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Guaraná is indigenous to the Brazilian Amazon where it has cultural and agroeconomic significance. However, its cultivation is constrained by a disease termed oversprouting of guaraná caused by Fusarium decemcellulare, with yield losses reaching as high as 100%. The disease can affect different parts of the plant, causing floral hypertrophy and hyperplasia, stem galls, and oversprouting of vegetative buds. To date, no study has been conducted characterizing the genetic diversity and population structure of this pathogen. Here, we report genetic diversity and genetic structure among 224 isolates from eight guaraná production areas of Amazonas State, Brazil, that were genotyped using a set of 10 inter-simple-sequence repeat (ISSR) markers. Despite moderate gene diversity (Hexp = 0.21 to 0.32), genotypic diversity was at or near maximum (223 multilocus genotypes among 224 isolates). Population genetic analysis of the 10 ISSR marker fragments with STRUCTURE software identified two populations designated C1 and C2 within the F. decemcellulare collection from the eight sites. Likewise, UPGMA hierarchical clustering and discriminant analysis of principal components of the strains from guaraná resolved these same two groups. Analysis of molecular variance demonstrated that 71% of genetic diversity occurred within the C1 and C2 populations. A pairwise comparison of sampling sites for both genetic populations revealed that 59 of 66 were differentiated from one another (P < 0.05), and high and significant gene flow was detected only between sampling sites assigned to the same genetic population. The presence of MAT1-1 and MAT1-2 strains, in conjunction with the high genotypic diversity and no significant linkage disequilibrium, suggests that each population of F. decemcellulare might be undergoing sexual reproduction. Isolation by distance was not observed (R2 = 0.02885, P > 0.05), which suggests that human-mediated movement of seedlings may have played a role in shaping the F. decemcellulare genetic structure in Amazonas State, Brazil.

Thirteen draft genome assemblies are presented for four Colletotrichum gloeosporioides complex species, namely, Colletotrichum aeschynomenes , Colletotrichum asianum , Colletotrichum fructicola , and Colletotrichum siamense , which were isolated from tropical tree hosts as endophytes.

Guaraná is indigenous to the Brazilian Amazon where it has cultural and agroeconomic significance. However, its cultivation is constrained by a disease termed oversprouting of guaraná caused by the

International audience; Scientific communication is facilitated by a data-driven, scientifically sound taxonomy that considers the end-user's needs and established successful practice. Previously (Geiser et al. 2013; Phytopathology 103:400-408. 2013), the Fusarium community voiced near unanimous support for a concept of Fusarium that represented a clade comprising all agriculturally and clinically important Fusarium species, including the F. solani Species Complex (FSSC). Subsequently, this concept was challenged by one research group (Lombard et al. 2015 Studies in Mycology 80: 189-245) who proposed dividing Fusarium into seven genera, including the FSSC as the genus Neocosmospora, with subsequent justification based on claims that the Geiser et al. (2013) concept of Fusarium is polyphyletic (Sandoval-Denis et al. 2018; Persoonia 41:109-129). Here we test this claim, and provide a phylogeny based on exonic nucleotide sequences of 19 orthologous protein-coding genes that strongly support the monophyly of Fusarium including the FSSC. We reassert the practical and scientific argument in support of a Fusarium that includes the FSSC and several other basal lineages, consistent with the longstanding use of this name among plant pathologists, medical mycologists, quarantine officials, regulatory agencies, students and researchers with a stake in its taxonomy. In recognition of this monophyly, 40 species recently described as Neocosmospora were recombined in Fusarium, and nine others were renamed Fusarium. Here the global Fusarium community voices strong support for the inclusion of the FSSC in Fusarium, as it remains the best scientific, nomenclatural and practical taxonomic option available.

The fungal species Metarhizium pingshaense, M. anisopliae, M. robertsii, and M. brunneum, a monophyletic group informally referred to as the PARB species complex, are well known facultative entomopathogens, including many commercialized strains used for biological pest control. Accurate and expedient species identification of Metarhizium isolates represents an important first step when addressing ecological as well as application-related questions involving these fungi. To this end, a species-specific multiplexed polymerase chain reaction (PCR) assay was developed for identification and discrimination among Metarhizium PARB complex species, based on unique sequence signature differences within the nuclear ribosomal intergenic spacer (rIGS) and nuclear intergenic spacer regions MzFG546 and MzIGS2. Species-specificities of the four primer pairs were assessed following a three-step approach including: (1) in silico verification of sequence signatures by BLASTN searches against publically available genome and amplicon sequence data, (2) corroboration of assay specificity and robustness by performing test PCR amplifications against a taxonomically curated reference strain collection of 68 Metarhizium strains representing 12 species, and (3) testing against a field collection of 19 unknown Metarhizium isolates from soil of a Swiss meadow. The specificity of these four primer pairs provide an efficient means to detect and discriminate PARB species in studies targeting ecological aspects of indigenous isolates, as well as efficacy, persistence and potential non-target effects of applied biocontrol strains.

Insect predators are the most important natural enemies of the crapemyrtle bark scale (CMBS) in the USA. Mycopesticides (biocontrol fungi) are considered an IPM tool to increase CMBS mortality; however, their impacts on CMBS predators are unknown. The objectives of this study were to assess the abundance and diversity of CMBS natural enemies in Louisiana; evaluate the impacts of mycopesticides on survival of CMBS predators by life stage; and determine if entomopathogenic spores delivered to crapemyrtles are transferred to predators under field conditions. The mycopesticides Ancora® (Isaria fumosorosea PFR97), BioCeres® (Beauveria bassiana ANT-03), and BotaniGard® (B. bassiana GHA) were tested against the coccinellids Chilocorus spp. and Hyperaspis bigeminata under laboratory and field conditions. Adults and larvae of the coccinellids were treated with each mycopesticide and survival recorded over a 14-day period. The most common natural enemies on CMBS infested trees were the coccinellids Chilocorus cacti, C. stigma, and Hyperaspis bigeminata (Coleoptera: Coccinellidae). In laboratory bioassays BotaniGard® reduced survival of adults and larvae of both genera by at least 57%. BioCeres® reduced the survival of Chilocorus spp. adults by 40% and Ancora® reduced survival of H. bigeminata larvae by 69%. Under field conditions, CMBS infestations were sprayed with the mycopesticides and coccinellids were collected every other day for a two-week period. Spores of the applied mycopesticides were recovered from the coccinellids; however, it is not known if infection occurred in the field trial or spores were delivered to CMBS infestations by the coccinellids. We conclude that mycopesticides negatively impacted the survival of coccinellids in laboratory trials, and coccinellids can transport pathogen spores under field conditions.

Scientific communication is facilitated by a data-driven, scientifically sound taxonomy that considers the end-user's needs and established successful practice. In 2013, the

Genomic sequence data indicate that certain fungi in the genus Metarhizium have the capacity to produce lysergic acid-derived ergot alkaloids, but accumulation of ergot alkaloids in these fungi has not been demonstrated previously. We assayed several Metarhizium species grown under different conditions for accumulation of ergot alkaloids. Isolates of M. brunneum and M. anisopliae accumulated the lysergic acid amides lysergic acid α-hydroxyethyl amide, ergine, and ergonovine on sucrose-yeast extract agar but not on two other tested media. Isolates of six other Metarhizium species did not accumulate ergot alkaloids on sucrose-yeast extract agar. Conidia of M. brunneum lacked detectable ergot alkaloids, and mycelia of this fungus secreted over 80% of their ergot alkaloid yield into the culture medium. Isolates of M. brunneum, M. flavoviride, M. robertsii, M. acridum, and M. anisopliae produced high concentrations of ergot alkaloids in infected larvae of the model insect Galleria mellonella, but larvae infected with M. pingshaense, M. album, M. majus, and M. guizhouense lacked detectable ergot alkaloids. Alkaloid concentrations were significantly higher when insects were alive (as opposed to killed by freezing or gas) at the time of inoculation with M. brunneum. Roots of corn and beans were inoculated with M. brunneum or M. flavoviride and global metabolomic analyses indicated that the inoculated roots were colonized, though no ergot alkaloids were detected. The data demonstrate that several Metarhizium species produce ergot alkaloids of the lysergic acid amide class and that production of ergot alkaloids is tightly regulated and associated with insect colonization. IMPORTANCE Our discovery of ergot alkaloids in fungi of the genus Metarhizium has agricultural and pharmaceutical implications. Ergot alkaloids produced by other fungi in the family Clavicipitaceae accumulate in forage grasses or grain crops; in this context they are considered toxins, though their presence also may deter or kill insect pests. Our data report ergot alkaloids in Metarhizium species and indicate a close association of ergot alkaloid accumulation with insect colonization. The lack of accumulation of alkaloids in spores of the fungi and in plants colonized by the fungi affirms the safety of using Metarhizium species as biocontrol agents. Ergot alkaloids produced by other fungi have been exploited to produce powerful pharmaceuticals. The class of ergot alkaloids discovered in Metarhizium species (lysergic acid amides) and their secretion into the growth medium make Metarhizium species a potential platform for future studies on ergot alkaloid synthesis and modification.

Globally distributed, soil associated Metarhizium species used in insect biological control are evidently facultatively sexual and obligately outcrossing, yet sexual morphs have not been observed for most species and corroboration that they recombine in nature remains limited. Community-wide genetic diversity of Metarhizium species among 480 soil isolates from 14 states of western USA was investigated to assess the contributions of clonality and recombination in determining each species’ population structure. Seven species, varying greater than 100-fold in relative abundance, were identified by phylogenetic analysis of 5′ EF1-α (5TEF), including M. robertsii (n = 372), M. guizhouense (n = 37), M. brunneum (n = 37), M. lepidiotae (n = 14), M. pemphigi (n = 11), M. anisopliae (n = 7) and M. pingshaense (n = 2). Analyses of composite multilocus genotypes integrating 5TEF sequences, multilocus microsatellites and mating type idiomorphs conducted on a subset of 239 isolates revealed that all species populations display pronounced clonal structure. Following clone-correction procedures to remove redundant clonal genotypes and collapse clonal lineages, each species’ population sample was determined to be composed of a dozen or fewer genetically unique individuals. Thus, the Metarhizium community inhabiting western USA is conservatively estimated to comprise as few as 34 distinct genetic individuals, with a single, geographically ubiquitous clonal lineage of M. robertsii constituting 45% of total isolates. M. robertsii was the only population determined to be in linkage equilibrium. However, the high proportion of private alleles differentiating most M. robertsii clonal lineages argues against contemporary panmixia, thus the recombination signal detected may be historical. Nevertheless, within M. robertsii, M. brunneum and M. guizhouense there are closely related genotypes of opposite mating type, which suggests that if recombination is contemporary, it likely occurs between closely related individuals. The restricted number of genetic individuals observed throughout western North American Metarhizium species may signify that these represent peripheral populations descended from limited numbers of founders among which there has been little recombination relative to the extent of clone expansion and within-clone genetic divergence.

Leafcutter ants propagate co-evolving fungi for food. The nearly 50 species of leafcutter ants (Atta, Acromyrmex) range from Argentina to the United States, with the greatest species diversity in southern South America. We elucidate the biogeography of fungi cultivated by leafcutter ants using DNA sequence and microsatellite-marker analyses of 474 cultivars collected across the leafcutter range. Fungal cultivars belong to two clades (Clade-A and Clade-B). The dominant and widespread Clade-A cultivars form three genotype clusters, with their relative prevalence corresponding to southern South America, northern South America, Central and North America. Admixture between Clade-A populations supports genetic exchange within a single species, Leucocoprinus gongylophorus. Some leafcutter species that cut grass as fungicultural substrate are specialized to cultivate Clade-B fungi, whereas leafcutters preferring dicot plants appear specialized on Clade-A fungi. Cultivar sharing between sympatric leafcutter species occurs frequently such that cultivars of Atta are not distinct from those of Acromyrmex. Leafcutters specialized on Clade-B fungi occur only in South America. Diversity of Clade-A fungi is greatest in South America, but minimal in Central and North America. Maximum cultivar diversity in South America is predicted by the Kusnezov-Fowler hypothesis that leafcutter ants originated in subtropical South America and only dicot-specialized leafcutter ants migrated out of South America, but the cultivar diversity becomes also compatible with a recently proposed hypothesis of a Central American origin by postulating that leafcutter ants acquired novel cultivars many times from other nonleafcutter fungus-growing ants during their migrations from Central America across South America. We evaluate these biogeographic hypotheses in the light of estimated dates for the origins of leafcutter ants and their cultivars.

The ending of dual nomenclatural systems for pleomorphic fungi in 2011 requires the reconciliation of competing names, ideally linked through culture based or molecular methods. The phylogenetic systematics of Hypocreales and its many genera have received extensive study in the last two decades, however resolution of competing names in Cordycipitaceae has not yet been addressed. Here we present a molecular phylogenetic investigation of Cordycipitaceae that enables identification of competing names in this family, and provides the basis upon which these names can be maintained or suppressed. The taxonomy presented here seeks to harmonize competing names by principles of priority, recognition of monophyletic groups, and the practical usage of affected taxa. In total, we propose maintaining nine generic names, Akanthomyces, Ascopolyporus, Beauveria, Cordyceps, Engyodontium, Gibellula, Hyperdermium, Parengyodontium, and Simplicillium and the rejection of eight generic names, Evlachovaea, Granulomanus, Isaria, Lecanicillium, Microhilum, Phytocordyceps, Synsterigmatocystis, and Torrubiella. Two new generic names, Hevansia and Blackwellomyces, and a new species, Beauveria blattidicola, are described. New combinations are also proposed in the genera Akanthomyces, Beauveria, Blackwellomyces, and Hevansia.

Feeding an increasing global population requires continued improvements in agricultural efficiency and productivity. Meeting estimated future production levels requires the adoption of practices that increase output without environmental degradation associated with external inputs to supplement nutrition or control pests. Enriching the community of microbes associated with plants in agricultural systems for those providing ecosystem services such as pest control is one possible component towards achieving sustainable productivity increases. In this review we explore the current state of knowledge for Hypocreales fungi used in biological control. Advances in understanding the field ecology, diversity and genetic determinants of host range and virulence of hypocrealean fungi provide the means to improve their efficacy.

The coffee berry borer Hypothenemus hampei (CBB) is a major pest of coffee, and the entomopathogenic fungus Beauveria bassiana (Bb) is used worldwide for its biological control. Commercial formulations of Bb are sprayed on coffee, but local isolates of Bb may also provide some level of natural control. We compared selected local Bb isolates from CBB-infested coffee fruits in Puerto Rico against the commercial strain, GHA, derived from Mycotrol®. Isolates were tested for their virulence toward CBB in vitro. Two local isolates and the commercial isolate were sprayed on coffee plants in the field, and percent CBB infected, percent fruits with CBB damage, and number of CBB per fruit were surveyed over eight weeks in three consecutive years. Genotypes of Bb isolates in the field were discriminated with microsatellites to determine if isolates persisted after application. Several local isolates and mixtures of isolates were as virulent in vitro as the commercial isolate. In the field, all isolates significantly reduced CBB damage; one local isolate was more successful than the others. Genetic structure of local Bb populations varied from field to field and from year to year. Local isolates persisted; the commercial isolate did not, except in one plot. The commercial isolate may not be adapted to the warm, humid environment of coffee farms. Local isolates and perhaps combinations of isolates can provide more effective control, although current regulations preclude their use.

Peramine is a non-ribosomal peptide-derived pyrrolopyrazine (PPZ)-containing molecule with anti-insect properties. Peramine is known to be produced by fungi from genus Epichloe, which form mutualistic endophytic associations with cool-season grass hosts. Peramine biosynthesis has been proposed to require only the two-module non-ribosomal peptide synthetase (NRPS) peramine synthetase (PerA), which is encoded by the 8.3 kb gene perA, though this has not been conclusively proven. Until recently, both peramine and perA were thought to be exclusive to fungi of genus Epichloe; however, a putative perA homologue was recently identified in the genome of the insect-pathogenic fungus Metarhizium rileyi. We use a heterologous expression system and a hydrophilic interaction chromatography-based analysis method to confirm that PerA is the only pathway-specific protein required for peramine biosynthesis. The perA homologue from M. rileyi (MR_perA) is shown to encode a functional peramine synthetase, establishing a precedent for distribution of perA orthologs beyond genus Epichloe. Furthermore, perA is part of a larger seven-gene PPZ cluster in M. rileyi, Metarhizium majus and the stalked-cup lichen fungus Cladonia grayi. These PPZ genes encode proteins predicted to derivatize peramine into more complex PPZ metabolites, with the orphaned perA gene of Epichloe spp. representing an example of reductive evolution.

Hirsutella (Ophiocordycipitaceae: Hypocreales) is a genus of insect, mite, and nematode pathogens with an asexual morph, which generally produce a mucilaginous cluster of one or several conidia on phialides that are basally subulate and taper to a fine neck. The generic name Hirsutella has been proposed for suppression in favour of Ophiocordyceps as a consequence of the ending of dual nomenclature for different morphs of pleomorphic fungi in 2011. Though the generic name is well established, geographically dispersed, and speciose, exceptionally few sequences are available in online databases. We examined 46 isolates of 23 Hirsutella species from the USA, curated by the USDA-ARS Collection of Entomopathogenic Fungal Cultures (ARSEF Culture Collection), that previously had not been molecularly characterized and produced a phylogeny of these organisms; we included previously published Hirsutella and Ophiocordyceps taxa. In producing the largest phylogeny of Hirsutella isolates so far, we provide: (1) context for discussing previously-hypothesized relationships; (2) evidence for revisions as taxonomic transitions move forward; and (3) available molecular data to be incorporated into further evolutionary studies of Ophiocordycipitaceae.

Summary Fungi in the genus Metarhizium are insect pathogens able to function in other niches, including soil and plant rhizosphere habitats. In agroecosystems, cropping and tillage practices influence soil fungal communities with unknown effects on the distribution of Metarhizium, whose presence can reduce populations of crop pests. We report results from a selective media survey of Metarhizium in soils sampled from a long-term experimental farming project in the mid-Atlantic region. Field plots under soybean cultivation produced higher numbers of Metarhizium colony-forming units (cfu) than corn or alfalfa. Plots managed organically and via chisel-till harboured higher numbers of Metarhizium cfu than no-till plots. Sequence typing of Metarhizium isolates revealed four species, with M. robertsii and M. brunneum predominating. The M. brunneum population was essentially fixed for a single clone as determined by multilocus microsatellite genotyping. In contrast, M. robertsii was found to contain significant diversity, with the majority of isolates distributed between two principal clades. Evidence for recombination was observed only in the most abundant clade. These findings illuminate multiple levels of Metarhizium diversity that can be used to inform strategies by which soil Metarhizium populations may be manipulated to exert downward pressure on pest insects and promote plant health.

Biological control of spittlebugs with Metarhizium (Hypocreales: Clavicipitaceae) in Brazilian sugarcane is an example of effective pest management. However, little is known about the richness, distribution and ecology of Metarhizium species in Brazilian agroecosystems. We investigated Metarhizium diversity within a collection of 96 Brazilian isolates from spittlebugs and other insects, strains used for spittlebug control and soil isolates from sugarcane and other field crops and pristine habitats. A multilocus phylogenetic analysis of 5′-TEF and nuclear intergenic loci MzFG543igs and MzIGS3 yielded robust support for current species limits of the two most abundant taxa, Metarhizium anisopliae and Metarhizium robertsii, and the resolution of two lineages that lie beyond currently recognized species limits in this complex. With a single exception, all isolates from insects belong to a single subclade of M. anisopliae. These data will serve as resources about Metarhizium biodiversity for insect biological control initiatives in South America.

Summary Moniliophithora roreri, causal agent of cacao frosty pod rot, is considered one of the most devastating pathogens of this crop. In this study,we report the development of and validation of 28 microsatellite loci from an enriched library. From these, ten loci were demonstrated tobe polymorphic in an Ecuadorian population composed of 27 isolates. The mean number of alleles per locus was 3.2, and the observedmean and expected heterozygosity were 0.03 and 0.43, respectively. The polymorphic microsatellites described herein may be used tostudy the genetic diversity as well as to comprehend other aspects of M. roreri population biology. 1 Introduction The hemibiotrophic basidiomycete Moniliophthora roreri (Cif.) H. C. Evans, Stalpers, Smason & Benny, infects only fruits ofneotropical trees from the genera Theobroma and Herrania.InTheobroma cacao L. fruit, the major component of chocolate, itcauses frosty pod rot disease, also known as moniliasis, an extremely destructive disease and currently the main yield-limitingfactor in several Latin American countries (Phillips-Mora 2003). Pod losses due to the disease are usually over 30%, but itmight reach 100% depending on factors such as disease management, favourable environmental conditions and time of intro-duction of the disease in a particular agroecological zone or site (Katip 1994; Krauss et al. 2003). It has been proposed thatM. roreri has its origin in the north-western region of Colombia and is now present in 11 Central and South Americancountries (Phillips-Mora et al. 2007). This disease poses a major threat to important cacao-producing areas in Brazil andCaribbean countries, such as Trinidad and Tobago, Dominican Republic and Haiti that are still free from the pathogen.It has been shown by AFLP/ISSR data that M. roreri is genetically variable and forms five genetic groups (Phillips-Mora etal. 2007), being two major ones: (i) the Bolivar group; comprising isolates from north of Santander in Colombia, all Peruvianand Venezuelan isolates, and 10 isolates from the periphery of Ecuador and (ii) the Co-West group, including, mainly, isolatesfrom western Colombia, central Ecuador and Central America. The other groups are all apparently endemic to Colombia(Co-East and Co-Central groups) or north-western Ecuador. The use of microsatellites or simple sequence repeats (SSRs) willallow broader inferences regarding the population biology of M. roreri. This knowledge is necessary to establish successfulintegrated management strategies for crop breeding. SSRs are considered suitable for genetic studies because they areco-dominantly inherited, allowing precise discrimination even of closely related individuals as well as allowing the heterozygotein diploid genomes to be distinguished. In addition, microsatellite analysis is inexpensive, highly reproducible and transfer-able across related species (Jarne and Lagoda 1996; Goul~ao and Oliveira 2001). SSRs have been the choice in populationgenetic studies of plant pathogens (Taylor et al. 1999), including pathogens of cacao such as M. perniciosa (Gramacho et al.2007), due to their high variability and statistical power (Reusch 2001). Like RAPDs, ISSRs are dominant markers, quick,cheap, easy to handle. Despite the lower reproducibility, ISSRs are applicable to screening information about the genetic vari-ation. However, limited statistical analysis and poor cross-experimental reproducibility represent some drawbacks of ISSRsmarkers, where the homozygous presence of a fragment is not distinguishable from its heterozygote. In this way, SSRs arevery powerful tools and should be more widely used for population studies of this fungus. The use of SSR allows researchersto address key questions about the population genetics of M. roreri, to cite a few practical applications recombination, mat-ing systems, host adaptation and population structure due to host adaptation. Thus, in this study, we characterized 28 micro-satellite loci for genetic analyses of populations of M. roreri that represent the first SSRs developed for this species.

In the USA, the development and field application of Beauveria brongniartii (Sacc.) Petch (Hypocreales: Clavicipitaceae) to control the invasive Asian longhorned beetle, Anoplophora glabripennis (Motschulsky) (Coleoptera: Cerambycidae) have been hampered because it was unknown whether this fungal species is native to North America. With the recent confirmation of the occurrence of B. brongniartii in North America there is renewed interest in this species, particularly as it is an effective pathogen of cerambycids in Japan.

Anamorphic Beauveria are cosmopolitan entomopathogenic fungi that parasitize a broad range of insect species in virtually all terrestrial habitats. A diversity survey of 189 exemplar strains of Beauveria from the RCEF culture collection, representative of its taxonomic diversity, geographic distribution and insect host range in China, was conducted based on a combination of DGGE genotyping and nucleotide sequence analysis of the Bloc nuclear intergenic region. The DGGE assays detected 42 electrophoretically distinct haplotypes, with each haplogroup including 1–13 individuals. Nucleotide sequence analysis established that all haplogroups were uniquely distinguished by one or more nucleotide differences and that isolates from the same DGGE haplogroup share sequence identity. A phylogenetic analysis inclusive of this Bloc haplotype diversity assigned the Chinese Beauveria strains to six species lineages corresponding to B. bassiana sensu lato. (Bals.) Vuill, B. brongniartii (Sacc.) Petch, B. australis S.A. Rehner & Humber, B. asiatica S.A. Rehner & Humber, B. pseudobassiana S.A. Rehner & Humber and B. caledonica Bissett & Widden. B. australis is reported for the first time in China. This study represents the first phylogenetic survey of Beauveria species diversity in China, and demonstrates a simple and effective screening strategy to facilitate the identification of Beauveria genotypes.

Entomopathogenic fungi in the genus Metarhizium are useful for biological control programmes against economically important arthropod pests worldwide. However, understanding the true diversity and ecology of these organisms is hampered by convergent morphologies between species. The application of molecular techniques has enabled greater resolution of species than allowed by morphology alone. In particular, the commonly used biocontrol agent M. anisopliae was found to be a species complex composed of nine species. This prior work was conducted with commonly used markers in fungal phylogenetics (BTUB, RPB1, RPB2 and TEF), which likely under-represent diversity in the M. anisopliae complex. Using sequence data from nuclear genomes of M. acridum and M. robertsii we identified regions of conserved gene synteny and developed primers to amplify intergenic regions of seven loci. Using ex-type and authenticated tissue specimens for species in the M. anisopliae complex, we demonstrate that sequence data derived from intergenic loci is more variable and phylogenetically informative than previously available markers. These new markers will facilitate investigations at or below the species level for the M. anisopliae complex. The method of marker development employed here should be extendable to any group with sufficiently divergent genome data available.

In this letter, we advocate recognizing the genus Fusarium as the sole name for a group that includes virtually all Fusarium species of importance in plant pathology, mycotoxicology, medicine, and basic research. This phylogenetically guided circumscription will free scientists from any obligation to use other genus names, including teleomorphs, for species nested within this clade, and preserve the application of the name Fusarium in the way it has been used for almost a century. Due to recent changes in the International Code of Nomenclature for algae, fungi, and plants, this is an urgent matter that requires community attention. The alternative is to break the longstanding concept of Fusarium into nine or more genera, and remove important taxa such as those in the F. solani species complex from the genus, a move we believe is unnecessary. Here we present taxonomic and nomenclatural proposals that will preserve established research connections and facilitate communication within and between research communities, and at the same time support strong scientific principles and good taxonomic practice Fil: Geiser, David M.. State University of Pennsylvania; Estados Unidos Fil: Aoki, Takayuki. National Institute of Agrobiological Sciences; Japón Fil: Bacon, Charles W.. Toxicology and Mycotoxin Research; Grecia Fil: Baker, Scott E.. Pacific Northwest National Laboratory; Estados Unidos Fil: Bhattacharyya, Madan K.. University of Iowa; Estados Unidos Fil: Chulze, Sofia Noemi. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Scandiani, María Mercedes. Laboratorio Agrícola Río Paraná. San Pedro; Argentina Fil: Ploetz, Randy C.. University of Florida; Estados Unidos Fil: Proctor, Robert H.. NCAUR-ARS-USDA; Estados Unidos Fil: Rehner, Stephen A.. United States Department of Agriculture; Estados Unidos Fil: Robert, Vincent A. R. G.. Fungal Biodiversity Center; Países Bajos Fil: Rooney, Alejandro P.. NCAUR-ARS-USDA; Estados Unidos Fil: Salleh, Baharuddin bin. University of Malaysia; Malasia Fil: Scauflaire, Jonathan. Université Catholique de Louvain; Bélgica Fil: Short, Dylan P. G.. University of California; Estados Unidos Fil: Steenkamp, Emma. University of Pretoria; Sudáfrica Fil: Suga, Haruhisa. Gifu University; Japón Fil: Summerell, Brett A.. The Royal Botanic Garden Sydney; Australia Fil: Sutton, Deanna A.. University of Texas; Estados Unidos Fil: Thrane, Ulf. Technical University of Denmark; Dinamarca Fil: Trail, Francis. Michigan State University; Estados Unidos Fil: Van Diepeningen, Anne. Fungal Biodiversity Center; Países Bajos Fil: VanEtten, Hans D.. University of Arizona; Estados Unidos Fil: Viljoen, Altus. University of Stellenbosch; Sudáfrica Fil: Waalwijk, Cees. Wageningen University; Países Bajos Fil: Ward, Todd J.. NCAUR-ARS-USDA. Pretoria; Sudáfrica Fil: Wingfield, Michael J.. University of Pretoria; Sudáfrica Fil: Xu, Jin-Rong. Purdue University; Estados Unidos Fil: Yang, Xiao-Bing. University of Iowa; Estados Unidos Fil: Yli-Mattila, Tapani. University of Turku; Finlandia Fil: Zhang, Ning. Rutgers University; Estados Unidos

An essential first step to elucidating the ecology and life histories of Metarhizium anisopliae-group species as entomopathogens, endophytes and soil-adapted fungi is the ability to define species limits and confidently infer a species phylogeny. In a multilocus phylogeny of the core Metarhizium anisopliae species complex, the majority of isolates sampled herein group within the currently defined limits of M. pingshaense, M. anisopliae, M. robertsii and M. brunneum, designated informally as the “PARB” clade. Multilocus phylogenetic analyses reveal pervasive congruent hierarchical structure among the genomic regions analyzed, which suggest that current PARB species delimitations likely encompass additional cryptic complexes. Further, the interpolation of isolates from different continents throughout each species lineage indicates periodic inter-continental dispersals. Although no PARB species has yet been confirmed to produce a sexual state, we demonstrate the mutually exclusive incidence of the MAT1 and MAT2 mating type idiomorphs among individuals in all PARB species. This configuration of mating type is diagnostic of a heterothallic, obligately outcrossing mating system, indicating the conservation of and ongoing potential for sexual reproduction in all PARB species. As one example of the utility of IGS markers, the commercially registered M. anisopliae strain F52, which is widely used for pest control in North America, Canada and Europe, is shown to be a member of the M. brunneum complex. While current PARB species delimitations evidently encompass cryptic partitions, formal recognition of segregate species should be approached cautiously until further evidence of their phylogenetic exclusivity, ecological distinctiveness or other unique attributes is demonstrated. Nevertheless, acknowledgment of these intraspecific partitions will provide a useful conceptual framework to guide future investigations of the community structure, phylogeography, population genetics, ecology and reproductive biology of this recent species radiation.

In fungi, stable diploid genome arrangements are rare. Here we present evidence from nuclear intergenic DNA sequencing, microsatellite genotyping, and configuration of the mating-type locus to demonstrate two independent origins of persistent diploid genome organization in the Metarhizium majus species complex. Most taxa in the complex are genotypically haploid, with individual isolates consistently displaying a single allele across all nuclear loci, as well as having a single mating-type locus. In contrast, individuals of M. majus and the clade designated here MGT1 are shown to be diploid, based on a consistent finding of heterozygosity and the presence of both MAT1 and MAT2 mating-type loci. In single locus phylogenies, nuclear intergenic alleles of M. majus and MGT1 each form monophyletic groups, indicating that diploidy in both taxa likely originated by the union of conspecific individuals. Sequence divergence in the APN2/MAT1-1-3 and APN2/MAT2-1 intergenic spacers indicate the two MAT loci are physically separated in the genomes of both diploid taxa, although the linkage relationship of the MAT loci to one another is unknown. The presence of both mating genes in a single nucleus suggests these diploid genomes may represent a mating event that failed to complete meiosis. Whether or not these isolates are able to complete the sexual cycle under any conditions and form ascospores remains an open question.

Summary We studied disease progression of, and host responses to, four species in the Metarhizium anisopliae complex expressing green fluorescent protein (GFP). We compared development and determined their relative levels of virulence against two susceptible arthropods, the cattle tick Rhipicephalus annulatus and the lepidopteran Galleria mellonella, and two resistant ticks, Hyalomma excavatum and Rhipicephalus sanguineus. Metarhizium brunneum Ma7 caused the greatest mortality of R. annulatus, Metarhizium robertsii ARSEF 2575 and Metarhizium pingshaense PPRC51 exhibited intermediate levels of virulence, and Metarhizium majus PPRC27 caused low mortality of cattle ticks. Conidia of all four species germinated on all hosts examined, but on resistant hosts, sustained hyphal growth was inhibited and GFP emission steadily and significantly decreased over time, suggesting a loss of fungal viability. Cuticle penetration was observed only for the three most virulent species infecting susceptible hosts. Cuticles of resistant and susceptible engorged female ticks showed significant increases in red autofluorescence at sites immediately under fungal hyphae. This is the first report (i) of tick mortality occurring after cuticle penetration but prior to haemocoel colonization and (ii) that resistant ticks do not support development of Metarhizium germlings on the outer surface of the cuticle. Whether reduced Metarhizium viability on resistant tick cuticles is due to antibiosis or limited nutrient availability is unknown.

Recent molecular phylogenetic studies have revealed the existence of at least 50 species of Morchella worldwide and demonstrated a high degree of continental endemism within the genus. Here we describe 19 phylogenetic species of Morchella from North America, 14 of which are new (M. diminutiva, M. virginiana, M. esculentoides, M. prava, M. cryptica, M. frustrata, M. populiphila, M. sextelata, M. septimelata, M. capitata, M. importuna, M. snyderi, M. brunnea and M. septentrionalis). Existing species names (M. rufobrunnea, M. tomentosa, M. punctipes and M. angusticeps) are applied to four phylogenetic species, and formal description of one species (M. sp. "Mel-8") is deferred pending study of additional material. Methods for assessing morphological features in Morchella are delineated, and a key to the known phylogenetic species of Morchella in North America is provided. Type studies of M. crassistipa, M. hotsonii, M. angusticeps and M. punctipes are provided. Morchella crassistipa is designated nomen dubium.

PubMedID: 22802394 Arguably more mycophiles hunt true morels (Morchella) during their brief fruiting season each spring in the northern hemisphere than any other wild edible fungus. Concerns about overharvesting by individual collectors and commercial enterprises make it essential that science-based management practices and conservation policies are developed to ensure the sustainability of commercial harvests and to protect and preserve morel species diversity. Therefore, the primary objectives of the present study were to: (i) investigate the utility of the ITS rDNA locus for identifying Morchella species, using phylogenetic species previously inferred from multilocus DNA sequence data as a reference; and (ii) clarify insufficiently identified sequences and determine whether the named sequences in GenBank were identified correctly. To this end, we generated 553 Morchella ITS rDNA sequences and downloaded 312 additional ones generated by other researchers from GenBank using emerencia and analyzed them phylogenetically. Three major findings emerged: (i) ITS rDNA sequences were useful in identifying 48/62 (77.4%) of the known phylospecies; however, they failed to identify 12 of the 22 species within the species-rich Elata Subclade and two closely related species in the Esculenta Clade; (ii) at least 66% of the named Morchella sequences in GenBank are misidentified; and (iii) ITS rDNA sequences of up to six putatively novel Morchella species were represented in GenBank. Recognizing the need for a dedicated Webaccessible reference database to facilitate the rapid identification of known and novel species, we constructed Morchella MLST (http://www.cbs.knaw. nl/morchella/), which can be queried with ITS rDNA sequences and those of the four other genes used in our prior multilocus molecular systematic studies of this charismatic genus. © 2012 by The Mycological Society of America.

The estimation of species diversity in fungal endophyte communities is based either on species counts or on the assignment of operational taxonomic units (OTUs). Consequently, the application of different species recognition criteria affects not only diversity estimates but also the ecological hypotheses that arise from those observations. The main objective of the study was to examine how the choice and number of genetic markers and species delimitation criteria influence biodiversity estimates. Here, we compare approaches to defining species boundaries in three dominant species complexes of tropical endophytes, specially Colletotrichum gloeosporioides agg., Pestalotiopsis microspora agg. and Trichoderma harzianum agg., from two Amazonian trees: Hevea brasiliensis and H. guianensis. Molecular tools were used to describe and compare the diversity of the different assemblages. Multilocus phylogenetic analyses [gpd, internal transcribed spacer (ITS) and tef1] and modern techniques for phylogenetic species delimitation were overlaid with ecological data to recognize putative species or OTUs. The results demonstrate that ITS alone generally underestimates the number of species predicted by other nuclear loci. These results question the use of ITS and arbitrary divergence thresholds for species delimitation.

Colletotrichum interacts with numerous plant species overtly as symptomatic pathogens and cryptically as asymptomatic endophytes. It is not known whether these contrasting ecological modes are optional strategies expressed by individual Colletotrichum species or whether a species' ecology is explicitly pathogenic or endophytic. We explored this question by inferring relationships among 77 C. gloeosporioides s.l. strains isolated from asymptomatic leaves and from anthracnose lesions on leaves and fruits of Theobroma cacao (cacao) and other plants from Panamá. ITS and 5'-tef1 were used to assess diversity and to delineate operational taxonomic units for multilocus phylogenetic analysis. The ITS and 5'-tef1 screens concordantly resolved four strongly supported lineages, clades A-D: Clade A includes the ex type of C. gloeosporioides, clade B includes the ex type ITS sequence of C. boninense, and clades C and D are unidentified. The ITS yielded limited resolution and support within all clades, in particular the C. gloeosporioides clade (A), the focal lineage dealt with in this study. In contrast the 5'-tef1 screen differentiated nine distinctive haplotype subgroups within the C. gloeosporioides clade that were concordant with phylogenetic terminals resolved in a five-locus nuclear phylogeny. Among these were two phylogenetic species associated with symptomatic infections specific to either cacao or mango and five phylogenetic species isolated principally as asymptomatic infections from cacao and other plant hosts. We formally describe two new species, C. tropicale and C. ignotum, that are frequent asymptomatic associates of cacao and other Neotropical plant species, and epitypify C. theobromicola, which is associated with foliar and fruit anthracnose lesions of cacao. Asymptomatic Colletotrichum strains isolated from cacao plants grown in China included six distinct C. gloeosporioides clade taxa, only one of which is known to occur in the Neotropics.

Coffee (Coffea arabica) plant tissues were surface-sterilized and fungal endophytes isolated using standard techniques, followed by DNA extraction and sequencing of the internal transcribed spacer region (ITS). A total of 843 fungal isolates were recovered and sequenced (Colombia, 267; Hawai'i, 393; Mexico, 109; Puerto Rico, 74) yielding 257 unique ITS genotypes (Colombia, 113; Hawai'i, 126; Mexico, 32; Puerto Rico, 40). The most abundant taxa were Colletotrichum, Fusarium, Penicillium, and Xylariaceae. Overall, 220 genotypes were detected in only one of the countries sampled; only two genotypes were found in all four countries. Endophytes were also isolated from Coffea canephora, Coffea congensis, Coffea liberica, Coffea macrocarpa, Coffea racemosa, and Coffea stenophylla in Hawai'i. The high biodiversity of fungal endophytes in coffee plants may indicate that most of these are “accidental tourists” with no role in the plant, in contrast to endophytes that could be defined as “influential passengers” and whose role in the plant has been elucidated. This study, the most comprehensive analysis of fungal endophytes associated with a single host species, demonstrates that coffee plants serve as a reservoir for a wide variety of fungal endophytes that can be isolated from various plant tissues, including the seed, and illustrates the different fungal communities encountered by C. arabica in different coffee-growing regions of the world.

The fungus Ascosphaera apis is a worldwide fungal pathogen of honey bees. To provide tools for understanding the dispersal history of this pathogen, strain differences in virulence, and host-pathogen interactions, we used the draft genome assembly of A. apis to develop microsatellite loci for this species. We present testing results for 25 scorable loci revealing two to eight alleles per locus in a survey of Maryland isolates of this fungus.

Although intensively investigated for biological control of insect pests, little is known about the ecology of the fungal entomopathogenic genus Beauveria in natural or agricultural habitats. In this study, we used molecular phylogenetic and genotypic information to infer species diversity, reproductive potential and genetic structure of Beauveria occurring within a single arable field and bordering hedgerow in Denmark. Isolates were sampled from cultivated field and hedgerow soils, from insects harbouring latent fungal infections, and from the phylloplanes of three plant species common in the hedgerow flora. A nuclear phylogeny of this local Beauveria assemblage resolved seven phylogenetic species, including (i) five phylogenetic species within Beauveria bassiana sensu stricto; (ii) Clade C, a taxonomically uncharacterized species that is morphologically indistinguishable but phylogenetically distant from B. bassiana s.s.; and (iii) Beauveria brongniartii. All seven species were present throughout the hedgerow habitat, including as infections in insects. Significantly, only B. bassiana s.s. phylogenetic species Eu_1 was isolated from tilled soils. Mating type polymerase chain reaction assays demonstrated that all five B. bassiana s.s. phylogenetic species possess bipolar outcrossing mating systems. Of these, only the Eu_1 population contained two mating types; however, a 31:2 skew in MAT1:MAT2 mating types suggests a low frequency of sexual reproduction in this population. The four remaining B. bassiana s.s. phylogenetic species were fixed for single mating types and these populations are evidently clonal. Multilocus microsatellite genotyping revealed polymorphism in all five phylogenetic species of B. bassiana s.s.; however, all show evidence of clonal genetic structure.

Fungal endophytes are quite common in nature and some of them have been shown to have adverse effects against insects, nematodes, and plant pathogens. Our research program is aimed at using fungal endophytes-mediated plant defense as a novel biological control mechanism against the coffee berry borer, the most devastating pest of coffee throughout the world. A survey of fungal endophytes in coffee plants from Hawaii, Colombia, Mexico, and Puerto Rico has revealed the presence of various genera of fungal entomopathogens, including Acremonium, Beauveria, Cladosporium, Clonostachys, and Paecilomyces. Two of these, B. bassiana and Clonostachys rosea, were tested against the coffee berry borer and were shown to be pathogenic. This paper reviews the possible mode of action of entomopathogenic fungal endophytes. Published by Elsevier Inc.

Enterobacter cloacae is a plant-beneficial bacterium that shows promise for suppression of damping-off of cucumber and other crops caused by Pythium ultimum . We have been using a mutational approach to determine the E. cloacae genes important in bacterial–plant and bacterial–pathogen interactions in the spermosphere and rhizosphere. E. cloacae M43 is a transposon mutant of E. cloacae 501R3 that was significantly impaired in colonization of seeds and roots of diverse crop plants. Strain M43 did not increase in population on cucumber, sunflower, and wheat seeds and was significantly reduced in growth on pea seeds relative to strain 501R3. Populations of M43 were also dramatically lower than those of strain 501R3 in cucumber, pea, sunflower, and wheat rhizosphere in 42 d experiments. Molecular characterization of M43 demonstrated that there was a single transposon insertion in the genome of this strain and that this insertion was in a region of the E. cloacae genome with a high degree of DNA sequence identity with aceF. aceF encodes the dihydrolipoamide acetyltransferase subunit of the pyruvate dehydrogenase complex (PDHC). Cell lysates from strain 501R3 grown on minimal medium plus 50 mM glycerol and 2 mM acetate contained 0.011±0.0036 U pyruvate dehydrogenase activity while cell lysates from M43 grown under identical conditions contained no detectable pyruvate dehydrogenase activity. Additionally, the nutritional use profile of M43 under aerobic and anaerobic conditions was as expected for an ace mutant. Experiments reported here strongly suggest a role for aceF and the PDHC in colonization of seeds and roots of diverse crop plants by E. cloacae .

The entomopathogenic fungus Beauveria bassiana was established in coffee seedlings after fungal spore suspensions were applied as foliar sprays, stem injections, or soil drenches. Direct injection yielded the highest post-inoculation recovery of endophytic B. bassiana. Establishment, based on percent recovery of B. bassiana, decreased as time post-inoculation increased in all treatments. Several other endophytes were isolated from the seedlings and could have negatively influenced establishment of B. bassiana. The recovery of B. bassiana from sites distant from the point of inoculation indicates that the fungus has the potential to move throughout the plant. Published by Elsevier Ltd on behalf of The British Mycological Society.

Cross-species transferability of 41 previously published simple sequence repeat (SSR) markers was assessed for 11 species of the entomopathogenic fungus Metarhizium. A collection of 65 Metarhizium strains including all 54 used in a recent phylogenetic revision of the genus were characterized. Between 15 and 34 polymorphic SSR markers produced scorable PCR amplicons in seven species, including M. anisopliae, M. brunneum, M. guizhouense, M. lepidiotae, M. majus, M. pingshaense, and M. robertsii. To provide genotyping tools for concurrent analysis of these seven species fifteen markers grouped in five multiplex pools were selected based on high allelic diversity and easy scorability of SSR chromatograms. ISSN:0022-2011 ISSN:1096-0805

The anamorph genus Metarhizium is composed of arthropod pathogens, several with broad geographic and host ranges. Members of the genus, including "M. anisopliae var. frigidum" nomen nudum and Metarhizium flavoviride, have been used as biological insecticides. In a recent revision of the genus the variety "M. anisopliae var. frigidum" was suggested to be a synonym of M. flavoviride based largely on ITS sequence phylogenetic analysis. In this study we conducted morphological evaluations and multigene phylogenetic analyses with EF-1alpha, RPB1 and RPB2 for strains of M. flavoviride and "M. anisopliae var. frigidum." Included in these evaluations were the ex-type of M. flavoviride var. flavoviride and what likely would be considered the "ex-type' of the invalidly published taxon "M. anisopliae var. frigidum". Based on morphological and molecular evidence we conclude that "M. anisopliae var. frigidum" is distinct from M. flavoviride and the taxon M. frigidum sp. nov. is described.

Insect societies face constant challenges from disease agents. Ants deploy diverse antimicrobial compounds against pathogens and the key sources are metapleural glands (MGs). Are MG products passively secreted and used indiscriminately or are they selectively used when ants are challenged by pathogens? In 26 species from five subfamilies, ants use foreleg movements to precisely groom the MG opening. In the absence of experimental infection, MG grooming rates are low and workers groom themselves after contacting the MGs. The derived leaf-cutter ants ( Atta and Acromyrmex ) also groom their fungal gardens, substrata (leaves), queens and nest-mates after MG grooming. Atta respond to a challenge by fungal conidia by increasing the rate of MG grooming, but do not do so when an inert powder is applied. This increase occurs in the first hour after a potential infection, after which it returns to baseline levels. Ants with open MGs produce more infrabuccal pellets (IP) than ants with sealed MGs and conidia within pellets from the former are less likely to germinate. Thus, ants selectively groom their MGs when disease agents are present, suggesting that they also selectively use their MG secretions, which has important implications for understanding the evolution of hygienic behaviour in social groups.

The genus Mycocepurus is a phylogenetically basal attine ant, so studies of its biology may provide insight into the evolution of behaviours associated with fungus‐growing that characterize the tribe Attini. Mycocepurus smithii from Puerto Rico produces sexual females from July to September, but no males were observed in 2 years of observations, confirming previous observations elsewhere. Colonies were founded between July and August and most nests were haplometrotic (85% of 74 nests). After excavating a tunnel and small chamber, a foundress queen inserted her fore wings into the ceiling and used the wing surfaces as a platform on which the incipient fungal garden was grown. Foundresses foraged for substrate to grow the fungus garden. Growth of incipient colonies was slow: the first workers emerged 2–5 months after colony founding and, after 8 months, colonies contained on average only a single worker.

An isolate of the nematode-trapping fun- gus Monacrosporium drechsleri was collected from cul- tures of the root-knot nematode Meloidogyne arenaria that had been maintained on tomato roots in green- house pots in Beltsville, Maryland. The plant-parasitic nematodes Heterodera glycines, Meloidogyne incognita and Pratylenchus zeae and the free-living nematodes Caenorhabditis elegans and Panagrellus redivivus were placed on colonies of M. drechsleri grown in Petri dishes to study ability of the isolate to trap various nematode hosts. None of the nematodes placed near adhesive knobs were motile within 1 d. To determine where M. drechsleri fits within the existing phylogeny of nematode-trapping fungi, the ITS1-ITS2 regions of rDNA and the nuclear gene EF1-a were sequenced for the new isolate of M. drechsleri, for the species M. parvicolle and M. lysipagum, and for an isolate of M. ellipsosporum distinct from the one listed in GenBank. Parsimony trees were constructed showing the closest molecular relative of M. drechsleri to be the newly sequenced isolate of M. ellipsosporum; the latter had a highly divergent sequence from the sequence re- corded in GenBank for a different isolate of M. ellip- sosporum. Unique, consistent and discrete morpho- logical characters are absent in these related taxa, so an independent molecular character should be con- sidered essential for their accurate identification.

The genus Metarhizium historically refers to green-spored asexual insect pathogenic fungi. Through culturing and molecular methods, Metarhizium has been linked to Metacordyceps sexual states. Historically fungal nomenclature has allowed separate names for the different life stages of pleomorphic fungi. However, with the move to one name for one fungus regardless of life stage, there is a need to determine which name is correct. For Metarhizium the situation is complicated by the fact that Metacordyceps sexual states are interspersed among additional asexual genera, including Pochonia, Nomuraea and Paecilomyces. Metarhizium has priority as the earliest available name, but delimiting the boundaries of this genus remains problematic. To clarify relationships among these taxa we have obtained representative material for each genus and established a molecular dataset of the protein-coding genes BTUB, RPB1, RPB2 and TEF. The resulting phylogeny supports Metarhizium combining the majority of species recognized in Metacordyceps as well as the green-spored Nomuraea species and those in the more recently described genus Chamaeleomyces. Pochonia is polyphyletic, and we restrict the definition of this genus to those species forming a monophyletic clade with P. chlamydosporia, and the excluded species are transferred to Metapochonia gen. nov. It is our hope that this unified concept of sexual and asexual states in Metarhizium will foster advances in communication and understanding the unique ecologies of the associated species.

Ophiocordycipitaceae is a diverse family comprising ecologically, economically, medicinally, and culturally important fungi. The family was recognized due to the polyphyly of the genus Cordyceps and the broad diversity of the mostly arthropod-pathogenic lineages of Hypocreales. The other two cordyceps-like families, Cordycipitaceae and Clavicipitaceae, will be revised taxonomically elsewhere. Historically, many species were placed in Cordyceps, but other genera have been described in this family as well, including several based on anamorphic features. Currently there are 24 generic names in use across both asexual and sexual life stages for species of Ophiocordycipitaceae. To reflect changes in Art. 59 in the International Code of Nomenclature for algae, fungi, and plants (ICN), we propose to protect and to suppress names within Ophiocordycipitaceae, and to present taxonomic revisions in the genus Tolypocladium, based on rigorous and extensively sampled molecular phylogenetic analyses. When approaching this task, we considered the principles of priority, monophyly, minimizing taxonomic revisions, and the practical utility of these fungi within the wider biological research community.

Pathogens can cause substantial mortality of amphibian eggs. If the timing of hatching is phenotypically plastic, embryos could escape from otherwise lethal infections by hatching early. We tested this with the arboreal eggs of red-eyed treefrogs, Agalychnis callidryas. A filamentous ascomycete (Dothideales: Phaeosphaeriaceae) was present on ∼7% of egg clutches collected from a pond in the rain forest in Panama and, when present, killed 40% of the eggs, on average. Inoculation experiments confirmed that the fungus attacked and killed healthy embryos, establishing that this fungus is a pathogen of A. callidryas eggs. As predicted from life history theory, embryos hatched earlier from both naturally infected and inoculated clutches than from fungus-free control clutches. Within infected clutches, live embryos in contact with fungal hyphae hatched before those embryos not in contact with the fungus. Accelerated hatching allowed embryos to survive that otherwise would have been killed, and tadpoles hatched from infected clutches were themselves uninfected. Red-eyed treefrog embryos also hatch early if attacked by predators, apparently in response to vibratory cues. Because fungal infection provides no vibratory stimuli, embryos must respond to different cues in fungus-induced hatching than in predator-induced hatching. The behavioral decision of when to hatch is complex and merits further investigation. Our study indicates that pathogens can influence the timing of life history transitions, as do other stage-specific risks.

Analyses of DNA have not been widely used to distinguish coral sibling species. The three members of the Montastraea annularis complex represent an important test case: they are widely studied and dominate Caribbean reefs, yet their taxonomic status remains unclear. Analysis of amplified fragment length polymorphisms (AFLPs) and a microsatellite locus, using DNA from sperm, showed that Montastraea faveolata is genetically distinct. One AFLP primer yielded a diagnostic product (880 bp in M. faveolata 920 bp in M. franksi and M. annularis) whose homology was established by DNA sequencing. A second primer revealed a 630 bp band that was fixed in M. faveolata, and rare in M. franksi and M. annularis; in this case homologies were confirmed by Southern hybridizations. A tetranucleotide microsatellite locus with several alleles exhibited strong frequency differences between M. faveolata and the other two taxa. We did not detect comparable differences between M. annularis and M. franksi with either AFLPs (12 primers screened) or the microsatellite locus. Comparisons of AFLP patterns obtained from DNA from sperm, somatic tissues, and zooxanthellae suggest that the technique routinely amplifies coral (animal) DNA. Thus analyses based on somatic tissues may be feasible, particularly after diagnostic differences have been established using sperm DNA.