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Teleomorph of Hypocrea crystalligena; a, b, d, f. Fresh stromata, (a. young, velutinous, b. with visible ostioles, f. (over-) mature). e, h, i. Dry stromata (e. fraction of d; i. showing white powder on surface). c. Surface of wet stroma showing hyaline ostioles. g. Ostiole in section showing periphyses and apical cells. j. Perithecium in section. k. Ostiole in face view. l. Surface of stroma in face view. m. Subperithecial tissue in section. n, o, p. Asci with ascospores, n, o in cotton blue/lactic acid.
Source publication
The new species Hypocrea crystalligena (Hypocreales, Ascomycota, Fungi) is described as a holomorph and characterized based on an integrated phenotypic and phylogenetic approach, using teleomorph and anamorph morphologies, culture studies and analyses of phylogenetic markers including internal transcribed spacer 1 and 2 (ITS1 and 2), two last intro...
Contexts in source publication
Context 1
... when fresh (FIG. 3a, b, d, f ) ca. 1-6(-8) diam, 0.5-2 mm high, single, gregarious to densely aggregated in large numbers, starting as white mycelium, becoming compacted, flat to pulvinate, light (yellowish-, ochraceous-to light reddish-)brown, 4A4, 5-6B5-6 to 6-7D5-6, with white margin, finely tomentose to velutinous; later glabrous, distinctly and thickly ...
Context 2
... when dry (FIG. 3e, h, i), (0.7-)1.5-3.5 (-4.7) mm long, (0.5-)1.2-3.0(-4.0) mm wide, (0.2-) 0.5-1.0(-1.7) mm high (n 5 30), unchanged or slightly darker in 3% KOH, flatter than fresh, pulvinate to discoid, frequently surface finely but distinctly verrucous or tubercular to wrinkled; imma- ture stromata velvety, short-haired, surface of mature stromata usually ...
Context 3
... dark (reddish-)brown 6EF6-8 to 7-8EF5- 8. Ostiolar area (ostioles plus dark margin) in face view, (24-)32-53(-63) mm (n 5 30), inconspicuous and only visible under high magnification or conspic- uous and semiglobose in some stromata, after addition of water ostioles generally appearing as light and hyaline dots on a bright reddish stroma surface (FIG. 3c). Ostioles in section (FIG. 3g, j) Peridium 8-13(-15) mm thick at base, (14-)15-20 (-22) mm at apex (n 5 15), yellowish brown to reddish brown, slightly paler than cortex, apically thickened and directly merging with (sub-)cortical layer, of refractive, glassy, laterally strongly com- pressed cells. Cortical layer (12-)15-22(-25) mm (n ...
Context 4
... 7-8EF5- 8. Ostiolar area (ostioles plus dark margin) in face view, (24-)32-53(-63) mm (n 5 30), inconspicuous and only visible under high magnification or conspic- uous and semiglobose in some stromata, after addition of water ostioles generally appearing as light and hyaline dots on a bright reddish stroma surface (FIG. 3c). Ostioles in section (FIG. 3g, j) Peridium 8-13(-15) mm thick at base, (14-)15-20 (-22) mm at apex (n 5 15), yellowish brown to reddish brown, slightly paler than cortex, apically thickened and directly merging with (sub-)cortical layer, of refractive, glassy, laterally strongly com- pressed cells. Cortical layer (12-)15-22(-25) mm (n 5 15) thick, reddish brown, ...
Context 5
... apically thickened and directly merging with (sub-)cortical layer, of refractive, glassy, laterally strongly com- pressed cells. Cortical layer (12-)15-22(-25) mm (n 5 15) thick, reddish brown, orange-brown in lactic acid, of small angular thick-walled glassy compressed cells of indistinct outline, (3-)5-10(-11) mm (n 5 15) diam in face view (FIG. 3l), 3-6(-7) mm (n 5 15) diam in vertical section (FIG. 3g, j). Hairs rare on mature stromata, ca. 10-20 mm long, subhyaline to reddish brown, apically rounded. Subcortical tissue thin, loose textura intricata of thin-walled hyaline hyphae ca. 2- 4.5 mm wide, parallel to the surface, branching and running vertical between perithecia, ...
Context 6
... layer, of refractive, glassy, laterally strongly com- pressed cells. Cortical layer (12-)15-22(-25) mm (n 5 15) thick, reddish brown, orange-brown in lactic acid, of small angular thick-walled glassy compressed cells of indistinct outline, (3-)5-10(-11) mm (n 5 15) diam in face view (FIG. 3l), 3-6(-7) mm (n 5 15) diam in vertical section (FIG. 3g, j). Hairs rare on mature stromata, ca. 10-20 mm long, subhyaline to reddish brown, apically rounded. Subcortical tissue thin, loose textura intricata of thin-walled hyaline hyphae ca. 2- 4.5 mm wide, parallel to the surface, branching and running vertical between perithecia, partly inter- mingled with subglobose to ellipsoidal hyaline ...
Context 7
... mature stromata, ca. 10-20 mm long, subhyaline to reddish brown, apically rounded. Subcortical tissue thin, loose textura intricata of thin-walled hyaline hyphae ca. 2- 4.5 mm wide, parallel to the surface, branching and running vertical between perithecia, partly inter- mingled with subglobose to ellipsoidal hyaline cells. Subperithecial tissue (FIG. 3m) dense, hyaline textura angularis to epidermoidea of isodiametric subglobose to angular thin-walled cells directly below perithecia, downward gradually merging into oblong segments of thick, mainly vertically oriented hyphae, becoming smaller toward the base of the stroma, no distinct layers discernible; cells (4-)12-44(-63) 3 ...
Context 8
... angularis to epidermoidea of isodiametric subglobose to angular thin-walled cells directly below perithecia, downward gradually merging into oblong segments of thick, mainly vertically oriented hyphae, becoming smaller toward the base of the stroma, no distinct layers discernible; cells (4-)12-44(-63) 3 (3.5-)6.5- 15.0(-18.5) mm (n 5 30). Asci (FIG. 3p) (68- Chaverri and Samuels (2003) and Druzhinina et al (2005), and all species that form ''lone lineages''. Species are named as in FIG. 2A. The gray scale indicates bootstrap support of nodes marked by circles; statistical support of unmarked nodes was lower than 60% of 500 bootstrap replicates. (-98) 3 (3.5-)4.0-4.8(-5.2) mm, ...
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... indicates bootstrap support of nodes marked by circles; statistical support of unmarked nodes was lower than 60% of 500 bootstrap replicates. (-98) 3 (3.5-)4.0-4.8(-5.2) mm, including a stipe (5-) 7-20(-28) mm (n 5 30) long, emerging in fascicles, cylindrical, stipe often with an inflated base, no croziers seen; apical ring minute. Ascospores (FIG. 3n-p) hyaline, dimorphic with usually slight difference in shape and size, finely verruculose, disarticulating inside asci, distal part ascospore 2.5- 3.2(-4.2) 3 2.5-3.0 mm (n 5 40, three specimens), subglobose, sometimes slightly tapered toward upper end, proximal part ascospore (2.5-)3.0-3.7(-4.5) 3 2.2-2.5(-2.7) mm (n 5 40, three ...
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... Phylogenetically Trichoderma hailarense is related to T. gamsii and T. neokoningii (Fig. 1) and does not meet the sp∃!(rpb2 99 ≅tef1 97 ) standard for T. gamsii or T. neokoningii. Morphologically, colonies of T. gamsii and T. neokoningii on PDA formed conidia sporadically or in hemispherical pustules and conidia of T. gamsii and T. neokoningii were ellipsoidal to oblong, smooth-walled (Jaklitsch et al. 2006). However, colonies of T. hailarense did not form conidia on PDA and conidia of T. hailarense on other media were obovoid, delicately roughened and easily distinguished from those of T. gamsii and T. neokoningii. ...
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... Another gene often used in fungal phylogenetic analyses is tef-1? ( Jaklitsch et al. 2006;Stenglein et al. 2010;Zhao et al. 2016He et al. 2017), which had the second highest PCR and sequencing success rates (Table 3). This gene has previously been regarded as the target DNA barcode in certain groups ( Geiser et al. 2004;Druzhinina et al. 2005;Li et al. 2013); however, our analyses showed that tef-1? the occur- rence of overlap between intra-and inter-species var- iation among the candidate genes (Figs. 1, 2 and 7) was the highest for this gene. ...
Russula is a worldwid genus which has a high species diversity . Aiming accurate and rapid species identification, candidate genes nLSU (28S), ITS, tef-1α, mtSSU, rpb1, and rpb2, were analysed as potential DNA barcodes. This analysis included 433 sequences from 38 well-circumscribed Russula species of eight subgenera. Two vital standards were analysed for success species identification using DNA barcodes, specifically inter- and intra-specific variations together with the success rates of PCR amplification and sequencing. Although the gap between inter- and intra-specific variations was narrow, ITS met the qualification standards for a target DNA barcode. Overlapping inter- and intra-specific pairwise distances were observed in nLSU, tef-1α, mtSSU, and rpb2. The success rates of PCR amplification and sequencing in mtSSU and rpb1 were lower than those of others. Gene combinations were also investigated for resolution of species recognition. ITS-rpb2 was suggested as the likely target DNA barcode for Russula, owing to the two viatal standards above. Since nLSU has the lowest minimum of inter-specific variation, and tef-1α has the highest overlap between intra- and inter-species variations among the candidate genes, they are disqualified from the selection for DNA barcode of Russula.
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... Secondly, it is useful when there is no clear means to match adults with immature specimens (Pegg et al. 2006;Ahrens et al. 2007;Shenoy et al. 2007). Thirdly, it is also helpful to discriminate species when morphological traits are ambiguous (Saunders 2005;Jaklitsch et al. 2006;Kumar et al. 2007). ...
In recent years, DNA barcoding has become quite popular for molecular identification of species because it is simple, quick and an affordable method. Present study was conducted to identify spiders of most abundant families, i.e. Salticidae and Lycosidae from citrus orchards in Sargodha district using DNA barcoding. A total of 160 specimens were subjected to DNA barcoding but, sequences up to 600 bp were recovered for 156 specimens. This molecular approach proved helpful to assign the exact taxon to those specimens which were misidentified through morphological characters in the study. We were succeeded to discriminate six species of Lycosidae and nine species of Salticidae through DNA barcoding. Results revealed the presence of clear barcode gap (discontinuity in intra- and inter-specific divergences) for members of both families. Furthermore, the maximum intra-specific divergence was less than NN (nearest neighbour) distance for all species. This suggested the reliability of DNA barcoding for spider's identification up to species level. We got 98% success in our study. It is concluded from present study that DNA barcoding is more reliable tool especially for immature spiders, when morphological characters are ambiguous.
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This chapter attempts to summarize the recently available and rapidly increasing amount of information in the literature about the occurrence and biodiversity of Trichoderma species in different ecological habitats. Members of the genus are common in soil and rhizosphere of plants in natural and agricultural fields and forests and on decaying wood. They are also occurring in the air, settled dust and different water-related habitats including marine environments and drinking water. Furthermore, certain species are known as endophytes of plants, colonizers of mushroom-related natural and artificial substrata and facultative pathogens of humans, demonstrating a high adaptability to various ecological niches.
... ITS barcodes of this new species were included in TrichOKEY. The same authors (Jaklitsch et al., 2006 ) described also the new holomorphic species H. crystalligena/ T. crystalligenum, a common European species with a white-spored Trichoderma anamorph based on an integrated approach including teleomorph and anamorph morphologies, culture characteristics and sequences of the ITS region and fragments of the tef1 and rpb2 genes. H. crystalligena was described as a separate evolutionary lineage with H. megalocitrina and H. psychrophila as its nearest neighbors, forming one phylogenetic clade with the H. pulvinata/H. ...
... citrina node. The ITS sequences characteristic to H. crystalligena were used to develop species specific DNA barcodes, which were incorporated into TrichOKEY, resulting in version 1.2 (Jaklitsch et al., 2006). In 2007, the upgraded TrichOKEY version 2 was supplemented with the species specific barcodes of T. pleuroticola and T. pleurotum (Fig. 3.1), two members of the Harzianum clade that are responsible for the green mould disease of the cultivated oyster mushroom , Pleurotus ostreatus (Komoń-Zelazowska et al., 2007). ...
Various Trichoderma and Hypocrea species have remarkable importance in industry, agriculture as well as clinical practice. The precise identification of isolates at the species level is crucial in all fields of Trichoderma research. Barcoding is a powerful tool for the identification of eukaryotic species, including fungi. In this chapter, TrichOKEY, a DNA barcode-based online programme developed for the identification of Trichoderma/Hypocrea species is presented and its practical applications are discussed.
... Phylogenetic reconstruction using the Bayesian approach was performed according to Jaklitsch et al. (2006), through tef1, using Mr Bayes v3.1.2. The best model proposed by Mr Modeltest v2.3 based on AIC was a HasegawaeKishinoeYano (HKY) þ gamma distribution (G) model. ...
Some species of Trichoderma have successfully been used in the commercial biological control of fungal pathogens, e.g., Sclerotinia sclerotiorum, an economically important pathogen of common beans (Phaseolus vulgaris L.). The objectives of the present study were (1) to provide molecular characterization of Trichoderma strains isolated from the Brazilian Cerrado; (2) to assess the metabolic profile of each strain by means of Biolog FF Microplates; and (3) to evaluate the ability of each strain to antagonize S. sclerotiorum via the production of cell wall-degrading enzymes (CWDEs), volatile antibiotics, and dual-culture tests. Among 21 isolates, we identified 42.86% as Trichoderma asperellum, 33.33% as Trichoderma harzianum, 14.29% as Trichoderma tomentosum, 4.76% as Trichoderma koningiopsis, and 4.76% as Trichoderma erinaceum. Trichoderma asperellum showed the highest CWDE activity. However, no species secreted a specific group of CWDEs. Trichoderma asperellum 364/01, T. asperellum 483/02, and T. asperellum 356/02 exhibited high and medium specific activities for key enzymes in the mycoparasitic process, but a low capacity for antagonism. We observed no significant correlation between CWDE and antagonism, or between metabolic profile and antagonism. The diversity of Trichoderma species, and in particular of T. harzianum, was clearly reflected in their metabolic profiles. Our findings indicate that the selection of Trichoderma candidates for biological control should be based primarily on the environmental fitness of competitive isolates and the target pathogen.
... The complete inventory of teleomorph-forming species in Central Europe indicated that there are 75 species (Jaklitsch, 2009Jaklitsch, , 2011). In addition to this, about 12 anamorphic species (for which no sexual stages are known) were reported in surveys of European soils and in taxonomic studies, as described by Wuczkowski et al. (2003), Kraus et al. (2004), Jaklitsch et al. (2006a, b), KomonZelazowska et al. (2007), Hagn et al. (2007), Migheli et al. (2009), Zachow et al. (2009) and Meincke et al. (2010. Thus, the total number of Hypocrea/ Trichoderma species identified so far in Europe may be about 100. ...
... Development of genus-specific primers. The genus-specific primers were developed based on the master alignment of ITS1 and 2 from 88 Hypocrea and Trichoderma reference strains (Druzhinina et al., 2005) complemented by the new species described since that time (Jaklitsch et al., 2005Jaklitsch et al., , 2006a Overton et al., 2006; Samuels et al., 2006b; Komon-Zelazowska et al., 2007; Jaklitsch et al., 2008; Jaklitsch, 2009 Jaklitsch, , 2011). All primers are reverse primers and are complementary to the forward primer ITS5 (59-GGAAGTAAAAGTCGTAACAAGG-39) (Table 2). ...
In this paper, we report on the in situ diversity of the mycotrophic fungus Trichoderma (teleomorph Hypocrea, Ascomycota, Dikarya) revealed by a taxon-specific metagenomic approach. We designed a set of genus-specific internal transcribed spacer (ITS)1 and ITS2 rRNA primers and constructed a clone library containing 411 molecular operational taxonomic units (MOTUs). The overall species composition in the soil of the two distinct ecosystems in the Danube floodplain consisted of 15 known species and two potentially novel taxa. The latter taxa accounted for only 1.5 % of all MOTUs, suggesting that almost no hidden or uncultivable Hypocrea/Trichoderma species are present at least in these temperate forest soils. The species were unevenly distributed in vertical soil profiles although no universal factors controlling the distribution of all of them (chemical soil properties, vegetation type and affinity to rhizosphere) were revealed. In vitro experiments simulating infrageneric interactions between the pairs of species that were detected in the same soil horizon showed a broad spectrum of reactions from very strong competition over neutral coexistence to the pronounced synergism. Our data suggest that only a relatively small portion of Hypocrea/Trichoderma species is adapted to soil as a habitat and that the interaction between these species should be considered in a screening for Hypocrea/Trichoderma as an agent(s) of biological control of pests.
... Hebert et al. 2004a, Hebert et al. 2004b, Greenstone et al. 2005, Smith et al. 2005, Barber and Boyce 2006, Meier et al. 2006, Kerr et al. 2007, Kumar et al. 2007, Pfenninger et al. 2007, Stahls and Savolainen 2008, Zhou et al. 2009). Meanwhile, other candidate genes, including Internal Transcribed Spacer (ITS), trnH-psbA intergenic spacer (trnH-psbA), Ribulose-bisphosphate carboxylase (rbcL) and Maturase K (matK) were analysed by different research groups (Jaklitsch et al. 2006, Evans et al. 2007, Ran et al. 2010, de Groot et al. 2011, Liu et al. 2011, Piredda et al. 2011, Yesson et al. 2011). Till recently, there are about 30 DNA barcode candidates are tested, and 4 to 8 of them are widely used for the identification of diversified taxonomic groups with a relatively good resolution. ...
... 10 g of soil samples (if necessary pulverized by means of a mortar and pestle, and passed through a 0.5 mm soil screen mesh to remove large debris and root fragments; [39] ) were suspended in 90 mL sterile distilled water and thoroughly mixed. A 10 mL aliquot was then used to prepare a series of dilutions in the range of 10 –1 to 10 –3 , and inoculated on to potato dextrose agar (PDA), malt extract agar (MEA) and synthetic low nutrient agar (SNA) [40], each supplemented with streptomycin (50 mg/L) to prevent bacterial growth. Three replicates were done for each medium and soil sample. ...
... Genomic DNA was extracted from mycelia grown on MEA with the Plant DNeasy Minikit (QIAgen GmbH, Hilden, Germany) according to the manufacturer's instructions. A region of nuclear DNA containing the ITS1 and two regions of the rRNA gene cluster, was amplified by PCR with the primer combinations SR6R and LR1 [40], and an approximately 1-kb portion of the gene encoding translation elongation factor 1-alpha (tef1) was amplified and sequenced using the primers EF1 (5'- ATGGGTAAGGA(A/G)GACAAGAC-3') and EF2 (GGA(G/A)GTACCA GT(G/C)ATCATGTT-3) as described by Jaklitsch et al. [40]. PCR products were purified with the QIAquick PCR Purification Kit (QIAgen) and were subjected to automated sequencing at MWG (Martinsried, Germany). ...
... Genomic DNA was extracted from mycelia grown on MEA with the Plant DNeasy Minikit (QIAgen GmbH, Hilden, Germany) according to the manufacturer's instructions. A region of nuclear DNA containing the ITS1 and two regions of the rRNA gene cluster, was amplified by PCR with the primer combinations SR6R and LR1 [40], and an approximately 1-kb portion of the gene encoding translation elongation factor 1-alpha (tef1) was amplified and sequenced using the primers EF1 (5'- ATGGGTAAGGA(A/G)GACAAGAC-3') and EF2 (GGA(G/A)GTACCA GT(G/C)ATCATGTT-3) as described by Jaklitsch et al. [40]. PCR products were purified with the QIAquick PCR Purification Kit (QIAgen) and were subjected to automated sequencing at MWG (Martinsried, Germany). ...
The southwestern highlands forests of Ethiopia are the origin of the coffee plant Coffea arabica. The production of coffee in this area is affected by tracheomycosis caused by a soil-born fungus Gibberella xylarioides. The use of endemic antagonistic strains of mycoparasitic Trichoderma species would be a nature conserving means to combat this disease. We have used molecular methods to reveal that the community of Trichoderma in the rhizosphere of C. arabica in its native forests is highly diverse and includes many putatively endemic species. Among others, the putative new species were particularly efficient to inhibit growth of G. xylarioides.
