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Encapsulation of Trichoderma harzianum Preserves Enzymatic Activity and Enhances the Potential for Biological Control.

ABSTRACT: Trichoderma harzianum is a biological control agent used against phytopathogens and biostimulation in agriculture. However, its efficacy can be affected by biotic and abiotic factors, and microencapsulation has been used to maximize the efficacy. The objective was to develop polymeric microparticles to encapsulate T. harzianum, to perform physicochemical characterization to evaluate its stability, to evaluate effects on the soil microbiota, antifungal activity in vitro and enzymatic activity. Size distribution of wet and dry microparticles was 2000 and 800 ?m, respectively. Scanning electron microscopy showed spherical morphology and encapsulation of T. harzianum. Photostability assays showed that encapsulation protected the fungus against ultraviolet radiation. The evaluation of the microbiota showed that the proportion of denitrifying bacteria increased when compared to the control. The T. harzianum encapsulation showed an improvement in the chitinolytic and cellulosic activity. In vitro tests showed that encapsulated fungus were able to provide a greater control of S. sclerotiorum.

SUBMITTER: Maruyama CR

PROVIDER: S-EPMC7110528 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Encapsulation of Trichoderma harzianum Preserves Enzymatic Activity and Enhances the Potential for Biological Control.

Maruyama Cintia Rodrigues CR Bilesky-José Natália N de Lima Renata R Fraceto Leonardo Fernandes LF

Frontiers in bioengineering and biotechnology 20200325

Trichoderma harzianum is a biological control agent used against phytopathogens and biostimulation in agriculture. However, its efficacy can be affected by biotic and abiotic factors, and microencapsulation has been used to maximize the efficacy. The objective was to develop polymeric microparticles to encapsulate T. harzianum, to perform physicochemical characterization to evaluate its stability, to evaluate effects on the soil microbiota, antifungal activity in vitro and e ...[more]

PMID: 32269991

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Project description:The Solanaceae family is generally known to be the third most economically important plant taxon, but also harbors a host of plant pathogens. Diseases like wilt and fruit rot of solanaceous crops cause huge yield losses in the field as well as in storage. In the present study, eight isolates of Trichoderma spp. were obtained from rhizospheric micro-flora of three solanaceous crops: tomato, brinjal, and chili plants, and were subsequently screened for pre-eminent biocontrol activity against three fungal (Fusarium oxysporum f. sp. lycopersicum, Colletotrichum gloeosporioides, and Rhizoctonia solani) and one bacterial (Ralstonia solanacearum) pathogen. Morphological, ITS, and tef1α marker-based molecular identification revealed eight isolates were different strains of Trichoderma. Seven isolates were distinguished as T. harzianum while one was identified as T. asperellum. In vitro antagonistic and biochemical assays indicated significant biocontrol activity governed by all eight isolates. Two fungal isolates, T. harzianum MC2 and T. harzianum NBG were further evaluated to decipher their best biological control activity. Preliminary insights into the secondary metabolic profile of both isolates were retrieved by liquid chromatography-mass spectrometry (LC-MS). Further, a field experiment was conducted with the isolates T. harzianum MC2 and T. harzianum NBG which successfully resulted in suppression of bacterial wilt disease in tomato. Which possibly confer biocontrol properties to the identified isolates. The efficacy of these two strains in suppressing bacterial wilt and promoting plant growth in the tomato crop was also tested in the field. The disease incidence was significantly reduced by 47.50% and yield incremented by 54.49% in plants treated in combination with both the bioagents. The results of scanning electron microscopy were also in consensus with the in planta results. The results altogether prove that T. harzianum MC2 and T. harzianum NBG are promising microbes for their prospective use in agricultural biopesticide formulations.

Project description:Trichoderma harzianum is a filamentous fungus well adapted to different ecological niches. Owing to its ability to antagonize a wide range of plant pathogens, it is used as a biological control agent in agriculture. Selected strains of T. harzianum are also able to increase the tolerance of plants to biotic and abiotic stresses. However, little is known about the regulatory elements of the T. harzianum transcriptional machinery and their role in the biocontrol by this species. We had previously reported the involvement of the transcription factor THCTF1 in the T. harzianum production of the secondary metabolite 6-pentyl-pyrone, an important volatile compound related to interspecies cross-talk. Here, we performed a subtractive hybridization to explore the genes regulated by THCTF1, allowing us to identify a multiprotein bridging factor 1 (mbf1) homolog. The gene from T. harzianum T34 was isolated and characterized, and the generated Thmbf1 overexpressing transformants were used to investigate the role of this gene in the biocontrol abilities of the fungus against two plant pathogens. The transformants showed a reduced antifungal activity against Fusarium oxysporum f. sp. lycopersici race 2 (FO) and Botrytis cinerea (BC) in confrontation assays on discontinuous medium, indicating that the Thmbf1 gene could affect T. harzianum production of volatile organic compounds (VOC) with antifungal activity. Moreover, cellophane and dialysis membrane assays indicated that Thmbf1 overexpression affected the production of low molecular weight secreted compounds with antifungal activity against FO. Intriguingly, no correlation in the expression profiles, either in rich or minimal medium, was observed between Thmbf1 and the master regulator gene cross-pathway control (cpc1). Greenhouse assays allowed us to evaluate the biocontrol potential of T. harzianum strains against BC and FO on susceptible tomato plants. The wild type strain T34 significantly reduced the necrotic leaf lesions caused by BC while plants treated with the Thmbf1-overexpressing transformants exhibited an increased susceptibility to this pathogen. The percentages of Fusarium wilt disease incidence and values of aboveground dry weight showed that T34 did not have biocontrol activity against FO, at least in the 'Moneymaker' tomato variety, and that Thmbf1 overexpression increased the incidence of this disease. Our results show that the Thmbf1 overexpression in T34 negatively affects its biocontrol mechanisms.

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Encapsulation of Trichoderma harzianum Preserves Enzymatic Activity and Enhances the Potential for Biological Control.

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Encapsulation of <i>Trichoderma harzianum</i> Preserves Enzymatic Activity and Enhances the Potential for Biological Control.

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