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MALDI-TOF MS analysis of peptide mixture from Trichoderma citrinoviride showing singly charged pseudomolecular adducts of Na + and K + around m/z 1800-2040.
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A cork oak endophytic strain of Trichoderma citrinoviride, previously selected for its antagonistic potential against various fungal pathogens involved in oak decline, was screened for the production of bioactive secondary metabolites. From liquid culture a mixture of polypeptide antibiotics (peptaibols) belonging to the paracelsin family was isola...
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Citations
... Its peptaibiotics profile has been tested in five works, against four different fungal phytopathogens Lorito et al., 1996;Rebuffat et al., 1995;Schirmbock et al., 1994) and one plant virus (Kai et al., 2018) (Table 1). Trichoderma asperellum (Alfaro-Vargas et al., 2022;Tamandegani et al., 2020), T. atroviride (Oh et al., 2002), T. cerinum (Khare et al., 2018), T. citronoviride (Maddau et al., 2009), T. effusum (Balázs et al., 2023), T. gamsii (Marik et al., 2018), T. koningiopsis (Marik et al., 2018), T. longibrachiatum (Balázs et al., 2023;Tamandegani et al., 2020), T. longibrachiatum f. bissettii (Balázs et al., 2023), T. pseudokoningii (Li et al., 2014;Luo et al., 2010;Shi et al., 2012;Song et al., 2006;Zhao et al., 2018), T. Reesei (Balázs et al., 2023;Marik et al., 2019), T. saturnisporum (Balázs et al., 2023), and T. virens (Viterbo et al., 2007) are the other species prospected in this field ( Table 1). Outside of that genus, Apiocrea spp. ...
... For each tested compound, or group of compounds, information is provided regarding its length (when applicable), source organism, target phytopathogens, effective dosage, mode of action (when reported by the authors), and the original reference. This table is a shortened version of Table S1 included in the Supplementary Data, where source (Li et al., 2014;Song et al., 2006;Zhang et al., 2022) Trichogin Kronen et al., 2001;Lam et al., 2021;Ritzau et al., 1997) Atroviridins Grigoriev et al., 2003;Kim et al., 2000) Harzianins (Grigoletto et al., 2020;Khare et al., 2018;Maddau et al., 2009;Marik et al., 2019, Marik et al., 2018Rodríguez et al., 2011;Tamandegani et al., 2020) (continued on next page) 2012). ...
... The extraction of peptaibiotics is usually performed by applying various organic solvents, like ethyl acetate (Maddau et al., 2009;Grigoletto et al., 2020;Liu et al., 2020), methanol (Marik et al., 2013), nbutanol Rebuffat et al., 1995;Yeo et al., 2002), among others, to the broth or mycelium of cultured strains. For instance, Katoch et al. (2019) and Viterbo et al. (2007) used acetonitrile:methanol:water (1:1:1) to extract both the polar and non-polar fractions from fungal myceliae, whereas Marik et al. (2018) used a chloroform:methanol 2:1 solution. ...
Agricultural systems are in need of low-cost, safe antibiotics to protect crops from pests and diseases. Peptaibiotics, a family of linear, membrane-active, amphipathic polypeptides, have been shown to exhibit antibacterial, antifungal, and antiviral activity, and to be inducers of plant resistance against a wide range of phytopathogens. Peptaibiotics belong to the new generation of alternatives to agrochemicals, aligned with the United Nations Sustainable Development Goals and the One Health approach toward ensuring global food security and safety. Despite that, these fungi-derived, non-ribosomal peptides remain surprisingly understudied, especially in agriculture, where only a small number has been tested against a reduced number of phytopathogens. This lack of adoption stems from peptaibiotics' poor water solubility and the difficulty to synthesize and purify them in vitro, which compromises their delivery and inclusion in formulations. In this review, we offer a comprehensive analysis of peptaibiotics' classification, biosynthesis, relevance to plant protection, and mode of action against phytopathogens, along with the techniques enabling researchers to extract, purify, and elucidate their structure, and the databases holding such valuable data. It is also discussed how chemical synthesis and ionic liquids could increase their solubility, how genetic engineering and epigenetics could boost in vitro production, and how omics can reduce screenings' workload through in silico selection of the best candidates. These strategies could turn peptaibiotics into effective, ultra-specific, biodegradable tools for phytopathogen control.
... 10 The search was performed in NCBI and ScienceDirect and using as search terms 'Trichoderma citrinoviride' in combination with 'toxic', 'pathogen' and 'virulence'. The search identified 10 relevant publications involving the species T. citrinoviride either in pathogenicity (Kuhls et al., 1999) or toxigenicity (i.e., production of compounds with antifeedant activity against aphids (Evidente et al., 2008(Evidente et al., , 2009, peptaibols (Maddau et al., 2009;Sadykova et al., 2015a,b;Castagnoli et al., 2018;Marik et al., 2019), terpens and other compounds with antimicrobial and cytotoxic activity (Hu et al., 2014;Liang et al., 2016a,b;Liu et al., 2020)). Based on the information retrieved from the literature search, the whole genome sequence (WGS) data of the production strain T. citrinoviride DSM 33578 was interrogated . ...
Following a request from the European Commission, the EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) was asked to deliver a scientific opinion on the safety and efficacy of the product Huvezym® neXo 100 G/L containing an endo-1,4-beta xylanase, an endo-1,4-beta-glucanase and a xyloglucan-specific-endo-beta-1,4-glucanase produced by a non-genetically modified strain of Trichoderma citrinoviride (DSM 33578) as a zootechnical additive for feed in all poultry species, ornamental birds and piglets (weaned and suckling). The information regarding the production strain did not allow to confirm its taxonomic identification. The batches used for the characterisation of the final formulations showed compliance with the minimum specifications of the additive in terms of enzyme activities but showed ratios between the enzymes lower than the ones specified for the additive. The FEEDAP Panel considered that the below-described conclusions would apply to the final formulations of the additive as per specifications with xylanase:glucanase, xylanase:xyloglucanase and glucanase:xyloglucanase ratios of 15, 15 and 1, respectively. Based on the data available, the Panel concluded that the additive is safe for the target species, consumers and the environment. Huvezym® neXo 100 G (granulated form) is neither skin corrosive nor eye irritant but should be considered a potential skin sensitiser. Huvezym® neXo 100 L (liquid) is neither skin corrosive nor sensitising and it is not an eye irritant. Due to lack of data, no conclusions can be drawn on the skin irritation of the final formulations of the additive. Due to the proteinaceous nature of the active substances, the additive is considered a respiratory sensitiser. The FEEDAP Panel concluded that the additive has the potential to be efficacious in chickens for fattening, chickens reared for laying and breeding, and all growing poultry and ornamental birds at the minimum intended level of 1,500 EPU, 100 CU and 100 XGU/kg complete feed. Owing to the lack of sufficient data, the Panel could not conclude on the efficacy of the additive for laying hens and weaned piglets.
... (Daniel and Rodrigues Filho, 2007). Trichoderma citrinoviride produced peptaibols secondary metabolites in the liquid culture, which was screened and analyzed by TLC, HPLC, MALDI-TOF MS, and nano-ESI-QTOF MS, and they showed strong antifungal activity against Biscogniauxia mediterranea and Artemisia salin (Maddau et al., 2009). ...
Abstract
The overpopulation and rapid expansion of urban lifestyles has accelerated global demand of food supply, which has triggered unprecedented destruction of forest land and created a heavy toll on the ecosystem. Each year a large portion of agricultural crops are destroyed by different kind of pest infestations, which ensures the use of chemical pesticides despite their toxic aftereffects. The fungi and fungal metabolites are playing a crucial as well as potential role in the biopesticide industry. The inter- and intraspecific competition, rapid hyphal growth, ability to absorb water and nutrients, and phytoremediation features are critical arsenals of fungal biocontrol agents. A number of secondary metabolites were recognized and certified as biocidal, nematocidal, and weedicidal agents. Here we have provided details of large number of fungal metabolites and fungi and discussed their interesting mechanism of action as biopesticide. This chapter has also highlighted the future-research aspects and commercialization requirements of fungal-biopesticides as an integral part of sustainable agriculture.
... Despite the numerous studies carried out, so far, oak decline continues to be the main pathological problem of oak forests especially in the Mediterranean region because of its complex aetiology and the resulting difficulties in defining suitable control strategies (Broberg, Doonan, Mundt, Denman, & McDonald, 2018;Denman et al., 2018;Denman, Plummer, Kirk, Peace, & McDonald, 2016;Fernandez-Escobar et al., 1999;Luque, Pera, & Parladé, 2008;Maddau et al., 2009). ...
The oak decline is known as one of the most destructive complex diseases causing high economic losses around the world, especially in Iran. The main objective of the present study was to investigate the possible role of bacteria as causative agents of oak decline in the Zagros forests of Iran. To do this, stem, root and leaf samples were taken from symptomatic Persian oak trees (Quercus brantii) in different zones of Zagros forests (Ilam Province, Iran). From 150 bacterial isolates, 20 showed pathogenicity against Geranium seedlings. Among 20 hypersensitivity test positive strains, four strains showed pathogenicity against oak saplings. Based on morphological and 16S rRNA gene sequence analysis, three strains were identified as Bacillus pumilus and one strain as non‐sporulating Gram‐negative Stenotrophomonas maltophilia. Pathogenicity studies of different B. pumilus and S. maltophilia strains revealed that they have potential to cause the disease in oak saplings and symptoms of disorder in Persian oak trees. To our knowledge, there are no previous records of B. pumilus and S. maltophilia causing decline on Fagaceous trees like Q. brantii. More detailed field and molecular studies are required to confirm the absolute role of such bacteria in occurrence of oak decline in Zagros forests.
... Varieties of peptaibols have been reported from Trichoderma spp. [26][27][28]. From the pharmacological perspective, peptaibols exhibit variety of bioactivities with claims of antibacterial, antifungal, anticancer, immunosuppressive, antimycoplasmic, antitrypanosomal and wound healing properties, while from agricultural perspective, their role in biocontrol and induced disease resistance has also been reported [27][28][29][30][31][32][33][34]. Thus, they have potential to be used in medicine, agriculture and industry [35,36]. ...
... [26][27][28]. From the pharmacological perspective, peptaibols exhibit variety of bioactivities with claims of antibacterial, antifungal, anticancer, immunosuppressive, antimycoplasmic, antitrypanosomal and wound healing properties, while from agricultural perspective, their role in biocontrol and induced disease resistance has also been reported [27][28][29][30][31][32][33][34]. Thus, they have potential to be used in medicine, agriculture and industry [35,36]. ...
... Most of the Trichoderma spp. including the endophytic ones are peptaibol producers [27]. These peptaibols exhibit a variety of indispensable bioactivities [61]. ...
Background:
Exploration of microbes isolated from north western Himalayas for bioactive natural products.
Results:
A strain of Trichoderma lixii (IIIM-B4) was isolated from Bacopa monnieri L. The ITS based rDNA gene sequence of strain IIIM-B4 displayed 99% sequence similarity with different Trichoderma harzianum species complex. The highest score was displayed for Hypocrea lixii strain FJ462763 followed by H. nigricans strain NBRC31285, Trichoderma lixii strain CBS 110080, T. afroharzianum strain CBS124620 and Trichoderma guizhouense BPI:GJS 08135 respectively. Position of T. lixii (IIIM-B4) in phylogenetic tree suggested separate identity of the strain. Microbial dynamics of T. lixii (IIIM-B4) was investigated for small peptides. Medium to long chain length peptaibols of 11 residue (Group A), 14 residue (Group B) and 17 residue (Group C) were identified using Matrix Assisted Laser Desorption/Ionization-Time of Flight (MALDI-TOF) mass spectrometer. Optimization is undeniably a desideratum for maximized production of desirable metabolites from microbial strain. Here optimization studies were carried out on T. lixii (IIIM-B4) using different growth media through Intact Cell Mass Spectrometry (ICMS). A multifold increase was obtained in production of 11 residue peptaibols using rose bengal medium. Out of these, one of them named as Tribacopin AV was isolated and sequenced through mass studied. It was found novel as having unique sequence Ac-Gly-Leu-Leu-Leu-Ala-Leu-Pro-Leu-Aib-Val-Gln-OH. It was found to have antifungal activity against Candida albicans (25 μg/mL MIC).
Conclusion:
In this study, we isolated a strain of T. lixii (IIIM-B4) producing medium and long chain peptaibols. One of them named as Tribacopin AV was found novel as having unique sequence Ac-Gly-Leu-Leu-Leu-Ala-Leu-Pro-Leu-Aib-Val-Gln-OH, which had antifungal properties.
... This extraction technique has been proven to be reproducible, quantitatively comparable, and affordable compared to other techniques, such as SFE and accelerated solvent extraction (ASE) [11][12][13][14]. Although the Naviglio Extractor ® has been employed on several matrices [13,[15][16][17], to our knowledge, its use on grape marc is yet to be explored. In addition, to be valuable, an extraction technology should not affect or should be able to retain the biological activities of the compounds extracted from that matrix, and therefore, its employment needs to be tested for this aspect. ...
The aim of this work is the evaluation of a green extraction technology to exploit winery waste byproducts. Specifically, a solid–liquid extraction technology (Naviglio Extractor®) was used to obtain polyphenolic antioxidants from the Cagnulari grape marc. The extract was then chemically characterized by spectrophotometric analysis, high-performance liquid chromatography, and mass spectrometry, revealing a total polyphenol content of 4.00 g/L ± 0.05, and the presence of anthocyanins, one of the most representative groups among the total polyphenols in grapes. To investigate potential biological activities of the extract, its ability to counteract hydrogen peroxide-induced oxidative stress and cell death was assessed in primary human endothelial cells. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test, used to assess potential extract cytotoxicity, failed to show any deleterious effect on cultured cells. Fluorescence measurements, attained with the intracellular reactive oxygen species (ROS) probe 2′,7′-dichlorodihydrofluorescein diacetate (H2DCF-DA), revealed a strong antioxidant potential of the marc extract on the used cells, as indicated by the inhibition of the hydrogen peroxide-induced ROS generation and the counteraction of the oxidative-induced cell death. Our results indicate the Naviglio extraction, as a green technology process, can be used to exploit wine waste to obtain antioxidants which can be used to produce enriched foods and nutraceuticals high in antioxidants.
... Likewise other Trichoderma spp., this fungus has been isolated from many different environments and substrates. These included: a cork oak in Italy (Maddau et al. 2009), decayed cyanobacterial bloom collected in an eutrophic lake in Saudi Arabia (Mohamed et al. 2014), potting media and different kind of cultivated and uncultivated soils in several countries (Al-Sadi et al. 2015), decayed wood from several mountain districts in Poland (Błaszczyk et al. 2016), sediments in Chinese wetlands (Saravanakumar et al. 2016), mycelium of the shiitake mushroom in Korea (Kim et al. 2016), and a soil sample taken in a common bean plantation in Spain (Rodríguez-González et al. 2017). In addition, human pathogenic aptitude has been also reported for T. citrinoviride isolates on immunocompromised patients (Kuhls et al. 1999). ...
From the green alga Cladophora sp. collected in Italy, the marine fungal strain A12 of Trichoderma citrinoviride was isolated, identified and characterized. LC-MS qTOF analysis was applied to perform a metabolic profile of the fungal culture. Chromatographic techniques and spectroscopic methods were used to isolate and characterize the major secondary metabolites produced by this strain in liquid culture. In particular, four known sorbicillinoids (trichodermanone C, spirosorbicillinol A, vertinolide and sorbicillin) were purified and identified, together with 2-phenylethanol and tyrosol. Moreover, metabolomic analysis allowed to detect small amounts of trichodimerol, rezishanone A, 2′,3′-dihydrosorbicillin and bisvertinol. For the first time a significant inhibitory effect on nitrite levels has been shown for trichodermanone C in lipopolysaccharide-stimulated J774A.1 macrophages.
... The mycelial extract of T. citrinoviride DAOM 242936 revealed a mixture of 20-residue peptaibols previously reported from the same species (Table 2; Maddau et al. 2009). The peptaibol-enriched extract was active in the fungal membrane assay we used at the highest concentration tested (100 µg mL -1 ; Table 6). ...
Buildings that have been flooded often have high concentrations of Trichoderma spores in the air while drying. Inhaled spores, spore and mycelial fragments contain large amounts of fungal glucan and natural products that contribute to the symptoms associated with indoor mold exposures. In this study, we considered both small molecules and peptaibol profiles of T. atroviride, T. koningiopsis, T. citrinoviride and T. harzianum strains obtained from damp buildings in Eastern Canada. Twenty-residue peptaibols and sorbicillin-derived metabolites (1-6) including a new structure, (R)-vertinolide (1), were characterized from T. citrinoviride. T. koningiopsis produced several koninginins (7-10), trikoningin KA V and the 11-residue lipopeptaibols, trikoningins KB I and KB II. T. atroviride biosynthesized a mixture of 19-residue trichorzianine-like peptaibols whereas T. harzianum produced 18-residue trichokindin-like peptaibols and the 11-residue harzianin HB I that was subsequently identified from the studied T. citrinoviride strain. Two α-pyrones, 6-pentyl-pyran-2-one (11) and an oxidized analog (12), were produced by both T. atroviride and T. harzianum. Aside from low molecular weight natural products, inhalation of Trichoderma spores and mycelial fragments may result in exposures to membrane disrupting peptaibols. This investigation contributes to a more comprehensive understanding of the toxigenic potential of fungi commonly found in water-damaged buildings.
... At the lower concentration tested (10 µg/mL), the peptide mixture was still highly active against all fungi except A. quercina and B. mediterranea with a percentage growth inhibition between 45.6 and 78.7 %. It was also highly toxic in an Artemia salina bioassay with a LC 50 value of 1.24 µg/mL [200]. ...
Background:
Fungi play an important role in terrestrial ecosystems interacting positively or negatively with plants. These interactions are complex and the outcomes are different depending on the fungal lifestyles, saprotrophic, mutualistic or pathogenic. Furthermore, fungi are well known for producing secondary metabolites, originating from different biosynthetic pathways, which possess biological properties of considerable biotechnological interest. Among the terrestrial ecosystems, temperate forests represent an enormous reservoir of fungal diversity. This review will highlight the goldmine of secondary metabolites produced by pathogenic and endophytic fungi of forest trees with focus on their biological activities.
Methods:
A structured search of bibliographic databases for peer-reviewed research literature was undertaken using a research discovery application providing access to a large and authoritative source of references. The papers selected were examined and the main results were reported and discussed.
Results:
Two hundred forthy-one papers were included in the review, outlined a large number of secondary metabolites produced by pathogenic and endophiltic fungi and their biological activities, including phytotoxic, antifungal, antioomycetes, antibacterial, brine shrimp lethality, mosquito biting deterrence and larvicidal, cytotoxic, antiproliferative and many other bioactivities.
Conclusion:
The findings of this review confirm the importance of secondary metabolites produced by pathogenic and endophytic fungi from forest plants growing in temperate regions as an excellent prospects to discover compounds with new bioactivities and mode of actions. In addition, the potential of some metabolites as a source of new drugs and biopesticides is underlined.
... Trichoderma species are the unique group of filamentous fungi that are highly competitive saprophytes, are rhizosphere dwelling and root colonizers, and are extensively studied as potential sources of biocontrol agents, enzymes (cell wall degrading enzymes, CWDEs), and bioactive peptides (Mukherjee et al. 2012;Maddau et al. 2009;Wiest et al. 2002;Chugh and Wallace, 2001). These fungi exhibit important roles in agriculture, medicine, and industry (Brotman et al. 2010;Schuster and Schmoll 2010). ...
Trichoderma is an anamorphic filamentous fungal genus with immense potential for production of small valuable secondary metabolites with indispensable biological activities. Microbial dynamics of a psychrotrophic strain Trichoderma velutinum ACR-P1, isolated from unexplored niches of the Shiwalik region, bestowed with rich biodiversity of microflora, was investigated for production of nonribosomal peptides (NRPs) by metabolite profiling by intact-cell mass spectrometry (ICMS) employing matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometer. Being the first report on NRPs production by T. velutinum, studies on optimization of growth conditions by Response Surface Methodology (RSM) for production of NRPs by ACR-P1 was carried out strategically. Multifold enhancement in the yield of NRPs belonging to subfamily SF4 with medium chain of amino acid residues having m/z 1437.9, 1453.9, and 1452.0 at pH 5.9 at 20 °C and of subfamily SF1 with long-chain amino acid residues having m/z 1770.2, 1784.2, 1800.1, 1802.1, and 1815.1 was achieved at pH 7.0 at 25 °C. Complexities of natural mixtures were thus considerably reduced under respective optimized culture conditions accelerating the production of novel microbial natural products by saving time and resources.
