Identification and evaluation of secondary metabolites by gas chromatography-mass spectrometry (GC-MS) in native strains of Trichoderma species

Abstract

Trichoderma species is a heterotrophic saprobion organism, which competes with other microorganisms, preventing them from developing. This fungus has mechanisms of action, among which are: production of antibiotics and metabolites, and hyperparasitism, which also promotes the induction of systemic resistance in plants. In this study, five native Trichoderma strains which were collected from agricultural land in Tamaulipas, Mexico were evaluated. The objective of this research was to identify the secondary metabolites produced by native strains of Trichoderma spp. under in vitro conditions and evaluation of the effect of these compounds on the growth of the pathogen Fusarium oxysporum and the germination of tomato seeds (Solanum esculentum). The production of Trichoderma's metabolites was performed by culturing in 150 ml of potato broth supplemented with sucrose (5%) and yeast extract (5%). Inoculation was performed by the addition of a 4 mm disk of active mycelium and allowed to grow at 25C ± 2°C and 12 h light/dark on a rotary shaker at 150 rpm for three weeks. Subsequently, the extraction of metabolites was performed using liquid: liquid phase which consisted of the filtrate and methylene chloride in a ratio of 1:3, and then the organic phase was recovered. The organic fraction was evaporated at 40°C with the aid of a rotary evaporator (Buchi®); the sample was recovered in 1 ml of methylene chloride. Analysis by GC-MS indicates that Trichoderma isolates produced 41 secondary metabolites of volatile and semi-volatile molecules and was observed, such that the number of compounds varies from the species analyzed and the collection site. The evaluation of the antagonistic activity of the extracts of Trichoderma spp. on F. oxysporum showed no significant differences between treatments (P=0.05); however, it was observed that pigmentation decreased in the mycelium of the pathogen. Regarding the evaluation of the effect of the extracts on the seeds of tomato, it was observed that the compounds of Trichoderma asperellum (TV1) induced the germination and the development of the seedling.

Trichoderma longibrachiatum (TG1) Enhances Wheat Seedlings Tolerance to Salt Stress and Resistance to Fusarium pseudograminearum

“…CWDEs such as chitinase, β-1,3-glucanases, and cellulase, for example, were not only essential features of the mycoparasitism for colonizing the pathogen but also had a significant antifungal effect on the pathogen. This result is in agreement with that of Yuef et al (2018) where the chitinase and glucanase activities of T. asperellum , T. virens , T. gamsii , and T. logibrachiatum inhibited 15% of mycelia growth of Phytophthora parasitica and 45% of F. oxysporum . The identification of biological control mechanisms that are highly effective against plant pathogens represents a tool that can be explored for plant disease management, especially in the case of important phytopathogens that are difficult to control, such as F. pseudograminearum .…”

Pathogenicity of Sclerotium rolfsii Isolates Causing Stem and Root Rot Disease of Cowpea Vigna unguiculata (L.) Walp and Management Using Trichoderma Species

“…A distinct inhibitory zone that was observed between T. virens, T. atroviride and S. rolfsii isolate and visible lysis of the pathogen mycelia in vitro may be due to the mechanism of antibiosis and production of secondary metabolites by the antagonists. This gives credence to the reports of Shoresh et al (2010) and Martínez-Padrón et al (2018) that the production of secondary metabolites enhances the ability of Trichoderma spp. to control plant pathogens.…”

Variability in metabolites produced by Talaromyces pinophilus SPJ22 cultured on different substrates

“…Different analytical techniques generally employed in metabolites studies include solid-phase microextraction (SPME), high-performance liquid chromatography (HPLC), nuclear magnetic resonance (NMR), and gas chromatography-mass spectrometry (GC-MS) [ 18 – 21 ]. Thus, the selection of a particular approach is informed by certain factors, including sample matrix, sample quantity, the concentration, and properties of the metabolites [ 22 ].…”

“…Thus, the selection of a particular approach is informed by certain factors, including sample matrix, sample quantity, the concentration, and properties of the metabolites [ 22 ]. GC-MS, the preferred technique for low polarity, volatile, and semi-volatile compounds [ 21 ], is one of the first techniques used in several disciplines for metabolite/metabolomics profiling [ 23 ]. Furthermore, GC coupled to high-resolution time-of-flight mass spectrometry (HRTOF-MS) is one of the recent innovations and robust metabolomic technologies, with improved mass resolutions and data collection rates [ 24 , 25 ].…”

“…Due to the involvement of T. pinophilus in various types of associations with plants and related pathogens and pests, coupled with their biological applications, particularly in crop protection and growth enhancement, the identification of bioactive metabolites with these important biological activities can aid in the development of novel biofungicides, biobactericides, biopesticides, biofertilisers, and biofuel, especially when they are primarily or only found in T. pinophilus [ 21 ]. This study aimed to examine the diversity in the essential metabolites produced by T. pinophilus from South African dairy feed in different media using GC-HRTOF-MS as well as classify the extracted compounds according to their chemical nature.…”