INTRODUCTION:

Trichoderma spp. are one of the most commonly isolated saprotrophic, filamentous fungi which are frequently found in soil. Green-spored ascomycetes that can be found all over the world. They can grow on wood, bark, and other fungi, illustrating their high opportunistic capacity and their adaptability to different ecological conditions¹. They can deal with such different environments as the rich and expanded habitat of a tropical rain forest as well as with the dark and sterile setting of a biotechnological fermentor or shake flask. Trichoderma is favoured by the presence of plant roots, where they can colonize easily. In addition to colonizing roots, Trichoderma spp. can attack various root parasitizes besides they can gain nutrition from other surrounding fungi².

Trichoderma belongs to Division: Ascomycota; Class: Sordariomycetes, Order: Hypocreales and Family Hypocreaceae. Trichoderma colonies spreading rapidly, at first thin, smooth-surfaced, translucent, later forming floccose tufts composed of conidiophores, conidia, and sterile hyphae, which when mature become yellow-green or remain white. Conidiophores erect and straggling, bearing branches and phialides irregularly or in verticils. Phialides born single or in a cluster, ovate to flask-shaped. Conidia globose, subglobose or elliptical, hyaline or green, and non-septate. Chlamydospores are present in most Trichoderma species, usually intercalary, globose, or elliptical, colourless with smooth-walled. Morphological identification of Trichoderma species depends on conidiophores. If conidiophores are long and thick with side branches mostly short, bearing crowded plump phialides then it is Trichoderma hamatum. On the other hand when conidiophores and their side branches are long with phialides not crowded include Trichoderma viride, Trichoderma koningii, Trichoderma harzianum, and others^2,3.

Trichoderma species were very common in soil and on woody products, it has been lengthily studied for its strongly enzymatic activities and their antagonistic action against several root-infecting fungi. Many Trichoderma strains do not have a sexual stage; however, they produce only asexual spores. For very few Trichoderma strains especially those that are considered for biocontrol purposes, the sexual stage is known. However, it but not extensively studied among the other strains. When the sexual stage is found, it is observed within the Ascomycetes in the genus Hypocrea. Recently, the taxa have gone from nine to about thirty-three Trichoderma spp. based on the differences in morphology, asexual sporulation apparatus, as well as molecular approaches³. Trichoderma spp. are highly successful colonizers of their habitats, which is reflected both by their efficient utilization of the substrate as well as their secretion capacity for antibiotics, enzymes, and other secondary metabolites of industrial value. Trichoderma spp. have gained great importance in plants biological control agents for controlling against many fungal spp. infecting plants⁴. Also, these species showed their capacity for the bioremediation of different organic and inorganic toxic pollutants⁵.

This review summarizes the various industrial applications and the biological control activity exerted by various Trichoderma spp., which could play a main role in the green economy era that aims at promoting environmental safeguarding and in turn human health.

Trichoderma spp. as bioremediation agents

A worldwide problem that greatly affects water and soil is the contamination by xenobiotic compounds. Xenobiotics are chemical substances that are foreign to the biological system such as drugs, naturally occurring compounds, and other environmental agents. There are different classes of xenobiotics such as pesticides, solvents, hydrocarbons, polychlorinated aromatics, as well as various pollutants including plastics, surfactants, and silicones, etc. It is crucial to get rid of these harmful pollutants requires specific which could be achieved by some chemical, physical, and/or biological techniques. The choice of a suitable method for the removal of these contaminants depends on some factors including the soil properties as well as the type of the contaminants⁶. Chemical reduction, ion exchange, precipitation, and evaporation recovery as well as electrochemical treatment are among the most commonly applied methods for the pollutants removal from different environments⁷. However, these conventional methods for pollutants removal have shown many drawbacks including the high price of the recovery process, high energy required during the treatment, incomplete pollutant removal, plus the production of other toxic by-products. Thus, the application of bioremediation could provide an alternative solution for the pollutants removal that is of lower cost compared to other remediation techniques, that only dispose of or stabilize the contaminants⁸.

Microorganisms existence in the soil is crucial. These microorganisms constitute the living part of the soil, also they are very important for the transformation and development of soil structure. Trichoderma spp. are fungi that are commonly found in soils and on some plant roots. Trichoderma sp. shows many benefits to the soil and plants as it attacks other fungal pathogens, and shows resistance to many agrochemicals and toxicants such as heavy metals, organometallic compounds, cyanide, and tannery effluents^9,10. Thus, Trichoderma sp. gains obvious importance among other fungus genera, and more studies are required to explore its role in the bioremediation of toxic pollutants.

Toxic metals (like arsenic cadmium, copper, manganese, mercury, and zinc) are being increasingly released into the environment. These result from the geochemical and industrial wastewater. Moreover, their applications in various products such as fertilizers, pesticides, tanning, wood preservatives as well as other products also result in the high release of toxic metals. These metals removal is not a simple process as they cannot be destroyed^8-11.

Many microorganisms show their ability to tolerate toxic metals, of which fungi show their capacity to get rid of these toxic metals as they show large microbial biomass¹². Microorganisms display various mechanisms to resist the metal presence. Among these mechanisms are cell membrane metal efflux¹³, phytochelatins which are glutathione-derived peptides¹¹, and intracellular chelation by metallothionein proteins¹⁴. Trichoderma sp. shows efficient soil colonization and biodegradation potential^4,15. Various Trichoderma isolates showed their ability to tolerate As, Ni, and Zn⁵. Other studies also reported that some Trichoderma sp. could tolerate more than one kind of metal⁸. Errasquin and Vazquez,⁸ reported that the ability of this fungal spp. to accumulate heavy metals from wastewater could be related to the presence of negatively charged groups found in various biopolymers on microbial cell wall resulting in the biosorption of toxic metal⁸. Additionally, some spp. of Trichoderma have been able to tolerate several toxic metals such as arsenic, cadmium, copper, and zinc through their bioaccumulation by energy-dependent metal influx^16,17.

Interestingly, Trichoderma spp. showed great importance in bioremediation of organic contaminants. Polycyclic aromatic hydrocarbons (PAHs) are potent environmental pollutants, and they are among the most hazardous organic, carcinogenic chemicals that are composed of three or more fused benzene rings with linear, angular, or cluster arrangements. Some soils are contaminated with these hazardous compounds due to several reasons such as industrial or sewage effluents, and past industrial activities like coal gasification or wood impregnation with creosotes¹⁸. It is well known that PAHs are hydrophobic pollutants that are sparingly soluble and strongly bound to the surrounding soil particles and organic matter. This reason makes PAHs are usually non-bioavailable while PAHs composed of five rings or more are so difficult to be degraded by microorganisms¹⁹. PAHs degradation can be mediated by various microorganisms. Some Trichoderma isolates were able to tolerate crude oil (COil), phenanthrene (PHE), and naphthalene (NAPH) through in vitro systems. There were no significant differences reported among Trichoderma strains when they were exposed to oil however variabilities were reported when these strains were exposed to different doses of either those oils²⁰. A previous study demonstrated the potential of Trichoderma ressei to promote plant growth in soil contaminated with diesel and this treatment provided an effective solution for soil contaminated with oil²¹.

It was also found that the extensive application of pesticides leads to the occurrence of agrochemicals in the environment in high amounts. Thus, the application of recalcitrant organic compounds in agricultural activities has become a source of contamination that in turn causes harmful consequences to human health. That is why the removal of pesticides from different environments gains great attention from scientists. The traditional methods for pesticide treatment such as adsorption onto active charcoal, flocculation, filtration, and precipitation, etc.) have many technical and economical disadvantages. Some studies developed new and economically viable techniques to remove these pesticides^22,23. However, the coupling of these physicochemical treatment processes with various biological treatments could result in an effective solution for the removal of the pesticide. In this process, the physicochemical step represents the pre-treatment process to help to improve the biodegradability of the effluent and/or to reduce toxicity²²and this was followed by the biological treatment. Fungi including Trichoderma are becoming recognized for their ability to efficiently biodegrade toxic contaminants^4,7,10. Different fungi including Trichoderma are very effective in removing toxic compounds because of the extracellular enzyme system that catalyzes the reactions responsible for degrading the toxic aromatic compounds. Fungi are also able to degrade various chemicals, including pesticide residues that exist in soil including DDT chlordane, and lindane^9,24.

The study reported by Katayama and Matsumura²⁵ showed that Trichoderma harzianum degraded some agrochemicals like DDT, dieldrin, endosulfan, pentachlorophenol, and pentachloronitrobenzene however it was not able to degrade hexachlorocyclohexane. Another study reported by²⁶ also showed that Trichoderma strain T32, a mutant of pesticide-tolerant Trichoderma, found in agrochemical contaminated soil showed its ability to degrade carbendazim. Tang et al.,²⁷ reported some proteins differentially expressed from Trichoderma in response to dichlorvos which is an organophosphate pesticide²⁷.

Trichoderma spp. factory for enzymes:

Filamentous fungi are valuable sources for industrial enzymes due to their high capacity for extracellular protein production²⁸. The replacement of gasoline with lignocellulosic ethanol has gained great importance in recent years. However, there is a major problem facing this process which is the high cost of hydrolyzing lignocellulose into fermentable sugars²⁹. Thus, finding less expensive sources of cellulolytic enzymes is required³⁰. Trichoderma reesei has a relatively small number of cellulases-encoding genes³¹. However, it is one of the efficient producers of cellulolytic enzymes^32,33. This spp. produces large amounts of cellulases and utilizes a wide variety of carbon sources that include pentose sugars as well^34,35. Moreover, other Trichoderma spp. including T. viride, T. koningii, and T. virens were able to produce cellulases³⁶. The cellulases produced by Trichoderma reesei, the biotechnological workhorse of the genus, are important industrial products, especially concerning the production of second-generation biofuels from cellulosic waste³⁶. Hence, Trichoderma spp. could be a very promising source of cellulolytic enzymes that could overcome this problem.

Trichoderma spp. ability to secreted amylases was also evaluated. High amylase activity was shown by T. viride. Also, T. harzianum, T. koningii and T. pseudokoningii were able to produce amylases^36,37. Starch is a glucose polymer, that contains two types of α-glucans: amylose and amylopectin. Amylose is a linear water-insoluble polymer of glucose that is joined by α-1, 4 glycosidic linkages^38,39. α-Amylases shows various applications at the industrial level in sugar, baking, brewing, paper, and textile industries⁴⁰. The major application of α-amylases in the sugar industry is shown in the production of high fructose corn syrups (HFCS), which are required in large quantities in the beverage industry where HFCS is used as sweeteners. Starch saccharification is an important process required for the production of HFCS^41,42.

Another study reported the ability of Trichoderma spp. to produce lipase enzymes. Many fungal spp. produce lipases are water-soluble enzymes that hydrolyze the esters of glycerol with preferably long-chain fatty acids. Lipases are very important in many industries including the detergent, food, pharmaceutical, and chemical industries⁴³. The microbial lipases gained great industrial importance due to their substrate specificity as well as their stability under different chemical and physical conditions⁴⁴. Among the tested T. spp., many spp. produced lipases in variable amounts. T. pseudokoningii showed the highest lipase activity followed by Trichoderma koningii, also T. harzianum and T. virens showed their ability to produce lipase enzyme^36,45.

Pectinases are one of the upcoming enzymes of the fruit and textile industries. These enzymes break down complex polysaccharides of plant tissues into simpler molecules like galacturonic acids. Pectinase enzymes were among the first enzymes that had been used at homes. However, their commercial importance was reported in 1930 especially in the preparation of fruit juices and wine preparations. Nowadays, pectinases gain high commercial importance, as these enzymes are responsible for the degradation of pectin (complex molecules that are found as structural polysaccharides in the plant cells). Pectinases represent an integral part of the wine, fruit juice, and textile industries^46-48. Moreover, they show many biotechnological applications. Pectinase activity was also detected for Trichoderma spp. The studies showed that T. virens, T. koningii, T. viride, and T. pseudokoningii produced pectinases with different amounts³⁶. Thus, Trichoderma spp. exhibit wide industrial applications due to the high hydrolytic enzymes secretary capacity.

Trichoderma spp. as biocontrol agents:

Efficient biocontrol strains Trichoderma spp. are being developed as promising biological fungicides, and their weaponry for this function also includes secondary metabolites with potential applications as novel antibiotics. Biological control, the use of antagonistic organisms that interfere with plant pathogens represent an ecological approach to overcome the problems caused by hazardous chemical pesticides applied in plant protection. The mycoparasite Trichoderma is an efficient biocontrol agent excreting extracellular chitinases, β-1-3 glucanases, and proteases. Studies have demonstrated that Trichoderma can ameliorate also plant performance in the presence of various abiotic stresses such as drought, salinity, and heavy metals. Understanding the molecular basis of the diverse modes of action Trichoderma can lead to better environmental-friendly control of plant diseases ^[49-54]. Abo-elyousr et al.,⁵⁵ reported that Trichoderma harzianum isolated from onion stalks showed antagonistic activity in vitro against the onion purple blotch pathogen Alternaria porri.

Trichoderma spp. are known to be strong opportunistic invaders, fast-growing, heavy spores, excreting cell wall lytic enzymes, and powerful antibiotic producers even under highly competitive environment for space, nutrients, and light⁵⁶. These properties collectively enabled Trichoderma to grow in different habitats. Gal-Hemed et al.⁵⁷ isolated different marine Trichoderma isolates and examined their potential as halotolerant bio-control agents and found them effective against Rhizoctonia solani inducing systemic defense responses in plants. Species of Trichoderma are effective in controlling the level of diseases caused by Fusarium oxysporum in many crop plants⁵⁸. However, the levels of disease control caused by Fusarium oxysporum were found to be variable within different strains of Trichoderma⁵⁹.

Trichoderma spp. as plant promotors:

Bio-fertilizers are one of the environmentally friendly alternatives to the chemical fertilizers that are being marketed to increase the soils fertility and the crops productivity and yield without causing harmful environmental effects^60,61. Bio-fertilizers are based on the microorganisms like Trichoderma spp. that can colonize the rhizosphere of the plants and promote plant growth and enhance their tolerance to the biotic and abiotic stresses via various mechanisms⁶². Trichoderma spp. are versatile opportunistic plant symbionts that can cause substantial changes in the metabolism of host plants, thereby increasing plant growth and activating plant defense to various diseases.

Endophytes in general and endophytic Trichoderma spp. have a well-established role as an economical and eco-friendly plant growth promoter, which leads to an increase in crop production^63,64. Trichoderma spp. are widely reported as plant growth promoters. Increased root and shoot biomass are the most common expression of growth promotion but changes in plant morphology and development are also reported. Growth promotion can be highly variable due to several limiting factors including crop type, growing conditions, inoculum rate, and formulation type. Many Trichoderma bio-inoculants are now commercially available with strain mixes becoming increasingly common due to their greater constancy of performance. Various mechanisms have been proposed to explain growth promotion including control of minor pathogens, enhanced nutrient uptake, increased carbohydrate metabolism and photosynthesis, and phytohormone synthesis. Trichoderma stimulates growth by influencing the balance of hormones such as indole acetic acid, gibberellic acid, and ethylene^55,57,^65-⁶⁸.

Trichoderma spp. can be a useful tool to improve the microbial community in the rhizosphere area to enhance plant growth and development. Kamaruzzaman et al.⁶⁹ found that Trichoderma harzianum (ST5) strains could stimulate early growth in peanut plants, potentially leading to the use of these strains as novel bio-promoter in agriculture with potential for increased crop yields.

Fortuitously, when crop plants’ roots are colonized by certain root endophytic fungi in the genus Trichoderma, this induces up-regulation of genes and pigments that improve the plants’ photosynthesis Fortuitously, when crop plants’ roots are colonized by certain root endophytic fungi in the genus Trichoderma, this induces up-regulation of genes and pigments that improve the plants’ photosynthesis.

Roots of maize treated with T. harzianum strain T-22 were about twice as long compared to untreated plants after several months from treatment⁴. Mayo et al.⁷⁰ reported that T. harzianum T019 induced the expression of plant defense-related genes and produced a higher level of ergosterol in treated bean plants, indicating its positive effects on plant growth and defense in the presence of the pathogen. Rao et al.⁷¹ suggested that treatment of legume seeds with T. viride induces systemic resistance by reprogramming defense mechanisms in these legumes. Saravanakumar et al.⁷², showed that Trichoderma induced systematic resistance (ISR) against Curvularia leaf spot in maize by increasing the expression of genes related to the jasmonate/ethylene signaling pathways.

Vinale et al.⁷³, isolated pyrone 6-pentyl-2H-pyran-2-one from different Trichoderma spp. (T. viride, T. atroviride, T. harzianum, and T. koningii) and has shown both in vivo and in vitro antifungal activities towards several plant pathogenic fungi. Harman et al.,⁴ isolated gliovirin from Trichoderma virens which has high activity against P. ultimum. Mari et al.⁷⁴, found that a strain of Trichoderma harzianum produced volatile antifungal substances. Vinale et al.⁷⁵, reported that Trichoderma can produce gluconic and citric acids that decrease the soil pH, enhance the solubilization of phosphates, micronutrients, and mineral components such as iron, magnesium, and manganese.

Several possible mechanisms have been suggested to explain this beneficial response of Trichoderma on plant growth, which include control of deleterious root microorganisms, direct production of growth-stimulating factors, such as plant hormones, and increase of nutrient uptakes through enhanced root growth. Trichoderma spp. are known to stimulate plant growth by increasing water and mineral uptakes as well as suppressing plant diseases^76,77. Root colonization by Trichoderma spp. causes substantial changes to the plant proteome and metabolome, resulting in improvements of the crop productivity and tolerability to abiotic and biotic stresses ^10,77. However, a lot of attention has been paid only to the physiological and biological reprogramming of plants triggered by dynamic interactions with Trichoderma spp., such as in Capsicum annuum⁷⁸, Helianthus annuus⁷⁹, Solanum lycopersicum⁸⁰, Cucumis sativus⁸¹ and Allium cepa⁸².

Recently, Soliman et al.⁸³, found that the inoculation of Trichoderma spp. enhanced the synthesis of proline, glutathione, proteins and increased the relative water content, in addition to increased membrane stability and reduced the generation of hydrogen peroxide in Cucurbita pepo seedlings under salt stress. The potential mechanism of Trichoderma in alleviating salt stress can be discussed in light of the following conclusions. Elkelish et al.⁸⁴, found that under salinity stress conditions, the Trichoderma spp. restore the uptake of essential elements like Mg²⁺ and block the Na uptake that was negatively accumulated under saline conditions. Many studies have reported that Trichoderma associated with plants subjected to salinity stress is triggered to synthesize higher amounts of plant growth regulators like α-naphthalene acetic acid (NAA), indole-3- acetic acid (IAA), and indole-3-butyric acid (IBA), cytokinin-like molecules that may aid in the mitigation of salinity stress⁸⁵. Furthermore, Trichoderma induces phytohormones like salicylic acid and jasmonic acid⁸⁶. The Trichoderma spp. have been found to lower the formation of abscisic acid during salinity stress and also facilitate the movement of cytokinins through the root-to-shoot system⁸⁷. Trichoderma is responsible for the synthesis of 1-aminocyclopropane-1-carboxylate (ACC) deaminase, the primary precursor to ethylene breakdown and it triggers induced systemic tolerance in plants⁸⁸. Brotman et al.,⁸⁹ confirmed the role of Trichoderma in the activation of the antioxidant machinery to recover the oxidized ascorbate to enhance the tolerance mechanism under saline conditions. Trichoderma treatment of salinity-stressed plants was shown to increase the GSH/GSSG ratio and act as a quencher for free radicals and cytotoxic compounds. Therefore, it aids in the cell's protection from oxidative damage⁹⁰. Brotman et al.,¹ concluded that, under salinity stress conditions, Trichoderma generates changes in host plants, and these changes are mainly associated with stress-related genes and proteins. The inoculation of plant seedlings with Trichoderma significantly induced the expression of MDAR, APX1, and GST genes and triggered the antioxidant machinery to act against salinity-induced oxidative stress.

Trichoderma：A new fungal source for Cyclosporin A

Cyclosporin A is a member of cyclic undecapeptides with anti-inflammatory, immunosuppressive, antifungal, and antiparasitic properties^91,92. It is used extensively in the prevention and treatment of graft-versus-host reactions in bone marrow transplantation and for the prevention of rejection of kidney, heart, and liver transplants⁹³. Cyclosporin A was initially developed as an antifungal antibiotic⁹⁴.

Cyclosporines are cyclic peptides that are composed of 11 amino acids, and cyclosporine A represents the major component of them. Cyclosporine A has a molecular weight of 1202.6 and is differentiated from remaining cyclosporines by the type of amino acid existing in carbon number 2. Cyclosporine A is a calcineurin inhibitor and has well-known reported biological activities that include having antiinflammatory, antiparasitic, immunosuppressive, and antimicrobial characteristics^92,95-97. Moreover, it has been applied extensively during bone marrow transplantation in preventing and treating graft-versus-host reactions, and for preventing organ rejection in transplants⁹⁸. Cyclosporine A was firstly isolated from Tolypocladium inflatum⁹⁹. Furthermore, cyclosporine A production was also reported by submerged fermentation using different fungal species such as Aspergillus terreus, Cladosporium columbinum, Penicillium fellutanum, Cylindrocarpon lucidum, and many Trichoderma specie^100-103.

Trichoderma polysporum (Linx ex Pers.) Rifai was reported as a producer of both Cyclosporin A (as the main component), and Cyclosporin C in submerged culture and were extracted therefrom using organic solvents^100,104. Similarly, Trichoderma harzianum, has produced cyclosporin A, and the amount of drug calculated was 44.06µg/mL on a medium composed of glucose, 5%; peptone, 1%; KH₂PO₄, 0.5%; KCL, 0.25% (w/v), and using butyl acetate for the extraction process, and high-performance liquid chromatography for detection¹⁰².

Being potent drugs, cyclosporines are always involved in continuous studies to discover new medical applications for them. Recently, many studies have reported the in vitro ability of cyclosporine to inhibit replication of several coronaviruses such as SARS and MERS. Especially the cyclosporine‐analog, alisporivir, which was reported as an inhibitor of SARS‐CoV‐2 in vitro. However, these studies lack clinical evidence due to the quick ending of both SARS and MERS epidemics. It should be noted that till now no evidence using cyclosporine has an extra risk for severe COVID‐19 in addition to the comorbidities such as hypertension, diabetes, obesity, and smoking which usually coexist in these patients¹⁰⁵.

CONCLUSION:

Fungi in general, and the genus Trichoderma in particular are generous promising sources of bioactive compounds^106-113. This review article has provided a brief overview of some important applications of the genus Trichoderma. Many challenges need to be addressed to develop commercially successful applications of Trichoderma. These include the improvement and enhancement of the capability of fungal strains belonging to this genus for production of lignocellulosic enzymes, cyclosporine, antifungal volatile compounds, and antibiotics in addition to their plants biocontrol and promoters efficacy under commercial condition, the development of high quality, economical methods of fermentation, the maintenance of cell viability and biocontrol efficacy in the formulated product, the identification of Trichoderma antagonists that exhibit a wide spectrum of activity against several different pathogens on different commodities. Also, Trichoderma spp. are effective in plant growth improvement through induction of defense mechanisms, leading to enhanced resistance of plants against pathogens. So, the detailed analysis of the broad-scale metabolites revealed an increase in certain classes of metabolites, which may explain the positive effects of Trichoderma spp. on plant growth promotion and improvement of their resistance against the pathogens.

CONFLICT OF INTEREST:

Authors declare there is no conflict of interest.

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Received on 29.05.2021 Modified on 02.07.2021

Res. J. Pharmacognosy and Phytochem. 2021; 13(3):149-157.

DOI: 10.52711/0975-4385.2021.00025