Significance of lytic enzymes from Trichoderma spp. in the biocontrol of fungal plant pathogens

The use of specific mycolytic soil microorganisms to control plant pathogens is an ecological approach to overcome the problems caused by standard chemical methods of plant protection. The ability to produce lytic enzymes is a widely distributed property of rhizosphere-competent fungi and bacteria. Due to the higher activity of Trichoderma spp. lytic enzymes as compared to the same class of enzymes from other microorganisms and plants, effort is being aimed at improving biocontrol agents and plants by introducing Trichoderma genes via genetic manipulations. An overview is presented of the data currently available on lytic enzymes from the mycoparasitic fungus Trichoderma. This revised version was published online in August 2006 with corrections to the Cover Date.     

Population genetics of fungal diseases of plants

Although parasitism is one of the most common lifestyles among eukaryotes, population genetics on parasites lag for behind those on free-living organisms. Yet, the advent of molecular markers offers great tools for studying important processes, such as dispersal, mating systems, adaptation to host and speciation. Here we highlight some studies that used molecular markers to address questions about the population genetics of fungal (including oomycetes) plant pathogens. We conclude that population genetics approaches have provided tremendous insights into the biology of a few fungal parasites and warrant more wide use in phytopathology. However, theoretical advances are badly needed to best apply the existing methods. Fungi are of prime interest not only because they are major parasites of plants and animals, but they also constitute tractable and highly useful models for understanding evolutionary processes. We hope that the emerging field of fungal evolution will attract more evolutionary biologists in the near future.     

Substrate docking and molecular dynamic simulation for prediction of fungal enzymes from Trichoderma species-assisted extraction of nanocellulose from oil palm leaves

Abstract

Fungi of the Trichoderma species are valued industrial enzymes in support of the ‘zero-waste’ technology to convert agro-industrial biomass into valuable products, i.e. nanocellulose (NC). In this study, an in silico approach using substrate docking and molecular dynamic (MD) simulation was used to predict the order of which the multilayers of cellulosic polymers, i.e. lignin, hemicellulose and cellulose in oil palm leaves (OPL) are degraded by fungal enzymes, endocellulase and exocellulase. The study aimed to establish the catalytic tendencies of the enzymes to optimally degrade the cellulosic components of OPL for high yield production of NC. Energy minimized endocellulase and exocellulase models revealed satisfactory scores of PROCHECK (90.0% and 91.2%), Verify3D (97.23% and 98.85%) and ERRAT (95.24% and 91.00%) assessments. Active site prediction by blind docking, COACH meta-server and multiple sequence alignment indicated the catalytic triads for endocellulase and exocellulase were Ser116–His205–Glu249 and Ser382–Arg124–Asp385, respectively. Binding energy of endocellulase docked with hemicellulose (?6.0 ? kcal mol^?1) was the most favourable followed by lignin (?5.6 ? kcal mol^?1) and cellulose (?4.4 ? kcal mol^?1). Exocellulase, contrarily, bonded favorably with lignin (?8.7 ? kcal mol^?1), closely followed by cellulose (?8.5 ? kcal mol^?1) and hemicellulose (?8.4 ? kcal mol^?1). MDs simulations showed that interactions of complexes, endocellulase–hemicellulose and the exocellulase–cellulose being the most stable. Thus, the findings of the study successfully identified the specific actions of sugar-acting enzymes for NC production.

Communicated by Ramaswamy H. Sarma     

The expression of extracellular fungal cell wall hydrolytic enzymes in different Trichoderma harzianum isolates correlates with their ability to control Pyrenochaeta lycopersici

Four isolates of Trichoderma harzianum (ThN3, Th11, Th12 and Th16) were selected for their ability to control the in vitro development of the tomato root pathogen Pyrenochaeta lycopersici. Analysis of the mechanisms involved in biocontrol showed that the formation of non-volatile metabolites appears to be one of those involved in biocontrol of P. lycopersici by all T. harzianum isolates tested. Nevertheless, the higher secretion of chitinases, both in number of isoenzymes and activity by the Th11 strain, correlated well with its higher ability to control this agent in laboratory and greenhouse experiments as compared to the other T. harzianum isolates tested. The secretion of beta-1,3-endoglucanases and/or proteases appeared to have less significance than endochitinases in the biological control of P. lycopersici.     

Lytic Enzymes of Trichoderma and Their Role in Plant Defense from Fungal Diseases: A Review

Lytic enzymes of mycoparasitic fungi of the genus Trichoderma, capable of suppressing a number of fungal phytopathogens that originate in air or soil, are reviewed. The topics analyzed include (1) regulation of production of chitinases, -1,3-glucanases, and proteases; (2) molecular and catalytic properties of purified enzymes; and (3) their in vitro ability to degrade cell walls and inhibit spore germination or germ-tube elongation in various phytopathogenic fungi. Among the results summarized are reports of cloning and expression of genes coding for certain lytic enzymes of Trichoderma spp. These genes are used for obtaining plant transgenes with increased resistance to fungal diseases and Trichoderma transformants that produce higher levels of one lytic enzyme (a chitinase or protease) and thereby exhibit a more pronounced ability to suppress phytopathogenic fungi.     

The role of adjuvant agents in treating fungal diseases

Invasive fungal infections (IFIs) remain a significant cause of morbidity and mortality despite the recent introduction of new antifungal medications. In this review, the available data on the use of adjuvant agents for the treatment of IFIs are discussed. Cytokines such as interferon-γ, colony-stimulating factors, granulocyte transfusions, and the monoclonal antibody efungumab may have in a role in the management of IFIs through augmentation of the host immune response, whereas pathogen-specific vaccines may help prevent infection. Pentraxin 3, an acute phase protein, may assist in the prevention and treatment of aspergillosis. Deferasirox, an iron chelator, is being investigated as an adjunctive therapy for the treatment of zygomycosis. Lactoferrin, an ironbinding protein, appears to have activity in Candida and Aspergillus infections, and omiganan may help prevent fungal catheter-related infections. Although none of these agents are currently approved for the treatment of IFIs, they may be involved in current and/or future treatment options when used in combination with antifungal drugs.     

The potential role of arbuscular mycorrhizal fungi in protecting endangered plants and habitats

Ecosystems worldwide are threatened with the extinction of plants and, at the same time, invasion by new species. Plant invasiveness and loss of species can be caused by similar but opposing pressures on the community structures. Arbuscular mycorrhizal fungi (AMF) can have multiple positive effects on plant growth, productivity, health, and stress relief. Many endangered species live in symbiosis with AMF. However, the list of the International Union for Conservation of Nature and Natural Resources (IUCN Red List of Threatened Species) indicates that the mycorrhizal status of most of the threatened species has not been assessed. Rare plants often occur in specialized and also endangered habitats and might utilize specialized or unique AMF. The specificity of any endangered plant to its AMF population has not been investigated. Because most of the current AMF isolates that are available colonize a broad range of plant species, selected inocula could be used to promote growth of endangered plants before the proper and more effective indigenous AMF are characterized. Application of AMF in field sites to protect endangered plants is hardly feasible due to the complexity of plant community structures and the large amount of fungal inocula needed. Endangered plants could, however, be grown as greenhouse cultures together with appropriate fungi, and, at the relevant developmental stage, they could be re-planted into native sites to prevent extinction and to preserve plant community ecology.     

Biological preparations with different mechanism of action for protecting potato against fungal diseases

Mycological analysis throughout the vegetation period of potato (Solanum tuberosum) made it possible to study in detail the structure of micromycete community, to determine typical dominant (frequency, more than 60%), typical common (frequency, 30 to 60%), typical rare (frequency, 10 to 30%), and casual (frequency, less than 10%) species and to estimate changes in the microorganism community caused by plant protection preparations with different mechanisms of action. It was shown that, as a result of occurrence of resistant forms, synthetic preparations against fungal pathogens of potato (such as TMTD, Ridomil gold MC, and Cupricol) were only slightly more effective than biological preparations (Trichodermin and AgroChit), with the former considerably changing the natural saprophytic mycological community. An increase in the soil pool of Trichoderma harzianum as a result of application of a biological preparation based on this antagonistic fungus correlated with its effectiveness against the soil pathogen Fusarium sp., which causes root rots. A chitosan-based elicitor preparation more effectively suppressed the development of early (Alternaria sp. and Macrosporium sp.) and late (Phytophthora sp.) blights of leaves and had a weaker effect on the soil microflora.     

Biological preparations with different mechanisms of action for protecting potato against fungal diseases

Mycological analysis throughout the vegetation period of potato (Solanum tuberosum) made it possible to study in detail the structure of the micromycete community, to determine typical dominant (frequency, more than 60%), typical common (frequency, 30 to 60%), typical rare (frequency, 10 to 30%), and casual (frequency, less than 10%) species and to estimate changes in the microorganism community caused by plant protection preparations with different mechanisms of action. It was shown that, as a result of occurrence of resistant forms, synthetic preparations against fungal pathogens of potato (such as TMTD, Ridomil gold MC, and Cupricol) were only slightly more effective than biological preparations (Trichodermin and AgroChit), with the former considerably changing the natural saprophytic mycological community. An increase in the soil pool of Trichoderma harzianum as a result of application of a biological preparation based on this antagonistic fungus correlated with its effectiveness against the soil pathogen Fusarium sp., which causes root rot. A chitosan-based elicitor preparation more effectively suppressed the development of early (Alternaria sp. and Macrosporium sp.) and late (Phytophthora sp.) blighting of leaves and had a weaker effect on soil microflora.     

Lytic enzymes in the autolysis of filamentous fungi

The degrees of autolysis attained by five different genera of filamentous fungi during an incubation period of 60 days, under the same culture conditions were: 87.3% for Penicillium oxalicum; 65.9% for Neurospora crassa; 62.7% for Polystictus versicolor; 51.7% for Aspergillus niger and 23.5% for Nectria galligena. N. crassa, A. niger and P. versicolor reached the end of the autolysis during this incubation period (60 days), whereas P. oxalicum and N. galligena did not.The excretion of the lytic enzymes -N-acetyl-glucosaminidase, -1–3 glucanase, chitinase, invertase and acid phosphatase into the culture medium during growth and autolysis was investigated. The excretion of these enzymes was consistent with the degree of autolysis reached, the maximum excretion belonging to P. oxalicum and the minimum to N. galligena. The N. crassa invertase was excreted into the culture liquid at levels very much higher than the other enzymes studied, and at levels very much higher than the invertases excreted by the other fungi.     

Mechanisms of action and biocontrol potential of Trichoderma against fungal plant diseases - A review

Plant diseases are among the major causes of the low productivity of crops, causing yield losses of up to 30%, heralding an enormous threat to global food security. Indiscriminate use of chemical-based fungicides for controlling fungal diseases has raised severe concerns about ecosystem health. Moreover, pathogens have become insensitive against these chemicals necessitating excessive use of chemicals for adequate control. The resulting accumulation of these chemicals in the food chain has provoked numerous health complications. For combating the adversaries of chemical-based fungicides, biological control of fungal pathogens is proposed as an eco-friendly alternative. Among various biological controls, Trichoderma-based biological control agents (BCAs) are widely used in agriculture for controlling soil-borne pathogens. These BCAs are commercialized and known as; stimulators of resistance in plants, growth enhancers, bio-fertilizers, and bio-pesticides. Biological management of plant pathogens has yielded valuable results in the sustainability of ecosystems and compelling improvements in the quality and quantity of agricultural produce. These BCAs exhibit potential against pathogens, remarkably improve photosynthesis, plant growth, and nutrient use efficiency for impressive crop yields. Despite these peculiarities, Trichoderma's mechanisms against pathogens and their growth promotional effects are not thoroughly investigated, hence formulating the prime objective of the current review. Along with these, Trichoderma-based fungicides marketed in different geographical locations are encompassed in this review. Finally, the knowledge gaps and future research directions for improving the efficacy of Trichoderma-based BCAs are discussed.     

Frequency and significance of fungal isolations from conjunctival sac and their role in ocular infections

Five hundred conjunctival swabs, from 150 males and 100 females with no history of ocular infections, were collected and cultured for the isolations of fungi. Eighty (16 %) of the total specimens yielded positive fungal isolations. The isolation rate was more from the males than the females subjects. Mycelial fungi were predominant than the yeast organisms. Aspergillus species were the commonest isolates with A. flavus taking the lead in the isolations being positive 16 of the total 24 Aspergillus species isolated. A variable rate of fungal isolations was observed in different months of the year. The percentage of the isolations increases after the local use of Efcorlin-N. Nineteen of the 20 eyes studied did not yield the same fungal species in the repeated samples.Presented at the Second All India Congress of Medical Microbiologists at Hydrabad from December 22–24, 1978.     

Stress metabolites and their role in coastal plants

The stress metabolites proline, glycine betaine and sorbitol were accumulated in the leaves of some angiosperms from sand dunes and shingle. Chloride, where it was measured, was not accumulated to high concentrations in leaves suggesting that these soils are not saline. Sand dunes and shingle soils have low water-holding capacity, so it is possible that solute accumulation was a response to drought which could be of adaptive significance. In sand dunes low water availability could be associated with increased leaf temperatures because of reduced transpiration rates and high soil temperatures. The role of stress metabolites in heat tolerance was considered. Proline, betaine, sorbitol and mannitol increased the heat stability of glutamine synthetase (GS) and glutamate: oxaloacetate aminotransferase from Ammophila arenaria. For GS the effect increased with solute concentration. The polyols were more effective at high temperatures. The heat stability of GS from the moss Tortula ruraliformis and the brown alga Fucus vesiculosus was increased by mannitol. The effect of the solutes was independent of plant species and type of enzyme. It is suggested that the accumulation of solutes may have ecological importance in protecting sand-dune plants from heat damage during periods of drought.