Evaluating species in Botryosphaeriales.

The Botryosphaeriales (Dothideomycetes) includes numerous endophytic, saprobic, and plant pathogenic species associated with a wide range of symptoms, most commonly on woody plants. In a recent phylogenetic treatment of 499 isolates in the culture collection (CBS) of the Westerdijk Institute, we evaluated the families and genera accommodated in this order of important fungi. The present study presents multigene phylogenetic analyses for an additional 230 isolates, using ITS, tef1, tub2, LSU and rpb2 loci, in combination with morphological data. Based on these data, 58 species are reduced to synonymy, and eight novel species are described. They include Diplodia afrocarpi (Afrocarpus, South Africa), Dothiorella diospyricola (Diospyros, South Africa), Lasiodiplodia acaciae (Acacia, Indonesia), Neofusicoccum podocarpi (Podocarpus, South Africa), N. rapaneae (Rapanea, South Africa), Phaeobotryon ulmi (Ulmus, Germany), Saccharata grevilleae (Grevillea, Australia) and S. hakeiphila (Hakea, Australia). The results have clarified the identity of numerous isolates that lacked Latin binomials or had been deposited under incorrect names in the CBS collection in the past. They also provide a solid foundation for more in-depth future studies on taxa in the order. Sequences of the tef1, tub2 and rpb2 genes proved to be the most reliable markers. At the species level, results showed that the most informative genes were inconsistent, but that a combination of four candidate barcodes (ITS, tef1, tub2 and rpb2) provided reliable resolution. Furthermore, given the large number of additional isolates included in this study, and newly generated multigene DNA datasets, several species could also be reduced to synonymy. The study illustrates the value of reassessing the identity of older collections in culture collections utilising modern taxonomic frameworks and methods. Citation: Zhang W, Groenewald JZ, Lombard L, et al. 2021. Evaluating species in Botryosphaeriales. Persoonia 46: 63-115. https://doi.org/10.3767/persoonia.2021.46.03.

Families of Diaporthales based on morphological and phylogenetic evidence.

Diaporthales is an important ascomycetous order comprising phytopathogenic, saprobic, and endophytic fungi, but interfamilial taxonomic relationships are still ambiguous. Despite its cosmopolitan distribution and high diversity with distinctive morphologies, this order has received relativelyiaceae, Macrohilaceae, Melanconidaceae, Pseudoplagiostomaceae, Schizoparmaceae, Stilbosporaceae and Sydowiellaceae. Taxonomic uncertainties among genera are also clarified and recurrent discrepancies in the taxonomic position of families within the Diaporthales are discussed. An updated outline and key to families and genera of the order is presented.

Phylogenetic revision of Camarosporium (Pleosporineae, Dothideomycetes) and allied genera.

A concatenated dataset of LSU, SSU, ITS and tef1 DNA sequence data was analysed to investigate the taxonomic position and phylogenetic relationships of the genus Camarosporium in Pleosporineae (Dothideomycetes). Newly generated sequences from camarosporium-like taxa collected from Europe (Italy) and Russia form a well-supported monophyletic clade within Pleosporineae. A new genus Camarosporidiella and a new family Camarosporidiellaceae are established to accommodate these taxa. Four new species, Neocamarosporium korfii, N. lamiacearum, N. salicorniicola and N. salsolae, constitute a strongly supported clade with several known taxa for which the new family, Neocamarosporiaceae, is introduced. The genus Staurosphaeria based on S. lycii is resurrected and epitypified, and shown to accommodate the recently introduced genus Hazslinszkyomyces in Coniothyriaceae with significant statistical support. Camarosporium quaternatum, the type species of Camarosporium and Camarosporomyces flavigena cluster together in a monophyletic clade with significant statistical support and sister to the Leptosphaeriaceae. To better resolve interfamilial/intergeneric level relationships and improve taxonomic understanding within Pleosporineae, we validate Camarosporiaceae to accommodate Camarosporium and Camarosporomyces. The latter taxa along with other species are described in this study.

A multiproxy approach to evaluate biocidal treatments on biodeteriorated majolica glazed tiles.

The Fishing House located on the grounds of the Marquis of Pombal Palace, Oeiras, Portugal, was built in the 18th century. During this epoch, Portuguese gardens, such as the one surrounding the Fishing House, were commonly ornamented with glazed wall tile claddings. Currently, some of these outdoor tile panels are covered with dark colored biofilms, contributing to undesirable aesthetic changes and eventually inducing chemical and physical damage to the tile surfaces. Phylogenetic analyses revealed that the investigated biofilms are mainly composed of green algae, cyanobacteria and dematiaceous fungi. With the aim of mitigating biodeterioration, four different biocides (TiO2 nanoparticles, Biotin® T, Preventol® RI 80 and Albilex Biostat® ) were applied in situ to the glazed wall tiles. Their efficacy was monitored by visual examination, epifluorescence microscopy and DNA-based analysis. Significant changes in the microbial community composition were observed 4 months after treatment with Preventol® RI 80 and Biotin® T. Although the original community was inactivated after these treatments, an early stage of re-colonization was detected 6 months after the biocide application. TiO2 nanoparticles showed promising results due to their self-cleaning effect, causing the detachment of the biofilm from the tile surface, which remained clean 6 and even 24 months after biocide application. © 2013 Society for Applied Microbiology and John Wiley & Sons Ltd.

Evaluation of culture-based techniques and 454 pyrosequencing for the analysis of fungal diversity in potting media and organic fertilizers.

AIMS: Potting media and organic fertilizers (OFs) are commonly used in agricultural systems. However, there is a lack of studies on the efficiency of culture-based techniques in assessing the level of fungal diversity in these products. A study was conducted to investigate the efficiency of seven culture-based techniques and pyrosequencing for characterizing fungal diversity in potting media and OFs. METHODS AND RESULTS: Fungal diversity was evaluated using serial dilution, direct plating and baiting with carrot slices, potato slices, radish seeds, cucumber seeds and cucumber cotyledons. Identity of all the isolates was confirmed on the basis of the internal transcribed spacer region of the ribosomal RNA (ITS rRNA) sequence data. The direct plating technique was found to be superior over other culture-based techniques in the number of fungal species detected. It was also found to be simple and the least time consuming technique. Comparing the efficiency of direct plating with 454 pyrosequencing revealed that pyrosequencing detected 12 and 15 times more fungal species from potting media and OFs respectively. Analysis revealed that there were differences between potting media and OFs in the dominant phyla, classes, orders, families, genera and species detected. Zygomycota (52%) and Chytridiomycota (60%) were the predominant phyla in potting media and OFs respectively. CONCLUSIONS: The superiority of pyrosequencing over cultural methods could be related to the ability to detect obligate fungi, slow growing fungi and fungi that exist at low population densities. SIGNIFICANCE AND IMPACT OF THE STUDY: The evaluated methods in this study, especially direct plating and pyrosequencing, may be used as tools to help detect and reduce movement of unwanted fungi between countries and regions.

A phylogenetic study of Dothiorella and Spencermartinsia species associated with woody plants in Iran, New Zealand, Portugal and Spain.

Dothiorella and Spencermartinsia are two botryosphaeriaceous genera with dark 2-celled conidia and found in parasitic, saprophytic or endophytic association with various woody host plants. Based on ITS and EF1-α sequence data and morphology, eight new species are described from Iran, New Zealand, Portugal and Spain. Of these, five species are placed in Dothiorella, namely D. iranica, D. parva, D. prunicola, D. sempervirentis and D. striata, and three species belong to Spencermartinsia named as S. citricola, S. mangiferae and S. plurivora. An identification key to the species of each genus is provided.

Phylogenetic lineages in the Botryosphaeriales: a systematic and evolutionary framework.

The order Botryosphaeriales represents several ecologically diverse fungal families that are commonly isolated as endophytes or pathogens from various woody hosts. The taxonomy of members of this order has been strongly influenced by sequence-based phylogenetics, and the abandonment of dual nomenclature. In this study, the phylogenetic relationships of the genera known from culture are evaluated based on DNA sequence data for six loci (SSU, LSU, ITS, EF1, BT, mtSSU). The results make it possible to recognise a total of six families. Other than the Botryosphaeriaceae (17 genera), Phyllostictaceae (Phyllosticta) and Planistromellaceae (Kellermania), newly introduced families include Aplosporellaceae (Aplosporella and Bagnisiella), Melanopsaceae (Melanops), and Saccharataceae (Saccharata). Furthermore, the evolution of morphological characters in the Botryosphaeriaceae were investigated via analysis of phylogeny-trait association. None of the traits presented a significant phylogenetic signal, suggesting that conidial and ascospore pigmentation, septation and appendages evolved more than once in the family. Molecular clock dating on radiations within the Botryosphaeriales based on estimated mutation rates of the rDNA SSU locus, suggests that the order originated in the Cretaceous period around 103 (45-188) mya, with most of the diversification in the Tertiary period. This coincides with important periods of radiation and spread of the main group of plants that these fungi infect, namely woody Angiosperms. The resulting host-associations and distribution could have influenced the diversification of these fungi. TAXONOMIC NOVELTIES: New families - Aplosporellaceae Slippers, Boissin & Crous, Melanopsaceae Phillips, Slippers, Boissin & Crous, Saccharataceae Slippers, Boissin & Crous.

The Botryosphaeriaceae: genera and species known from culture.

In this paper we give an account of the genera and species in the Botryosphaeriaceae. We consider morphological characters alone as inadequate to define genera or identify species, given the confusion it has repeatedly introduced in the past, their variation during development, and inevitable overlap as representation grows. Thus it seems likely that all of the older taxa linked to the Botryosphaeriaceae, and for which cultures or DNA sequence data are not available, cannot be linked to the species in this family that are known from culture. Such older taxa will have to be disregarded for future use unless they are epitypified. We therefore focus this paper on the 17 genera that can now be recognised phylogenetically, which concentrates on the species that are presently known from culture. Included is a historical overview of the family, the morphological features that define the genera and species and detailed descriptions of the 17 genera and 110 species. Keys to the genera and species are also provided. Phylogenetic relationships of the genera are given in a multi-locus tree based on combined SSU, ITS, LSU, EF1-α and ß-tubulin sequences. The morphological descriptions are supplemented by phylogenetic trees (ITS alone or ITS + EF1-α) for the species in each genus. TAXONOMIC NOVELTIES: New species - Neofusicoccum batangarum Begoude, Jol. Roux & Slippers. New combinations - Botryosphaeria fabicerciana (S.F. Chen, D. Pavlic, M.J. Wingf. & X.D. Zhou) A.J.L. Phillips & A. Alves, Botryosphaeria ramosa (Pavlic, T.I. Burgess, M.J. Wingf.) A.J.L. Phillips & A. Alves, Cophinforma atrovirens (Mehl & Slippers) A. Alves & A.J.L. Phillips, Cophinforma mamane (D.E. Gardner) A.J.L. Phillips & A. Alves, Dothiorella pretoriensis (Jami, Gryzenh., Slippers & M.J. Wingf.) Abdollahz. & A.J.L. Phillips, Dothiorella thailandica (D.Q. Dai., J.K. Liu & K.D. Hyde) Abdollahz., A.J.L. Phillips & A. Alves, Dothiorella uruguayensis (C.A. Pérez, Blanchette, Slippers & M.J. Wingf.) Abdollahz. & A.J.L. Phillips, Lasiodiplodia lignicola (Ariyawansa, J.K. Liu & K.D. Hyde) A.J.L. Phillips, A. Alves & Abdollahz., Neoscytalidium hyalinum (C.K. Campb. & J.L. Mulder) A.J.L. Phillips, Groenewald & Crous, Sphaeropsis citrigena (A.J.L. Phillips, P.R. Johnst. & Pennycook) A.J.L. Phillips & A. Alves, Sphaeropsis eucalypticola (Doilom, J.K. Liu, & K.D. Hyde) A.J.L. Phillips, Sphaeropsis porosa (Van Niekerk & Crous) A.J.L. Phillips & A. Alves. Epitypification (basionym) - Sphaeria sapinea Fries. Neotypifications (basionyms) - Botryodiplodia theobromae Pat., Physalospora agaves Henn, Sphaeria atrovirens var. visci Alb. & Schwein.

Resolving the Diplodia complex on apple and other Rosaceae hosts.

Diplodia species are known as pathogens on many woody hosts, including fruit trees, worldwide. In this study a collection of Diplodia isolates obtained mostly from apple and other Rosaceae hosts were identified based on morphological characters and DNA sequence data from ITS and EF1-α loci. The results show that the diversity of species associated with twig and branch cankers and fruit rot of apples is larger than previously recognised. Four species were identified, namely D. seriata and D. malorum (which is here reinstated for isolates with D. mutila-like conidia). Diplodia intermedia sp. nov. is closely related to D. seriata, and D. bulgarica sp. nov. is morphologically and phylogenetically distinct from all Diplodia species reported from apples.

Fungal Planet description sheets: 128-153.

Novel species of microfungi described in the present study include the following from Australia: Catenulostroma corymbiae from Corymbia, Devriesia stirlingiae from Stirlingia, Penidiella carpentariae from Carpentaria, Phaeococcomyces eucalypti from Eucalyptus, Phialophora livistonae from Livistona, Phyllosticta aristolochiicola from Aristolochia, Clitopilus austroprunulus on sclerophyll forest litter of Eucalyptus regnans and Toxicocladosporium posoqueriae from Posoqueria. Several species are also described from South Africa, namely: Ceramothyrium podocarpi from Podocarpus, Cercospora chrysanthemoides from Chrysanthemoides, Devriesia shakazului from Aloe, Penidiella drakensbergensis from Protea, Strelitziana cliviae from Clivia and Zasmidium syzygii from Syzygium. Other species include Bipolaris microstegii from Microstegium and Synchaetomella acerina from Acer (USA), Brunneiapiospora austropalmicola from Rhopalostylis (New Zealand), Calonectria pentaseptata from Eucalyptus and Macadamia (Vietnam), Ceramothyrium melastoma from Melastoma (Indonesia), Collembolispora aristata from stream foam (Czech Republic), Devriesia imbrexigena from glazed decorative tiles (Portugal), Microcyclospora rhoicola from Rhus (Canada), Seiridium phylicae from Phylica (Tristan de Cunha, Inaccessible Island), Passalora lobeliae-fistulosis from Lobelia (Brazil) and Zymoseptoria verkleyi from Poa (The Netherlands). Valsalnicola represents a new ascomycete genus from Alnus (Austria) and Parapenidiella a new hyphomycete genus from Eucalyptus (Australia). Morphological and culture characteristics along with ITS DNA barcodes are also provided.

First Report of Diplodia seriata Causing Shoot Blight and Cankers of Cotoneaster salicifolius in Bulgaria.

Shoot and branch blight was observed on shrubs of Cotoneaster salicifolius in the region of Plovdiv, Bulgaria in autumn and winter surveys during 2005 and 2006. Scattered pycnida were found on the withered branches, which bore sunken cankers with marginal bark cracks. Fungal colonies obtained from affected host tissue developed on potato dextrose agar (PDA) at 24 to 25°C in the dark. Colonies grew rapidly, were floccose, initially white, becoming grayish brown, and finally went from gray to black. Dark brown, unilocular pycnida formed after 2 to 3 weeks in culture. Conidia that formed at the tips of cylindrical, percurrently, proliferating, conidiogenous cells were ovoid, straight, pale brown to dark brown, internally verruculose, aseptate or occasionally with a single median septum. The mean conidial dimensions were 20 × 12 µm, the extreme range was 15 to 25 × 11 to 15 µm. These characters correspond to Diplodia seriata De Not., which is the anamorph of "Botryosphaeria" obtusa (2). Identity was confirmed from the nucleotide sequences of the internal transcribed spacer (ITS) from the rRNA repeat as described elsewhere (1). BLAST searches in GenBank showed 100% identity of isolates with reference sequences of D. seriata including that from the ex-epitype culture. The ITS sequence of a representative isolate (CAP337) has been deposited in GenBank (Accession No. EU483658). Pathogenicity was tested by inserting 3-mm-diameter mycelia plugs from 7-day-old PDA cultures into scalpel-wounded mature apple fruits (cv. Granny Smith) and wounded shoots of C. salicifolius. Sterile PDA plugs were placed into similarly made wounds of control specimens. There were three replicates for each treatment and the inoculated wounds were subsequently covered with Parafilm. Small necrotic lesions occurred on the apple fruits 3 to 5 days after inoculation and after 10 to 12 days on the shoots. Within 4 to 5 weeks, all apple fruits were entirely rotten, shrunken, and covered with pycnida, whereas the shoots withered and typical blight symptoms developed above the inoculation sites. The pathogen was reisolated from all inoculated samples but not from any control treatment. To our knowledge, this is the first report of D. seriata on C. salicifolius in Bulgaria. It is likely that this pathogen will be found increasingly more frequently because of the intensive introduction of decorative Rosaceae species into urban districts. References: (1) A. Alves et al. Mycologia 96:598, 2004. (2) A. J. L. Phillips et al. Fungal Divers. 25:141, 2007.

First Report of Canker Disease Caused by Botryosphaeria parva on Cork Oak Trees in Italy.

A survey was carried out in the spring of 2003 to study the fungi associated with declining trees in a cork oak (Quercus suber L.) forest located in Sassari Province, Sardinia, Italy (40°52'N, 9°01'E) at an altitude of 150 m (above sea level). Several isolates obtained from live twigs and branches showing sunken necrotic bark lesions were identified as Fusicoccum parvum Pennycook & Samuels (teleomorph Botryosphaeria parva Pennycook & Samuels). Neither pycnidia nor ascomata were observed on the symptomatic samples collected. On potato dextrose agar (PDA) at 25°C, the isolates developed an aerial and compact mycelium, initially white but becoming gray after 4 to 6 days, and produced pycnidia after 1 month on sterile cork oak twigs placed on the surface of PDA. Conidia from culture were hyaline, ellipsoidal to fusiform, externally smooth, thin walled, nonseptate, 12 to 19 (15.5) × 5.5 to 8.5 (6.5) µm, with length/width ratio of 2.4 ± 0.1 (mean ± standard error). Identity was confirmed by analysis of the nucleotide sequences of the internal transcribed spacer (ITS) from the rRNA repeat and the translation elongation factor 1-alpha (EF1-α), as done elsewhere (1,4). BLAST searches at GenBank showed a high identity with reference sequences (ITS: >99%; EF1-α: 100%). Representative sequences of both regions were deposited at GenBank (ITS: Accession No. DQ487157; EF1-α: Accession No. DQ487158). Pathogenicity tests were carried out on seven 2-year-old cork oak seedlings maintained in a greenhouse at 14 to 26°C with the B. parva strain CBS 119937 obtained in this study. A mycelial plug (3 to 4 mm2) taken from the margin of an actively growing colony on PDA was put in a shallow wound made by a scalpel on the basal part of the stem of each seedling. Sterile PDA plugs were placed into similar wounds on three control seedlings. The inoculation points were wrapped in Parafilm to retain moisture for 1 week. After 4 weeks, all seedlings inoculated with B. parva died and showed a collapse of the stem cortical tissues associated with dark brown discolorations and vascular necrosis measuring 10.9 ± 0.4 cm. No symptoms were visible in the control seedlings. The pathogen was reisolated from all the inoculated seedlings, thus fulfilling Koch's postulates. The results confirm the virulence of this fungus and point to its possible involvement in the aetiology of cork oak decline. B. parva is a cosmopolitan, plurivorous pathogen causing disease in several hosts of economic importance, such as grapevine (3), kiwi (2), and Eucalyptus spp. trees (1). To our knowledge, this is the first report of B. parva causing canker disease on cork oak trees. References: (1) A. Gezahgne et al. S. Afr. J. Bot. 70:241, 2004. (2) S. R. Pennycook and G. J. Samuels. Mycotaxon 24:445, 1985. (3) A. J. L. Phillips. Phytopathol. Mediterr. 41:3, 2002. (4) B. Slippers et al. Mycologia 96:83, 2004.

Botryosphaeria and Related Taxa Causing Oak Canker in Southwestern Spain.

Although root disease caused by Phytophthora cinnamomi is considered to be the most frequent and damaging disease associated with Quercus spp. decline in southwestern Spain, cankers commonly are observed on branches of declining Mediterranean Quercus spp. in this region. In surveys carried out in eight declining Quercus forests, strips of necrotic inner bark were common on diseased branches. Botryosphaeria stevensii, B. dothidea, and Diplodia sarmentorum consistently were isolated from these branches. Isolates of all three species caused cankers when inoculated onto excised Quercus branches. Inoculations on healthy branches in the field also induced canker development, but only B. stevensii caused lesions that girdled and killed the branches. The optimum temperature for in vitro growth of B. stevensii and B. dothidea was above 25°C, with slow growth at 35°C. In contrast, D. sarmentorum had an optimum temperature for growth of about 21°C, and did not grow at 35°C. The common occurrence and wide distribution of these pathogens, their association with cankers, and their ability to infect Quercus spp. suggest that they may contribute to the Quercus spp. decline in southwestern Spain.

Plectosphaerella species associated with root and collar rots of horticultural crops in southern Italy.

Plectosphaerella cucumerina, most frequently encountered in its Plectosporium state, is well known as a pathogen of several plant species causing fruit, root and collar rot, and collapse. It is considered to pose a serious threat to melon (Cucumis melo) production in Italy. In the present study, an intensive sampling of diseased cucurbits as well as tomato and bell pepper was done and the fungal pathogens present on them were isolated. Phylogenetic relationships of the isolates were determined through a study of ribosomal RNA gene sequences (ITS cluster and D1/D2 domain of the 28S rRNA gene). Combining morphological, culture and molecular data, six species were distinguished. One of these (Pa. cucumerina) is already known. Four new species are described as Plectosphaerella citrullae, Pa. pauciseptata, Pa. plurivora and Pa. ramiseptata. Acremonium cucurbitacearum is shown to be a synonym of Nodulisporium melonis and is transferred to Plectosphaerella as Plectosphaerella melonis comb. nov. A further three known species of Plectosporium are recombined in Plectosphaerella.

Resolving the Diaporthe species occurring on soybean in Croatia.

Diaporthe (anamorph = Phomopsis) species are plant pathogens and endophytes on a wide range of hosts including economically important crops. At least four Diaporthe taxa occur on soybean and they are responsible for serious diseases and significant yield losses. Although several studies have extensively described the culture and morphological characters of these pathogens, their taxonomy has not been fully resolved. Diaporthe and Phomopsis isolates were obtained from soybean and other plant hosts throughout Croatia. Phylogenetic relationships were determined through analyses of partial translation elongation factor 1-alpha (EF1-α) gene and ITS nrDNA sequence data. By combining morphological and molecular data, four species could be distinguished on soybeans in Croatia. Diaporthe phaseolorum is described in this study and its synonyms are discussed. Diaporthe phaseolorum var. caulivora is raised to species status and the name Diaporthe caulivora is introduced to accommodate it. A species previously known as Phomopsis sp. 9 from earlier studies on sunflower, grapevine, rooibos and hydrangea is reported for the first time on soybean, and is formally described as Diaporthe novem. The well-known soybean pathogen Phomopsis longicolla was also collected in the present study and was transferred to Diaporthe longicolla comb. nov. The presence of these species on herbaceous hosts raises once more the relevance of weeds as reservoirs for pathogens of economically important plants.

Phylogeny and morphology of four new species of Lasiodiplodia from Iran.

Four new species of Lasiodiplodia; L. citricola, L. gilanensis, L. hormozganensis and L. iraniensis from various tree species in Iran are described and illustrated. The ITS and partial translation elongation factor-1α sequence data were analysed to investigate their phylogenetic relationships with other closely related species and genera. The four new species formed well-supported clades within Lasiodiplodia and were morphologically distinct from all other known species.

Barriopsis iraniana and Phaeobotryon cupressi: two new species of the Botryosphaeriaceae from trees in Iran.

Species in the Botryosphaeriaceae are well known as pathogens and saprobes of woody hosts, but little is known about the species that occur in Iran. In a recent survey of this family in Iran two fungi with diplodia-like anamorphs were isolated from various tree hosts. These two fungi were fully characterised in terms of morphology of the anamorphs in culture, and sequences of the ITS1/ITS2 regions of the ribosomal DNA operon and partial sequences of the translation elongation factor 1-alpha. Phylogenetic analyses placed them within a clade consisting of Barriopsis and Phaeobotryon species, but they were clearly distinct from known species in these genera. Therefore, they are described here as two new species, namely Barriopsis iraniana on Citrus, Mangifera and Olea, and Phaeobotryon cupressi on Cupressus sempervirens.

A class-wide phylogenetic assessment of Dothideomycetes.

We present a comprehensive phylogeny derived from 5 genes, nucSSU, nucLSU rDNA, TEF1, RPB1 and RPB2, for 356 isolates and 41 families (six newly described in this volume) in Dothideomycetes. All currently accepted orders in the class are represented for the first time in addition to numerous previously unplaced lineages. Subclass Pleosporomycetidae is expanded to include the aquatic order Jahnulales. An ancestral reconstruction of basic nutritional modes supports numerous transitions from saprobic life histories to plant associated and lichenised modes and a transition from terrestrial to aquatic habitats are confirmed. Finally, a genomic comparison of 6 dothideomycete genomes with other fungi finds a high level of unique protein associated with the class, supporting its delineation as a separate taxon.

Resolving the phylogenetic and taxonomic status of dark-spored teleomorph genera in the Botryosphaeriaceae.

Species in the Botryosphaeriaceae are common plant pathogens and saprobes found on a variety of mainly woody hosts. Teleomorphs typically have hyaline, aseptate ascospores. However, some have been reported with brown ascospores and their taxonomic status is uncertain. A multi-gene approach (SSU, ITS, LSU, EF1-alpha and beta-tubulin) was used to resolve the correct phylogenetic position of the dark-spored 'Botryosphaeria' teleomorphs and related asexual species. Neodeightonia and Phaeobotryon are reinstated for species with brown ascospores that are either 1-septate (Neodeightonia) or 2-septate (Phaeobotryon). Phaeobotryosphaeria is reinstated for species with brown, aseptate ascospores that bear an apiculus at either end. The status of Sphaeropsis is clarified and shown to be the anamorph of Phaeobotryosphaeria. Two new genera, namely Barriopsis for species having brown, aseptate ascospores without apiculi and Spencermartinsia for species having brown, 1-septate ascospores with an apiculus at either end are introduced. Species of Dothiorella have brown, 1-septate ascospores and differ from Spencermartinsia in the absence of apiculi. These six genera can also be distinguished from one another based on morphological characters of their anamorphs. Although previously placed in the Botryosphaeriaceae, Dothidotthia, was shown to belong in the Pleosporales, and the new family Dothidotthiaceae is introduced to accommodate it.

Morphology and phylogeny of Botryosphaeria dothidea causing fruit rot of olives.

The taxonomic position of the causal agent of fruit rot of olives was determined from fresh collections of the fungus from central Greece. In culture it formed two types of conidia, namely fusiform, hyaline, aseptate conidia typical of the genus Fusicoccum, and dark-walled, ovoid, ellipsoid or fusiform, 1-2 septate conidia that are not typically observed in Fusicoccum. A phylogenetic analysis based on ITS and EF1-alpha sequences placed the fungus within the same clade as Fusicoccum aesculi, which is the anamorph of Botryosphaeria dothidea, and the type of the genus Fusicoccum.