. overall, the parsimony analysis yielded less well-supported nodes compared to the distance analysis. the dermateaceae was well-supported ...
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... parsimonyuninformative characters and 133 parsimony-informative characters. the maximum of 1000 equally most parsimonious trees were retained (tl = 367; CI = 0.638; rI = 0.928; rC = 0.591), the first of which is presented in Fig. 1. the overall topology was identical between the distance tree (data not shown) and the presented parsimony tree ( Fig. 1) with some minor rearrangements of terminal clades in the different families. overall, the parsimony analysis yielded less well-supported nodes compared to the distance analysis. the dermateaceae was well-supported in both analyses, whereas the Phacidiaceae was only supported in the distance analysis. the Darkera clade itself is ...
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... the dermateaceae was well-supported in both analyses, whereas the Phacidiaceae was only supported in the distance analysis. the Darkera clade itself is well-supported in both analyses, although the deeper structure of the sub-clades of the Phacidiaceae collapses into a basal polytomy in the parsimony analysis (see strict consensus branches in Fig. 1). the Phyllostictaceae is well-supported in both analyses, whereas the Botryosphaeriaceae is strongly supported in the distance analysis (98 % bootstrap support) but less so in the parsimony analysis (76 % bootstrap support). the lSu phylogeny based on the current dataset alone does not provide a well-supported topology for the ...
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... a secondary invader, which proved to be rather uncommonly encountered. With its unilocular conidiomata, and pigmented, appendaged conidia, it is somewhat reminiscent of Harknessia (Crous CrouS ET AL. Macrophomina (Sarr et al. 2014). Phylogenetically however, it proved to be allied to genera in the Tiarosporella complex in the Botryosphaeriaceae (Fig. 1), which was quite ...
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... paludosa occurs rather commonly in Germany on Carex spp., Eriophorum polystachium and Trichophorum cespitosum (= Scirpus caespitosus) (Sutton 1980, Nag raj 1993, and is obviously widely distributed in europe. It is also known to occur in Canada and the uSa (Nag raj 1993). the present collection closely matches the morphology of the holotype, and is also from Germany, where this taxon occurs commonly on Carex, Eriophorum andTrichophorum (Sutton 1980, Nag raj 1993). ...
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... the epitypification of Tiarosporella, based on T. paludosa, allowed us to separate this genus from its close allies in the Botryosphaeriaceae that actually form a subclade (Fig. 2), representing several genera with conidial appendages. this subclade includes genera such as Botryobambusa (see liu et al. 2012 fig. 11, though appendage overlooked by the authors), and two new genera, namely Marasasiomyces, and Eutiarosporella. Eutiarosporella is morphologically similar to Marasasiomyces (long necked, hairy conidiomata, and holoblastic conidiogenesis), except that it forms conidiomata in clusters, which is not the case in Marasasiomyces. the latter ...

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... Subsequently, Phillips et al. (2013) provided detailed descriptions and keys to 17 genera and 110 species known from culture at that time. A revision of the Tiarosporella complex resulted in the addition of four new genera (Crous et al. 2015), while Alanphillipsia and Sardiniella were introduced by Crous et al. (2013) and Linaldeddu et al. (2016a), respectively. Yang et al. (2017) introduced Oblongo collomyces and reduced Spencermartinsia to synonymy with Dothiorella. ...
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... Based on ITS and LSU sequence phylogeny, Crous et al. (2015a) introduced Eutiarosporella as a new genus for tiarosporella-like fungi with long-necked conidiomata and holoblastic conidiogenesis (A.J.L. Phillips). ...
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... Eutiarosporella was introduced by Crous et al. (2015) and is typified by Eutiarosporella tritici (B. Sutton & Marasas) Crous on Triticum aestivum from South Africa. ...
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... TEF1␣ was sequenced for Darkera species to examine interspecific and intraspecific variation because it is a promising candidate as a universal secondary barcode and is represented by several Darkera spp. sequences (Crous et al. 2015b;Stielow et al. 2015). DNA was amplified using a PCR master mix consisting of 0.5 L 2 M dNTPs, 0.04 L 20 mol/L forward primer, 0.04 L 20 mol/L reverse primer, 1 L 10× Titanium Taq buffer (Clontech, Mountain View, Calif.), 0.1 L 50× Titanium Taq enzyme (Clontech), 1 L of DNA template, and 7.3 L sterile purified water per reaction (Allain-Boulé et al. 2004). ...
... Nag Raj (1993) examined the D. parca holotype (DAOM 145413b, as Tiarosporella parca) and recorded larger conidial dimensions than those observed by Whitney et al. (1975): (29-)35-43 m × 9-12 m versus (20-)23-40 m × 4-6(-7) m. Strains identified by Crous et al. (2015b) as D. parca from asymptomatic Picea obovata needles in Siberia have conidial dimensions closer to those described by Whitney et al. (1975) than Nag Raj (1993. Crous et al. (2015b) described D. picea from Picea abies needles in Finland, Norway, and Switzerland; D. picea is characterized by large conidia with dimensions almost exactly those of the Darkera cf. ...
... Strains identified by Crous et al. (2015b) as D. parca from asymptomatic Picea obovata needles in Siberia have conidial dimensions closer to those described by Whitney et al. (1975) than Nag Raj (1993. Crous et al. (2015b) described D. picea from Picea abies needles in Finland, Norway, and Switzerland; D. picea is characterized by large conidia with dimensions almost exactly those of the Darkera cf. parca strains isolated in this study ( Fig. 10; Supplementary data, Table S1). ...
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... The Botryosphaeriaceae family comprises 23 phylogenetically well-defined genera and over one hundred species known from culture of which many recognized as aggressive fungal pathogens of economically important plants around the world (Phillips et al. 2013;Hyde et al. 2014;Crous et al. 2015;Dissanayake et al. 2016). Several species belonging to the genera Botryosphaeria, Diplodia, Dothiorella, Lasiodiplodia, Neofusicoccum and Spencermartinsia have been extensively studied for their ability to produce a wide array of secondary metabolites some of which potentially involved in the pathogenesis process (Andolfi et al. 2011(Andolfi et al. , 2012Cimmino, Maddau et al. 2017;Cimmino, Cinelli et al. 2017;Masi et al. 2016Masi et al. , 2018Reveglia, Savocchia, Billones-Baaijens, Cimmino et al. 2018;Reveglia, Savocchia, Billones-Baaijens, Masi et al. 2018). ...
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... Recent molecular studies on Dothideomycetes have revealed hidden lineages and prompted revision of several families in this class (Crous et al. 2015, Guatimosim et al. 2015, Knapp et al. 2015, Jaklitsch & Voglmayr 2016, Van Nieuwenhuijzen et al. 2016. Although multiple molecular systematic studies, mainly of pathogenic fungi of woody plants , Slippers et al. 2013, Alves et al. 2014, Fan et al. 2015, Trakunyingcharoen et al. 2015, have generated a robust phylogeny for Botryosphaeriales, many additional new lineages in this group, including endophytes and saprophytes, have been discovered from various niches (Thambugala et al. 2014, Crous et al. 2015, Wyka & Broders 2016, Osorio et al. 2017, Yang et al. 2017. Drawing on many recent collections, our study has revealed previously unrecognised diversity within Lophiotremataceae and its relatives and the potential taxonomic importance of the asexual morphs in defining families. ...
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The family Lophiotremataceae (Pleosporales, Dothideomycetes) is taxonomically revised on the basis of morphological observations and phylogenetic analyses of sequences of nuclear rDNA SSU, ITS, and LSU regions and tef1 and rpb2 genes. A total of 208 sequences were generated from species of Lophiotremataceae and its relatives. According to phylogenetic analyses, Lophiotremataceae encompasses the genus Lophiotrema and five new genera: Atrocalyx, Crassimassarina, Cryptoclypeus, Galeaticarpa, and Pseudocryptoclypeus. These genera are characterised by ascomata with or without a slit-like ostiole and pycnidial conidiomata. Three new families, Aquasubmersaceae, Cryptocoryneaceae, and Hermatomycetaceae, are proposed. Two genera previously recognised as members of Lophiotremataceae, namely, Aquasubmersa having ascomata with a papillate ostiolar neck and pycnidial conidiomata and Hermatomyces possessing sporodochial conidiomata and dimorphic (lenticular and cylindrical) conidia, are included in Aquasubmersaceae and Hermatomycetaceae, respectively. Cryptocoryneum, characterised by the presence of stromatic sporodochia, cheiroid conidia, and conidial arms developed downward from the cap cells, is placed in Cryptocoryneaceae. Two new genera, Antealophiotrema and Pseudolophiotrema, are established, but their familial placements remain unresolved. Antealophiotrema bears ascomata morphologically similar to those of Lophiotrema, but is differentiated from the latter by having ascomata with a well-developed peridium and a monodictys-like asexual morph. Pseudolophiotrema is also similar to Lophiotrema, but can be distinguished by ascomata with a thin peridium. A total of three new families, seven new genera, eight new species, and two new combinations are described and illustrated.