Plant Growth Promotion and Biocontrol of Pythium ultimum by Saline Tolerant Trichoderma Isolates under Salinity Stress

This present study evaluates three isolates of Trichoderma as plant growth promoting or biological control agents: Trichoderma aggressivum f. sp. europaeum, Trichoderma saturnisporum, and the marine isolate obtained from Posidonia oceanica, Trichoderma longibrachiatum. The purpose is to contribute to an overall reduction in pesticide residues in the fruit and the environment and to a decrease in chemical fertilizers, the excess of which aggravates one of the most serious abiotic stresses, salinity. The tolerance of the different isolates to increasing concentrations of sodium chloride was evaluated in vitro, as well as their antagonistic capacity against Pythium ultimum. The plant growth promoting capacity and effects of Trichoderma strains on the severity of P. ultimum on melon seedlings under saline conditions were also analysed. The results reveal that the three isolates of Trichoderma, regardless of their origin, alleviate the stress produced by salinity, resulting in larger plants with an air-dry weight percentage above 80% in saline stress conditions for T. longibrachiatum, or an increase in root-dry weight close to 50% when T. aggressivum f. sp. europaeum was applied. Likewise, the three isolates showed antagonistic activity against P. ultimum, reducing the incidence of the disease, with the highest response found for T. longibrachiatum. Biological control of P. ultimum by T. aggressivum f. sp. europaeum and T. saturnisporum is reported for the first time, reducing disease severity by 62.96% and 51.85%, respectively. This is the first description of T. aggressivum f. sp. europaeum as a biological control agent and growth promoter. The application of these isolates can be of enormous benefit to horticultural crops, in both seedbeds and greenhouses

Fungi in Danish soils under organic and conventional farming

A multi-soil study was conducted in Denmark including 29 sites, 8 classified as ‘Organic’, 11 as ‘Conventional with manure and synthetic fertilisers’ and 10 as ‘Conventional with synthetic fertilisers’. The variability of fungal abundance within the three farming systems and the long-term effects of different farming systems on fungal propagules in soil were evaluated. Fungal abundance showed large variations within all three farming systems and this variability reduced the possibility to obtain general conclusions on fungal composition in soils under different farming systems. This was illustrated by the results on total propagule numbers of filamentous fungi and yeasts. Penicillium spp. and Gliocladium roseum were more abundant under organic than conventional farming, while Trichoderma spp. were most abundant in conventionally farmed soils with synthetic fertilisers. These results were not altered after adjusting for possible differences in basic soil properties like total-C and N, extractable P, CEC, base saturation and soil density. The paper discusses whether the differences in fungal abundance are characteristics of a farming system itself or associated with certain management factors being more prevalent in one farming system than the other

Biocontrol of root and crown rot in tomatoes under greenhouse conditions using Trichoderma harzianum and Paenibacillus lentimorbus. Additional effect of solarization

Indexación: ScieloTrichoderma harzianum 650 (Th650) and Paenebacillus lentimorbus 629 (Pl629) selected earlier for their ability to control Rhizoctonia solani, Fusarium solani and F. oxysporum in vitro, were applied alone or combined with solarization (summer assay) and/or with methyl bromide (MeBr) (summer and winter assays) to a soil with a high inoculum level, for the control of tomato root rot caused by the complex F. oxysporum f. sp. lycopersici - Pyrenochaeta lycopersici - Rhizoctonia solani. Evaluations were also performed independently for root damage caused by P. lycopersici, and also for R. solani in the summer assay. MeBr decreased tomato root damage caused by the complex from 88.7% to 21.2% and from 78.4% to 35.7% in the summer and in the winter assay, respectively. None of the bio-controllers could replace MeBr in the winter assay, but Th650 and Pl629 reduced root damage caused by this complex in the summer assay. Treatments with bio-controllers were improved by their combination with solarization in this season. Independent evaluations showed that the positive control of Th650 towards R. solani and the lack of effect on P. lycopersici correlates well with the endochitinase pattern expressed by Th650 in response to these phytopathogens. Root damage caused by R. solani can be controlled at a similar level as it does MeBr in summer assays, thus representing an alternative to the use of this chemical fungicide for the control of this phytopathogen.Financial support: Fondecyt 1990785

Antagonistic capacities of Trichoderma species and their mass multiplication with agricultural wastes

El objetivo de esta investigación fue aislar y caracterizar cepas de Trichoderma nativas de Misiones (Argentina)explorando sus capacidades antagónicas y su multiplicación masiva utilizando diferentes residuos agroindustriales.Quince cepas nativas de Trichoderma spp. fueron aisladas de muestras de suelo. Estos aislamientos secaracterizaron mediante observaciones morfológicas y moleculares basados en secuencias de ADN de la regiónespaciadora transcrita interna del ADNr. Las cepas de Trichoderma spp. fueron identificadas como T. koningiopsis,T. harzianum, T. pleuroticola y T. brevicompactum. Estas cepas mostraron actividades antagónicas in vitro contraAlternaria sp., Fusarium sp. y Botrytis sp.. T. koningiopsis LBM 090, LBM 091, LBM 092 y LBM 098, T. pleuroticolaLBM 097 y T. harzianum LBM 096 presentaron una inhibición del crecimiento micelial mayor del 50% y un índicede antagonismo entre 3 y 4 contra los fitopatógenos ensayados. La cáscara de arroz y el pulido del arroz fueronlas combinaciones más adecuadas para la multiplicación de T. harzianum LBM 096.The aim of this research was to isolate and characterize Trichoderma native strains from Misiones (Argentina) exploring their antagonistic capacities to phytopatogens fungi and their mass multiplication using different agricultural wastes. Fifteen native strains of Trichoderma spp. were isolated from soil samples. These isolates were characterized via morphological observations and molecular phylogenetic analysis based on DNA sequences of the rDNA internal transcribed spacer region. The Trichoderma native strains were identified as T. koningiopsis, T. harzianum, T. pleuroticola and T. brevicompactum. All strains showed antagonistic activities in vitro against Alternaria sp., Fusarium sp. and Botrytis sp. T. koningiopsis LBM 090, LBM 091, LBM 092, and LBM 098 strains, T. pleuroticola LBM 097 and T. harzianum LBM 096 presented radial mycelial growth inhibition higher than 50% and antagonism index between 3 and 4 against the phytopathogens assayed. Among the different substrate sources evaluated, rice husk and rice polishing were the most suitable combination for mass multiplication of T. harzianum LBM 096.Fil: Sadañoski, Marcela Alejandra. Universidad Nacional de Misiones. Facultad de Ciencias Exactas Químicas y Naturales. Departamento de Bioquímica Clínica. Laboratorio de Biotecnología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; ArgentinaFil: Gutierrez Brower, Gimena. Universidad Nacional de Misiones. Facultad de Ciencias Exactas Químicas y Naturales. Departamento de Bioquímica Clínica. Laboratorio de Biotecnología Molecular; ArgentinaFil: Castrillo, María Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentina. Universidad Nacional de Misiones. Facultad de Ciencias Exactas Químicas y Naturales. Departamento de Bioquímica Clínica. Laboratorio de Biotecnología Molecular; ArgentinaFil: Lopez, Ana Clara. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentina. Universidad Nacional de Misiones. Facultad de Ciencias Exactas Químicas y Naturales. Departamento de Bioquímica Clínica. Laboratorio de Biotecnología Molecular; ArgentinaFil: Ojeda, Paola. Universidad Nacional de Misiones. Facultad de Ciencias Exactas Químicas y Naturales. Departamento de Bioquímica Clínica. Laboratorio de Biotecnología Molecular; ArgentinaFil: Zapata, Pedro Dario. Universidad Nacional de Misiones. Facultad de Ciencias Exactas Químicas y Naturales. Departamento de Bioquímica Clínica. Laboratorio de Biotecnología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; ArgentinaFil: Villalba, Laura Lidia. Universidad Nacional de Misiones. Facultad de Ciencias Exactas Químicas y Naturales. Departamento de Bioquímica Clínica. Laboratorio de Biotecnología Molecular; ArgentinaFil: Otegui, Monica Beatriz. Universidad Nacional de Misiones. Facultad de Ciencias Exactas Químicas y Naturales. Departamento de Bioquímica Clínica. Laboratorio de Biotecnología Molecular; Argentin

Mutagenesis of Trichoderma reesei endoglucanase I: impact of expression host on activity and stability at elevated temperatures.

BackgroundTrichoderma reesei is a key cellulase source for economically saccharifying cellulosic biomass for the production of biofuels. Lignocellulose hydrolysis at temperatures above the optimum temperature of T. reesei cellulases (~50°C) could provide many significant advantages, including reduced viscosity at high-solids loadings, lower risk of microbial contamination during saccharification, greater compatibility with high-temperature biomass pretreatment, and faster rates of hydrolysis. These potential advantages motivate efforts to engineer T. reesei cellulases that can hydrolyze lignocellulose at temperatures ranging from 60-70°C.ResultsA B-factor guided approach for improving thermostability was used to engineer variants of endoglucanase I (Cel7B) from T. reesei (TrEGI) that are able to hydrolyze cellulosic substrates more rapidly than the recombinant wild-type TrEGI at temperatures ranging from 50-70°C. When expressed in T. reesei, TrEGI variant G230A/D113S/D115T (G230A/D113S/D115T Tr_TrEGI) had a higher apparent melting temperature (3°C increase in Tm) and improved half-life at 60°C (t1/2 = 161 hr) than the recombinant (T. reesei host) wild-type TrEGI (t1/2 = 74 hr at 60°C, Tr_TrEGI). Furthermore, G230A/D113S/D115T Tr_TrEGI showed 2-fold improved activity compared to Tr_TrEGI at 65°C on solid cellulosic substrates, and was as efficient in hydrolyzing cellulose at 60°C as Tr_TrEGI was at 50°C. The activities and stabilities of the recombinant TrEGI enzymes followed similar trends but differed significantly in magnitude depending on the expression host (Escherichia coli cell-free, Saccharomyces cerevisiae, Neurospora crassa, or T. reesei). Compared to N.crassa-expressed TrEGI, S. cerevisiae-expressed TrEGI showed inferior activity and stability, which was attributed to the lack of cyclization of the N-terminal glutamine in Sc_TrEGI and not to differences in glycosylation. N-terminal pyroglutamate formation in TrEGI expressed in S. cerevisiae was found to be essential in elevating its activity and stability to levels similar to the T. reesei or N. crassa-expressed enzyme, highlighting the importance of this ubiquitous modification in GH7 enzymes.ConclusionStructure-guided evolution of T. reesei EGI was used to engineer enzymes with increased thermal stability and activity on solid cellulosic substrates. Production of TrEGI enzymes in four hosts highlighted the impact of the expression host and the role of N-terminal pyroglutamate formation on the activity and stability of TrEGI enzymes

Fungal cellulase; production and applications: minireview

Cellulose is the most abundant biomaterial derived from the living organisms on the earth; plant is the major contributor to the cellulose pool present in the biosphere. Cellulose is used in variety of applications ranging from nanomaterials to biofuel production. For biofuel production, cellulose has first to be broken-down into its building blocks; β-D-glucosyl unit which subsequently can be fermented to different product such as ethanol, acetic acids, among others. Cellulase is the enzymatic system, which degrades cellulose chains to glucose monomers. Cellulase is a group of three enzymes endoglucanase, exoglucanases and β-glucosidases which act together to hydrolyze cellulose to glucose units. Cellulases are found in bacteria, fungi, plants, and some animals. Fungi are the preferred source of cellulase for industrial applications since they secrete large quantities of cellulase to culture medium. Despite a remarkable number of fungi found to produce cellulase enzymes, few have been extensively investigated because they produce large quantities of these enzymes extracellularly. In this mini-review, the production of cellulase from fungi and the parameters affecting cellulase production are discussed briefly on light of recent publications. Furthermore, potential applications of cellulase enzymes are highlighted

Commercial biocontrol agents reveal contrasting comportments against two mycotoxigenic fungi in cereals: Fusarium Graminearum and Fusarium Verticillioides

The aim of this study was to investigate the impact of commercialized biological control agents (BCAs) against two major mycotoxigenic fungi in cereals, Fusarium graminearum and Fusarium verticillioides, which are trichothecene and fumonisin producers, respectively. With these objectives in mind, three commercial BCAs were selected with contrasting uses and microorganism types (T. asperellum, S. griseoviridis, P. oligandrum) and a culture medium was identified to develop an optimized dual culture bioassay method. Their comportment was examined in dual culture bioassay in vitro with both fusaria to determine growth and mycotoxin production kinetics. Antagonist activity and variable levels or patterns of mycotoxinogenesis inhibition were observed depending on the microorganism type of BCA or on the culture conditions (e.g., different nutritional sources), suggesting that contrasting biocontrol mechanisms are involved. S. griseoviridis leads to a growth inhibition zone where the pathogen mycelium structure is altered, suggesting the diffusion of antimicrobial compounds. In contrast, T. asperellum and P. oligandrum are able to grow faster than the pathogen. T. asperellum showed the capacity to degrade pathogenic mycelia, involving chitinolytic activities. In dual culture bioassay with F. graminearum, this BCA reduced the growth and mycotoxin concentration by 48% and 72%, respectively, and by 78% and 72% in dual culture bioassay against F. verticillioides. P. oligandrum progressed over the pathogen colony, suggesting a close type of interaction such as mycoparasitism, as confirmed by microscopic observation. In dual culture bioassay with F. graminearum, P. oligandrum reduced the growth and mycotoxin concentration by 79% and 93%, respectively. In the dual culture bioassay with F. verticillioides, P. oligandrum reduced the growth and mycotoxin concentration by 49% and 56%, respectively. In vitro dual culture bioassay with different culture media as well as the nutritional phenotyping of different microorganisms made it possible to explore the path of nutritional competition in order to explain part of the observed inhibition by BCAs