Trichoderma species are widely used as biofungicides for the control of fungal plant pathogens. Several studies have been performed to identify the main genes and compounds involved in Trichoderma–plant–microbial pathogen cross-talks. However, there is not much information about the exact mechanism of this profitable interaction. Peptaibols secreted mainly by Trichoderma species are linear, 5–20 amino acid residue long, non-ribosomally synthesized peptides rich in α-amino isobutyric acid, which seem to be effective in Trichoderma–plant pathogenic fungus interactions. In the present study, reversed phase (RP) high-performance liquid chromatography (HPLC) coupled with electrospray ionization (ESI) mass spectrometry (MS) was used to detect peptaibol profiles of Trichoderma strains during interactions with fungal plant pathogens. MS investigations of the crude extracts deriving from in vitro confrontations of Trichoderma asperellum and T. longibrachiatum with different plant pathogenic fungi (Fusarium moniliforme, F. culmorum, F. graminearum, F. oxysporum species complex, Alternaria solani and Rhizoctonia solani) were performed to get a better insight into the role of these non-ribosomal antimicrobial peptides. The results revealed an increase in the total amount of peptaibols produced during the interactions, as well as some differences in the peptaibol profiles between the confrontational and control tests. Detection of the expression level of the peptaibol synthetase tex1 by qRT-PCR showed a significant increase in T. asperellum/R. solani interaction in comparison to the control. In conclusion, the interaction with plant pathogens highly influenced the peptaibol production of the examined Trichoderma strains.
Five Iranian Trichoderma isolates from species T. viride, T. viridescens, T. asperellum, T. longibrachiatumand T. citrinoviride -selected from the Fungal Collection of the Bu Ali Sina University, Hamedan, Iran -were investigated for their peptaibol production. All examined isolates showed remarkable antibacterial activities during the screening of their extracts for peptaibol content with a Micrococcus luteus test culture. HPLC-ESI-IT MS was used for identification and elucidation of the amino acid sequences of peptaibols. The detected peptaibol compounds contain 20 or 18 amino acid residues and belong to the trichobrachin and trichotoxin groups of peptaibols, respectively. T. longibrachiatum and T. citrinoviride produced trichobrachins, while trichotoxins could be detected in T. viride, T. viridescens and T. asperellum. Out of 37 sequences detetermined, 26 proved to be new, yet undescribed compounds, while others were identified as previously reported trichotoxins (trichotoxin A-50s and T5D2) and trichobrachins (longibrachins AI, AII, AIII, BII and BIII). Compounds within the two groups of detected peptaibols differed from each other only by a single or just a few amino acid changes.
This research was conducted to monitor the aflatoxigenic fungi and aflatoxin contamination of walnut in the Hamedan province. For this purpose, 40 samples were analyzed. Aspergillus, Alternaria, Rhizopus, Cladosporium, Fusarium, yeast, and some different bacteria were isolated from walnuts. Aspergillus is the most frequent genus. Aspergillus flavus was predominantly isolated. HPLC was used for evaluation of aflatoxin contamination of walnut samples. Aflatoxins G1 (AFG1), B1 (AFB1), G2 (AFG2), and B2 (AFB2) were produced by 20 isolates. AFG1 and AFB1 were being predominant at concentration ranges of 1.7-18.2 and 0-8.2 ngg, respectively. Highest levels were found in one sample that was highly contaminated with Aspergillus flavus/Aspergillus parasiticus. Methyl beta cyclodextrin also was performed for detection of aflatoxigenic Aspergillus isolates. The results showed that only 31.6% (p < 0.05) of A. flavus and A. parasiticus isolates were able to produce aflatoxin. A significant difference was shown between shielded and unshielded walnut in aflatoxin contamination. The content of aflatoxin in most of the walnut samples did not reach to maximum tolerable limit for aflatoxin B1 in EU standard (p > 0.05). Thus, systematic and continues monitoring of walnuts is recommended.
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