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Trichoderma species are among the most common fungi frequently isolated as saprotrophs from free soil, soil litter, dead wood, and the rhizosphere of different crops. Four sets of species-specific primers were designed from the tef1 and rpb2 genes, in order to identify Trichoderma asperellum ( tef1 gene), T. longibrachiatum ( tef1 gene), T. virens ( tef1 gene), and T. harzianum ( rpb2 gene). Here, we report the development of a multiplex PCR assay to detect and distinguish each of these four most common Trichoderma species-viz., T. asperellum, T. harzianum, T. longibrachiatum, and T. virens-simultaneously in a single reaction through their distinct amplicons of 507, 824, 452, and 330 bp, respectively. The developed multiplex PCR technique will provide a rapid, simple, and reliable alternative to conventional methods and a new site for identification of different species of Trichoderma in a single reaction. [ABSTRACT FROM AUTHOR]

Stromata of Trichoderma species having green ascospores were collected in various regions of China. Based on morphology of the sexual and asexual morph, culture characteristics, and sequence analyses of rpb2 and tef1 genes, 17 species with green ascospores were identified. Among them, Trichoderma rosulatum, T. rufobrunneum and T. stipitatum are described as new species, and seven other species are reported for the first time from China. Trichoderma rosulatum produces small bright yellow or pale greenish stromata with dense dark green ostioles and gliocladium-like conidiophores, shows a close relationship to T. thelephoricola, and belongs to the Chlorospora clade. Trichoderma rufobrunneum, which typically forms reddish brown stromata, is recognised as a member of the Harzianum clade. Trichoderma stipitatum is characterised by turbinate, pale yellow to nearly orange stromata and verticillium-like conidiophores; it forms a distinct, independent lineage with strong bootstrap support in the phylogenetic trees. The distinctions between the new species and their close relatives are discussed, and their phylogenetic positions are explored. [ABSTRACT FROM AUTHOR]

Collections of Trichoderma having hyaline ascospores from different areas of China were examined. Using combined analyses of morphological data, culture characters and phylogenetic information based on rDNA sequences of partial nuc translation elongation factor 1-α encoding gene (TEF1-α) and the gene encoding the second largest nuc RNA polymerase subunit (RPB2), three new species, Trichoderma applanatum, T. oligosporum and T. sinoluteum, were discovered and are described. Trichoderma applanatum produces continuous flat to pulvinate, white to cream stromata with dense orange or pale brown ostioles, and simple acremonium-like to verticillium-like conidiophores, belongs to the Hypocreanum clade and is closely related to T. decipiens. Trichoderma oligosporum forms reddish brown stromata with a downy surface, hyaline conidia and gliocladium-like conidiophores, and is closely related to but distinct from T. crystalligenum in the Psychrophila clade. Trichoderma sinoluteum, as a member of the Polysporum clade, is characterized by pale yellow stromata, white pustulate conidiomata, pachybasium-like conidiophores, and hyaline conidia. Differences between the new species and their close relatives are discussed. [ABSTRACT FROM AUTHOR]

A new species of the genus Orbilia, O. yuanensis (Orbiliales, Orbiliomycetes), was collected from Yunnan Province (China) and Taiwan. The fungus differs from previously known Orbilia species in that it has rod-shaped ascospores with a tear-shaped spore body. An anamorph that was obtained from ascospore isolates is characterized as producing long, curved macroconidia and much smaller, partly helicoid microconidia. Morphologically, this anamorph fits the polyphyletic genus Anguillospora. Based on a sequence analysis of the internal transcribed spacer region (ITS) gained from two isolates, O. yuanensis nested near species of the Orbilia luteorubella aggregate, although at a rather high distance. To date, there have been two reports regarding a connection between Orbilia species and their Anguillospora anamorph. The connection between O. yuanensis and its Anguillospora anamorph discussed here is the third published report. In this paper, both teleomorph and anamorph are illustrated and described. [ABSTRACT FROM AUTHOR]

Host jumps by microbial symbionts are often associated with bursts of species diversification driven by the exploitation of new adaptive zones. The objective of this study was to infer the evolution of habitat preference (decaying plants, soil, living fungi, and living plants), and nutrition mode (saprotrophy and mycoparasitism) in the fungal genus Trichoderma to elucidate possible interkingdom host jumps and shifts in ecology. Host and ecological role shifts were inferred by phylogenetic analyses and ancestral character reconstructions. The results support several interkingdom host jumps and also show that the preference for a particular habitat was gained or lost multiple times. Diversification analysis revealed that mycoparasitism is associated with accelerated speciation rates, which then suggests that this trait may be linked to the high number of species in Trichoderma. In this study it was also possible to infer the cryptic roles that endophytes or soil inhabitants play in their hosts by evaluating their closest relatives and determining their most recent ancestors. Findings from this study may have implications for understanding certain evolutionary processes such as species radiations in some hyperdiverse groups of fungi, and for more applied fields such as the discovery and development of novel biological control strategies. [ABSTRACT FROM AUTHOR]

Aims The aim was to isolate, identify and characterize endophytes from pigeon pea ( Cajanus cajan [ L.] Millsp.), as novel producer of cajanol and its in vitro cytotoxicity assay. Methods and Results Isolation, identification and characterization of novel endophytes producing cajanol from the roots of pigeon pea were investigated. The endophytes were identified as Hypocrea lixii by morphological and molecular methods. Cajanol produced by endophytes were quantified by liquid chromatography-tandem mass spectrometry ( LC- MS/ MS). R-18 produced the highest levels of cajanol (322·4 ± 10·6 μg l−1 or 102·8 ± 6·9 μg g−1 dry weight of mycelium) after incubation for 7 days. The cytotoxicity towards human lung carcinoma cells ( A549) of fungal cajanol was investigated in vitro. Conclusions First, a novel endophyte Hypocrea lixii, producing anticancer agent cajanol, was isolated from the host pigeon pea ( Cajanus cajan [ L.] Millsp.). Fungal cajanol possessed stronger cytotoxicity activity towards A549 cells in time- and dose-dependent manners. Significance and Impact of the Study This endophyte is a potential handle for scientific and commercial exploitation, and it could provide a promising alterative approach for large-scale production of cajanol to satisfy new anticancer drug development. [ABSTRACT FROM AUTHOR]

Commercial exploitation of lignocellulose for biotechnological production of fuels and commodity chemicals requires efficient-usually enzymatic-saccharification of the highly recalcitrant insoluble substrate. A key characteristic of cellulose conversion is that the actual hydrolysis of the polysaccharide chains is intrinsically entangled with physical disruption of substrate morphology and structure. This 'substrate deconstruction' by cellulase activity is a slow, yet markedly dynamic process that occurs at different length scales from and above the nanometer range. Little is currently known about the role of progressive substrate deconstruction on hydrolysis efficiency. Application of advanced visualization techniques to the characterization of enzymatic degradation of different celluloses has provided important new insights, at the requisite nano-scale resolution and down to the level of single enzyme molecules, into cellulase activity on the cellulose surface. Using true in situ imaging, dynamic features of enzyme action and substrate deconstruction were portrayed at different morphological levels of the cellulose, thus providing new suggestions and interpretations of rate-determining factors. Here, we review the milestones achieved through visualization, the methods which significantly promoted the field, compare suitable (model) substrates, and identify limiting factors, challenges and future tasks. Biotechnol. Bioeng. 2013; 110: 1529-1549. © 2013 Wiley Periodicals, Inc. [ABSTRACT FROM AUTHOR]

Xylan is an important part of plant biomass and represents a renewable raw material for biorefineries. Contrary to cellulose, the structure of hemicellulose is quite complex. Therefore, the biodegradation of xylan needs the cooperation of many enzymes. For industrial production of xylanase multienzyme complexes (cocktails) and selected monocomponent xylanases, different Trichoderma reesei mutants and recombinants are used. T. reesei QM 6a (wild-type parent of best existing mutants) was selected as a starting material in the 1960s when the modern in-depth analytical methods were not yet in use. Therefore, screening of fungi genetically close to T. reesei in biodegradation of xylan may have a scientific value. Fifteen different strains from Trichoderma section Longibrachiatum have been tested for extracellular xylan-degrading enzyme production on three carbon sources (wheat straw, corn fiber, and eucalyptus wood) in shake flask cultivation. The enzyme activities were evaluated by traditional colorimetric enzyme assays and by HPLC and matrix assisted laser desorption/ionization time-of-flight mass spectrometry. Degradation of xylan was studied on four different xylan-rich model substrates. T. reesei CPK 155, Trichoderma parareesei TUB F-2535, and Trichoderma gracile TUB F-2543 isolates were equally good or better in degradation of the wheat arabinoxylan (WAX) and corn fiber alcohol insoluble solids as hydolysis substrates than the well-known T. reesei QM 6a and RUT C30 strains. Though Trichoderma saturnisporum ATCC 18903 gave relatively low volumetric enzyme activities by traditional colorimetric assays, it could release quite large amount of hydrolysis products (mono- and oligosaccharides) from WAX. Therefore, these fungi may be potential candidates for further experiments. Enzyme production on wheat straw and corn fiber carbon sources was more effective than on eucalyptus wood. [ABSTRACT FROM AUTHOR]

A new species of Trichoderma, Trichoderma guizhouense, isolated from soil in Guizhou Province, is described based on morphology and phylogenetic analyses. This species exhibits characteristic Trichoderma morphology but is distinct from related species based on characters of phialides and conidia. Two DNA markers, the translation elongation factor 1 alpha ( tef1) and RNA polymerase II subunit b ( rpb2) were used for phylogenetic analyses. The correlation between morphological and molecular-based clustering demonstrated two studied isolates are a new species. [ABSTRACT FROM AUTHOR]

Certain species of the filamentous fungal genus Trichoderma (e.g. Trichoderma longibrachiatum and Trichoderma citrinoviride) are among the emerging clinical pathogens and also the most common species in the indoor space of mould-damaged buildings. The molecules involved in its pathology are not known. In the present study, we report that 0.5-2.6 wt% of the T. longibrachiatum mycelial biomass consisted of thermostable secondary metabolites mitochondriotoxic to mammalian cells. These were identified by LC/ MS as one 11-residue and eight 20-residue peptaibols, AcAib-Asn-Leu/Ile-Leu/Ile-Aib-Pro-Leu/Ile-Leu/Ile-Aib-Pro-Leuol/Ileol (1175 Da) and AcAib-Ala-Aib-Ala-Aib-Ala/Aib-Gln-Aib-Val/Iva-Aib-Gly-Leu/Ile-Aib-Pro-Val/Iva-Aib-Val/Iva/Aib-Gln/Glu-Gln-Pheol(1936-1965 Da) (Aib, α-aminoisobutyric acid; Ac, acetyl; Ileol, isoleucinol; Iva, isovaline; Leuol, leucinol; Pheol, phenylalaninol). The toxic effects on boar sperm cells depended on these peptaibols, named trilongins. The trilongins formed voltage dependent, Na+/ K+ permeable channels in biomembranes. The permeability ratios for Na+ ions, relative to K+, of the 11-residue trilongin channel (0.95 : 1) and the 20-residue trilongin channel (0.8 : 1) were higher than those of alamethicin. The combined 11-residue and 20-residue trilongins generated channels that remained in an open state for a longer time than those formed by either one of the peptaibols alone. Corresponding synergy was observed in toxicokinetics. With 11-residue and 20-residue trilongins combined 1 : 2 w/w, an effective median concentration ( EC50) of 0.6 μg· mL−1 was reached within 30 min, and the EC50 shifted down to 0.2 μg· mL−1 upon extended exposure. By contrast, with 11-residue or 20-residue trilonging separately in 30 min of exposure, the EC50 values were 15 and 3 μg· mL−1, respectively, and shifted down to 1.5 and 0.4 μg· mL−1 upon extended exposure. This is the first report on ion-channel forming peptaibols with synergistic toxicity from T. longibrachiatum strains isolated from clinical samples. Database Nucleotide sequence data have been deposited in the GenBank database under accession numbers HQ593512 and HQ593513, . [ABSTRACT FROM AUTHOR]

The Longibrachiatum Clade of Trichoderma is revised. Eight new species are described ( T. aethiopicum, T. capillare, T. flagellatum, T. gillesii, T. gracile, T. pinnatum, T. saturnisporopsis, T. solani). The twenty-one species known to belong to the Longibrachiatum Clade are included in a synoptic key. Trichoderma parareesei and T. effusum are redescribed based on new collections or additional observations. Hypocrea teleomorphs are reported for T. gillesii and T. pinnatum. Previously described species are annotated. [ABSTRACT FROM AUTHOR]

Monoconidial cultures of 33 isolates of Trichoderma from Buenos Aires Province, Argentina were characterized on the basis of twenty eight morphological, physiological and biochemical features. All of them were screened for proteinase, endochitinase and β-1,3 glucanase activity. Universally primed PCR (UP-PCR) and inter-simple sequence repeat (ISSR) techniques were used to examine the genetic variability among isolates, which resulted in 127 bands for the total number of isolates. These results were subjected to numerical analysis revealing 20 haplotypes grouped in five clusters. The ability of Trichoderma isolates to antogonize soil-borne fungal plant pathogens using a dual culture assay was done against five fungal species: Alternaria sp., Bipolaris sorokiniana, Fusarium graminearum, F. solani, and Pyricularia oryzae. The highest inhibition values (85% RI) were obtained against B. sorokiniana and P. oryzae. Three isolates of T. harzianum named as FCCT2, FCCT3 and FCCT9 were capable of causing a high growth inhibition on four of the fungal species assayed, which was in agreement with their higher extracellular hydrolytic activity. Our results suggest that these isolates have the potential to be effective agents for biocontrol of cereal and tomato fungal pathogens. [ABSTRACT FROM AUTHOR]

A member of the nematophagous anamorph genus Arthrobotrys was isolated from its teleomorph Orbilia blumenaviensis comb. nov. (= Orbilia fici), a species closely related to O. auricolor but deviating in having lanceolate paraphyses. In the presence of nematodes, the anamorph forms three-dimensional adhesive networks. A trimorphism in its conidia was observed which vary in shape and number of septa. In the first isolate, two types of heteropolar conidia were obtained. These differ markedly from the type strain of A. vermicola, which forms mainly homopolar (fusoid) conidia. In a further ascospore isolate of O. blumenaviensis, however, mainly fusoid conidia referable to A. vermicola occurred. Combining morphological and phylogenetic analysis, we conclude that these isolates with differently shaped conidia and/or differences in the ITS rDNA (ca. 2-4%) belong to a single anamorph species, A. vermicola. Both teleomorph and anamorph are illustrated and described. [ABSTRACT FROM AUTHOR]

The oak platypodid, Platypus quercivorus (Murayama), is the most destructive pest of Quercus and related fagaceous species in Japan. To find an effective biocontrol agent, we determined the effectiveness of entomopathogenic microorganisms against P. quercivorus. P. quercivorus-infested Oak ( Quercus serrata, Q. crispula and Q. acutissima) logs were obtained from various regions in Japan, and 28 isolates of microorganisms were isolated from P. quercivorus larvae from those logs. Using molecular methods, the microbes were then classified into 18 species. After a bioassay, we found that three fungi, Lecanicillium sp., Isaria tenuipes, Beauveria bassiana, and the bacterium Serratia marcescens had the greatest potential as biological control agents against P. quercivorus. Another bioassay was then performed to determine the effect of spore or cell densities on the pathogenicity of the four microorganisms, and optimum spore or cell densities for the microbes were determined to be 10 and 10/ml. Based on the results and other criteria, we tentatively selected B. bassiana as the best candidate for future use against P. quercivorus larvae. [ABSTRACT FROM AUTHOR]

A new anamorphic fungus is described based on four isolates from ascospores of Orbilia aff. luteorubella. This fungus differs from previously known Orbilia anamorphs in producing inversely pyramidal, unicellular conidia with several protuberances at their distal end. Conidia produce 1-7 prominent denticles that emerge from a node at the conidiophore apex. Conidiogenesis is holoblastic. Because phylogenetic analysis indicated greater than 90% ITS sequence similarities among the four isolates they are treated here as a single species. In the sequence analysis of the internal transcribed spacer region (ITS) these isolates and other sequences identified as O. aff. luteorubella were nested within Orbilia and formed a clade with 99% bootstrap support. This clade is separated from nematode-trapping species of Orbilia. Based on both morphological and molecular analyses, we propose a new genus, Pseudotripoconidium. [ABSTRACT FROM AUTHOR]

Fungi of the genus Trichoderma are soilborne, green-spored ascomycetes that can be found all over the world. They have been studied with respect to various characteristics and applications and are known as successful colonizers of their habitats, efficiently fighting their competitors. Once established, they launch their potent degradative machinery for decomposition of the often heterogeneous substrate at hand. Therefore, distribution and phylogeny, defense mechanisms, beneficial as well as deleterious interaction with hosts, enzyme production and secretion, sexual development, and response to environmental conditions such as nutrients and light have been studied in great detail with many species of this genus, thus rendering Trichoderma one of the best studied fungi with the genome of three species currently available. Efficient biocontrol strains of the genus are being developed as promising biological fungicides, and their weaponry for this function also includes secondary metabolites with potential applications as novel antibiotics. The cellulases produced by Trichoderma reesei, the biotechnological workhorse of the genus, are important industrial products, especially with respect to production of second generation biofuels from cellulosic waste. Genetic engineering not only led to significant improvements in industrial processes but also to intriguing insights into the biology of these fungi and is now complemented by the availability of a sexual cycle in T. reesei/ Hypocrea jecorina, which significantly facilitates both industrial and basic research. This review aims to give a broad overview on the qualities and versatility of the best studied Trichoderma species and to highlight intriguing findings as well as promising applications. [ABSTRACT FROM AUTHOR]

This investigation aimed to assess whether MALDI-TOF MS analysis of the proteome could be applied to the study of Trichoderma, a fungal genus selected because it includes many species and is phylogenetically well defined. We also investigated whether MALDI-TOF MS analysis of peptide mass fingerprints would reveal apomorphies that could be useful in diagnosing species in this genus. One hundred and twenty nine morphologically and genetically well-characterized strains of Hypocrea and Trichoderma, belonging to 25 species in 8 phylogenetic clades, were analyzed by MALDI-TOF MS mass spectrometry. The resulting peak lists of individual samples were submitted to single-linkage cluster analysis to produce a taxonomic tree and were compared to ITS and tef1 sequences from GenBank. SuperSpectra™ for the 13 most relevant species of Trichoderma were computed. The results confirmed roughly previously defined clades and sections. With the exceptions of T. saturnisporum (Longibrachiatum Clade) and T. harzianum (Harzianum Clade), strains of individual species clustered very closely. T. polysporum clustered distantly from all other groups. The MALDI-TOF MS analysis accurately reflected the phylogenetic classification reported in recent publications, and, in most cases, strains identified by DNA sequence analysis clustered together by MALDI-TOF MS. The resolution of MALDI-TOF MS, as performed here, was roughly equivalent to ITS rDNA. The MALDI-TOF MS technique analyzes peptides and represents a rough equivalent to sequencing, making this method a useful adjunct for determination of species limits. It also allows simple, reliable, and quick species identification, thus representing a valid alternative to gene sequencing for species diagnosis of Trichoderma and other fungal taxa. [ABSTRACT FROM AUTHOR]

Background: Trichoderma reesei, a mitosporic green mould, was recognized during the WW II based on a single isolate from the Solomon Islands and since then used in industry for production of cellulases. It is believed to be an anamorph (asexual stage) of the common pantropical ascomycete Hypocrea jecorina. Methodology/Principal Findings: We combined molecular evolutionary analysis and multiple methods of phenotype profiling in order to reveal the genetic relationship of T. reesei to H. jecorina. The resulting data show that the isolates which were previously identified as H. jecorina by means of morphophysiology and ITS1 and 2 (rRNA gene cluster) barcode in fact comprise several species: i) H. jecorina/T. reesei sensu stricto which contains most of the teleomorphs (sexual stages) found on dead wood and the wild-type strain of T. reesei QM 6a; ii) T. parareesei nom. prov., which contains all strains isolated as anamorphs from soil; iii) and two other hypothetical new species for which only one or two isolates are available. In silico tests for recombination and in vitro mating experiments revealed a history of sexual reproduction for H. jecorina and confirmed clonality for T. parareesei nom. prov. Isolates of both species were consistently found worldwide in pantropical climatic zone. Ecophysiological comparison of H. jecorina and T. parareesei nom. prov. revealed striking differences in carbon source utilization, conidiation intensity, photosensitivity and mycoparasitism, thus suggesting adaptation to different ecological niches with the high opportunistic potential for T. parareesei nom. prov. Conclusions: Our data prove that T. reesei belongs to a holomorph H. jecorina and displays a history of worldwide gene flow. We also show that its nearest genetic neighbour - T. parareesei nom. prov., is a cryptic phylogenetic agamospecies which inhabits the same biogeographic zone. These two species thus provide a so far rare example of sympatric speciation within saprotrophic fungi, with divergent ecophysiological adaptations and reproductive strategies. [ABSTRACT FROM AUTHOR]

Sapstain fungi, which reduce the value of pine wood, were isolated from an industrial wood yard in north-eastern Germany. The predominant wood discolouring species on the industrial wood yard and in the forests of the investigated region was Sphaeropsis sapinea (syn. Diplodia pinea), but Ophiostoma minus was also found. These fungi were challenged with antagonistic micro-organisms in laboratory trials and in field experiments. Amongst the tested microorganisms only strains of filamentous fungi ( Trichoderma sp. and Phlebiopsis gigantea) could control the sapstain fungi on pine wood blocks efficiently. Although P. gigantea was unable to inhibit stain formation completely, the wood was bleached by this fungus in later incubation stages. In two field trials, sapstain on pine wood logs was successfully retarded for a period of 10 and 12 weeks, respectively, with a white sporulating mutant of Trichoderma harzianum. Thus, the concept using filamentous fungi as antagonists against sapstain developed under laboratory conditions also proved to be valid under natural conditions in forest eco-systems. [ABSTRACT FROM AUTHOR]

Abstract  We report the effects of some bisorbicillinoids isolated from biomass of the fungus Trichoderma citrinoviride on settling and feeding preference of the aphid Schizaphis graminum. Purification of the fungal metabolites was carried out by a combination of column chromatography and thin-layer chromatography using direct and reverse phases. Chemical identification was performed by spectroscopic methods including nuclear magnetic resonance and mass spectrometry. The identified bisorbicillinoids appeared to be bislongiquinolide, its 16,17-dihydro derivative, trichodimerol, and dihydrotrichodimerol. A feeding preference test with alate morphs of S. graminum was used to identify the active fractions. Among the four bisorbicillinoids, dihydrotrichodimerol and bislongiquinolide influenced aphid feeding preference, restraining specimens from settling on leaves treated with metabolites. Taste neurons sensitive to these compounds, particularly to bislongiquinolide, were located on tarsi of the S. graminum alate morphs. [ABSTRACT FROM AUTHOR]

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Copyright of Canadian Journal of Microbiology is the property of Canadian Science Publishing and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Hypocrea/Trichoderma is a genus of soil-borne or wood-decaying fungi containing members important to mankind as producers of industrial enzymes and biocontrol agents against plant pathogens, but also as opportunistic pathogens of immunocompromised humans and animals, while others can cause damage to cultivated mushroom. With the recent advent of a reliable, BarCode-aided identification system for all known taxa of Trichoderma and Hypocrea, it became now possible to study some of the biological fundamentals of the diversity in this fungal genus in more detail. In this article, we will therefore review recent progress in (1) the understanding of the geographic distribution of individual taxa; (2) mechanisms of speciation leading to development of mushroom diseases and facultative human mycoses; and (3) the possible correlation of specific traits of secondary metabolism and molecular phylogeny. [ABSTRACT FROM AUTHOR]

The Brevicompactum clade is recognized as a separate lineage in Trichoderma/ Hypocrea. This includes T. brevicompactum and the new species T. arundinaceum, T. turrialbense, T. protrudens and Hypocrea rodmanii. The closest relative of the Brevicompactum clade is the Lutea clade. With the exception of H. rodmanii, all members of this clade produce the simple trichothecene-type toxins harzianum A or trichodermin. All members of the clade produce peptaibiotics, including alamethicins. Strains previously reported as T. harzianum (ATCC 90237), T. viride (NRRL 3199) or Hypocrea sp. (F000527, CBS 113214) to produce trichothecenes are reidentified as T. arundinaceum. The Brevicompactum clade is not closely related to species that have biological application. [ABSTRACT FROM AUTHOR]

The protologues are reviewed of all 227 fungal names with epithets that refer to New Zealand, and orthographic corrections are indicated for those epithets that do not comply with the requirements of the International code of Botanical Nomenclature (ICBN). One hundred and seventy-one of the names were "new" (i.e., basionyms, nomina nova, nomina nuda); 21% of them had epithets that failed to comply with ICBN orthographic requirements. Fifty-six of the names were recombinations, 30% of which had epithets that did not match the orthography of their basionyms. A set of standard orthographies is suggested, and the appropriateness of their use is discussed. [ABSTRACT FROM AUTHOR]

The ribosomal Internal Transcribed Spacer (ITS) regions of the two recognized species of Coccidioides were studied using a reference set of strains that had been previously identified with species defining microsatellite polymorphisms. Unambiguous identification of the two species proved to be possible by amplifying and sequencing the ITS region. PCR-reactions are sensitive to amplification conditions requiring their careful optimization. Stable amplification and sequencing was achieved with primers ITS3 and 4, enabling species diagnosis. Alternatively, Restriction Fragment Length Polymorphism (RFLP) of the entire ITS region using an annealing temperature of 52°C with the restriction enzymes BsrI and XcmI can also distinguish the species. Three strains typifying the species, Glenospora meteuropaea, G. metamericana and Geotrichum louisianoideum, were analyzed and found to be conspecific with C. posadasii. Although these species have nomenclatural priority over C. posadasii, the latter will be proposed for conservation as it has been included in the US select agent list. In addition, Coccidioides immitis is neotypified in this report. Results of antifungal susceptibility testing did not reveal differences between the two species. [ABSTRACT FROM AUTHOR]

Abstract??Seventy-nineTrichodermastrains were isolated from soil taken from 28 commercial plantations ofAgave tequilanacv. ?Azul? in the State of Jalisco,Mexico. Nine of these isolates produced nonvolatile metabolites that completely inhibited the growth ofThielaviopsis paradoxaon potato dextrose agar plates. These isolates were identified asTrichoderma longibrachiatumon the basis of their morphology and DNA sequence analysis of two genes (ITS rDNA and translation elongation factor EF-1?). Mycoparasitism ofTh. paradoxabyT. longibrachiatumstrains in dual cultures was examined by scanning electron microscopy. TheTrichodermahyphae grew alongside theTh. paradoxahyphae, but penetration ofThielaviopsishyphae byTrichodermawas no apparent. Aleurioconidia ofTh. paradoxawere parasitized byTrichoderma. Both hyphae and aleurioconidia ofTh. paradoxalost turgor pressure, wrinkled, collapsed and finally disintegrated. In liquid cultures, all nineTrichodermaisolates produced proteases, ?-1,3-glucanases and chitinases that would be responsible for the degradation ofThielaviopsishyphae. These results demonstrate that the modes of action ofT. longibrachiatuminvolved againstTh. paradoxa in vitroexperiments are mycoparasitism and the production of nonvolatile toxic metabolites. [ABSTRACT FROM AUTHOR]

This survey is concerned only with filamentous fungi living in the soil layer. The observed diversity of soil fungi largely depends on the method used and the numbers of isolates obtained. Particle-plating usually yields higher numbers of taxa than dilution plating. The Centraalbureau voor Schimmelcultures (CBS) preserves a great diversity of soil fungi. The CBS database contained 2,210 species of soil fungi in 2001, an estimated 70% of the known species available in culture. Thus, the current estimate for described culturable soil fungi is approximately 3,150 species, many of which have a cosmopolitan distribution. Adding the ca. 150 spp. of nonculturable Glomerales results in 3,300 species of currently known soil fungi. Molecular studies in such groups as Fusarium, Trichoderma, Penicillium, and Aspergillus are finding a number of more narrowly distributed cryptic species. Thus the number of species of soil fungi is expected to be considerably higher than the 3300 species currently known. [ABSTRACT FROM AUTHOR]

Toward a better understanding of the biochemical events that lead to biocontrol of plant pathogenic fungi by Hypocrea/Trichoderma spp., we investigated the importance of carbon catabolite (de)repression and cellulase formation in the antagonization of Pythium ultimum by Hypocrea jecorina ( Trichoderma reesei) on agar plates and in planta. Hypocrea jecorina QM9414 could antagonize and overgrow P. ultimum but not Rhizoctonia solani in plate confrontation tests, and provided significant protection of zucchini plants against P. ultimum blight in planta. A carbon catabolite derepressed cre1 mutant of H. jecorina antagonized P. ultimum on plates more actively and increased the survival rates of P. ultimum-inoculated zucchini plants in comparison with strain QM9414. A H. jecorina mutant impaired in cellulase induction could also antagonize P. ultimum on plates and provided the same level of protection of zucchini plants against P. ultimum as strain QM9414 did. We conclude that cellulase formation is dispensable for biocontrol of P. ultimum, whereas carbon catabolite derepression increases the antagonistic ability by apparently acting on other target genes. [ABSTRACT FROM AUTHOR]

Umbelopsis ramanniana is a well-known species in this genus. A characteristic morphological feature of this fungus is the remarkable variation in the sporangiospore shape, which implies the genetic variations occur in the nucleotide sequences of the internal transcribed spacer (ITS) regions of the nuclear ribosomal DNA (nrDNA) in the U. ramanniana isolates. The relationship between the variations of the sequences of the nrDNA ITS regions and those of the sporangiospore morphology was investigated for 12 isolates of U. ramanniana collected in Europe. Neighbor-joining and parsimony analyses on the sequences suggested that these isolates split into three groups. Precise examination of the morphology showed that the isolates of those respective groups were different from each other in their sporangiospore shape. The present study implies at least three intraspecific groups exist in U. ramanniana and that the variations in the nucleotide sequences of the nrDNA ITS regions correlate well with those in the sporangiospore shape in these intraspecific groups. [ABSTRACT FROM AUTHOR]

The interaction between Trichoderma pseudokoningii (Rifai) 511, 2212, 741A, 741B and 453 and the arbuscular mycorrhizal fungi Glomus mosseae (Nicol. & Gerd.) Gerdemann & Trappe BEG12 and Gigaspora rosea Nicolson & Schenck BEG9 were studied in vitro and in greenhouse experiments. All T. pseudokoningii strains inhibited the germination of G. mosseae and Gi. rosea except the strain 453, which did not affect the germination of Gi. rosea. Soluble exudates and volatile substances produced by all T. pseudokoningii strains inhibited the spore germination of G. mosseae. The germination of Gi. rosea spores was inhibited by the soluble exudates produced by T. pseudokoningii 2212 and 511, whereas T. pseudokoningii 714A and 714B inhibited the germination of Gi. rosea spores by the production of volatile substances. The strains of T. pseudokoningii did not affect dry matter and percentage of root length colonization of soybean inoculated with G. mosseae, except T. pseudokoningii 2212, which inhibited both parameters. However, all T. pseudokoningii strains decreased the shoot dry matter and the percentage of AM root length colonization of soybean inoculated with Gi. rosea. The saprotrophic fungi tested seem to affect AM colonization of root by effects on the presymbiotic phase of the AM fungi. No influence of AM fungi on the number of CFUs of T. pseudokoningii was found. The effect of saprotrophic fungi on AM fungal development and function varied with the strain of the saprotrophic species tested. [ABSTRACT FROM AUTHOR]

Hypocrea phyllostachydis was collected from the bamboo species Phyllostachys bambusoides in southwestern France (Dept. Pyréneés Atlantiques). It can be distinguished from other morphologically similar species by the small subglobose or broadly ellipsoidal conidia and small ascospores. Conidiophores of the Trichoderma state of H. phyllostachydis do not branch in a pyramidal fashion, as is typical of most species of Trichoderma. Rather, it has an irregular branching pattern, with a long central axis and relatively short lateral secondary branches. A key to species of Hypocrea with green ascospores from France is presented. [ABSTRACT FROM AUTHOR]

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Trichoderma species are filamentous fungi that were previously considered to be culture contaminants. We report 2 well-documented cases of invasive Trichoderma infections, and we comprehensively review the literature on this topic. Trichoderma species are mainly responsible for continuous ambulatory peritoneal dialysis-associated peritonitis (7 cases) and invasive infections in immunocompromised patients (9 cases) with a hematologic malignancy or solid-organ transplant. Definitive diagnosis is difficult to achieve because of the lack of specific diagnosis tools. Species identification can benefit from a molecular approach. Trichoderma longibrachiatum is the most common species involved in these infections. Regardless of the type of infection, the prognosis was poor, with 8 deaths among 18 cases. This may be partially because of the resistance of these organisms to the majority of available antifungal agents, including amphotericin B. Trichoderma species now should be added to the growing list of emerging filamentous fungal pathogens. [ABSTRACT FROM AUTHOR]

Hypocrea patella is reevaluated. Its Trichoderma anamorph is described and the phylogenetic position of the species is determined through sequences of the ITS regions of rDNA. It is sister to a clade that includes Trichoderma longibrachiatum/H. schweinitzii. Hypocrea patella f. tropica is accepted for a Costa Rican collection. Hypocrea neorufa and its Trichoderma anamorph are described. Its phylogenetic position is determined by sequences of the ITS region of rDNA and the protein-coding translation-elongation factor (EF-1α). It is derived from within a clade that includes T. viride/H. rufa, T. atroviride/H. atroviridis, T. koningii/H. koningii and T. asperellum. [ABSTRACT FROM AUTHOR]

Molecular approaches including internal transcribed spacer (ITS) sequences of ribosomal DNA, universal primer polymerase chain reaction (UP-PCR) fingerprinting, and DNA–DNA hybridization were used to study the genetic relatedness of species within Trichoderma sect. Pachybasium. In the analysis of ITS and 5.8S sequences of ribosomal DNA, parsimony analysis demonstrated that forty-one strains were distributed into five main groups supported by high bootstrap values. The species of Trichoderma sect. Pachybasium were clustered into groups I, II, and IV, with the strains of Trichoderma fasculatum and Trichoderma strictipile forming a separate branch, an independent group V. Some species within each group showed nearly identical sequence differences (fewer than 1–3 bp). UP-PCR and DNA–DNA hybridization were further used to clarify the genetic relatedness of these species with highly similar ITS sequences. Highly similar or identical UP-PCR profiles and high values of DNA complementarity (>70%) were observed among some species, Trichoderma hamatum and Trichoderma pubescens; Trichoderma croceum,Trichoderma polysporum and Trichoderma album; Trichoderma crassum and Trichoderma flavofuscum; and Trichoderma strictipile and Trichoderma fasciculatum. Although every species can be differentiated morphologically, the species showed highly similar molecular characteristics in the above cases, indicating that they could be conspecific. However, in some cases (Trichoderma longipile, T. crassum and T. flavofuscum; Trichoderma fertile and Trichoderma minutisporum; Trichoderma tomentosum, Trichoderma inhamatum and Trichoderma harzianum) there were discriminative patterns of UP-PCR and (or) low levels (<50%) of DNA–DNA hybridization; even their ITS sequences were similar, suggesting a closely phylogenetic relationship.Key words: fingerprinting, ITS sequencing, DNA–DNA hybridization, taxonomy, Trichoderma.Nous avons eu recours à des approches moléculaires incluant les séquences d'espaceur interne transcrit (ITS) d'ADN ribosomal, la caractérisation par réaction de polymérase en chaîne à amorce universelle (UP-PCR), et l'hybridation ADN–ADN, afin d'étudier la parenté génétique d'espèces appartenant à Trichoderma sect. Pachybasium. Dans l'analyse de séquences d'ADN ribosomal d'ITS et de 5,8S, une analyse par parcimonie a démontré que quarante-et-une souches pouvaient être classées dans cinq groupes principaux soutenus par des valeurs de bootstrap élevées. Les espèces de Trichoderma sect. Pachybasium ont été rassemblées dans les groupes I, II et IV, alors que les souches de Trichoderma fasculatum et Trichoderma strictipile ont formé un embranchement distinct, un groupe V indépendant. Certaines espèces à l'intérieur de chaque groupe affichaient des différences de séquences quasiment identiques (moins de 1–3 pb). Les analyses de UP-PCR et d'hybridation se sont rajoutées afin de clarifier la parenté génétique des ces espèces ayant des séquences ITS très semblables. Nous avons observé des profils de UP-PCR fort semblables ou identiques ainsi que des valeurs élevées de complémentarité d'ADN (>70%) parmi certaines espèces, notamment Trichoderma hamatum et Trichoderma pubescens; Trichoderma croceum,Trichoderma polysporum et Trichoderma album; Trichoderma crassum etTrichoderma flavofuscum; et finalement Trichoderma strictipile et Trichoderma fasciculatum. Bien que chacune des espèces pouvaient être différentiées selon la morphologie, les espèces mentionnées ci-haut affichaient des caractéristiques moléculaires fortement semblables, ce qui indique qu'elles pourraient être conspécifiques. Cependant, dans certains cas (Trichoderma longipile, T. crassum et T. flavofuscum; Trichoderma fertile et Trichoderma minutisporum; Trichoderma tomentosum, Trichoderma inhamatum et Trichoderma harzianum) nous avons obtenu des patrons de UP-PCR discriminants et/ou de bas niveaux (<50%) d'hybridation ADN–ADN, même si leurs séquences ITS étaient proches, ce qui signalait l'existence d'un lien phylogénétique rapproché.Mots clés : caractérisation, séquençage d'ITS, hybridation ADN–ADN, taxonomie, Trichoderma.[Traduit par la Rédaction] [ABSTRACT FROM AUTHOR]

Phylogenetic relationships within the genus Cladonia, including Cladina (Cladoniaceae, Lecanoromycetes), were reconstructed based upon simultaneous analyses of DNA sequences and morphological and chemical data. We used sequences from the internal transcribed spacer 1 (ITS1), the 5.8 rDNA gene, and the internal transcribed spacer 2 (ITS2) of the nuclear rDNA gene cluster, and partial sequences from the protein-coding β-tubulin gene. The analyses included 235 specimens of 168 taxa representing all currently recognized sections of Cladonia and Cladina and the outgroup genera Cladia, Pycnothelia, and Ramalea. Analyses were performed using optimization alignment with three different parameter values. The results of all analyses support the inclusion of Cladina in Cladonia. The current sectional division of Cladonia was not supported, and a new provisional classification for the genus is proposed. [Copyright &y& Elsevier]

Colletotrichum species are plant pathogens, saprobes, and endophytes in many economically important hosts. Many studies have investigated the diversity and pathogenicity of Colletotrichum species in common ornamentals, fruits, and vegetables. However, Colletotrichum species occurring in aquatic plants are not well known. During the investigation of the diversity of endophytic fungi in aquatic plants in southwest China, 66 Colletotrichum isolates were obtained from aquatic plants there, and 26 of them were selected for sequencing and analyses of actin (ACT), chitin synthase (CHS-1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), the internal transcribed spacer (ITS) region, and β-tubulin (TUB2) genomic regions. Based on morphological characterization and multi-locus phylogenetic analyses, 13 Colletotrichum species were recognized, namely, C. baiyuense sp. nov., C. casaense sp. nov., C. demersi sp. nov., C. dianense sp. nov., C. fructicola, C. garzense sp. nov., C. jiangxiense, C. karstii, C. philoxeroidis sp. nov., C. spicati sp. nov., C. tengchongense sp. nov., C. vulgaris sp. nov., C. wuxuhaiense sp. nov. Two species complexes, the C. boninense species complex and C. gloeosporioides species complex, were found to be associated with aquatic plants. Pathogenicity tests revealed a broad diversity in pathogenicity and aggressiveness among the eight new Colletotrichum species. [ABSTRACT FROM AUTHOR]

Pinus sylvestris bark represents a rich source of active compounds with antifungal, antibacterial, and antioxidant properties. The current study aimed to evaluate the antifungal potential of P. sylvestris bark against Botryosphaeria dothidea, Dothiorella sarmentorum, and Neofusicoccum parvum (Botryosphaeriaceae) through its chemical (water extracts) and biological (Trichoderma spp. isolated from the bark) components. The water bark extracts were prepared at two temperatures (80 and 120 °C) and pH regimes (7 and 9). The presence of bark extracts (30%) caused inhibition of mycelial growth of B. dothidea and D. sarmentorum for 39 to 44% and 53 to 60%, respectively. Moreover, we studied the antagonistic effect of three Trichoderma isolates originating from the pine bark. Trichoderma spp. reduced growth of B. dothidea by 67%–85%, D. sarmentorum by 63%–75% and N. parvum by 55%–62%. Microscopic examination confirmed typical mycoparasitism manifestations (coiling, parallel growth, hook-like structures). The isolates produced cellulase, β-glucosidase and N-acetyl-β-glucosaminidase. The volatile blend detected the emission of several volatile compounds with antimicrobial activity, including nonanoic acid, cubenene, cis-α-bergamotene, hexanedioic acid, and verticillol. The present study confirmed in vitro potential of P. sylvestris bark extracts and Trichoderma spp. against the Botryosphaeriaceae. The study is an important step towards the use of environmentally friendly methods of Botryosphaeriaceae disease control. [ABSTRACT FROM AUTHOR]

Soil microorganisms are one of the main indicators used for assessing the stability of the soil ecosystem, the metabolism in the soil, and its fertility. The most important are the active soil microorganisms and the influence of the fertilizer applied to the soil on the abundance of these microorganisms. We aimed to investigate how the applied organic fertilizers affect the most active soil microorganisms, which determine the soil fertility and stability. Fungi, yeast-like fungi abundance, and abundance of three physiological groups of bacteria were analyzed: non-symbiotic diazotrophic, organotrophic, and mineral nitrogen assimilating. This study is valuable because relatively few similar studies have been performed on infertile Lithuanian soils. The first results of a long-term study were obtained. The results show the effect of fertilizers on trends in the changes of microorganism community diversity; however, more analysis is needed to assess the impact of organic fertilizers on the most active soil microorganisms. Therefore, the investigation was continued. The results of the 2020 quantitative analysis of culturable soil microorganisms show that the highest abundance of organotrophic and non-symbiotic diazotrophic bacteria were recorded during the summer season. Meanwhile, the abundance of bacteria assimilating mineral nitrogen and fungi was higher in autumn. Agrochemical parameters were determined at the beginning of the experiment. The highest concentration of Nmin in the soil was determined after fertilizing the plants with the combination of granulated poultry manure (N170) + biological substance Azotobacter spp. The yield of barley was calculated. It was found that the highest yield of spring barley in 2020 was obtained by fertilizing the experimental field with organic in combination with mineral fertilizers. [ABSTRACT FROM AUTHOR]

A new species, Colletotrichum menglaense, isolated from air in Mengla, Xishuangbanna, Yunnan Province, China, was characterized and described combining morphological characteristics and multigene phylogenetic analysis. Morphologically, it is characterized by oblong, sometimes slightly constricted, micro-guttulate conidia and simple obovoid to ellipsoidal appressoria. Phylogenetic analysis of the ITS, ACT, CHS, and GAPDH sequences showed that C. menglaense belongs to the C. gloeosporioides complex. The pathogenicity of C. menglaense on fruits of several crop plants, including strawberry, orange, grape, tomato, and blueberry, was tested and confirmed by the re-isolation of C. menglaense. [ABSTRACT FROM AUTHOR]