Trichoderma harzianum isolates have been broadly used for biocontrol of plant diseases caused by fungi. Cladosporium herbarum is a common saprophyte and seed borne fungus, which is easy to manipulate under controlled conditions. It was chosen as a model to test the effectivity of seed treatments with T. harzianum. Common bean seeds (cv. Pérola) contaminated with C. herbarum were treated with conidial suspension (CS) and autoclaved filtrate (AF) of five isolates of T. harzianum and subsequently submitted to health and germination tests. The proportion of normal seedlings formed, the length of roots, hypocotyls and leaves, and total plantlet length, total plantlet biomass, root mass ratio (RMR), stem mass ratio (SMR), leaf mass ratio (LMR), aerial part/root system ratio (AP/RS) and leaf area were also evaluated. Isolates CEN289 and CEN290 (CS and AF) provided 66 to 77% of supression of C. herbarum on seeds and a higher number of normal seedlings as compared with control. It also yielded a higher total biomass of plantlets. Moreover treatment with isolates CEN289 and CEN290 increased root and stem length in the experiments with CS. Such results indicate the potential of T. harzianum for seed treatment and suggest that it should be further tested as control for seed borne fungal diseases and as a plant growth promoter. The better performance found for CEN289 and CEN290 confirms the variability in terms of biocontrol activity among strains of T. harzianum.

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.

Estudou-se a interação entre Dicyma pulvinata e F. macrosporum ao microscópio eletrônico de varredura. Esporos de D. pulvinata germinaram na superfície das lesões induzidas por F. macrosporum em plantas de seringueira (Hevea brasiliensis), infectadas artificialmente, fixadas 8 h após a inoculação do antagonista. Aparentemente, os tubos germinativos se alongaram em direção ao patógeno. O contato íntimo entre o hiperparasita e o patógeno foi verificado em amostras fixadas 24 h após a aplicação de D. pulvinata. Ao término do processo, os esporos de F. macrosporum aparentemente invadidos pelo antagonista mostraram-se desintegrados e esvaziados de seu conteúdo. D. pulvinata cresceu sobre as lesões, sobrepondo totalmente o patógeno. Seis dias após a aplicação, conidióforos do fungo antagonista foram observados emergindo das estruturas do patógeno, produzindo esporos em grande quantidade. Verificou-se, também, um possível envolvimento de enzimas hidrolíticas na associação antagonística entre D. pulvinata e o patógeno. Estas informações podem contribuir para elucidar o modo de ação de D. pulvinata, um potencial agente de controle biológico para o mal das folhas da seringueira.

The interaction between Dicyma pulvinata and Fusicladium macrosporum was studied by scanning electron microscopy. Spores of D. pulvinata germinated on the surface of F. macrosporum lesions induced on artificially infected rubber plants were fixed 8 h after inoculation. D. pulvinata germ tubes seemed to elongate toward F. macrosporum. Close contact between the antagonistic fungus and F. macrosporum spores was verified 24 h after application of D. pulvinata. At the end of the process, spores of F. macrosporum seemed to have disintegrated and to be devoid of content. The hyperparasite grew completely over the pathogen. Six to seven days after application of the antagonistic fungus, D. pulvinata conidiophores were observed emerging from F. macrosporum structures with profuse sporulation. Studies have also shown the possibility of D. pulvinata producing hydrolytic enzymes, which could be associated with the control of plant pathogens. This information may help to elucidate some of the modes of action of D. pulvinata, a potential biological control agent for South American leaf blight of Hevea rubber plant.

Quarenta e nove isolados brasileiros de Dicyma pulvinata foram analisados quanto a características morfológicas, análises de RFLP, RAPD e AFLP. Este fungo é um micoparasita do Microcyclus ulei, o agente causal da mais importante doença da seringueira (Hevea brasiliensis), o mal-das-folhas. Estes isolados foram comparados com isolados indianos obtidos a partir de lesões de Cercosporidium sp., com um isolado proveniente da França obtido a partir de lesões de Cladosporium fulvum e com um isolado de D. ampullifera proveniente de Papua-Nova Guiné. A análise de parâmetros morfológicos confirmou a identificação dos isolados brasileiros. As representações gráficas das matrizes de distância de cada marcador molecular mostraram resultados semelhantes. Os isolados de D. pulvinata obtidos de M. ulei mostraram-se intimamente relacionados. Por outro lado, os isolados - referência, incluídos nestes estudos, mostraram-se bastante dispersos. De acordo com a análise de variância molecular, considerando todos os marcadores, os isolados de D. pulvinata obtidos de M. ulei, apresentaram diferença significativa entre isolados procedentes de áreas de cultivo de seringueira favoráveis à ocorrência do mal-das-folhas (regiões norte e nordeste do Brasil) e aqueles de áreas de escape da doença (Mato Grosso).

Forty-nine Brazilian Dicyma pulvinata isolates were examined by morphological traits and RFLP, RAPD and AFLP analyses. This fungus is a mycoparasite of Microcyclus ulei, the causal agent of the most devastating rubber (Hevea brasiliensis) disease, known as "South American Leaf Blight" (SALB). These isolates were compared with an Indian isolate from Cercosporidium sp., and a French isolate from Cladosporium fulvum. They were also compared with Dicyma ampullifera from Papua New Guinea. The morphological parameters analyzed confirmed the identification of the Brazilian isolates. The graphic representations of the distance matrices of each molecular marker showed similar results. Dicyma pulvinata isolates from M. ulei were closely related, whereas the reference isolates examined were dispersed. Among the D. pulvinata isolates obtained from M. ulei, a significant pairwise distance was obtained, for all the molecular markers, between the isolates from the areas favorable to the occurrence of SALB (North and Northeast of Brazil) and the region of escape for the disease (Mato Grosso State).

Isolados de Alternaria spp. patogênicos ao fedegoso (Senna obtusifolia) foram estudados quanto aos aspectos morfológicos e padrões isoenzimáticos, em gel de poliacrilamida. Com base nas características culturais, morfologia de colônias e morfometria de conídios, verificou-se que 11 dos 13 isolados pertenciam à espécie A. cassiae e os outros dois à A. alternata. Os resultados da análise eletroforética corroboraram as informações obtidas por meio de critérios morfológicos. Portanto, esta técnica tem potencial para ser usada na separação destas espécies de Alternaria. A ocorrência de mais de uma espécie deste gênero fúngico em S. obtusifolia foi constatada, ampliando o número de patógenos a serem avaliados no programa de desenvolvimento de um bioherbicida para esta espécie de planta daninha.

Alternaria spp. isolates pathogenic to sicklepod (Senna obtusifolia) were characterized based on morphological characteristics and isozyme patterns. Colonial morphology and conidial shape showed that 11 of the 13 isolates belong to A. cassiae. The other two isolates were identified as A. alternata. Isoenzyme patterns confirmed the results of morphological criteria. The occurrence of more than one species of this fungus infecting S. obtusifolia was found, widening the range of suitable pathogens to be evaluated in bioherbicide programs for sicklepod control.