-The objective of this work was to evaluate isolates of Trichoderma harzianum regarding biocontrol of common bean seed-borne pathogens, plant growth promotion, and rhizosphere competence. Five isolates of T. harzianum were evaluated and compared with commercial isolate (Ecotrich), Carboxin+Thiram, and an absolute control. Bean seeds of the cultivar Jalo Precoce, contaminated with Aspergillus, Cladosporium, and Sclerotinia sclerotiorum, were microbiolized with antagonists, and seed health tests were carried out. Isolates were evaluated on autoclaved substrate and in field conditions. Ten days after sowing (DAS), plant length was measured. To test rhizosphere competence, isolates were applied in boxes containing autoclaved washed sand, and root colonization was evaluated at 10 DAS, using five plants per box. The most effective isolates in the seed health tests were: CEN287 and CEN289 to control Aspergillus; the commercial isolate to control Cladosporium; and CEN287 and CEN316 to control S. sclerotiorum. Isolates CEN289 and CEN290 promoted bean growth in greenhouse and field. Seed treatment with T. harzianum reduces the incidence of Aspergillus, Cladosporium, and S. sclerotiorum in 'Jalo Precoce' common bean seeds.

Biological control of seed-borne pathogens has shown to enhance germination and physiological quality of seeds. The objectives of this study were to evaluate the in vitro antagonistic effect of five Trichoderma harzianum isolates (CEN287, CEN288, CEN289, CEN290, and CEN316) against Fusarium oxysporum f. sp. phaseoli (Foxy) and test its potential use in seed treatment. Initially, pathogen and antagonists were grown in paired cultures at 25ºC, from which samples were assessed using scanning electron microscopy (SEM). Then, clean or Foxy-infected seeds were treated with conidial suspension of the antagonists. Percent of Foxy-infected seeds and normal seedlings were evaluated at seven and nine days of incubation, respectively. All but one Trichoderma isolate (CEN290) inhibited Foxy mycelial growth. SEM analysis revealed that only one Trichoderma isolate (CEN287) showed parasitic interaction with Foxy. Two isolates (CEN287 and CEN316) significantly reduced the Foxy incidence and enhanced seed germination, though less effective than the fungicide mixture (carboxin + thiram). A principal component analysis indicated the importance of volatile metabolites in reducing Foxy incidence on common bean seeds. CEN287 Trichoderma harzianum isolate formed a single group due to its increase in germination rate of Foxyinfected seeds.

RESUMOEste trabalho objetivou avaliar seis isolados de Trichoderma harzianum no controle de Fusarium oxysporum f.sp. phaseoli em sementes, e seu efeito no crescimento inicial do feijoeiro comum. Os isolados foram inicialmente confrontados in vitro com o patógeno em testes de cultura pareada e antibiose a 25ºC. Sementes sadias e artificialmente infectadas pelo patógeno foram microbiolizadas com 2 mL de suspensões dos antagonistas (2,5 x 10 8 conídios mL -1 ) para cada 100 g de sementes, em rolo de papel germtest a 20 e 25°C. Os percentuais de incidência do patógeno e de plântulas normais foram avaliados aos sete e nove dias, respectivamente. Em casa de vegetação, os isolados foram aplicados a 5 x 10 9 conídios 500 g -1 de substrato autoclavado, com avaliação do comprimento das raízes e parte aérea das plantas 11 dias após o semeio (12 plantas tratamento -1 ). Todos os isolados apresentaram antagonismo in vitro contra o patógeno. Os isolados CEN202, CEN234, CEN238, CEN240 foram superiores à testemunha no controle de F. oxysporum em sementes, reduzindo entre 35 e 51% da incidência do patógeno e proporcionando entre 73 e 81% de plântulas normais. O comprimento total das plantas com tratamento CEN239 (37,43 cm) foi superior aos demais, cujo comprimento variou entre 27,84 e 33,95 cm. Palavras-chave: Phaseolus vulgaris, controle biológico, patologia de sementes, supressão de crescimento. ABSTRACT Control of Fusarium oxysporum f.sp phaseoli in vitro and on seeds and growth promotion of common bean in early stages by Trichoderma harzianumThe objective of this study was to evaluate the antagonistic capacity of six isolates of Trichoderma harzianum against Fusarium oxysporum f.sp. phaseoli on infected seeds and its effect on the early stages of growth in bean plants. The isolates were also evaluated in dual culture assays and under in vitro antibiosis at 25ºC. Healthy and infected seeds were treated with 2 mL 100 g -1 seeds of the antagonist suspensions (2.5 x 10 8 conidia mL -1 ) for healthy evaluation using blotter method at 20 and 25°C. The percentages of pathogen incidence and normal seedlings were evaluated at seven and nine days, respectively. The isolates were applied at 5 x 10 9 conidia 500 g -1 of sterilized substrate in greenhouse. After 11 days, roots and shoots of the bean plants were measured (12 plants treatment -1 ). Every isolate of T. harzianum presented in vitro antagonism against the pathogen. The strains CEN202, CEN234, CEN238 and CEN240 offered better control of F. oxysporum on seeds than the positive check, causing a reduction between 35 and 51% in pathogen incidence and 73-81% of normal seedling emergence. The total length of the plants treated with CEN239 (37.43 cm) was longer than the other treatments, with length varying between 27.84 and 33.95 cm.

A partir de 40 amostras de solo, provenientes dos estados do Rio Grande do Sul, Minas Gerais, São Paulo, Bahia, Goiás e Tocantins, uma coleção de 230 isolados monospóricos de Trichoderma spp. foi estabelecida, usando o meio seletivo TSM. Com o objetivo de selecionarem-se isolados com potencial para controle biológico de doenças, foram conduzidos testes de pareamento de culturas em meio BDA, a 20 ºC para Sclerotinia sclerotiorum e a 25 ºC para Fusarium solani f.sp. phaseoli. Antagonismo contra os dois patógenos foi observado em 10% dos isolados. Avaliações ao microscópio eletrônico de varredura de sete isolados selecionados in vitro mostraram que nem todos promoveram o hiperparasitismo dos patógenos, sugerindo a existência de outros mecanismos de antagonismo, como antibiose ou competição.

RESUMOAdditional keywords: plant pathogens, antagonistic fungus, biological control ABSTRACT method. Most selected isolates had an inhibitory effect on the mycelial growth of S. rolfsii and V. dahlia, compared to controls. Except for nonvolatile metabolite production tests, inhibitory action against V. dahliae was highest. Even the thermostable non-volatile metabolites showed inhibitory effect against both tested pathogens, evidencing that the secondary metabolites of the antagonist have a significant effect on these pathogenic species.Os fungos pertencentes ao gênero Trichoderma estão entre os mais estudados e utilizados como agentes de biocontrole de doenças vegetais em todo o mundo. Esses microrganismos de vida livre e altamente interativos no solo, nas superfícies radiculares e no interior dos tecidos vegetais (21) apresentam atividade antagônica contra fungos fitopatogênicos. A atividade de biocontrole pode ocorrer indiretamente, competindo por espaço e nutrientes, modificando as condições ambientais, produzindo antibióticos, inativando as enzimas do patógeno ou, diretamente, mediante o micoparasitismo (5, 12), que envolve eventos sequenciais complexos, incluindo reconhecimento, penetração e subsequente morte do hospedeiro (19).Muitas espécies de Trichoderma são reconhecidamente produtoras de metabólitos secundários, voláteis e não-voláteis, os quais apresentam amplo espectro de ação antimicrobiana. É notável o envolvimento de linhagens dessas espécies fúngicas também na ativação de mecanismos de defesa da planta e na regulação do crescimento vegetal (29), além de atuarem na complexa interação entre microrganismos no ambiente (28). Dentre os metabólitos produzidos por Trichoderma spp. citam-se: ácido harziânico, alameticinas, antraquinonas, azafilonas, daucanas, harzialactonas, bisorbicillinoides, butenolides, tricholina, glisopreninas, ácido heptelídico, gliovirina, pironas, tricotecenos, isocianatos, trichosetina, viridina, peptaiboles, entre outros (18,22). Tais metabólitos, cuja produção é atribuída a vários genes, são excretados nas fases de crescimento e esporulação e estão entre os produtos naturais mais importantes utilizados para inibir o crescimento de microrganismos (3,30).Verticillium dahliae Kleb é um fungo de solo que apresenta ampla distribuição geográfica, sendo capaz de infectar diversas espécies de plantas economicamente importantes como ornamentais, olerícolas e frutíferas (20). Esse patógeno pode induzir sintomas foliares e vasculares com declínio ou morte prematura da planta (11). No Brasil tem sido relatado em tomate, berinjela, jiló, algodão, morango, quiabo

Fifty four Trichoderma strains were isolated from soil samples collected from garlic and onion crops in eight different sites in Brazil and were identified using phylogenetic analysis based on combined ITS region, tef1-α, cal, act and rpb2 sequences. The genetic variability of the recovered Trichoderma species was analysed by AFLP and their phenotypic variability determined using MALDI-TOF. The strain clusters from both typing techniques coincided with the taxonomic determinations made from phylogenetic analysis. The phylogenetic analysis showed the occurrence of Trichoderma asperellum, Trichoderma asperelloides, Trichoderma afroharzianum, Trichoderma hamatum, Trichoderma lentiforme, Trichoderma koningiopsis, Trichoderma longibrachiatum and Trichoderma erinaceum, in the soil samples. We also identified and describe two new Trichoderma species, both in the harzianum clade of section Pachybasium, which we have named Trichoderma azevedoi sp. nov. and Trichoderma peberdyi sp. nov. The examined strains of both T. azevedoi (three strains) and T. peberdyi (12 strains) display significant genotypic and phenotypic variability, but form monophyletic clades with strong bootstrap and posterior probability support and are morphologically distinct from their respective most closely related species.

No abstract

Antibiosis is the mechanism by which certain microorganisms respond to the presence of others, secreting compounds or metabolites capable of inhibiting or impeding their development. The crude extract of Trichoderma contains a mixture of secondary compounds, which may show antibiotic effect, and has been used for the prospect of this fungus for biological control and other industrial purposes. Faced with the increasing demand of agriculture for ecologically compatible alternatives for the management of diseases, this work aimed to investigate the spectrum of action of NonVolatile Metabolites (NVMs) of Trichoderma isolates against different plant pathogenic fungi. The antagonistic potential of NVMs was evaluated through the incorporation method of the filtered liquid extract in PDA medium. The assays showed that all the NVMs produced inhibited the fungus Sclerotinia sclerotiorum similarly. On the other hand, strains CEN1245 and CEN1274, both belonging to the species Trichoderma brevicompactum, showed broad spectrum against Sclerotium rolfsii, Colletotrichum gloesporioides, Verticillium dahliae, Fusarium oxysporum and Cylindrocladium sp. The present study describes isolates producing non-volatile metabolites with broad spectrum of antifungal action, as well as pathogen-specific. The Trichoderma spp. NVMs obtained from different soil samples cultivated with vegetables, cassava and maize were efficient in inhibiting plant pathogenic fungi belonging to other patossystems, such as forest or fruit, which could increase their potential application in biological control of plant diseases. In addition, these antagonistic fungi should be studied in greater depth for the identification of bioactive molecules of industrial interest or in commercial formulations of products for biological control of plant pathogens.Keywords: secondary metabolites, antagonism, inhibition of mycelial growth, plant pathogenic fungi. Potencial antifúngico de extratos brutos de Trichoderma spp.Resumo: Antibiose é um mecanismo pelo qual certos microrganismos respondem à presença de outros, secretando compostos ou metabólitos capazes de inibir ou impedir o seu desenvolvimento. O extrato bruto de Trichoderma contém uma mistura de compostos secundários e tem sido utilizado na prospecção deste fungo para o controle biológico e demais fins industriais. Diante da crescente demanda da agricultura por alternativas ecologicamente compatíveis para o manejo de doenças, este trabalho teve como objetivo investigar o espectro de ação de Metabólitos Não Voláteis (MNVs), produzidos por isolados de Trichoderma, contra diferentes fungos fitopatogênicos. O potencial antagônico dos MNVs foi avaliado através do método de incorporação do extrato líquido filtrado em meio BDA. Os ensaios mostraram que todos os MNVs produzidos inibiram de forma semelhante o fungo Sclerotinia sclerotiorum. Por outro lado, os isolados CEN1245 e CEN1274, ambos Trichoderma brevicompactum, mostraram um amplo espectro de ação, atuando contra Sclerotium rolfsii, Colletotrichum gloespori...