Trichoderma hamatum Increases Productivity, Glucosinolate Content and Antioxidant Potential of Different Leafy Brassica Vegetables

Brassica crops include important vegetables known as “superfoods” due to the content of phytochemicals of great interest to human health, such as glucosinolates (GSLs) and antioxidant compounds. On the other hand, Trichoderma is a genus of filamentous fungi that includes several species described as biostimulants and/or biological control agents in agriculture. In a previous work, an endophytic strain of Trichoderma hamatum was isolated from kale roots (Brassica oleracea var. acephala), describing its ability to induce systemic resistance in its host plant. In the present work, some of the main leafy Brassica crops (kale, cabbage, leaf rape and turnip greens) have been root-inoculated with T. hamatum, having the aim to verify the possible capacity of the fungus as a biostimulant in productivity as well as the foliar content of GSLs and its antioxidant potential, in order to improve these “superfoods”. The results reported, for the first time, an increase in the productivity of kale (55%), cabbage (36%) and turnip greens (46%) by T. hamatum root inoculation. Furthermore, fungal inoculation reported a significant increase in the content of total GSLs in cabbage and turnip greens, mainly of the GSLs sinigrin and gluconapin, respectively, along with an increase in their antioxidant capacity. Therefore, T. hamatum could be a good agricultural biostimulant in leafy Brassica crops, increasing the content of GSLs and antioxidant potential of great food and health interest.

A Genomic and Transcriptomic Study on the DDT-Resistant Trichoderma hamatum FBL 587: First Genetic Data into Mycoremediation Strategies for DDT-Polluted Sites

Trichoderma hamatum FBL 587 isolated from DDT-contaminated agricultural soils stands out as a remarkable strain with DDT-resistance and the ability to enhance DDT degradation process in soil. Here, whole genome sequencing and RNA-Seq studies for T. hamatum FBL 587 under exposure to DDT were performed. In the 38.9 Mb-genome of T. hamatum FBL 587, 10,944 protein-coding genes were predicted and annotated, including those of relevance to mycoremediation such as production of secondary metabolites and siderophores. The genome-scale transcriptional responses of T. hamatum FBL 587 to DDT exposure showed 1706 upregulated genes, some of which were putatively involved in the cellular translocation and degradation of DDT. With regards to DDT removal capacity, it was found upregulation of metabolizing enzymes such as P450s, and potentially of downstream DDT-transforming enzymes such as epoxide hydrolases, FAD-dependent monooxygenases, glycosyl- and glutathione-transferases. Based on transcriptional responses, the DDT degradation pathway could include transmembrane transporters of DDT, antioxidant enzymes for oxidative stress due to DDT exposure, as well as lipases and biosurfactants for the enhanced solubility of DDT. Our study provides the first genomic and transcriptomic data on T. hamatum FBL 587 under exposure to DDT, which are a base for a better understanding of mycoremediation strategies for DDT-polluted sites.

In Vitro Antibacterial, Antifungal, Nematocidal and Growth Promoting Activities of Trichoderma hamatum FB10 and Its Secondary Metabolites

Microbial natural biocides have attracted much more attention in recent years in order to avoid the unrestricted use of chemical biocides in the environment. The aim of this study is to analyze the antibacterial and antifungal activities of secondary metabolites and growth promoting, nematicidal, and soil enzyme activity mediated by Trichoderma hamatum FB10. The bactericidal and fungicidal activities were performed using cell-free extract. Results revealed that the selected strain exert antibacterial activity against Acidovorax avenae, Erutimacarafavora, and Xanthomonas campestris. The selected fungal strain FB10 showed antagonistic activity against fungal pathogens such as, S. sclerotiorum, Rhizoctonia solani, Alternaria radicina, Alternaria citri, and Alternaria dauci. Among the bacterial pathogens, A. avenae showed least MIC (30 ± 2.5 µg/mL) and MBC (70 ± 1.25 µg/mL) values. T. hamatum FB10 strain synthesized bioactive volatile secondary metabolite, which effectively inhibited the growth of bacteria and fungi and indicated the presence of 6-pentyl-alpha-pyrone as the major compound (67.05%). The secondary metabolite synthesized by T. hamatum FB10 showed nematicidal activity against M. incognita eggs. Egg hatch inhibition was 78 ± 2.6% and juvenile stage mortality rate was 89 ± 2.5% when the strain FB10 was treated with nematode. The cell free extract of T. hamatum FB10 showed protease, amylase, cellulase, chitinase, glucanase activities. T. hamatum FB10 inoculated with green gram increased 11% plant height, compared to the control. The fresh weight of the experimental group inoculated with T. hamatum FB10 increased 33.6% more compared to the control group. The green gram seedlings inoculated with T. hamatum FB10 increased 18% more dry weight than control group. Soil enzymes such as, urease, phosphatase, catalase and saccharase were improved in the soil inoculated with T. hamatum FB10. These biochemical components play potent role in soil fertility, energy conversion, and in soil organic matter conversion.

Null Hyper-Parasitism, a Threat for Successful Biological Control Management

Null hyper-parasitism, a new term is coined by the author to define the hyper-parasitism by a microbial agent on another hyper-parasite. The novel phenomenon of null hyper-parasitism was discovered in the in vitro and in vivo experimentation, where the bio-control agent Trichoderma hamatum hyper-parasitic on Sclerotium rolfsii, a foot rot pathogen of groundnut, was hyper-parasitiosed by a microbial strain of Aspergillus niger and Bacillus thermophillus. Here A. niger and B. thermophillus as null hyper-parasite nullified the bio-control action of hyper-parasite Trichoderma hamatum on Sclerotium rolfsii. The in vivo experimentation suggest that such type of null hyper-parasitism exist in soil ecosystem, may be to maintain the natural microbial equilibrium and extinction of a microbial species from nature due to presence of hyper-parasite and its antagonistic or bio-control activity as evident in the above case of parasite/pathogen S. rolfsii, its hyper-parasite T. hamatum and null hypersite A. niger and B. Thermophillus. Now, therefore the success of the bio-control or hyper-parasitism of soil borne fungal plant pathogen by Trichoderma sp may be dependent on the non- existence of null hyper-parasite in the soil ecosystem where the hyperparasite has to be used.

Ability of Trichoderma hamatum Isolated from Plastics-Polluted Environments to Attack Petroleum-Based, Synthetic Polymer Films

Microorganisms colonizing plastic waste material collected in composting-, landfill-, and anaerobic digestion plants were isolated to obtain novel strains maximally adapted to the degradation of plastics due to long-term contact with plastic polymers. Twenty-six bacterial strains were isolated and identified by the 16 S rRNA method, and eighteen strains of yeasts and fungi using 18 S rRNA and the internal transcribed spacer ITS sequencing of the 18 S rRNA gene. In selected strains, the ability to degrade linear low-density polyethylene (LLDPE), low-density polyethylene (LDPE), polystyrene (PS), and polyvinyl chloride (PVC) was tested in aerobic liquid-medium cultures. An oxidative, two-step pretreatment of LLDPE and LDPE using γ- or UV-irradiation followed by a high-temperature treatment was carried out, and the pretreated plastics were also included in the degradation experiments. The respective weight losses after biodegradation by Trichoderma hamatum were: virgin and γ/T90-pretreated LLDPE (2.2 ± 1.2 and 3.9 ± 0.5%), virgin and UV/T60-pretreated LDPE (0.5 ± 0.4 and 1.3 ± 0.4%), and virgin PS (0.9 ± 0.4%). The Fourier transform infrared spectroscopy (FTIR) analysis showed that during the treatment of pretreated LLDPE, T. hamatum attacked low molecular weight LLDPE oligomers, reducing the functional groups (carbonyl C = O), which was paralleled by a slight increase of the molar mass of pretreated LLDPE and a decrease of the dispersity index, as demonstrated by gel permeation chromatography (GPC). Thermogravimetric analysis (TGA) highlighted the formation of functional groups on LLDPE due to polymer pretreatment that favored fungal attack at the polymer surface. The results provide insight into microbial consortia that spontaneously colonize the surface of plastics in various environments and their capability to attack plastic polymers.