ABSTRACT The knowledge of the geographical distribution of fungi is essential to promote the understanding of global and national fungal diversity patterns. In this study, we provide a literature-based checklist of Aspergillus, Penicillium, and Talaromyces species recorded in the biomes of the Brazilian Atlantic Forest and the Brazilian Pampa. Only Penicillium digitatum was reported for the Pampa biome. For the Atlantic Forest, a total of 169 species (68 Aspergillus, 79 Penicillium, and 22 Talaromyces) are reported, of which 20 are typified with specimens from this biome, including the novel species Penicillium nordestinense (section Lanata-Divaricata) that is described in this study. Penicillium nordestinense is phylogenetically related to the invalidly described species Penicillium setosum, and this species is validated here. Soil was the most common substrate from where species were reported. Several reported species are well known in biotechnological processes. Penicilliopsis zonata and Sclerocleista ornata, species previously treated in Aspergillus, are recorded. This checklist reflects the limited knowledge of fungal species in tropical environments, such as the Brazilian Atlantic Forest and Pampa biomes. This information is a good framework for understanding the Brazilian diversity of Aspergillus, Penicillium, and Talaromyces and provides data for future fungal biogeographical studies in tropical environments.

RESUMO Objetivou-se com este trabalho otimizar a produção de enzimas celulolíticas por hps C. gloeosporioideshps URM 7080 isolado de lesões foliares de Manihot esculenta, sintomáticas de Antracnose. Utilizou-se farelo de entrecasca de mandioca (MA) e palma forrageira triturada (PA) como substratos para fermentação submersa nas concentrações 0,5, 0,75 e 1%, pH inicial 5,0, 6,0 e 7,0; agitações 80, 110 e 140 rpm e temperaturas de 28, 32 e 36 °C em experimento fatorial fracionário 25 com 32 ensaios + 4 repetições no ponto central. Os melhores parâmetros para produção de cada enzima isolada foram: 1% de MA, 0,5% de PA, 28° C, com o pH inicial 5,0 para FPAse (Celulase total) e pH 7,0 para CMCase (Endoglucanase); Para βG (β-glucosidase), pH inicial 5,0 à 36° C, e concentração de 0,5 de PA e agitação de 140 rpm para as três enzimas em conjunto.

ABSTRACT The aim of the present study was to optimize the production of cellulolytic enzymes by C. gloeosporioides URM 7080 isolated from leaf lesions in Manihot esculenta showing symptoms of anthracnose. Cassava bran (MA) and crushed forage cactus (PA) were used as substrates for submerged fermentation at the concentrations of 0.5%, 0.75% and 1%; initial pH of 5.0, 6.0 and 7.0; agitation at 80, 110 and 140 rpm, and temperatures of 28, 32 and 36°C, in a 25 fractional factorial experiment with 32 trials + 4 replicates at the central point. The best parameters for the production of each isolated enzyme were: 1% MA, 0.5% PA, 28°C, and initial pH of 5.0 for FPase (total cellulase) and pH of 7.0 for CMCase (endoglucanase); initial pH of 5.0 at 36°C for βG (β-glucosidase), and concentration of 0.5% PA at 140 rpm for the three enzymes together.

ABSTRACT A literature-based checklist of species of Aspergillus, Penicillium, and Talaromyces recorded in the Brazilian tropical dry forest (Caatinga), the largest tropical dry forest region in South America, is provided. A total of 130 species (60 Aspergillus, 57 Penicillium, and 13 Talaromyces) are reported. Soil was the most common substrate, with 122 species records. Various reported species are well known in biotechnological processes. This checklist reflects the limited knowledge of fungal species in tropical dry environments. These data provide a good starting point for biogeographical studies on species of Aspergillus, Penicillium, and Talaromyces in dry environments worldwide. In addition, the new species Penicillium vascosobrinhous is introduced, an endophytic fungus isolated from cactus of the Caatinga forest in Brazil.

RESUMO Austropuccinia psidii é o agente etiológico de uma das principais doenças do jambeiro-do-pará: a ferrugem-amarela. Em uma ocorrência dessa doença foi constatada a ação de um fungo hiperparasito sobre uredossoro de A. psidii. A partir de folhas lesionadas, dois isolados de Fusarium foram caracterizados por análise morfológica e molecular. Os resultados revelaram que esses organismos pertencem a dois complexos de espécies: Fusarium fujikuroi (FFSC) e Fusarium solani (FSSC). A ocorrência desses fungos micoparasitas é uma forma de controle biológico natural para esta ferrugem e o seu estudo contribuirá para o manejo da ferrugem amarela.

ABSTRACT Austropuccinia psidii is the etiologic agent of one of the major diseases affecting “jambeiro-do-pará”: yellow rust. During the incidence of yellow rust, the action of a hyperparasitc fungus on uredosore of A. psidii was observed. From injured leaves, two Fusarium isolates were characterized based on morphological and molecular analysis. Results evidenced that these organisms belong to two species complexes: Fusarium fujikuroi (FFSC) and Fusarium solani (FSSC). Occurrence of these mycoparasite fungi is a form of natural biological control for this rust and the study of these fungi will contribute to the management of yellow rust.

Abstract Aspergillus fumigatus is an opportunistic saprobe fungus that accounts for 90% of cases of pulmonary aspergillosis in immunosuppressed patients and is known for its angiotropism. When it reaches the respiratory tract, A. fumigatus interacts with structural components and blood vessels of the lungs, such as elastin. To understand the effect of this structural component, we examined the effect of elastin on the production and development of the biofilm of A. fumigatus. In RPMI containing 10 mg/mL of elastin, a significant increase (absorbance p < 0.0001; dry weight p < 0.0001) in the production of biofilm was observed in comparison to when RPMI was used alone, reaching a maximum growth of 18.8 mg (dry weight) of biofilm in 72 h. In addition, elastin stimulates the production (p = 0.0042) of extracellular matrix (ECM) and decreases (p = 0.005) the hydrophobicity during the development of the biofilm. These results suggest that elastin plays an important role in the growth of A. fumigatus and that it participates in the formation of thick biofilm.

RESUMO A partir de coletas realizadas em plantios comerciais de mandioca (Manihot esculenta Crantz) em 11 municípios do Estado da Paraíba, obtiveram-se 27 isolados dos fungos Colletotrichum gloeosporoides f. sp. manihotis Henn (Penn.) e Fusarium oxysporum Schlent, fitopatogênicos à cultura. Desses isolados foi obtida a produção enzimática do complexo celulolítico, sob fermentação submersa à 120 rpm e 30 °C durante 7 dias, tendo CMC (Carboximetilcelulose) como fonte de carbono. As atividades enzimáticas foram quantificadas por meio do grau de hidrólise de tiras de papel de filtro (FPAse) e de CMC (CMCase). Utilizou-se a liberação de Paranitrofenol para quantificação das atividades da β-glucosidase (βG). As atividades mais elevadas observadas foram: 23,13 e 1,51 U/ml para FPAse e CMCase pelos isolados de C. gloeosporioides URM 7124 e 7125. Para βG, 0,0014 U/mL, pelo isolado 13.

ABSTRACT Material collected from commercial crops of cassava (Manihot esculenta Crantz) in 11 municipalities in the state of Paraíba included 27 isolates of the fungi Colletotrichum gloeosporoides f. sp. manihotis Henn (Penn.) and Fusarium oxysporum Schlent., which are pathogenic to the culture. From these isolates, cellulolytic enzyme complex production was obtained under submerged fermentation at 120 rpm and 30 °C for 7 days, with CMC (Carboxymethyl Cellulose) as carbon source. Enzyme activities were quantified by means of the degree of hydrolysis of strips of filter paper (FPAse) and CMC (CMCase). The release of Paranitrofenol was used to quantify the activities of β-glucosidase (βG). The highest activities were: 23.13 and 1.51 U/ml for FPase and CMCase by the isolates of C. gloeosporioides URM 7124 and 7125. For βG, 0.0014 U/ml by isolate 13.

ResumenEl suelo es un sistema biológico complejo, que desempeña un papel fundamental en las plantas y los animales, especialmente en los bosques secos como la Caatinga. Los hongos del suelo, tales como Aspergillus y Penicillium, pueden ser utilizados como bioindicadores para la conservación de la biodiversidad. El objetivo de este estudio fue aislar e identificar las especies de Aspergillus y Penicillium del suelo, en los municipios de Ibimirim y Tupanatinga en el Parque Nacional Catimbau. Cinco colecciones se llevaron a cabo en cada área durante la estación seca de 2012, un total de 25 muestras de suelos por área. Los hongos fueron aislados mediante la suspensión en agua destilada estéril y se sembraron en medio de cultivo Agar Sabouraud más Cloranfenicol y Rosa de Bengala, y también en el medio Agar Dicloran Glicerol. Los aislamientos fueron identificados en el Laboratorio de Colección de Hongos y se confirmaron por secuenciación del espaciador transcrito interno de ADN. Un total de 42 especies fueron identificadas, 22 de ellas pertenecientes al género Aspergillus y 20 al género Penicillium. Los aislamientos de Penicillium mostraron una distribución uniforme en Tupanatinga con índices de uniformidad entre 0.92 y 0.88 en Ibimirim. Entre los aislamientos de Aspergillus el valor encontrado en Tupanatinga (0.85) fue muy similar al encontrado en Ibimirim (0.86). Se observó una gran diversidad y bajo predominio de hongos en las muestras de suelo. Estos resultados contribuyen a la estimación de la diversidad de hongos en ambientes secos, especialmente en la Caatinga, donde la diversidad es decreciente en los suelos que han sufrido alteraciones.

Abstract Soil is a complex biological system that plays a key role for plants and animals, especially in dry forests such as the Caatinga.Fungi from soils, such as Aspergillus and Penicillium, can be used as bioindicators for biodiversity conservation. The aim of this study was to isolate and identify species of Aspergillus and Penicillium in soil, from the municipalities of Tupanatinga and Ibimirim, with dry forests, in the Catimbau National Park. Five collections were performed in each area during the drought season of 2012, totaling 25 soil samples per area. Fungi were isolated by suspending soil samples in sterile distilled water and plating on Sabouraud Agar media plus Chloramphenicol and Rose Bengal, and Glycerol Dicloran Agar. Isolates were identified by morphological taxonomy in the Culture Collection Laboratory and confirmed by sequencing of the Internal Transcribed Spacer of rDNA. A total of 42 species were identified, of which 22 belong to the genus Aspergillus and 20 to Penicillium. Penicillium isolates showed uniform distribution from the collecting area in Tupanatinga, and the evenness indices found were 0.92 and 0.88 in Tupanatinga and Ibimirim, respectively. Among isolates of Aspergillus evenness, the value found in Tupanatinga (0.85) was very close to that found in Ibimirim (0.86). High diversity and low dominance of fungi in soil samples was observed. These results contributed to the estimation of fungal diversity in dry environments of the Caatinga, where diversity is decreasing in soils that have undergone disturbance. Rev. Biol. Trop. 64 (1): 45-53. Epub 2016 March 01.

The entomopathogenic fungi of the genus Metarhizium have several subtilisin-like proteases that are involved in pathogenesis and these have been used to investigate genes that are differentially expressed in response to different growth conditions. The identification and characterization of these proteases can provide insight into how the fungus is capable of infecting a wide variety of insects and adapt to different substrates. In addition, the pr1A gene has been used for the genetic improvement of strains used in pest control. In this study we used quantitative RT-PCR to assess the relative expression levels of the pr1A gene in M. anisopliae and M. acridum during growth in different culture conditions and during infection of the sugar cane borer, Diatraea saccharalis Fabricius. We also carried out a pathogenicity test to assess the virulence of both species against D. saccharalis and correlated the results with the pattern of pr1A gene expression. This analysis revealed that, in both species, the pr1A gene was differentially expressed under the growth conditions studied and during the pathogenic process. M. anisopliae showed higher expression of pr1A in all conditions examined, when compared to M. acridum. Furthermore, M. anisopliae showed a greater potential to control D. saccharalis. Taken together, our results suggest that these species have developed different strategies to adapt to different growing conditions.

A mandioca é utilizada na alimentação humana, animal e na indústria. No Estado da Paraíba, é explorada comercialmente no Litoral, na microrregião do Brejo Paraibano e como cultura de subsistência no interior. Tendo em vista a ausência de estudos epidemiológicos da cultura no Estado, objetivou-se com este trabalho avaliar o progresso da severidade e da incidência da mancha parda (Cercosporidium henningsii), mancha branca (Phaeoramularia manihotis) e antracnose (Colletotrichum gloeosporioides) em condições de infecção natural em campo ao longo de um ano, no Sítio Geraldo, situado no município de Alagoa Nova-PB, Brejo paraibano. Os dados para os estudos epidemiológicos foram registrados no 18º dia de cada mês, no período de setembro de 2012 a agosto de 2013. Para avaliar a incidência, calculou-se a porcentagem de plantas apresentando sintomas. A severidade foi quantificada a partir das observações das lesões nos tecidos foliares com o emprego de escala diagramática, escala descritiva e índice de Índice de Mackinney. Os dados da curva de progresso foram submetidos à análise de correlação de Pearson em relação aos dados meteorológicos mensais. Calculou-se também a área abaixo da curva de progresso da doença (AACPD). Dentre as plantas selecionadas 95%, 17% e 8,5% apresentaram os sintomas de mancha parda, mancha branca e antracnose, respectivamente. As lesões destas doenças foram observadas na ordem de 13,3%; 4,7% e 3,3% de extensão nos tecidos aéreos. Os valores de incidência e severidade das três doenças aumentaram gradativamente no decorrer do período de avaliação, atingindo o resultado final de 61,02; 32,45 e 15,28 para AACPD. As taxas de progresso da severidade mensais mais elevadas foram de 0,441 para mancha parda em setembro, 0,197 para mancha branca e 0,192 para antracnose, em novembro e dezembro. Houve correlação positiva entre o índice pluviométrico e o progresso das doenças e negativa em relação à diminuição das temperaturas médias mensais para as três enfermidades.

The cassava is used in human and animal food, as well as in the industry. In the state of Paraíba, it is commercially explored on the Coast, in the microregion of "Brejo Paraibano" and as subsistence crop in inner regions of the state. In view of the lack of epidemiological studies of this crop in Paraiba State, the aim of this study was to evaluate the severity progress and the incidence of brown leaf spot (Cercosporidium henningsii), anthracnose (Colletotrichum gloeosporioides) and white leaf spot (Phaeoramularia manihotis) under conditions of natural infection in the field over one year at the small farm Geraldo, in the city of Alagoa Nova-PB, "Brejo Paraibano". The data used for the epidemiological studies were recorded on the 18th day of every month, from September 2012 t oAugust 2013. Toassess the incidence, the percentage of plant sex hibiting symptoms was calculated. Disease severity was quantified based on observations of lesions in leaf tissues by using a diagrammatic scale, descriptive scale and the Mackinneyindex. The disease progresscurve data were subjected to Pearson correlation analysis relative to the monthly meteorological data. The area under the disease progresscurve (AUDPC) was also calculated. Of the selected plants, 95%, 17% and 8.5% showed symptoms of brown leaf spot, white leaf spot and anthracnose, respectively. The lesions of these diseases were observed in the order of 13.3%, 4.7% and 3.3% of extension in shoot tissues. The values of incidence and severity of the three diseases gradually increased over the evaluation period, reaching the final results of 61.02, 32.45 and 15.28 for AUDPC. The highest monthlyrates of severity progress were 0.441 for brown leaf spotin September, 0.197 for white leaf spotand 0.192 for anthracnose in November and December. There was a positive correlation between the pluviometric indexand the disease progressand a negative correlation for the decline in the monthly average temperatures for the three diseases.

O controle microbiano consiste na utilização racional de patógenos, visando à manutenção da população de insetos em equilíbrio no ambiente. Metarhizium anisopliae é a espécie mais estudada e utilizada no controle biológico de insetos. A variabilidade genética dos fungos entomopatogênicos pode ser considerada uma das principais vantagens no controle microbiano de insetos e pode ser detectada por meio de marcadores moleculares, como ITS-RFLP, ISSR e ISSP. Esses marcadores são usados para a caracterização inter e intraespecífica de Metarhizium e outros fungos entomopatogênicos e poderão auxiliar na compreensão da diversidade genética e da estrutura das populações destes fungos. A persistência de M. anisopliae no solo e seu possível efeito na estrutura da comunidade microbiana deste solo são características importantes e pouco estudadas, que devem ser consideradas no processo de seleção de isolados para o controle biológico de insetos.

Microbial control of insects is based on the rational use of pathogens to maintain environmentally balanced pest population levels, and Metarhizium anisopliae has been the most studied and most utilized fungal species for that purpose. The natural genetic variability of entomopathogenic fungi is considered one of the principal advantages of microbial insect control. The inter- and intraspecific variability and the genetic diversity and population structures of Metarhizium and other entomopathogenic fungi have been examined using ITS-RFLP, ISSR, and ISSP molecular markers. The persistence of M. anisopliae in the soil and its possible effects on the structures of resident microbial communities must be considered when selecting isolates for biological insect control.

Objetivando-se levantar e quantificar a incidência de doenças da mandioca em plantios comerciais no Estado da Paraíba, realizaram-se coletas em 21 municípios de quatro microrregiões do Estado no primeiro semestre de 2011. As doenças detectadas foram mancha parda (Cercosporidium henningsii), mancha branca (Phaeoramularia manihotis), antracnose (Colletotrichum gloeosporioides f. sp. manihotis) e podridão mole (Fusarium oxysporum). A incidência apresentou-se com variação, sendo os maiores índices constatados nos municípios das microrregiões do Curimataú Ocidental, Sapé e Brejo paraibano. A mancha parda ocorreu em todos os municípios avaliados, sendo seguida da mancha branca, cuja porcentagem de plantas atacadas foi inferior. A antracnose foi detectada em cerca de 70% dos municípios avaliados, porém com incidência reduzida. A maior incidência da podridão causada por F. oxysporum foi observada na cidade de Imaculada, microrregião de Serra do Teixeira.

To survey and quantify the incidence of the diseases affecting cassava (Manihot esculenta) in commercial crops of the state of Paraiba - Brazil, samplings were conducted in 21 municipalities at four microregions of the state during the first semester of 2011. The detected diseases were brown leaf spot (Cercosporidium henningsii), white leaf spot (Phaeoramularia manihotis), anthracnose (Colletotrichum gloeosporioides f. sp. manihotis) and root rot (Fusarium oxysporum). The incidence varied, and the highest indexes were found for the municipalities of the microregions of Curimataú Ocidental, Sapé and Brejo Paraibano. Brown leaf spot occurred in all evaluated municipalities, followed by white leaf spot, which showed inferior percentage of affected plants. Anthracnose was detected for around 70% of the evaluated municipalities but had reduced incidence. The highest incidence of root rot caused by F. oxysporum was observed for the city Imaculada, microregion of the Serra do Teixeira.

In order to assess the effectiveness of Metarhizium anisopliae var. anisopliae (Metschnikoff) Sorokin isolates in controlling the sugarcane root spittlebug Mahanarva fimbriolata (Stal) (Hemiptera: Cercopidae), nine isolates obtained from a single geographical region were studied. 'Confirmed cumulative' and 'corrected cumulative' spittlebug mortality rates were measured for each of the isolates. Based on the confirmed mortality curve, the isolates URM5946, URM5951 and URM6033 were considered to be potentially the most effective in a biological control program for M. fimbriolata.

Estudos foram realizados para analisar a caracterização genética usando marcadores de RFLP-ITS e ISSP e a amplicação do gene de patogenicidade cap20 por PCR em isolados de Colletotrichum gloeosporioides de diferentes hospedeiros. Primers para o gene cap20 foram construídos a partir de seqüências selecionadas do GenBank (National Center of Biotechnology Information, http://www.ncbi.nlm.nih.gov) com o programa Primer 3. A análise dos dendrogramas revelou que o marcador RFLP-ITS foi mais informativo em separar as espécies de Colletotrichum, e que o primer EI1 evidenciou maior diversidade genética. A amplificação do DNA dos isolados de Colletotrichum para o gene cap20 com os primers P1 e P2 indicou que este gene pode apresentar variações dentro de C. gloeosporioides relacionada ao hospedeiro, e que também está presente em outras espécies de Colletotrichum.

Studies were performed to analyze the genetic characterization using RFLP-ITS and Intron (primer EI1) markers and the amplification of the cap20 pathogenicity gene by PCR in Colletotrichum gloeosporioides isolates of different hosts plant. The genetic variability was accessed using RFLP-ITS and Intron markers and grouping by UPGMA method. Primers to cap20 gene were constructed using selected sequences of the GenBank (National Center of Biotechnology Information, http://www.ncbi.nlm.nih.gov) with the Primer 3 program. The dendrograms analysis showed that the RFLP-ITS marker was more informative to separate the Colletotrichum sp, and that primer EI1 demonstrated greater genetic diversity. The amplification of the DNA of the Colletotrichum isolates to the cap20 gene with primers P1 and P2 indicated that this gene could present variations into C. gloeosporioides related with the host, and also that it was present in other Colletotrichum sp.

Quatro isolados de Sclerotinia sclerotiorum, foram testados quanto à patogenicidade em plantas de feijão, variedade IPA-10 sendo que todos se mostraram patogênicos. Foram avaliados o controle biológico e químico in vitro, utilizando-se oito isolados de Trichoderma e um de Ulocladium atrum, e o controle químico in vitro, com os fungicidas Tiofanato metílico, Iprodione e Carbendazim. Com exceção de U. atrum todos os isolados dos antagonistas mostraram potencial antagônico contra S. sclerotiorum, destacando-se o isolado 3601 como o de melhor desempenho. No controle químico, Tiofanato metílico foi o mais eficiente, sendo este fungicida e o isolado 3601 comparados in vivo em casa-de-vegetação. Foram observadas diferenças estatísticas entre os tratamentos, sendo que a aplicação do fungicida e do antagonista antes da introdução do patógeno foi mais eficiente, com redução do percentual de incidência em 32,94% e 37,04%, respectivamente.

Four isolates of Sclerotinia sclerotiorum were tested for pathogenicity in IPA-10 variety bean plants (Phaseolus vulgaris L.), and all were pathogenic. Biological control in vitro was evaluated using eight isolates of Trichoderma spp. and, one of Ulocladium atrum. Chemical control in vitro with fungicides Thiophanate methyl, Iprodione and Carbendazim was also tested. Except U. atrum, all Trichoderma isolates showed antagonistic potential against S. sclerotiorum, where isolate 3601 presented the best performance. Thiophanate methyl chemical control was the most efficient. This fungicide and isolate 3601were compared in vivo in greenhouse. There was statistical difference between the treatments, and the application of fungicide and antagonist before the pathogen was the most efficient approach, reducing the percentage of pathogenicity to 32.94% and 37.04%, respectively.

O fungo Fusarium solani (teleomorfo Haematonectria haematococca) apresenta uma expressiva importância na agricultura por ser considerado patógeno para várias culturas de interesse econômico causando doença conhecida por podridão das raízes, além de ser patógeno aos animais e ao homem, provocando nestes últimos, micoses superficiais e sistêmicas. A complexidade associada a sua identificação correta através de métodos tradicionais justifica os esforços de usar marcadores moleculares para caracterização dos isolados. Neste trabalho, três métodos baseados na tecnologia da PCR (um por ribotipagem por PCR e dois por impressão genética por PCR) foram utilizados para investigar a variabilidade molecular de dezoito isolados de F. solani de quatro Estados brasileiros, coletados de diferentes substratos. A análise genética revelou a variabilidade intraespecífica dos isolados de F. solani, sem qualquer correlação para a origem geográfica e substrato. Seu polimorfismo foi observado até mesmo na seqüência conservada do locus do rDNA, e o marcador SPAR (GTG)5 mostrou o mais alto polimorfismo. Em conjunto, estes resultados poderão auxiliar nos estudos da relação entre variabilidade do perfil genético de isolados e os fenótipos de resistência de determinados cultivares às doenças provocadas pelo fungo, orientando programas de melhoramento vegetal.

Fusarium solani fungus (teleomorph Haematonectria haematococca) is of relevance for agriculture, producing a disease that causes significant losses for many cultivars. Moreover, F. solani is an opportunistic pathogen to animals and humans. The complexity associated to its correct identification by traditional methods justifies the efforts of using molecular markers for isolates characterization. In this work, three PCR-based methods (one PCR-ribotyping and two PCR-fingerprinting) were used to investigate the molecular variability of eighteen F. solani isolates from four Brazilian States, collected from different substrates. Genetic analysis revealed the intraspecific variability within the F. solani isolates, without any correlation to their geographical origin and substrate. Its polymorphism was observed even in the very conserved sequence of rDNA locus, and the SPAR marker (GTG)5 showed the highest polymorphism. Together, those results may contribute to understand the relation between fungal genetic variability and cultivars resistance phenotypes to fungal-caused diseases, helping plant-breeding programs.