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Representative NMR spectrum of the methanol extract of the endometabolome of Fusarium verticillioides in confrontation with Streptomyces AV05
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The Streptomyces sp. strain AV05 isolated from an organic amendment was found to impact both growth and fumonisin production of Fusarium verticillioides during in vitro direct confrontation. In order to investigate the interactions between the Streptomyces sp. strain AV05 and F. verticillioides, a metabolomic approach was used. The study of the end...
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... one amine listed in Table 1 were identified as key markers and were validated with the use of a Wilcoxon test. The structure of discriminating metabolites was searched against the Human Metabolome Database (HMDB, version 4.0) and home database. The representative NMR spectrum of methanol extract of Fusarium cultivated with bacteria is shown in Fig. ...
Context 2
... order to understand the mechanism of inhibition of fumonisin production by the strain AV05, the extraction of the endometabolites was performed on 5-day-old cultures of Streptomyces strain AV05 and F. verticillioides in two conditions: when cultivated alone and in confrontation. The metabolomic approach was used to examine changes in the metabolome of the fungal strain during the confrontation assay and attempt to identify over or under expressed metabolites in F. verticillioides in the presence of the Streptomyces strain (Frändberg et al. 2000; Joo 2005). Results of the NMR analysis of the endometabolomes of Fusarium and Streptomyces ...
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... Recently, antibiotics, stress inducers, and biocontrol microorganisms have been largely used for the prevention of plant diseases. The main biocontrol microorganisms are bacteria, actinomycetes, fungi, and viruses [1], which can either secrete antimicrobial substances or perform niche competition to promote plant growth and induce plant systemic resistance [2]. Microbial biological control agents are effective alternatives to synthetic chemicals [3]. ...
Streptomyces alfalfa strain 11F has inhibitory effects on many phytopathogenic fungi and improves the establishment and biomass yield of switchgrass. However, the antagonistic effects of strain 11F on Fusarium wilt of watermelon and its secondary metabolites that contribute to its biocontrol activity are poorly understood. We evaluated the antagonistic and growth-promoting effects of strain 11F and conducted a transcriptome analysis to identify the metabolites contributing to antifungal activity. Strain 11F had marked inhibitory effects on six fungal pathogens. The incidence of Fusarium wilt of watermelon seedlings was decreased by 46.02%, while watermelon seedling growth was promoted, as indicated by plant height (8.7%), fresh weight (23.1%), and dry weight (60.0%). Clean RNA-sequencing data were annotated with 7553 functional genes. The 2582 differentially expressed genes (DEGs) detected in the Control vs. Case 2 comparison were divided into 42 subcategories of the biological process, cellular component, and molecular function Gene Ontology categories. Seven hundred and forty functional genes (55.47% of the DEGs) were assigned to Kyoto Encyclopedia of Genes and Genomes metabolic pathways, reflecting the complexity of the strain 11F metabolic regulatory system. The expression level of the gene phzF, which encodes an enzyme essential for phenazine-1-carboxylic acid (PCA) synthesis, was downregulated 3.7-fold between the 24 h and 48 h fermentation time points, suggesting that strain 11F can produce phenazine compounds. A phenazine compound from 11F was isolated and identified as phenazine-1-carboxamide (PCN), which contributed to the antagonistic activity against Fusarium oxysporum f. sp. niveum. PCA was speculated to be the synthetic precursor of PCN. The downregulation in phzF expression might be associated with the decrease in PCA accumulation and the increase in PCN synthesis in strain 11F from 24 to 48 h. Streptomyces alfalfae 11F protects watermelon seedlings from Fusarium wilt of watermelon and promotes seedling growth. The transcriptome analysis of strain 11F provides insights into the synthesis of PCN, which has antifungal activity against F. oxysporum f. sp. niveum of watermelon.
... These strategies consist of growing different microorganisms in the same confined environment, allowing them to interact with each other [74]. Such interactions may even alter the core metabolism of the cells, for example, by increasing the production of ergosterol [75]. In addition to a yield increase in common molecules, these types of strategies can increase the yield of previously unidentified or undetected molecules, facilitating their characterization [76]. ...
Antibiotics are a staple in current medicine for the therapy of infectious diseases. However, their extensive use and misuse, combined with the high adaptability of bacteria, has dangerously increased the incidence of multi-drug-resistant (MDR) bacteria. This makes the treatment of infections challenging, especially when MDR bacteria form biofilms. The most recent antibiotics entering the market have very similar modes of action to the existing ones, so bacteria rapidly catch up to those as well. As such, it is very important to adopt effective measures to avoid the development of antibiotic resistance by pathogenic bacteria, but also to perform bioprospecting of new molecules from diverse sources to expand the arsenal of drugs that are available to fight these infectious bacteria. Filamentous fungi have a large and vastly unexplored secondary metabolome and are rich in bioactive molecules that can be potential novel antimicrobial drugs. Their production can be challenging, as the associated biosynthetic pathways may not be active under standard culture conditions. New techniques involving metabolic and genetic engineering can help boost antibiotic production. This study aims to review the bioprospection of fungi to produce new drugs to face the growing problem of MDR bacteria and biofilm-associated infections.
... Streptomyces spp. not only inhibited the growth of F. verticillioides, but also reduced FBs production by interfering with the fungal metabolic pathways, particularly mycotoxigenesis (Nguyen et al., 2020). Strub et al. (2021) showed that Streptomyces sp. ...
... The biocontrol strain significantly promoted plant growth even in the presence of F. verticillioides. In agreement with Nguyen et al. (2020), Streptomyces spp. inhibited F. verticillioides growth and reduced FBs production affecting mycotoxigenesis. ...
Maize (Zea mays L.) is one of the most important cereals around the world since it is widely used for food and feed. This cereal is relevant to the economy of several countries due to its volume of production and exportation. During pre-harvest stages this crop is susceptible to infection by species of Aspergillus section Flavi and Fusarium and prone to mycotoxin contamination, mainly aflatoxins and fumonisins. Additionally, the climate change scenario predicted for next decades, including increases in the global temperature, CO2 levels and periods of hydric stress, would contribute to higher levels of mycotoxins worldwide. In this context, there is a need to find successful, feasible and eco-friendly tools for an effective mycotoxin mitigation in order to guarantee food security and safety. This review summarises the current situation of aflatoxin and fumonisin contamination in maize, highlighting the importance of the evaluation and implementation of biocontrol strategies at pre-harvest stage in the frame of an integrated control management to reduce the impact of mycotoxin accumulation under the risk of climate change expected for the next years.
... After undergoing in vitro confrontation culture, the metabolome changes of Fusarium verticillioides were much greater than those of Streptomyces sp. Compared with monoculture, many metabolites of Fusarium verticillioides were overproduced under resistant conditions compared to Streptomyces sp., especially 16 proteinogenic amino acids, inosine, and uridine, which means that the corresponding rate of protein synthesis would be slowed down, resulting in slower growth and less toxigenesis of Fusarium verticillioides [49]. Both the environment and the pH affect the biosynthesis of mycotoxins from Fusarium verticillioides. ...
Metabolomics is an essential method to study the dynamic changes of metabolic networks and products using modern analytical techniques, as well as reveal the life phenomena and their inherent laws. Currently, more and more attention has been paid to the development of metabolic histochemistry in the fungus field. This paper reviews the application of metabolomics in fungal research from five aspects: identification, response to stress, metabolite discovery, metabolism engineering, and fungal interactions with plants.
... A possibility for the reduction of fumonisins is the inhibition of fungal growth by strain ST03. In agreement with Nguyen et al. (2020), Streptomyces spp. not only inhibit the growth of F. verticillioides, but also reduce fumonisin production via interfering with the fungal metabolic pathways, particularly mycotoxigenesis. ...
Driven by climate change, Fusarium ear rot (FER) caused by Fusarium verticillioides occurs frequently in maize worldwide. In parallel, legislative regulations and increasing environmental awareness have spurred research on alternative FER biocontrol strategies. A promising group of bacterial control agents is Streptomyces species due to their metabolic versatility. However, insights into the molecular modes of action of these biocontrol agents are often lacking. This study aims at unraveling the biocontrol efficacy of Streptomyces rhizobacterial strains against F. verticillioides. We first assessed the direct antagonism of four Streptomyces strains ST02, ST03, ST07, and ST08. Then, a profile of 16 genes associated with intrinsic plant defense signaling was assessed in maize plants. Both in vitro and in vivo data showed that the biocontrol strain ST03 perfectly suppressed the growth of F. verticillioides. High inhibition efficacy was also observed for extracellular compounds in the supernatant secreted by this strain. Especially, for maize cobs, the biocontrol strain ST03 not only inhibited the proliferation of F. verticillioides but also significantly repressed fungal fumonisin production 7 days after inoculation. On maize plants, the direct antagonism was confirmed by a significant reduction of the fungal DNA level in soils when co-applied with F. verticillioides and strain ST03. In terms of its action on plants, strain ST03 induced downregulation of auxin responsive genes (AUX1, ARF1, and ARF2) and gibberellic acid (GA)-related gene AN1 even in the absence of F. verticillioides at early time points. In leaves, the biocontrol strain induced the expression of genes related to salicylic acid (SA), and 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA)-mediated pathways, and pathogenesis-related proteins in the presence or absence of the pathogen. Interestingly, the biocontrol strain significantly promoted plant growth even in the presence of F. verticillioides. All of which demonstrated that the Streptomyces strain ST03 is a promising FER biocontrol and a growth-promoting candidate.
... Antagonistic activity in vitro. The antagonistic activity of the 22 strains of streptomycetes against the isolation of F. oxysporum GL1 is shown in Table 2, where we can also see, for the streptomycetes in the soil of the states of Hidalgo and Aguascalientes, (Goredema et al., 2020;Nguyen et al., 2020;Tlemsani et al., 2020). Earlier results with Streptomyces sp. ...
... CA y Streptomyces sp. GCAL-9 son cercanos a los resultados reportados para otros actinomicetos sobre F. oxysporum (PI de 58.1%), lo que confirma la capacidad de algunas especies del género Streptomyces para inhibir a F. oxysporum (Goredema et al., 2020;Nguyen et al., 2020;Tlemsani et al., 2020). Resultados previos con Streptomyces sp. ...
em>Fusarium oxysporum causa la pudrición del cormo en gladiolo provocando pérdidas de hasta el 100%. Se seleccionaron aislados de Fusarium a partir de cormos infectados, se identificó morfológica y molecularmente y se seleccionó un aislado a partir de prueba de patogenicidad. Se seleccionó entre 22 aislados de estreptomicetos una cepa que presentó una actividad antagonista del 40% contra Fusarium . Se obtuvo el Extracto Bioactivo (EB) mediante Fermentación en Fase Sólida y se determinó la concentración mínima inhibitoria (MIC) y concentración mínima letal (MLC) por el método de microdilución. Se obtuvo una MIC para el EB de 0.19 mg mL-1 y una MLC de 0.38 mg mL-1, que se confirmó con un ensayo de germinación de microconidios a 8 h, mostrando un porcentaje de inhibición del 17 y 98% para ¼ y ½ de la MIC. Se evaluó el efecto del EB a 1 y 2 MIC’s de concentración contra la pudrición en cormos de gladiolo infectados, obteniendo un efecto protector en los cormos al mantener su dureza después de 15 días, en comparación con el fungicida Carbendazim. Los resultados indican a Streptomyces sp., como un potencial agente de control biológico contra F. oxysporum .
... It allows the microorganisms to initiate chemical interactions and results in the production of NPs that are not observed in pure-strain cultures such as pheromones, defence molecules or metabolites related to symbiotic associations, therefore revealing hidden biosynthetic pathways (Harwani et al., 2018;Jones and Wang, 2018;Nai and Meyer, 2018;Tan et al., 2019;Tshikantwa et al., 2018;Qian et al., 2019). Such alteration in the specialized metabolisms can be attributed to alteration in core metabolisms by co-culture (Cheng et al., 2013;Nguyen et al., 2019). In addition, metabolite exchanges, metabolisation and/or detoxification of defence molecules (Shang et al., 2017;Vallet et al., 2017) may also occur yielding the induction of chemical novelty (Gebreselassie and Antoniewicz, 2015). ...
... In NP research, metabolomics is considered as the large-scale analysis of metabolites of a given organism during various physiological states which can be interpreted from a holistic manner (Sumner et al., 2003). However, only few papers have adopted this approach (Bertrand et al., 2013c;Bohni et al., 2016;Combès et al., 2012;Nguyen et al., 2019;Shen et al., 2019;Traxler et al., 2013;Wu et al., 2015;Xu et al., 2018), indicating that such strategy is not yet fully adopted in NPs chemistry. ...
... Nowadays, chemical profiling is mostly achieved by liquid chromatography coupled to high resolution mass spectrometry (LC-HRMS) or mass spectrometry imaging (MS-imaging) (Bertrand et al., 2014b). In the reported cases, NMR is mostly used to reveal core metabolism alterations (Cheng et al., 2013;Martínez-Buitrago et al., 2019;Nguyen et al., 2019). Very recently, a 2D NMR fingerprint strategy (Buedenbender et al., 2018) has been reported to highlight metabolic alteration thus indicating the induction of polyketides, peptides and aromatic compounds base on specific observed resonances (Buedenbender et al., 2019). ...
Natural products (NPs) are considered as a cornerstone for the generation of bioactive leads in drug discovery programs. However, one of the major limitations of NP drug discovery program is “rediscovery” of known compounds, thereby hindering the rate of drug discovery efficiency. Therefore, in recent years, to overcome these limitations, a great deal of attention has been drawn towards understanding the role of microorganisms’ co-culture in inducing novel chemical entities. Such induction could be related to activation of genes which might be silent or expressed at very low levels (below detection limit) in pure-strain cultures under normal laboratory conditions. In this review, chemical diversity of compounds isolated from microbial co-cultures, is discussed. For this purpose, chemodiversity has been represented as a chemical-structure network based on the “Tanimoto Structural Similarity Index”. This highlights the huge structural diversity induced by microbial co-culture. In addition, the current trends in microbial co-culture research are highlighted. Finally, the current challenges (1 - induction monitoring, 2 - reproducibility, 3 - growth time effect and 4 - up-scaling for isolation purposes) are discussed. The information in this review will support researchers to design microbial co-culture strategies for future research efforts. In addition, guidelines for co-culture induction reporting are also provided to strengthen future reporting in this NP field.
... A significant number of fungal metabolic pathways (i.e. membrane component, protein and lipid biosynthesis) have been impacted by the actinobacteria (Nguyen et al., 2019). The alteration of metabolome was reported to be associated with functions and expression of one or several genes involved in metabolic pathways for F. verticillioides (Boudreau et al., 2013) and others fungi (Lorenz and Fink, 2001;Wang et al., 2003;Sheridan et al., 2014). ...
... We also observed the same results in a previous study (Nguyen et al., 2018). In another study (Nguyen et al., 2019), the metabolome of F. verticillioides and Streptomyces strain AV05 were investigated when cultivated alone and in co-culture. Interesting metabolites differently expressed were identified, especially for F. verticillioides. ...
... Transmembrane transport in BP category to have been affected to a lesser extent. Accumulation of ergosterol that was found in the endometabolome of Fusarium confronted to Streptomyces AV05 in a previous metabolomics study of the confrontation between the 2 strains (Nguyen et al., 2019) was in agreement with these observations. Both over and under expressions of 'phospholipid-translocating ATPase' are representative of the effect of Streptomyces presence on the fungal strain. ...
The actinobacteria Streptomyces sp. AV05 appears to be a potential biocontrol agent (BCA) against mycotoxigenic fungi. It was found to significantly inhibit F. verticillioides growth and mycotoxin production during their co-cultivation. F. verticillioides growth was durably affected while the decrease of the toxin production levels was reversible, suggesting different BCA actions. The study of both transcriptomes brought useful information on the microbial interaction. RNA-seq data indicated that the dual interaction modified genetic expression of both microorganisms as 18.5 % of the genes were differentially expressed for the fungus against 3.8 % for the actinobacteria. Fungal differentially expressed genes (DEGs) were equally up and down regulated while bacterial ones were mainly upregulated. We especially focused the analysis of DEGs on fungal defense reaction to bacterial attack. For example, if this potential BCA implements a strategy of antibiosis with the over expression of ‘siderophore-interacting protein’ linked to the production of bacteriocins, the fungus in a state of stress is able to adapt its metabolism by up-regulation of amidase. It could correspond to the induction of resistance gene clusters and suggest a detoxification process. Moreover fumonisins-related pathway appears underexpressed in the presence of Streptomyces that explain the reduction of fumonisin accumulation observed.
Co-cultivation is an effective method of inducing the production of specialized metabolites (SMs) in microbial strains. By mimicking the ecological interactions that take place in natural environment, this approach enables to trigger the biosynthesis of molecules which are not formed under monoculture conditions. Importantly, microbial co-cultivation may lead to the discovery of novel chemical entities of pharmaceutical interest. The experimental efforts aimed at the induction of SMs are greatly facilitated by computational techniques. The aim of this overview is to highlight the relevance of computational methods for the investigation of SM induction via microbial co-cultivation. The concepts related to the induction of SMs in microbial co-cultures are briefly introduced by addressing four areas associated with the SM induction workflows, namely the detection of SMs formed exclusively under co-culture conditions, the annotation of induced SMs, the identification of SM producer strains, and the optimization of fermentation conditions. The computational infrastructure associated with these areas, including the tools of multivariate data analysis, molecular networking, genome mining and mathematical optimization, is discussed in relation to the experimental results described in recent literature. The perspective on the future developments in the field, mainly in relation to the microbiome-related research, is also provided.
