Diagrammatic scale for quantifying severity of brown leaf spot on Carya illinoinensis.

Brown leaf spot disease, caused by the fungus Ragnhildiana diffusa, is responsible for losses in pecan orchards and quantification is essential in order to conduct epidemiological studies to establish levels of productivity losses, and assess control strategies. Therefore, the aim of this study was to develop and validate a diagrammatic scale for assessing the severity of brown leaf spot in pecan. 240 symptomatic leaflets were collected from adult trees in Anta Gorda, in the Brazilian state of Rio Grande do Sul, and used to assess the current percentage severity of the disease. The scale was based on the minimum and maximum observed severity of the disease, with six graduations defined according to the Weber-Fechner Visual Stimulus Law. The scale therefore consisted of eight levels (1.5, 4.6, 10.6, 17.3, 39.5, 65.1, 77.5, and 98%). The estimates of eight evaluators provided a basis for validating the scale. They made estimates of disease severity on 40 symptomatic digitalized pecan leaflets first, without using the scale and then referring to it. The scale provided greater precision, accuracy, and reproducibility of estimates and should result in more reliable epidemiological studies.

Dispersion of the soybean root rot by Cycloneda sanguinea (Coleoptera: Coccinellidae).

The dispersion of pathogenic microorganisms consists of the transport of pathogens from their source to inoculate a new host. Agricultural and economic importance of the Soybean root rot justifies studying this disease, especially the role of insects as dispersers. The aim of this study was to evaluate the role of the ladybird beetle, Cycloneda sanguinea Linnaeus (Coleoptera: Coccinellidae) in the dispersion of pathogens that cause Soybean root rot. Three pathogen species, Macrophomina phaseolina (Tassi) (Sphaeropsidales: Botryosphaeriaceae), Fusarium incarnatum-equiseti species complex (FIESC), and F. commune (Skovgaard) O'Donnell & Nirenberg were isolated from the midgut of ladybird beetles and cultured. Macrophomina phaseolina was identified by morphology while for the other two species, DNA was sequenced. The DNA extracted was amplified in the Internal Transcriber Spacer (ITS) region, sequenced and compared to voucher sequences deposited in the GenBank. Sequences of nucleotide ITS1-5.8 S were identified in the regions of rDNA-ITS4 ribosomal DNA. This is the first report of Macrophomina phaseolina (Tassi) (Sphaeropsidales: Botryosphaeriaceae), Fusarium incarnatum-equiseti species complex (FIESC), and F. commune (Skovgaard) O'Donnell & Nirenberg, being dispersed by C. sanguinea in Brazilian soybean fields.

Fusarium oxysporum dispersion by larvae of Cyclocephala modesta , Dyscinetus gagates and Diloboderus abderus in Brazil / Disperção de Fusarium oxysporum por larvas de Cyclocephala modesta, Dyscinetus gagates e Diloboderus abderus no Brasil

ABSTRACT: The scientific and economic importance of soybean root rot justifies the study of this disease, especially the role of insects as dispersers in the spread of root pathogens. The objective was to identify and evaluate qualitatively and quantitatively the role of Cyclocephala modesta, Dyscinetus gagates and Diloboderus abderus (Coleoptera: Melolonthidae ) in the dispersion of root pathogens in cultivated fields. The fungi were identified to the species level with DNA extraction and sequencing of these organisms within Melolonthidae larvae . Larvae were dissected and separated into the parts: mouth parts, prothorax, cuticle and digestive tract (alimentary canal). The Internal Transcribed Spacer region of the rRNA was amplified and compared to sequences deposited in GenBank. This is the first record of dispersion Fusarium oxysporum by Cyclocephala modesta, Dyscinetus gagates and Diloboderus abderus (Coleoptera: Melolonthidae ) under soybean in Brazil.

RESUMO: A importância científica e econômica da podridão radicular da soja justifica o estudo desta doença, especialmente o papel dos insetos como dispersores na disseminação de patógenos radiculares. O objetivo foi identificar e avaliar qualitativa e quantitativamente o papel de Cyclocephala modesta , Dyscinetus gagates e Diloboderus abderus (Coleoptera: Melolonthidae ) na dispersão de patógenos radiculares em campos cultivados. Os fungos foram identificados ao nível de espécie com a extração de DNA e sequenciamento desses organismos dentro de larvas Melolonthidae . As larvas foram dissecadas e separadas em partes: peças bucais, protórax, cutícula e trato digestivo (canal alimentar). A região ITS do RNA ribossômico foi amplificada e comparada com sequências depositadas no GenBank. Este é o primeiro registro da dispersão de Fusarium oxysporum por Cyclocephala modesta , Dyscinetus gagates e Diloboderus abderus (Coleoptera: Melolonthidae ) em cultura de soja no Brasil.

First report of the occurrence of Ophiocordyceps melolonthae (Ascomycota: Hypocreales: Ophiocordycipitaceae) in larvae of Diloboderus abderus Sturm (Coleoptera: Melolonthidae) in Brazil / Primeiro registro da ocorrência de Ophiocordyceps melolonthae (Ascomycota: Hypocreales: Ophiocordycipitaceae) em larvas de Diloboderus abderus Sturm (Coleoptera: Melolonthidae) no Brasil

This note is the first report on the infection of Diloboderus abderus Sturm (Coleoptera: Melolonthidae) larvae by the fungusOphiocordyceps melolonthae (Hypocreales: Ophiocordycipitaceae) in subtropical Brazil. Identification was made possible by extraction and sequencing of the fungal DNA that was covering the larvae's mouthparts, prothorax, cuticle, and digestive tract (alimentary canal). Amplification, sequencing and comparison of the ITS region of the ribosomal DNA with voucher sequences of GenBank were performed and were 95% similar to Ophiocordyceps melolonthae. The fungus is an entomopathogen which attacks Melolonthidae larvae, having scientific and economic importance because of the need for increased knowledge on its distribution and on alternatives for biological control of white grubs.

Esta nota é o primeiro registro da infecção de larvas de Diloboderus abderus Sturm pelo fungoOphiocordyceps melolonthae na região subtropical do Brasil. A identificação foi possível graças è extração e sequenciamento do DNA do fungo que cobria o aparelho bucal, protórax, cutícula e aparelho digestivo (canal alimentar) das larvas. Amplificação, sequenciamento e comparação da região ITS com sequências voucher do GenBank foram realizados, mostrando 95% de similaridade com Ophiocordyceps melolonthae. O fungo é um entomopatógeno que ataca larvas Melolonthidae, tendo importância científica e econômica devido è necessidade de aumento do conhecimento sobre sua distribuição e de alternativas de controle biológico de corós.

Banana leaf and glucose mineralization and soil organic matter in microhabitats of banana plantations under long-term pesticide use.

Soil organic matter (SOM) and microbial activity are key components of soil quality and sustainability. In the humid tropics of Costa Rica 3 pesticide regimes were studied-fungicide (low input); fungicide and herbicide (medium input); and fungicide, herbicide, and nematicide (high input)-under continuous banana cultivation for 5 yr (young) or 20 yr (old) in 3 microhabitats-nematicide ring around plants, litter pile of harvested banana, and bare area between litter pile and nematicide ring. Soil samples were incubated sequentially in the laboratory: unamended, amended with glucose, and amended with ground banana leaves. Soil organic matter varied with microhabitat, being greatest in the litter pile, where microbes had the greatest basal respiration with ground banana leaf, whereas microbes in the nematicide ring had the greatest respiration with glucose. These results suggest that soil microbes adapt to specific microhabitats. Young banana plantations had similar SOM compared with old plantations, but the former had greater basal microbial respiration in unamended and in glucose-amended soil and greater first-order mineralization rates in glucose-amended soil, thus indicating soil biological quality decline over time. High pesticide input did not decrease microbial activity or mineralization rate in surface soil. In conclusion, microbial activity in tropical volcanic soil is highly adaptable to organic and inorganic inputs.

Extratos de alho e alecrim na indução de faseolina em feijoeiro e fungitoxicidade sobre Colletotrichum lindemuthianum / Garlic and rosemary extracts in the induction of phaseollin in beans and fungitoxicity on Colletotrichum lindemuthianum

Extratos vegetais podem auxiliar no controle de doenças de plantas, por sua atividade antimicrobiana ou pela ativação de mecanismos de defesa. Neste trabalho, objetivou-se avaliar o efeito dos extratos aquosos de alho e alecrim autoclavados e não autoclavados sobre o crescimento micelial de Colletotrichum lindemuthianum, a produção de faseolina em feijoeiro e a severidade de antracnose em campo. O fungo foi incubado em meio de cultura contendo os extratos, e o crescimento foi mensurado por 18 dias. Para avaliação de faseolina, os extratos foram aplicados sobre hipocótilos de feijão, a extração foi feita em etanol e a leitura foi realizada em espectrofotômetro. A severidade da antracnose foi avaliada em plantas de feijão pulverizadas com os extratos e inoculadas com o fungo. Para o extrato de alho, a maior redução (57,6 por cento) no crescimento micelial foi com a dose de 3,0 por cento do extrato não autoclavado. Já para o extrato de alecrim, a maior redução (18,6 por cento) foi com o extrato autoclavado. A autoclavagem não teve efeito sobre a indução de faseolina. À campo, os extratos não reduziram significativamente a severidade de antracnose. O extrato de alho não autoclavado mostrou-se mais eficiente na redução do crescimento de C. lindemuthianum, enquanto que o de alecrim, autoclavado ou não, foi mais efetivo na indução de faseolina.

Plant extracts may assist in controlling plants diseases due to its antimicrobial activity and activation of defense mechanisms. This study aimed to evaluate the effect of aqueous extracts of garlic and rosemary, autoclaved or non autoclaved, on the mycelial growth of Colletotrichum lindemuthianum, production of phaseollin by beans and severity of anthracnose in the field. The fungus was incubated in medium containing the extracts and its growth measured for 18 days. For phaseollin evaluation the extracts were applied on bean hypocotyls, the phytoalexin extracted in ethanol and read in spectrophotometer. The severity of anthracnose was evaluated in bean plants sprayed with the extracts and inoculated with the fungus. For the garlic extract, the greater reduction (57,6 percent) in micelial growth was at 3 percent in the non autoclaved extract, as for the rosemary extract, the greater reduction (18,6 percent) was in the autoclaved extract. Autoclaving had no effect on the induction of phaseollin. At field conditions, the extracts did not reduce significantly the severity of anthracnose. The non autoclaved garlic extract showed to be more efficient in reducing the mycelial growth of C. lindemuthianum, while rosemary, autoclaved or not, proved to be more effective in the induction of phaseollin.

Variabilidade genética na região its do rDNA de isolados de trichoderma spp. (Biocontrolador) e Fusarium oxysporum f. sp. Chrysanthemi / Genetic variability in rDNA ITS region of Trichoderma spp. (biocontrole agent) and Fusarium oxysporum f. sp. chrysanthemi isolates

A análise de características morfológicas e culturais podem não ser suficientes para uma caracterização precisa das espécies de Trichoderma e Fusarium. Objetivou-se, neste trabalho, caracterizar a região do Espaço Interno Transcrito (ITS) do rDNA dos isolados UFSMT15.1, UFSMT16 e UFSMT17 de Trichoderma spp. utilizados no biocontrole de Fusarium oxysporum f. sp. chrysanthemi (isolado) UFSMF6. A extração de DNA de cada isolado foi realizada a partir de micélio produzido em meio líquido Batata-Dextrose. As amostras de DNA genômico foram submetidas à Reação em Cadeia da Polimerase (PCR) com os oligonucleotídeos iniciadores universais ITS1 e ITS4 e o produto gerado foi sequenciado. Os fragmentos gerados pela amplificação da PCR foram tratados com as enzimas de restrição HaeIII, HinfI e MboI. As regiões ITS1, ITS2 e 5.8S do rDNA desses isolados fúngicos foram amplificadas com sucesso. A região ITS dos isolados UFSMT15.1, UFSMT16 e UFSMT17 de Trichoderma e o isolado UFSMF6 de Fusarium apresentaram uma banda simples com um fragmento de aproximadamente 600 pares de base (pb). As enzimas de restrição HaeIII, HinfI e MboI geraram polimorfismo de bandas entre os isolados. Com base nas análises da sequência de DNA, os isolados UFSMT15.1, UFSMT16, UFSMT17 e UFSMF6 apresentaram maior similaridade com as espécies Trichoderma koningiopsis, Hypocrea virens, Hypocrea lixii e Fusarium oxysporum, respectivamente.

The analysis of morphological and cultural characteristics may not enough for the characterization of the species of Trichoderma and Fusarium. The aim of this work was to characterize the Internal Transcribed Spacer (ITS) region of the rDNA of UFSMT15.1, UFSMT16 and UFSMT17 isolates of Trichoderma spp. used in the biocontrol of Fusarium oxysporum f. sp. chrysanthemi UFSMF6. DNA extraction of each isolate was accomplished starting from hyphae produced in liquid medium Potato-Dextrose-Agar. The samples of genomic DNA were submitted to the Polymerase Chain Reaction (PCR) with the primers ITS1 and ITS4, and the product was sequenced. The fragments generated from PCR amplification were digested with the restriction enzymes HaeIII, HinfI and MboI. The ITS region of the Trichoderma's isolates UFSMT15.1, UFSMT16 and UFSMT17 and the Fusarium UFSMF6 isolate showed a simple band with a fragment with 600 base pairs (bp) approximately. The restriction enzymes HaeIII, HinfI and MboI generated band polymorphism among the isolates. The isolates UFSMT15.1, UFSMT16, UFSMT17 and UFSMF6 showed high similarity whit Trichoderma koningiopsis, Hypocrea virens, Hypocrea lixii and Fusarium oxysporum species, respectively.

Superação de dormência na qualidade de sementes e mudas: influência na produção de Senna multijuga (L. C. Rich) Irwin & Barneby / Overcoming of dormancy in the quality of seeds and seedlings: influence in the production of Senna multijuga (L.C. Rich) Irwin & Barneby

O objetivo deste trabalho foi determinar a qualidade fisiológica e sanitária de sementes de Senna multijuga(L. C. Rich.) Irwin & Barneby relacionada aos métodos de superação de dormência e à interferência na produção de mudas. As sementes foram submetidas aos seguintes métodos: imersão em água fervente, as sementes foram imersas em água, com temperatura de 100°C, até esfriar, por 24 horas; escarificação ácida, onde as sementes foram imersas em ácido sulfúrico (H2SO4) a 90%, por 10 e 20 minutos, e testemunha (sem tratamento). Foram realizados os testes de sanidade, germinação, tetrazólio e avaliação da qualidade das mudas. O delineamento experimental foi inteiramente casualizado. Para a avaliação da germinação foi utilizado um esquema fatorial (4 X 2), com quatro métodos de superação de dormência X dois fotoperíodos, para os substratos rolo-de-papel e vermiculita. A escarificação ácida constituiu-se no método mais eficiente para a superação da dormência das sementes de Senna multijuga. Penicillium sp. e Aspergillus sp. tiveram sua incidência aumentada quando o tegumento foi danificado pela escarificação ácida por 20 minutos. O controle de Fusarium spp. aumentou gradativamente com o aumento do tempo de exposição ao ácido sulfúrico.

The objective of this work was to determine the physiological and sanitary quality of Senna multijugaseeds related to the methods of dormancy overcoming and the influence on the seedlings quality. The seeds had been submitted to the following methods :immersion in water fervent, the seeds had been immersed in water, at 100°C, until cooling for 24 hours; acid escarification, where the seeds had been immersed in sulfuric acid (H2SO4) 90%, for 10 and 20 minutes, and control (without treatment). The tests of sanity, germination and tetrazolium had been carried through, and were evaluated the quality of the seedlings. For the germination of the evaluation an factorial project was used (4 X 2), with four methods of dormancy X two light periods, for substrate paper and vermiculite. The acid scarification consisted in the method most efficient for the dormancy of Senna multijugaseeds. Penicillium sp. and Aspergillus sp. they had for its increased incidence when the seed coast was damaged by the acid scarification for 20 minutes. The control of Fusarium spp. increases gradual with the increase of the time of sulfuric acid exposition.

Presença dos gêneros Trichoderma e Fusarium em solo rizosférico e não rizosférico cultivado com tomateiro e pepineiro, em horta e estufa / Presence of the genus Trichoderma and Fusarium in rhizosphere and bulk soil cultivated with tomato and cucumber, in vegetable garden and greenhouse

Para a elaboração adequada de programas de biocontrole de patógenos de solo, é necessário conhecer a distribuição de patógenos e antagonistas nesse ambiente. O objetivo deste trabalho foi estudar a presença dos gêneros Trichoderma e Fusarium em solo rizosférico e não-rizosférico, cultivado com tomateiro e pepineiro, em horta e estufa, e identificar os isolados de Fusarium spp. patogênicos às culturas. Para isso, em horta e estufa, foram realizadas 40 amostragens de solo rizosférico (tomateiro e pepineiro) e 20 de solo não-rizosférico. As suspensões dos solos foram diluídas, incubadas em meio BDA e os fungos identificados. Posteriormente, foi realizado o teste de patogenicidade para o tomateiro e o pepineiro, com os isolados de Fusarium spp. obtidos dos solos coletados. Em estufa, para o tomateiro e para o pepineiro, o número de pontos de amostragem com a presença de Trichoderma spp. em solo rizosférico foi significativamente maior (95 e 45 por cento, respectivamente) do que em solo não-rizosférico (10 por cento). Neste ambiente, ocorreu diferença na presença de Trichoderma spp. e Fusarium spp., sendo encontrados, respectivamente, em 10 e 55 por cento dos pontos de amostragem. No teste de patogenicidade, cinco isolados de Fusarium oxysporum do tomateiro e seis do pepineiro foram patogênicos às respectivas culturas. Em estufa, Trichoderma spp. ocorre com maior freqüência na rizosfera, enquanto Fusarium spp. está distribuído no solo, e a maioria dos isolados de Fusarium spp. não é patogênica ao tomateiro nem ao pepineiro.

For the adequate elaboration of biocontrol programs of soil borne pathogens, it is necessary to understand the distribution of pathogens and antagonists in this environment. This research was aimed at studing the presence of the fungi Trichoderma and Fusarium in rhizosphere and bulk soil, cultivated with tomato and cucumber, in vegetable garden and greenhouse, and to quantify the isolates of Fusarium spp. pathogenic to the crops. For that, in a vegetable garden and a greenhouse, 40 samples of rhizosphere (cucumber and tomato) and 20 of bulk soil were taken,. The soil suspensions were diluted and plated in PDA media and the fungi identified. After that, the test of pathogenicity of the isolates for tomato and cucumber found in the samples, was performed. For tomato and cucumber, in greenhouse, the number of sampling points with the presence of Trichoderma spp. in the rhizosphere (95 e 45 percent, respectively) was significantly greater than in the bulk soil (10 percent). In the bulk soil, the presence of Trichoderma spp. and Fusarium spp. differed significantly, being found, respectively, in 10 and 55 percent of the sampling points. In the pathogenicity test, 5 isolates of Fusarium oxysporum of tomato and 6 of cucumber were pathogenic to the respective crops. In greenhouse, Trichoderma spp. occurs more frequently in the rhizosphere, whereas Fusarium spp. is distributed in the soil and the majority of the Fusarium spp. isolates are neither pathogenic to tomato nor to cucumber.

Seleção de antagonistas fúngicos a Fusarium solani e Fusarium oxysporum em substrato comercial para mudas / Selection of fungi antagonistic to Fusarium solani and Fusarium oxysporum in commercial substrate for seedlings

Testes in vitro são geralmente utilizados para a seleção inicial de agentes de biocontrole contra fungos de solo, faltando metodologias que utilizem solo e/ou substrato. O objetivo deste trabalho foi realizar a seleção massal de isolados fúngicos antagônicos a F. solani e F. oxysporum em substrato comercial para mudas. Foram realizados dois experimentos com os patógenos F. solani e F. oxysporum e com 98 possíveis antagonistas fúngicos, dos gêneros Penicillium claviforme, Penicillium, Aspergillus e Cladosporium. A suspensão dos patógenos foi inserida no substrato, em copos plásticos, sendo acrescentada, cinco dias depois, a suspensão dos demais fungos. Avaliou-se o número de unidades formadoras de colônia de F. solani e F. oxysporum por grama de substrato após nove dias. Dos 98 isolados utilizados contra F. solani, 43 por cento não diferiram da testemunha e 57 por cento reduziram o seu desenvolvimento em substrato, sendo que os três melhores isolados fúngicos foram do gênero Penicillium claviforme. Os três isolados de Penicillium claviforme selecionados para F. solani também foram eficientes para F. oxysporum.

Tests in vitro are usually used for the initial selection of biocontrol agents against soil fungi, lacking methodologies using soil and/or substrate. The objective of this research was to accomplish the mass selection of fungi isolates antagonistic to F. solani and F. oxysporum in commercial substrate for seedlings. Two experiments were conducted, with the pathogens F. solani and F. oxysporum, and 98 possible antagonistic fungi of the genera Penicillium claviforme, Penicillium, Aspergillus and Cladosporium. The suspension of the pathogens was inoculated in the substrate, in plastic cups, and the suspension of the other fungi was added five days later. The number of colony-forming unit of F. solani and F. oxysporum/g of substrate was counted after nine days. Of the 98 isolates used against F. solani, 43 percent did not differ from the control, and 57 percent reduced its development in the substrate, with the three best isolates belonging to the genus Penicillium claviforme. The three isolates of Penicillium claviforme selected for F. solani were also efficient against F. oxysporum.

Doença Cria Giz Ascosphaera apis (Maassen ex Claussen) Olive & Spiltoir em abelhas Apis mellifera L. na Depressão Central do Rio Grande do Sul / Chalk brood disease Ascosphaera apis (Maassen ex Claussen) Olive & Spiltoir in the honey bee Apis mellifera L. in Central Depression of Rio Grande do Sul

A ocorrência da doença apícola "cria giz", em colônias de abelhas Apis mellifera no apiário do Departamento de Zootecnia da Universidade Federal de Santa Maria (UFSM), RS, Brasil, foi constatada em 2004. Foram coletadas amostras de crias de abelhas duras e mumificadas em favos falhados nas colônias com pequena população. Essas amostras foram analisadas no Laboratório de Fitopatologia do Departamento de Defesa Fitossanitária da UFSM, onde se observou a presença de estruturas fúngicas com formação típica do fungo Ascosphaera apis, conhecido como causador da doença "cria giz". Até então não existiam notificações da presença dessa doença na região central do Estado do Rio Grande do Sul, o que pode caracterizar um avanço dessa enfermidade para o interior, a partir de regiões próximas à fronteira com o Uruguai e a Argentina.

The occurrence of chalk brood disease in colonies of the honey bee Apis mellifera located at the apiary of the Department of Zootecnia of the Federal University of Santa Maria (UFSM), Brazil, was evidenced in 2004. Hard and mummified honey bee larvae were collected from failed combs found in poorly populated colonies. These samples were analyzed in the Laboratory of Phytopathology, at the Plant Protection Department (UFSM), where the presence of fungal structures typical of Ascosphaera apis were observed. This fungus is known to cause chalk-brood disease. This is the first record of the presence of this disease in the central region of the State of Rio Grande do Sul, Brazil, characterizing the dissemination of the disease coming from the border with Uruguay and Argentina.

Degradation and binding of atrazine in surface and subsurface soils.

Understanding the dissipation rates of chemicals in unsaturated and saturated zones of subsurface soils will help determine if reductions of concentrations to acceptable levels will occur. Chemical properties and microbial biomass and activity were determined for the surface (0-15 cm), lower root (50-105 cm), and vadose (175-220 cm) zones in a Huntington silty clay loam (Fluventic Hapludoll) collected from an agricultural field near Piketon, OH. The rates of sorption, mineralization, and transformation (formation of bound residues and metabolites) of atrazine were determined. Microbial activity was estimated from the mineralization of (14)C-benzoate. We observed decreased levels of nutrients (total organic carbon, N, and P) and microbial biomass with depth, while activity as measured with benzoate metabolism was higher in the vadose zone than in either the surface or the root zones. Sorption coefficients (K(f)) declined from 8.17 in the surface to 3.31 in the vadose zone. Sorption was positively correlated with organic C content. Rates of atrazine mineralization and bound residues formation were, respectively, 12-2.3-fold lower in the vadose than in the surface soil. Estimated half-lives of atrazine ranged from 77 to 101 days in the surface soil, but increased to over 900 days in the subsurface soils. The decreased dissipation of atrazine with increasing depth in the profile is the result of decreased microbial activity toward atrazine, measured either as total biomass or as populations of atrazine-degrading microorganisms. The combination of reduced dissipation and low sorption indicates that there is potential for atrazine movement in the subsurface soils.