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, 6: Hypocrea neorufa, stromata. 7–10: Hypocrea patella f. patella. 11, 12: Hypocrea patella f. tropica. , 6 from G.J.S. 96-143; 7, 9, 10 from G.J.S. 96-146; 8 from isotype of H. patella BPI 631626; 11, 12 from G.J.S. 96-198. Scale bars: 5, 7–9, 11 = 2 mm; 6, 10, 12 = 1 mm.
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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...
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Trichoderma isolates were collected from moist soils near a water source in different areas of China. ITS sequences were submitted to MIST (Multiloci Identification System for Trichoderma ) and meets the Trichoderma [ITS 76 ] standard. Combined analyses of phylogenetic analyses of both phylograms ( tef1 -α and rpb2 ) and morphological characteristi...
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... All the fungal species were cultured on 50 mL of PDA at 28 • C for 96-120 h in the dark. Then, Genomic DNA was extracted from the collected fungal mycelium using a spin column method (Qiamp mini kit, Qiagen GmbH, Germany) with slight modifications as previously described [51]. The PCR amplification reaction of the purified DNA samples was performed in a total volume of 25 µL mixture as previously described [52]. ...
Trichoderma species are known as excellent biocontrol agents against soil-borne pathogens that cause considerable crop losses. Eight strains of Trichoderma were isolated from five Egyptian regions. They identified based on translation elongation factor-1α (TEF1) sequencing as four different Trichoderma species: Trichoderma asperellum, Trichoderma harzianum, Trichoderma viride, and Trichoderma longibrachiatum. Optimal growth conditions (temperature and media), and the phosphate solubilization capability of Trichoderma strains were evaluated in vitro. Further, the ability of these strains to antagonize Fusarium solani, Macrophomina phaseolina, and Fusarium graminearum was also evaluated. The results revealed that Trichoderma harzianum (Th6) exhibited the highest antagonistic ability against F. solani, M. phaseolina and F. graminearum with inhibition rates of 71.42%, 72.97%, and 84.61%, respectively. Trichoderma viride (Tv8) exhibited the lowest antagonism against the same pathogens with inhibition rates of 50%, 64% and 69.23%, respectively. Simple-sequence repeats (SSRs) and random amplified polymorphic DNA (RAPD) markers were used to evaluate the genetic variability of the Trichoderma strains. The results revealed that of 45 RAPD amplified bands, 36 bands (80%) were polymorphic and of SSRs amplified 36 bands, 31 bands (86.11%) were polymorphic. The amplification of calmodulin and β-1,3-endoglucanase was noted at 500 bp and 230 bp, respectively. Data indicated that T. viride (Tv8) had the highest phosphate solubilization index (10.0 mm), while T. harzianum (Th6) had the lowest phosphate solubilization index (4.0 mm). In conclusion, T. harzianum (Th6) had the highest antagonistic activity in dual culture assay along with the growth rate; while T. viride (Tv8) had the highest phosphate solubilization activity. There are still gaps in obtaining new formulations, selecting potent Trichoderma strains to confirm disease control in planta. For improving Trichoderma recommendation in the organic agricultural system and sustaining the fertility of the soil, the field application of highly antagonistic biocontrol agents in different types of soil and plant species will be the first approach toward bio-pesticide treatments along with bio-fertilizer inoculation.
... DNA from Trichoderma strains was extracted, according to previously described methods [17]. Two gene regions: internal transcribed spacers (ITS1, 5.8S, ITS2 rDNA), and translation elongation factor (EF-1a) were amplified by PCR using the primers ITS1 and ITS4 for ITS [18], and the primers EF1-728F [19] and TEF1 rev [20] for EF-1a. ...
... Morphological structure was observed through 40X objective with ZEISS Axio Scope microscopy. Characterization was made follow taxonomic clade described in the international website repositories for every kind of funguses into studio[13],[14],[15]. ...
... For species identification, DNA was isolated from 7-day-old Trichoderma cultures growing in 100 ml of potato dextrose broth (PDB) following the methods of Dodd et al., 2002. Portions of the translation elongation factor alpha-1 (TEF1) gene were amplified with primers EF1-728F and TEF1 rev following the protocol in Samuels et al. (2006a). ...
The cocoa tree, Theobroma cacao L., suffers large yield losses in Aceh Indonesia due to the disease black pod rot, caused by Phytophthora spp. Despite having the largest area under cacao production in Sumatra, farmers in the Aceh region have low overall production because of losses to insect pests and black pod rot. Trichoderma spp. were isolated from the roots and leaves of cacao trees and screened as potential biological control agents. Isolates used in the study were T. asperellum isolates T2 and T4, T. longibrachiatum isolates T15 and T16, and T. virens isolates T1 and Tv. T1, T2, T4, and Tv completely colonized and destroyed Phythophthora tropicalis and Phytophthora palmivora mycelium in precolonized plate assays. All six isolates reduced P. tropicalis, but none reduced the growth of P. palmivora in dual plate assays. Phytophthora growth was suppressed on MIN media amended with sterile heat inactivated Trichoderma culture filtrates, with Tv best suppressing growth of both Phytophthora spp. T. virens isolate Tv was the only isolate observed coiling around P. tropicalis mycelium and disrupted the formation of P. palmivora sporangia. Of all six isolates, only Tv reduced P. palmivora lesion expansion in a detached pod assay, reducing severity by 71%. Tv also reduced P. palmivora infection on seedlings when applied aerially at 1x106 and 1x108 conidia/ml, by 19% and 59% respectively. T. virens isolate Tv is a mycoparasite, antagonizes Phytophthora in a dual plate assay, and shows antibiosis against Phytophthora spp., suggesting that multiple modes of action contribute to its ability to limit Phytophthora lesion expansion on cacao pods and seedlings.
... The entire dried mycelial mat was then placed in a 1.5 mL Eppendorf tube for immediate DNA extraction. Extraction of the genomic DNA was performed as reported previously (Dodd et al. 2002). Polymerase chain reaction for amplification of translation elongation factor (tef1) was performed as reported previously (Samuels et al. 2006b). ...
Endophytic strains of Trichoderma species can be used as an alternative to chemicals to control vascular streak dieback (VSD) disease of cacao. Of 21 Trichoderma isolated from Theobroma cacao (cacao) in Indonesia, 19 were identified as Trichoderma asperellum. Four isolates of this species (ART-4, ART-5, ART-6 and ART-8) were reintroduced into young cocoa seedlings by root inoculation and after 4 weeks all were recovered from roots and stems, while ART-4 and ART-5 were recovered from leaves as well. Spraying seedlings pre-inoculated with T. asperellum ART-4, ART-5 and ART-6 with mycelium of the VSD pathogen Ceratobasidium theobromae resulted in no apparent VSD symptoms on the leaves. Those seedlings pre-inoculated with ART-8 showed 8.9 % incidence of VSD symptoms on the leaves when compared to a 20.4 % incidence of VSD on positive control leaves, 12 weeks after inoculation. The same isolates were also reintroduced into 3-month-old cocoa seedlings via the connecting site following shoot grafting, and after 12 weeks all isolates were recovered from stem and leaves. Seedlings grafted with buds infected by VSD and treated by ART-4 showed no VSD symptoms on their leaves 12 weeks after grafting and inoculation. However, those treated with ART-5, ART-6 and ART-8 showed 33.3 %, 50.0 % and 56.0 % incidence of VSD symptoms on their leaves, respectively, compared to an 88.9 % incidence of VSD on positive control leaves. Therefore, the study results demonstrates for the first time the potential of T. asperellum isolates to control VSD disease on cacao.
... . Samuels et al. 2002;Dodd et al. 2002;Wuczkowski et al. 2003;Holmes et al. 2004, Zhang et al. 2005Hoyos-Carvajal et al. 2009;Druzhinina et al. 2010aDruzhinina et al. , 2010bBelayneh Mulaw et al. 2010 . ( Kullnig-Gradinger et al. 2002;Chaverri et al. 2003;Bissett et al. 2003 . ...
Fungi belonging to genus Trichoderma are well known for their effectiveness in the biological control of
different plant pathogenic fungi, inducing systemic resistance in crops and for promoting plant growth.
Phylogeny of 81 Trichoderma isolates (with emphasis on some biocontrol strains) belonging to six species obtained from paddy fields in Mazandaran province, was studied by partial sequence analyses of the translation elongation factor 1-α (tef1α) gene. Data were analyzed by Neighbor-Joining, Maximum
Parsimony and Minimum Evolution methods. For each analysis, one consensus tree was used out of all
generated phylograms. Phylogenetic trees of the 41 Trichoderma isolates inferred by all methods and
programs were similar. Results showed that 14 tef1α haplotypes can be recognized for T. harzianum (the
most frequent species in rice fields) isolates grouped into five clades. Therefore, this complex species
presented the highest intraspecific diversity when compared to other species in this study. In contrast, only two haplotypes were recognized for T. virens, the second most prevalent species. Furthermore, sequence analyses revealed three and two haplotypes for T. atroviride and T. hamatum, respectively. Results indicated that biological control strains represented various haplotypes and placed in different clades along with other native isolates. Consequently, a specific tef1α haplotype related to antagonism could not be distinguished. In addition, there was no correlation between tef1α haplotype and the geographic location and thus different species and haplotypes were isolated from the same sampling sites.
... The extraction of genomic DNA was performed as reported previously (Dodd et al. 2002) using either the DNeasy Plant Mini Kit (QIAgen GmbH, Hilden, Germany) or Puregene Genomic DNA Isolation Kit (Gentra Systems, Minneapolis, Minnesota). A part of the translation elongation factor 1 alpha (tef1) was amplified using the primers EF1728F (Carbone & Kohn 1999) and TEF1 rev (Samuels et al. 2002) or TEF1LLErev (Jaklitsch et al. 2005). ...
Trichoderma viridescens is recognised as a species complex. Multigene analyses based on the translation elongation factor 1-alpha encoding gene (tef1), a part of the rpb2 gene, encoding the second largest RNA polymerase subunit and the larger subunit of ATP citrate lyase (acl1) reveals 13 phylogenetic species with little or no phenotypic differentiation. This is the first use of acl1 in Trichoderma phylogenetics. The typification of T. viridescens s.str. is clarified and Hypocrea viridescens is replaced by the new name T. paraviridescens. Besides these two species, eleven are phylogenetically recognised and T. olivascens, T. viridarium, T. virilente, T. trixiae, T. viridialbum, T. appalachiense, T. neosinense, T. composticola, T. nothescens and T. sempervirentis are formally described and illustrated. Several species produce yellow diffusing pigment on cornmeal dextrose agar, particularly after storage at 15 °C, while T. olivascens is characterised by the formation of an olivaceous pigment. The results are compared with earlier publications on this group of species.
... References: Dodd et al. (2002), Jaklitsch (2011). Trichoderma novae-zelandiae (Samuels & Petrini) Jaklitsch & Voglmayr, ...
... Repres. sequences: tef1: AY937428; rpb2: n.a.. Reference: Dodd et al. (2002). ...
Unitary nomenclature demands the use of a single name for pleomorphic fungi determined according to priority. For this reason combinations in Trichoderma are here provided for 46 species for which such a combination is lacking. Although many more such species are known, only those are included here that are dealt with in more recent papers and where some DNA data are available in GenBank, even if erroneous; for other species it is strongly recommended to consult databases like Index Fungorum or MycoBank. Information on types is provided for most species, and representative cultures, GenBank accessions for tef1 and rpb2 , and important references are given for all species.
... Total DNA was extracted using the Ultraclean TM Microbial DNA isolation Kit (MBio, Solana Beach, CA) according the manufacturer's instructions. DNA sequences of the 18S rRNA gene were obtained from the GenBank database, and amplification of the ITS regions and a part of the TEF gene was preformed as described by Houbraken et al. (2011) and Dodd et al. (2002), respectively. The ITS and TEF dataset was combined and maximum likelihood analysis was performed using RAXML version 7.2.8. ...
Paecilomyces lilacinus was described more than a century ago and is a commonly occurring fungus in soil. However, in the last decade this fungus has been increasingly found as the causal agent of infections in man and other vertebrates. Most cases of disease are described from patients with compromised immune systems or intraocular lens implants. In this study, we compared clinical isolates with strains isolated from soil, insects and nematodes using 18S rRNA gene, internal transcribed spacer (ITS) and partial translation elongation factor 1-α (TEF) sequences. Our data show that P. lilacinus is not related to Paecilomyces, represented by the well-known thermophilic and often pathogenic Paecilomyces variotii. The new genus name Purpureocillium is proposed for P. lilacinus and the new combination Purpureocillium lilacinum is made here. Furthermore, the examined Purpureocillium lilacinum isolated grouped in two clades based on ITS and partial TEF sequences. The ITS and TEF sequences of the Purpureocillium lilacinum isolates used for biocontrol of nematode pests are identical to those causing infections in (immunocompromised) humans. The use of high concentrations of Purpureocillium lilacinum spores for biocontrol poses a health risk in immunocompromised humans and more research is needed to determine the pathogenicity factors of Purpureocillium lilacinum.
... Se colectó el micelio y se transfirió a un tubo de centrifuga de 1.5 ml. Posteriormente el ADN se extrajo empleando un kit comercial PureGene con el protocolo descrito para tejidos de plantas (Dodd et al., 2002 ...
A survey was conducted to determine the species of Trichoderma from the soil of commercial plantations of tequila agave (Agave tequilana) in the Los Altos Sur region located in Jalisco State, Mexico in order to study them as biocontrol agents against one of the plant pathogens that attack the tequila agave. Sixty four cultures of Trichoderma were identified morphologically and some of them by DNA sequencing analyses of the internal transcribed space (ITS), translation elongation factor 1 alpha (tef1), actin (act), and calmodulin (cal). Morphologically, 60 isolates (93.7%) were identified as T. longibrachiatum and ITS sequence data for 25 of these cultures confirmed their identity. Three cultures (4.6%) were identified as T. harzianum and DNA sequences of ITS, tef1, act and cal confirmed the result. One culture (1.5%) was identified morphologically and by ITS sequence data as T. reesei. Antagonism against Thielaviopsis paradoxa was evaluated for several isolates of each species of Trichoderma. Isolates of T. longibrachiatum, T. viridescens and T. reesei produced significantly higher inhibition against Th. paradoxa than T. harzianum.
