Inoculation with a microbial consortium increases soil microbial diversity and improves agronomic traits of tomato under water and nitrogen deficiency.

Microbial-based biostimulants, functioning as biotic and abiotic stress protectants and growth enhancers, are becoming increasingly important in agriculture also in the context of climate change. The search for new products that can help reduce chemical inputs under a variety of field conditions is the new challenge. In this study, we tested whether the combination of two microbial growth enhancers with complementary modes of action, Azotobacter chroococcum 76A and Trichoderma afroharzianum T22, could facilitate tomato adaptation to a 30% reduction of optimal water and nitrogen requirements. The microbial inoculum increased tomato yield (+48.5%) under optimal water and nutrient conditions. In addition, the microbial application improved leaf water potential under stress conditions (+9.5%), decreased the overall leaf temperature (-4.6%), and increased shoot fresh weight (+15%), indicating that this consortium could act as a positive regulator of plant water relations under limited water and nitrogen availability. A significant increase in microbial populations in the rhizosphere with applications of A. chroococcum 76A and T. afroharzianum T22 under stress conditions, suggested that these inoculants could enhance soil microbial abundance, including the abundance of native beneficial microorganisms. Sampling time, limited water and nitrogen regimes and microbial inoculations all affected bacterial and fungal populations in the rhizospheric soil. Overall, these results indicated that the selected microbial consortium could function as plant growth enhancer and stress protectant, possibly by triggering adaptation mechanisms via functional changes in the soil microbial diversity and relative abundance.

Bioformulations with Beneficial Microbial Consortia, a Bioactive Compound and Plant Biopolymers Modulate Sweet Basil Productivity, Photosynthetic Activity and Metabolites.

Increasing attention is being given to the development of innovative formulations to substitute the use of synthetic chemicals to improve agricultural production and resource use efficiency. Alternatives can include biological products containing beneficial microorganisms and bioactive metabolites able to inhibit plant pathogens, induce systemic resistance and promote plant growth. The efficacy of such bioformulations can be increased by the addition of polymers as adjuvants or carriers. Trichoderma afroharzianum T22, Azotobacter chroococcum 76A and 6-pentyl-α-pyrone (6PP; a Trichoderma secondary metabolite) were administrated singularly or in a consortium, with or without a carboxymethyl cellulose-based biopolymer (BP), and tested on sweet basil (Ocimum basilicum L.) grown in a protected greenhouse. The effect of the treatments on basil yield, photosynthetic activity and secondary metabolites production was assessed. Photosynthetic efficiency was augmented by the applications of the bioformulations. The applications to the rhizosphere with BP + 6PP and BP + T22 + 76A increased the total fresh weight of basil by 26.3% and 23.6%, respectively. Untargeted LC-MS qTOF analysis demonstrated that the plant metabolome was significantly modified by the treatments. Quantification of the profiles for the major phenolic acids indicated that the treatment with the T22 + 76A consortium increased rosmarinic acid content by 110%. The use of innovative bioformulations containing microbes, their metabolites and a biopolymer was found to modulate the cultivation of fresh basil by improving yield and quality, thus providing the opportunity to develop farming systems with minimal impact on the environmental footprint from the agricultural production process.

Temperature Differentially Influences the Capacity of Trichoderma Species to Induce Plant Defense Responses in Tomato Against Insect Pests.

Species of the ecological opportunistic, avirulent fungus, Trichoderma are widely used in agriculture for their ability to protect crops from the attack of pathogenic fungi and for plant growth promotion activity. Recently, it has been shown that they may also have complementary properties that enhance plant defense barriers against insects. However, the use of these fungi is somewhat undermined by their variable level of biocontrol activity, which is influenced by environmental conditions. Understanding the source of this variability is essential for its profitable and wide use in plant protection. Here, we focus on the impact of temperature on Trichoderma afroharzianum T22, Trichoderma atroviride P1, and the defense response induced in tomato by insects. The in vitro development of these two strains was differentially influenced by temperature, and the observed pattern was consistent with temperature-dependent levels of resistance induced by them in tomato plants against the aphid, Macrosiphum euphorbiae, and the noctuid moth, Spodoptera littoralis. Tomato plants treated with T. afroharzianum T22 exhibited enhanced resistance toward both insect pests at 25°C, while T. atroviride P1 proved to be more effective at 20°C. The comparison of plant transcriptomic profiles generated by the two Trichoderma species allowed the identification of specific defense genes involved in the observed response, and a selected group was used to assess, by real-time quantitative reverse transcription PCR (qRT-PCR), the differential gene expression in Trichoderma-treated tomato plants subjected to the two temperature regimens that significantly affected fungal biological performance. These results will help pave the way toward a rational selection of the most suitable Trichoderma isolates for field applications, in order to best face the challenges imposed by local environmental conditions and by extreme climatic shifts due to global warming.

Antimicrobial activity of harzianic acid against Staphylococcus pseudintermedius.

The emerging concern about the increase of antibiotic resistance has encouraged research efforts to develop effective alternatives to counteract bacterial infections. Herein, we studied a new perspective to therapeutic treatment against Staphylococcus pseudintermedius, an opportunistic pathogen documented as the major cause of skin, ear, and post-operative bacterial infections in dogs and cats. Antimicrobial activity of secondary metabolites produced by selected microbial strains belonging to Trichoderma, Talaromyces, Clonostachys and Coniothyrium fungal genera has been tested against S. pseudintermedius. Several extracts, particularly those obtained from Trichoderma harzianum E45 and ET45, showed a significant antimicrobial activity towards S. pseudintermedius methicillin-resistant (MRSP) and methicillin-susceptible (MSSP) strains. Bioassay-guided fractionation of E45 and ET45 extracts allowed to isolate harzianic acid as the major compound responsible for biological activities (e.g. antimicrobial, antibiofilm formation and biofilm disaggregation).

Application of Trichoderma harzianum, 6-pentyl-α-pyrone and Plant Biopolymer Formulations Modulate Plant Metabolism and Fruit Quality of Plum Tomatoes.

Many Trichoderma are successfully used to improve agriculture productivity due to their capacity for biocontrol and to stimulate plant growth and tolerance to abiotic stress. This research elucidates the effect of applications with Trichoderma harzianum strain T22 (T22), or biopolymer (BP) alone or in combination (BP + T22 or BP + 6-pentyl-α-pyrone (6PP); a Trichoderma secondary metabolite) on the crop performance, nutritional and functional quality of greenhouse tomato (Solanum lycopersicum L. cultivar Pixel). T22 elicited significant increases in total yield (+40.1%) compared to untreated tomato. The content of lycopene, an important antioxidant compound in tomatoes, significantly increased upon treatment with T22 (+ 49%), BP + T22 (+ 40%) and BP + 6PP (+ 52%) compared to the control. T22 treatments significantly increased the content of asparagine (+37%), GABA (+87%) and MEA (+102%) over the control; whereas BP alone strongly increased GABA (+105%) and MEA (+85%). The synthesis of these compounds implies that tomato plants are able to reuse the photorespiratory amino acids and ammonium for producing useful metabolites and reduce the pressure of photorespiration on plant metabolism, thus optimizing photosynthesis and growth. Finally, these metabolites exert many beneficial effects for human health, thus enhancing the premium quality of plum tomatoes.

Diplotaxis tenuifolia (L.) DC. Yield and Quality as Influenced by Cropping Season, Protein Hydrolysates, and Trichoderma Applications.

Increasing attention is being given to plant biostimulants as a sustainable farming practice aimed to enhance vegetable crop performance. This research was conducted on greenhouse-grown perennial wall rocket (Diplotaxis tenuifolia (L.) DC.), comparing three biostimulant treatments (legume-derived protein hydrolysates, Trichoderma harzianum T22, and protein hydrolysates + Trichoderma harzianum T22) plus an untreated control, in a factorial combination with three cropping seasons (autumn-winter, winter, winter-spring). Measurements were performed on leaf yield components, colorimetric indicators, mineral composition, bioactive compounds, and antioxidant activity. Leaf marketable yield and mean weight, as well as plant dry weight, showed the highest values in winter crop cycle. Biostimulant treatments resulted in 18.4% and 26.4% increase in leaf yield and number of leaves per rosette, respectively, compared to the untreated control. Protein hydrolysates led to the highest plant dry weight (+34.7% compared to the control). Soil plant analysis development (SPAD) index as well as NO3, PO4, SO4, and Ca contents were influenced more during the winter-spring season than the winter cropping season. The winter production season resulted in a 19.8% increase in the leaf lipophilic antioxidant activity, whereas the hydrophilic antioxidant activity was 34.9% higher during the winter-spring season. SPAD index was the highest with protein hydrolysates + Trichoderma applications, which also increased the colorimetric parameters compared to the untreated control. The treatment with protein hydrolysates + Trichoderma enhanced N, PO4, Mg, and Na contents, compared to both biostimulants applied singly and to the untreated control. Both biostimulants applied alone or the protein hydrolysates + Trichoderma combination led to the increase of the lipophilic and hydrophilic antioxidant activity, as well as ascorbic acid and chlorophyll b, compared to the untreated control. The present research revealed that protein hydrolysates and Trichoderma single applications, and even more their combination in the case of some nutrients content, represent an effective tool for enhancing the yield and the quality attributes of perennial wall rocket produced under the perspective of sustainable crop system.

Methyl t-butyl ether-degrading bacteria for bioremediation and biocontrol purposes.

A total of fifteen potential methyl t-butyl ether (MtBE)-degrading bacterial strains were isolated from contaminated soil. They have been identified as belonging to the genera Bacillus, Pseudomonas, Kocuria, Janibacter, Starkeya, Bosea, Mycolicibacterium, and Rhodovarius. Bacillus aryabhattai R1B, S. novella R8b, and M. mucogenicum R8i were able to grow using MtBE as carbon source, exhibiting different growth behavior and contaminant degradation ability. Their biocontrol ability was tested against various fungal pathogens. Both S. novella R8b and B. aryabhattai were effective in reducing the development of necrotic areas on leaves within 48 hours from Botritys cinerea and Alternaria alternata inoculation. Whereas, M. mucogenicum effectively controlled B. cinerea after 72 hours. Similar results were achieved using Pythium ultimum, in which the application of isolated bacteria increased seed germination. Only M. mucogenicum elicited tomato plants resistance against B. cinerea. This is the first report describing the occurrence of bioremediation and biocontrol activities in M. mucogenicum, B. aryabhattai and S. novella species. The production of maculosin and its antibiotic activity against Rhizoctonia solani has been reported for first time from S. novella. Our results highlight the importance of multidisciplinary approaches to achieve a consistent selection of bacterial strains useful for plant protection and bioremediation purposes.