Antifungal Secondary Metabolites Research Articles

Bacillus spp. as Bio-factories for Antifungal Secondary Metabolites: Innovation Beyond Whole Organism Formulations.

Several fungi act as parasites for crops causing huge annual crop losses at both pre- and post-harvest stages. For years, chemical fungicides were the solution; however, their wide use has caused environmental contamination and human health problems. For this reason, the use of biofungicides has been in practice as a green solution against fungal phytopathogens. In the context of a more sustainable agriculture, microbial biofungicides have the largest share among the commercial biocontrol products that are available in the market. Precisely, the genus Bacillus has been largely studied for the management of plant pathogenic fungi because they offer a chemically diverse arsenal of antifungal secondary metabolites, which have spawned a heightened industrial engrossment of it as a biopesticide. In this sense, it is indispensable to know the wide arsenal that Bacillus genus has to apply these products for sustainable agriculture. Having this idea in our minds, in this review, secondary metabolites from Bacillus having antifungal activity are chemically and structurally described giving details of their action against several phytopathogens. Knowing the current status of Bacillus secreted antifungals is the base for the goal to apply these in agriculture and it is addressed in depth in the second part of this review.

Identification of Putative Elicitors From Plant Root Exudates Responsible for PsoR Activation in Plant-Beneficial Pseudomonas spp. by Docking and Molecular Dynamics Simulation Approaches to Decipher Plant-Microbe Interaction.

Plants and rhizobacteria are coexisting since the beginning, but the exact mechanism of communication between them remains enigmatic. The PsoR protein of plant-beneficial Pseudomonas spp., a group of root-associated bacteria, is known to produce a range of antifungal and insecticidal secondary metabolites like 2,4-diacetyl phloroglucinol (DAPG), pyrrolnitrin, and chitinase making them great biocontrol agents and thus helping in plant growth promotion. To better understand the inter-kingdom signaling between plants and plant growth-promoting rhizobacteria (PGPR), the interaction of PsoR with various root exudates was investigated computationally. For this, we first modeled the PsoR protein and confirmed it using the Ramachandran plot. A total of 59 different low molecular weight phytochemicals, secreted as root exudates by plants, were identified by extensive text mining. They were virtually screened with the PsoR protein by molecular docking. Based on the lowest binding energy, ranging from −7.1 to −6.3 kcal mol−1, the top five exudates were chosen. To analyze the stability of the docked protein–ligand complex, a molecular dynamics (MD) simulation of 100 nanoseconds was done. Two root exudates, saponarin and 2-benzoxazolinone (BOA), showed suitable binding with PsoR by forming hydrogen, hydrophobic, and Van der Waals interactions. To confirm the MD simulation results, RMSF, RG, SASA, and interaction energy were calculated. This computational study first time reports that saponarin and 2-BOA, predominantly present in the root exudates of barley and wheat, respectively, demonstrate effective binding with the modeled PsoR protein and are likely of showing cross-kingdom interactions.

Mono-/Bis-Alkenoic Acid Derivatives From an Endophytic Fungus Scopulariopsis candelabrum and Their Antifungal Activity

During a screening for antifungal secondary metabolites, six new mono-/bis-alkenoic acid derivatives (2–7) and one known alkenoic acid derivative (1) were isolated from an endophytic fungi Scopulariopsis candelabrum. Their chemical structures were identified by 1H-NMR, 13C-NMR, 2D NMR, and high-resolution mass spectrometry, as well as comparisons with previously reported literatures. Among them, fusariumesters C‒F (2–5) are bis-alkenoic acid derivatives dimerized by an ester bond, while acetylfusaridioic acid A (6) and fusaridioic acid D (7) are alkenoic acid monomers. All the isolates were submitted to an antifungal assay against Candida albicans and the corn pathogen Exserohilum turcicum using the filter paper agar diffusion method. As a result, only compound 1 decorating with β-lactone ring turned out to be active against these two tested fungi. The broth microdilution assay against Candida albicans showed the minimum inhibitory concentration (MIC) value of 1 to be 20 μg/ml, while the minimum inhibitory concentration value of the positive control (naystatin) was 10 μg/ml. And the half maximal inhibitory concentration (IC50) value (21.23 μg/ml) of 1 against Exserohilum turcicum was determined by analyzing its inhibition effect on the mycelial growth, using cycloheximide (IC50 = 46.70 μg/ml) as the positive control.

Optimization of growth parameters for enhancing antifungal secondary metabolites of Talaromyces islandicus VSGF1 against drug resistant Candida spp.

Optimization of growth parameters for enhancing antifungal secondary metabolites of Talaromyces islandicus VSGF1 against drug resistant Candida spp.

Development of Chitosan-Based Nanoemulsion Gel Containing Microbial Secondary Metabolite with Effective Antifungal Activity: In vitro and in vivo Characterizations.

PurposeMicrobial resistance to antibiotics is one of the most important public health concerns of the 21st century. We isolated, purified, and structurally elucidated antifungal secondary metabolites from red soil microbes and encapsulated them into chitosan (CS)-based nanoemulsion (NE) gel (NEG).MethodsThree compounds were isolated and purified of which only one compound (Pure 2) showed potent antifungal activity (MFC: 8–132 µg/mL), which was also significantly (P<0.05) more efficient than fluconazole (MFC: 32–132 µg/mL). Pure 2 was structurally elucidated using 1D- and 2D-NMR before its incorporation into NEG. The formulations were prepared by high-speed homogenization technique. Physicochemical and pharmacological characterizations of formulations (ie, droplet size, PDI, zeta potential, drug content, viscosity, SEM, FTIR, spreadability, in vitro drug release, ex vivo permeation, in vitro antifungal and in vivo antifungal activities) were assessed.ResultsNMR analyses identified the compound as a derivative of phthalic acid ester (PAE). The optimized formulations displayed a droplet size <100 nm, -ve zeta potential, and PDI <0.45. The drug content was within the official limit of pharmacopeia (ie, 100±10%). Insignificant changes (P>0.05) in the viscosity of the formulations stored were observed. The morphology of the formulations indicated mesh-like structure. The FTIR study indicated that there were no interactions between the drug and other ingredients of the formulations. Optimum spreadability was observed in all formulations. NEG released 75.3±1.12% of Pure 2 after 12 hrs while NE released 85.33±1.88% of the compound. The skin permeation of F2 (71.15±1.28%) was significantly different (P<0.05) from F3 (81.80±1.91%) in rabbits. Complete and apparently safe recovery from the fungal infection was achieved in rabbits treated topically with Pure 2-loaded NEGs.ConclusionHence, the NEG-loaded PAE isolated from Pseudomonas fluorescens represents a possible alternative for the treatment of fungal infections as compared to available therapies.

Anti-Fungal Efficacy and Secondary Metabolite Analysis of the Methanolic Extract from Colpomenia peregrina towards Opportunistic Fungal Pathogens

The present study illustrates the antifungal efficacy of methanolic extract from marine brown seaweed Colopomenia peregrina gathered from Leepuram coast, South India, towards opportunistic fungal pathogens comprising of dermatophytes, non-dermatophytes, and yeasts. The opportunistic fungal pathogens used in the study are Aspergillus flavus (ATCC 27692), Aspergillus fumigatus (ATCC 19673), Microsporum gypseum (ATCC 24102), Cryptococcus neoformans (ATCC 14116), and Candida albicans (ATCC14053) which are commonly responsible for nosocomial infections. The NMR analysis revealed the presence of various chemical shifts showing the presence of protons containing Hydroxyl, Methoxy, Methyl groups, and –COO-CH2 groups.The presence of phytochemicals from the extract of seaweed confirmed the nutritional profile. The results revealed greater efficacy of methanolic extract towards Aspergillus fumigatus, Microsporum gypseum, Cryptococcus neoformans, and lower activity with Aspergillus flavus and Candida albicans compared with the standard anti-fungal fluconazole.

Antifungal metabolite profiling in Chaetomium globosum potential strain Cg2 effective against Bipolaris sorokiniana

The fungus reported to be a potential antagonist of various soil and seed borne plant pathogens. C. globosum mycoparasitizes and produce antifungal metabolites which suppress the growth of the pathogenic fungi. Despite sufficient knowledge available on secondary metabolites of C. globosum and their wide biological activities, limited literatures are available on the polar active constituents and their potential use as antifungal agent. The study was carried out to evaluate chemical composition of secondary metabolites from C. globosum when it interacts with Bipolaris sorokiniana (2017-2020). The volatile compounds identified through GC-MS revealed that, C. globosum strain Cg2 produced a variety of antifungal secondary metabolites, i.e. octadecene trans-limonene oxide, dodecane, tetracosonal, heptacosanol and octadecanoic acid which may be involved in the antagonism. Similarly, UPLC-QToF-ESIMS analysis of chloroform soluble fraction of C. globosum (Cg2), B. sorokiniana (BS112) and their interaction (Cg2-BS112) were undertaken to identify non-volatile metabolites.These metabolites were identified as Chaetomugilin A, D, E, F, Globoxanthone A, Chaetoviridin A, B, E, Chaetoglobin B, Chaetoquadrin A, Chaetocochin B and F, Chaetoglobosin Q and N.The work indicates that the biocontrol agent C. globosum showed high antifungal metabolite production thereby antagonising B. sorokininana pathogen. The obtained data will greatly enrich current C. globosum metabolomic information and provide a good foundation for better understanding of biocontrol mechanism of C. globosum against plant pathogens and facilitating widespread application in the field of bio-control.

Transcription in fungal conidia before dormancy produces phenotypically variable conidia that maximize survival in different environments.

Fungi produce millions of clonal asexual conidia (spores) that remain dormant until favourable conditions occur. Conidia contain abundant stable messenger RNAs but the mechanisms underlying the production of these transcripts and their composition and functions are unknown. Here, we report that the conidia of three filamentous fungal species (Aspergillus nidulans, Aspergillus fumigatus, Talaromyces marneffei) are transcriptionally active and can synthesize mRNAs. We find that transcription in fully developed conidia is modulated in response to changes in the environment until conidia leave the developmental structure. Environment-specific transcriptional responses can alter conidial content (mRNAs, proteins and secondary metabolites) and change gene expression when dormancy is broken. Conidial transcription affects the fitness and capabilities of fungal cells after germination, including stress and antifungal drug (azole) resistance, mycotoxin and secondary metabolite production and virulence. The transcriptional variation that we characterize in fungal conidia explains how genetically identical conidia mature into phenotypically variable conidia. We find that fungal conidia prepare for the future by synthesizing and storing transcripts according to environmental conditions present before dormancy.

Antifungal Secondary Metabolites Produced by the Fungal Endophytes: Chemical Diversity and Potential Use in the Development of Biopesticides.

Plant diseases caused by phytopathogenic fungi can lead to huge losses in the agricultural fields and therefore remain a continuous threat to the global food security. Chemical-based fungicides contributed significantly in securing crop production. However, indiscriminate application of fungicides has led to increased chemical resistance and potential risks to human health and environment. Thus, there is an urgent need for searching for new bioactive natural products and developing them into new biopesticides. Fungal endophytes, microorganisms that reside in the fresh tissues of living plants, are regarded as untapped sources of novel natural products for exploitation in agriculture and/or medicine. Chemical examination of endophytic fungi has yielded enormous antifungal natural products with potential use in the development of biopesticides. This review summarizes a total of 132 antifungal metabolites isolated from fungal endophytes in the past two decades. The emphasis is on the unique chemical diversity of these metabolic products, together with their relevant antifungal properties. Moreover, some “star molecules,” such as griseofulvin and trichothecene, as well as their synthetic derivatives that possess high potential as candidates of new natural fungicides, are also presented herein.

English

Macrophomina phaseolina is a highly destructive pathogen of more than 500 plant species. It is difficult to eradicate it through chemical means as no patented fungicide is available against this pathogen. Biological control is the possible alternative method for its suitable management. The present study was carried out to evaluate the biocontrol potential of five Trichoderma spp. against M. phaseolina and the possible mechanisms of action. Identifications of all the Trichoderma spp. viz. T. hamatum, T. harzianum, T. koningii, T. longipile and T. viride were confirmed on molecular basis by using two universal primer pairs namely ITS and EF1. Their biocontrol potential was evaluated in dual culture plate method where T. viride showed the highest inhibitory efficacy (63%) against M. phaseolina. T. koningii, T. hamatum and T. longipile showed akin effects by arresting growth of the pathogen by 46–47% followed by T. harzianum (28%). To find out the mechanisms of action, secondary extrolites of the best biocontrol fungus T. viride were tested against the pathogenic genomic DNA where all the concentrations partially degraded DNA bands after 24 h of incubation and a complete DNA band disappearance was noted after 48 h incubation. In addition, T. viride culture filtrates were partitioned with chloroform and ethyl acetate and subjected to GC-MS analysis for identification of potential antifungal constituents. The most abundant identified volatile compounds in the two organic solvent fractions were 9,12-octadecadienoic acid (Z,Z)- (44.54%), n-hexadecanoic acid (24.02%), hexadecanoic acid, 2-hydroxy-1-(hydroxymethyl) ethyl ester (14.25%), 9-tricosene, (Z)- (10.43%) and [1,1'-bicyclopropyl]-2-octanoic acid, 2'-hexyl-, methyl ester (10.43%). To conclude, T. viride was the best biocontrol agent against M. phaseolina and acts against the pathogen by DNA disintegration and production of antifungal secondary metabolites. © 2021 Friends Science Publishers

Evaluation of the production of antifungal metabolites against Colletotrichum gloeosporioides in Streptomyces 5.1 by random mutagenesis

Streptomyces 5.1 is a bacterium isolated from rice soils in the south of the Tolima department (Colombia). This microorganism is characterized by its antagonistic activity against rubber tree phytopathogens like Colletotrichum gloeosporioides, the causal agent of leaf anthracnose. The antifungal activity of this Streptomyces isolate has been associated with secondary metabolites production. However, the identity of those metabolites is unknown because its purification and identification have not been possible through classic chemical studies. Therefore, aiming to contribute in the study of the secondary metabolites produced by 5.1 from a molecular approach, this research seeks to identify -preliminarily- the genomic fingerprint changes associated with the production of antifungal secondary metabolites produced by Streptomyces 5.1 through the evaluation of a mutant library of 5.1 obtained by random mutagenesis using controlled ultraviolet light exposure. The antifungal activity of obtained mutants was evaluated using Colletotrichum gloeosporioides (C1) fungus as a biosensor, isolated by the Biotechnology Institute of Universidad Nacional de Colombia. In this way, the library of mutants of 5.1, initially formed by 300 isolations, was classified into two phenotypic groups of interest: enhanced mutants (1 isolate) and null mutants (11 isolates) of secondary metabolites. The genomic changes in both groups were analyzed by obtaining the genomic profile of the isolates using Repetitive Extragenic Palindromic (Rep-PCR). The obtained profiles evidenced the presence of one additional band in the enhanced mutant, and the absence of a specific band in the non-producing mutants, both in comparison with the original strain. These bands are proposed for a future sequencing study which will define their role in the production process of metabolites with antifungal activity in Streptomyces 5.1.

Transcriptional Changes in Potato Sprouts upon Interaction with Rhizoctonia solani Indicate Pathogen-Induced Interference in the Defence Pathways of Potato.

Rhizoctonia solani is the causer of black scurf disease on potatoes and is responsible for high economical losses in global agriculture. In order to increase the limited knowledge of the plants’ molecular response to this pathogen, we inoculated potatoes with R. solani AG3-PT isolate Ben3 and carried out RNA sequencing with total RNA extracted from potato sprouts at three and eight days post inoculation (dpi). In this dual RNA-sequencing experiment, the necrotrophic lifestyle of R. solani AG3-PT during early phases of interaction with its host has already been characterised. Here the potato plants’ comprehensive transcriptional response to inoculation with R. solani AG3 was evaluated for the first time based on significantly different expressed plant genes extracted with DESeq analysis. Overall, 1640 genes were differentially expressed, comparing control (−Rs) and with R. solani AG3-PT isolate Ben3 inoculated plants (+Rs). Genes involved in the production of anti-fungal proteins and secondary metabolites with antifungal properties were significantly up regulated upon inoculation with R. solani. Gene ontology (GO) terms involved in the regulation of hormone levels (i.e., ethylene (ET) and jasmonic acid (JA) at 3 dpi and salicylic acid (SA) and JA response pathways at 8 dpi) were significantly enriched. Contrastingly, the GO term “response to abiotic stimulus” was down regulated at both time points analysed. These results may support future breeding efforts toward the development of cultivars with higher resistance level to black scurf disease or the development of new control strategies.

ANTIFUNGAL ACTIVITY OF ACTINOBACTERIA WITH A POTENTIAL TO INHIBIT RICE BLAST FUNGUS MAGNAPORTHE ORYZAE (ANAMORPH PYRICULARIA ORYZAE)

Objective: The aims of the present study were to screen the actinobacteria with high potential ability to produce secondary metabolites that have inhibitory activity against plant pathogenic fungi, Magnaporthe oryzae. Production of secondary metabolites was analysis by thin-layer chromatography and bioautography assay.&#x0D; Methods: Screening and selection of potential Streptomyces sp. morphological, cultural, physiological, and biochemical characterization of the screened isolate was carried out. Antifungal compound was confirmed by bioautography assay.&#x0D; Results: Bioautography method use in this study was found to be antifungal fraction from the crude extract. Antifungal secondary metabolites can be readily located on the plates by observing clear zones where active compounds inhibit fungal growth.&#x0D; Conclusion: The bioautography assay shows that this isolates can produce antifungal compound. Therefore, this isolate proves to be a promising microbe which can be further studied for its applications a biocontrol agent against rice blast fungi.

Anti-fungal secondary metabolites and hydrolytic enzymes from rhizospheric bacteria in crop protection: a review

Plant growth promoting rhizobacteria (PGPR) residing in soil rhizosphere provide enormous beneficial effects to a plant host producing diverse secondary metabolites and enzymes useful for plant growth and protection. Siderophores, antibiotics, volatile compounds and hydrolytic enzymes are the major molecules secreted by the PGPRs, which have substantial antifungal properties and can provide plant protection. These compounds are responsible for the lysis and hyperparasitism of antagonists against deleterious fungal pathogens. Siderophore-producing PGPRs function by depriving the pathogen of iron nutrition. Antibiotics have been reported to be involved in the suppression of different fungal pathogens by inducing fungistasis, inhibition of spore germination, lysis of fungal mycelia. The PGPRs also secrete a wide range of low molecular weight volatile organic compounds (VOCs) that inhibit mycelial growth, sporulation, germination of phytophathogenic fungi, etc. Hydrolytic enzymes, mostly chitinase, protease and cellulose, lyse the cell wall of fungi. Therefore, plant growth-promoting rhizobacteria can be considered as an effective, eco-friendly, and sustainable replacement to the chemical fungicides. There are many PGPRs that perform very well in controlled conditions but not in field conditions, and hence the commercializing of hese products is not easy. Development of formulations with increased shelf life, a broad spectrum of action and consistent performance under field conditions can pave the way for commercializing the PGPRs at a faster rate.&#x0D; Journal of Bangladesh Academy of Sciences, Vol. 44, No. 2, 69-84, 2020

Chemical Ecology of Streptomyces albidoflavus Strain A10 Associated with Carpenter Ant Camponotus vagus.

Antibiotics produced by symbiotic microorganisms were previously shown to be of crucial importance for ecological communities, including ants. Previous works on ant–actinobacteria symbiosis are mainly focused on farming ants, which use antifungal microbial secondary metabolites to control pathogens in their fungal gardens. In this work, we studied microorganisms associated with carpenter ant Camponotus vagus. Pronounced antifungal activity of isolated actinobacteria strain A10 was found to be facilitated by biosynthesis of the antimycin A complex, consisting of small hydrophobic depsipeptides with high antimicrobial and cytotoxic activity. The actinomycete strain A10 was identified as Streptomyces albidoflavus. We studied the antagonistic activity of strain A10 against several entomopathogenic microorganisms. The antifungal activity of this strain potentially indicates a defensive symbiosis with the host ant, producing antimycins to protect carpenter ants against infections. The nature of this ant-microbe association however remains to be established.

Fungal-Associated Molecules Induce Key Genes Involved in the Biosynthesis of the Antifungal Secondary Metabolites Nunamycin and Nunapeptin in the Biocontrol Strain Pseudomonas fluorescens In5.

Pseudomonas fluorescens In5 synthesizes the antifungal cyclic lipopeptides (CLPs) nunamycin and nunapeptin, which are similar in structure and genetic organization to the pseudomonas-derived phytotoxins syringomycin and syringopeptin. Regulation of syringomycin and syringopeptin is dependent on the two-component global regulatory system GacS-GacA and the SalA, SyrF, and SyrG transcription factors, which activate syringomycin synthesis in response to plant signal molecules. Previously, we demonstrated that a specific transcription factor, NunF, positively regulates the synthesis of nunamycin and nunapeptin in P. fluorescens In5 and that the nunF gene is upregulated by fungal-associated molecules. This study focused on further unravelling the complex regulation governing CLP synthesis in P. fluorescens In5. Promoter fusions were used to show that the specific activator NunF is dependent on the global regulator of secondary metabolism GacA and is regulated by fungal-associated molecules and low temperatures. In contrast, GacA is stimulated by plant signal molecules leading to the hypothesis that P. fluorescens is a hyphosphere-associated bacterium carrying transcription factor genes that respond to signals indicating the presence of fungi and oomycetes. Based on these findings, we present a model for how synthesis of nunamycin and nunapeptin is regulated by fungal- and oomycete-associated molecules.IMPORTANCE Cyclic lipopeptide (CLP) synthesis gene clusters in pseudomonads display a high degree of synteny, and the structures of the peptides synthesized are very similar. Accordingly, the genomic island encoding the synthesis of syringomycin and syringopeptin in P. syringae pv. syringae closely resembles that of P. fluorescens In5, which contains genes coding for synthesis of the antifungal and anti-oomycete peptides nunamycin and nunapeptin, respectively. However, the regulation of syringomycin and syringopeptin synthesis is different from that of nunamycin and nunapeptin synthesis. While CLP synthesis in the plant pathogen P. syringae pv. syringae is induced by plant signal molecules, such compounds do not significantly influence synthesis of nunamycin and nunapeptin in P. fluorescens In5. Instead, fungal-associated molecules positively regulate antifungal peptide synthesis in P. fluorescens In5, while the synthesis of the global regulator GacA in P. fluorescens In5 is positively regulated by plant signal molecules but not fungal-associated molecules.

Sarocladium zeae is a systemic endophyte of wheat and an effective biocontrol agent against Fusarium head blight

Fusarium head blight (FHB) causes severe economic impacts by reducing yield and quality of small grain cereals, and poses health risks to both humans and animals via the accumulation of mycotoxins such as deoxynivalenol (DON). The use of endophytic fungi as potential biological control agents is an underexplored method for reducing the impact of FHB. There are several mechanisms through which endophytic fungi may affect biological control, including the production of antifungal secondary metabolites, resource competition with pathogens, and stimulation of innate plant defense responses. We determined that Sarocladium zeae, a known endophyte of corn that produces secondary metabolites inhibitory to Fusarium graminearum, is also able to colonize wheat extensively. Strains of S. zeae differ in their colonization ability, but S. zeae NRRL 34560 was shown to be a systemic endophyte of wheat, successfully colonizing the majority of internal plant organs and surviving within the plant through its life cycle. When allowed to pre-colonize wheat ahead of inoculation with F. graminearum, this strain significantly reduced FHB symptoms (57.9% reduction in area under the disease progress curve) and DON content in harvested wheat heads (61.2% reduction). While these protective effects may arise from multiple simultaneously acting mechanisms, we demonstrate that plant hormones related to defense signaling respond to the presence of S. zeae, indicating that defense priming may be an important mechanism leading to protection in this system.

Antifungal Secondary Metabolites Isolated from Mangrove Rhizosphere Soil-Derived Penicillium Fungi

Fungi isolated from mangrove rhizosphere soil of the South China Sea were investigated for the production of potential antifungal metabolites. With 28 fungal isolates, the strains Penicillium javanicum HK1-23 and P. janthinellum HK1-6 showed significant antifungal activities. A bioassay-guided investigation of the two fungal strains led to the isolation of two secondary metabolites, brefeldin A and penicillic acid, with high yields of 143 and 423 mg L−1, respectively. Penicillic acid showed potent antifungal activities toward Rhizoctonia solani and R. cerealis, with 67.5% and 76% growth inhibition, respectively, at 50 μg mL−1. Brefeldin A showed strong activity toward R. cerealis, with 56.4% growth inhibition at 50 μg mL−1. The research highlights the importance of exploring microbes from mangrove rhizosphere soil for the identification of bioactive metabolites for future fungicide development.

Biology and applications of Clonostachys rosea.

Clonostachys rosea is a promising saprophytic filamentous fungus that belongs to phylum Ascomycota. Clonostachys rosea is widespread around the world and exists in many kinds of habitats, with the highest frequency in soil. As an excellent mycoparasite, C. rosea exhibits strong biological control ability against numerous fungal plant pathogens, nematodes and insects. These behaviours are based on the activation of multiple mechanisms such as secreted cell-wall-degrading enzymes, production of antifungal secondary metabolites and induction of plant defence systems. Besides having significant biocontrol activity, C. rosea also functions in the biodegradation of plastic waste, biotransformation of bioactive compounds, as a bioenergy sources and in fermentation. This mini review summarizes information about the biology and various applications of C. rosea and expands on its possible uses.

Mollisiaceae: An overlooked lineage of diverse endophytes.

Mollisia is a taxonomically neglected discomycete genus (Helotiales, Leotiomycetes) of commonly encountered saprotrophs on decaying plant tissues throughout temperate regions. The combination of indistinct morphological characters, more than 700 names in the literature, and lack of reference DNA sequences presents a major challenge when working with Mollisia. Unidentified endophytes, including strains that produced antifungal or antiinsectan secondary metabolites, were isolated from conifer needles in New Brunswick and placed with uncertainty in Phialocephala and Mollisia, necessitating a more comprehensive treatment of these genera. In this study, morphology and multigene phylogenetic analyses were used to explore the taxonomy of Mollisiaceae, including Mollisia, Phialocephala, and related genera, using new field collections, herbarium specimens, and accessioned cultures and sequences. The phylogeny of Mollisiaceae was reconstructed and compared using the nuc internal transcribed spacer rDNA (ITS) barcode and partial sequences of the 28S nuc rDNA (LSU) gene, largest subunit of RNA polymerase II (RPB1), DNA topoisomerase I (TOP1), and the hypothetical protein Lipin/Ned1/Smp2 (LNS2). The results show that endophytism is common throughout the Mollisiaceae lineage in a diverse range of hosts but is infrequently attributed to Mollisia because of a paucity of reference sequences. Generic boundaries within Mollisiaceae are poorly resolved and based on phylogenetic evidence the family included species placed in Acephala, Acidomelania, Barrenia, Bispora, Cheirospora, Cystodendron, Fuscosclera, Hysteronaevia, Loramyces, Mollisia, Neopyrenopeziza, Obtectodiscus, Ombrophila, Patellariopsis, Phialocephala, Pulvinata, Tapesia (=Mollisia), and Trimmatostroma. Taxonomic novelties included the description of five novel Mollisia species and five novel Phialocephala species and the synonymy of Fuscosclera with Phialocephala, Acidomelania with Mollisia, and Loramycetaceae with Mollisiaceae.