Fungos endofíticos associados a acículas de Pinus taeda

O presente trabalho objetivou estudar os fungos endofíticos em acículas de árvores jovens de Pinus taeda L. e avaliar o efeito da posição de coleta na árvore. As amostras foram coletadas em duas alturas (30-50 cm e 100-130 cm acima do solo) e nas quatro posições cardeais (norte, sul, leste e oeste), em plantas com 18 meses de idade, localizadas em Colombo, PR, Brasil. As acículas foram submetidas a assepsia e fragmentos com 10 mm de comprimento foram plaqueados em meio BDA e incubados a 28 °C, sob fotofase de 12 h, por 15 dias. Para a identificação, as estruturas reprodutivas dos fungos foram produzidas pelo método do microcultivo. Foram isolados e identificados dezessete gêneros: Alternaria, Aspergillus, Cladosporium, Colletotrichum, Coniothyrium, Diplodia, Drechslera, Hansfordia, Monocillium, Nodulisporium, Panidio, Papulaspora, Pestalotiopsis, Phialophora, Pithomyces, Rhizoctonia e Xylaria Alguns morfotipos sem identificação foram Mycelia sterilia e fungos demaciáceos. O número de isolados da altura 30-50 cm foi significativamente maior que na outra altura. Não foi observada diferença significativa no número de isolados entre as posições cardeais de uma mesma altura. Diferenças significativas foram observadas entre os gêneros isolados e Xylaria foi o gênero mais frequente.

Detection, transmission, pathogenicity and chemical treatment of fungi in Ceiba speciosa seeds

The aim of this work was to evaluate fungus association, transmission and pathogenicity, besides chemical seed treatment in Ceiba speciosa seeds from different regions of southern Brazil. Seven seed samples were used to do the germination test, fungus detection by blotter test and potato-dextrose-agar (PDA), fungus transmission and pathogenicity tests; besides, chemical seed treatments were tested. Germination ranged from 0 to 59,5%. The following fungi were associated in the seeds: Fusarium sp., Alternaria sp., Colletotrichum sp., Curvularia sp. and Pestalotia sp.; in addition, Fusarium sp. was found in all the samples. Alternaria sp. and Fusarium sp. were transmitted by seeds. The isolates of Alternaria sp., Colletotrichum sp. and Fusarium sp., were pathogenic to seedlings and seeds. The seed treatment with methyl tiophanate and the combination captan + methyl tiophanate reduced Fusarium sp. incidence.

Semi-selective medium for Fusarium graminearum detection in seed samples

Fungi of the genus Fusarium cause a variety of difficult to control diseases in different crops, including winter cereals and maize. Among the species of this genus Fusarium graminearum deserves attention. The aim of this work was to develop a semi-selective medium to study this fungus. In several experiments, substrates for fungal growth were tested, including fungicides and antibiotics such as iprodiona, nystatin and triadimenol, and the antibacterial agents streptomycin and neomycin sulfate. Five seed samples of wheat, barley, oat, black beans and soybeans for F. graminearum detection by using the media Nash and Snyder agar (NSA), Segalin & Reis agar (SRA) and one-quarter dextrose agar (1/4PDA; potato 50g; dextrose 5g and agar 20g), either unsupplemented or supplemented with various concentrations of the antimicrobial agents cited above. The selected components and concentrations (g.L-1) of the proposed medium, Segalin & Reis agar (SRA-FG), were: iprodiona 0.05; nystatin 0,025; triadimenol 0.015; neomycin sulfate 0.05; and streptomycin sulfate, 0.3 added of ¼ potato sucrose agar. In the isolation from seeds of cited plant species, the sensitivity of this medium was similar to that of NSA but with de advantage of maintaining the colony morphological aspects similar to those observed in potato-dextrose-agar medium.

Atividade antifúngica de extratos vegetais sobre o desenvolvimento de fitopatógenos

A formação de uma consciência ecológica e a busca pela preservação do meio ambiente tem gerado a necessidade de testar produtos naturais, visando um controle alternativo de fitopatógenos. Desta forma, objetivou-se avaliar o potencial de dez extratos aquosos sobre o desenvolvimento in vitro de fungos fitopatogênicos. Foram conduzidos seis ensaios experimentais em delineamento inteiramente casualizado, com 11 tratamentos e 10 repetições para cada ensaio. Os tratamentos constaram dos extratos bruto aquosos de alho, arruda, canela, cravo-da-índia, cavalinha, eucalipto, hortelã, jabuticaba, melão de são caetano e nim na concentração de 20%, mais a testemunha (somente BDA). Os ensaios foram realizados com os fungos Aspergillus sp., Penicillium sp., Cercosporakikuchii, Colletotrichum sp., Fusarium solani e Phomopsis sp. Os extratos foram filtrados em papel wathman nº 1, colocados em banho maria a 65ºC, durante 1 hora, incorporados em meio BDA e após vertido em placas de Petri, transferiu-se discos de micélio dos patógenos (0,3 cm de diâmetro). Foi analisado o crescimento micelial da colônia, a porcentagem de inibição e a taxa de crescimento dos fungos. Observou-se que os meios de cultura contendo os extratos de cravo-da-índia, alho e canela apresentaram maior atividade antifúngica sobre os fitopatógenos, quando comparados aos demais extratos utilizados, destacando o extrato de cravo-da-índia, que inibiu completamente o desenvolvimento de todos os fitopatógenos testados.

Chitosan and fungicides on postharvest control of Guignardia citricarpa and on quality of 'Pêra Rio' oranges

Citrus fruits are affected by the black spot disease caused by the fungus Guignardia citricarpa. Chitosan can be used as covering for fruits and may delay the ripening process and inhibit the growth of some fungi. Thus, the control of citrus black spot using chitosan and the fungicides thiabendazole and imazalil was assessed in addition to the physicochemical quality of 'Pêra Rio' oranges. The oranges were immersed into chitosan, thiabendazole or imazalil, and in chitosan mixed with both fungicides. The fruits were then stored at 25 °C, 80% RH, for 7 days and, after this storage period, subjected to physicochemical analyses. Chitosan in association with the fungicides reduced black spot in 'Pêra Rio' oranges and delayed the change in the orange skin colour from green to yellow during the postharvest storage. Total soluble solids, titratable acidity, pH, ascorbic acid content and ratio were not influenced by the treatments. Thus, chitosan applied with the fungicides thiabendazole and imazalil showed potential to control the development of black spot lesions on 'Pêra Rio' oranges during the postharvest period.

Genetic variability of Puccinia triticina Eriks. in Brazil

Studies on the genetic variability of Puccinia triticina in inoculum collected in Brazil started in 1941 with Vallega (20). The pioneering work in Brazil dates from 1949 (16) at "Instituto Agronômico do Sul", Ministry of Agriculture (MA), in Pelotas, Rio Grande do Sul State (RS), and continued after 1975 at Embrapa Wheat in Passo Fundo, RS. In 2002, analyses for the identification of P. triticina races continued at OR Seed breeding, simultaneously to Embrapa's program, both in Passo Fundo. The investigators involved in the identification of races in Brazil were Ady Raul da Silva in Pelotas (MA), Eliza Coelho in Pelotas (MA) and in Passo Fundo (Embrapa), Amarilis Labes Barcellos in Pelotas (MA) and in Passo Fundo (Embrapa and OR), Camila Turra in Passo Fundo (OR) and Marcia Chaves in Passo Fundo (Embrapa). From 1979 to 2010 growing season, 59 races were determined, according to the differentiation based on the expression of each Lr resistance gene. On average, one to three new races are detected per year. Research has focused on the use of vertical resistance; however, lately some institutes have searched more durable resistance, of the adult-plant type (horizontal, less race-specific). The uninterrupted monitoring of the wheat rust pathogenic population in Brazil during so many decades allowed the understanding of the evolution and virulence of races. The use of international nomenclature adopted by some programs has allowed the comparison of the fungus variability in Brazil with that in other countries, especially where frontiers are not barriers for spore transportation, confirmed by the occurrence of the same races all over one region.

Net melon resistance to Didymella bryoniae according to grafting and potassium levels

Chemical control of the fungus Didymella bryoniae, the causal agent of the disease gummy stem blight in melon, is frequently inefficient; thus, alternatives such as grafting and nutrition must be studied. Rootstocks and potassium levels were tested aimed at controlling this disease in net melon under protected environment. The melon hybrid 'Bônus II', ungrafted and grafted onto 'Dinero' melon and 'Strong Tosa' pumpkin rootstocks, was cultivated and inoculated by using the toothpick insertion method with 7-mm mycelial disks from the isolate D. bryoniae Dbr 37; for control, only toothpick insertion was used. The plants were subjected to the following potassium levels: 0, 62.5, 125, 187.5, 250 mg L-1. Grafted 'Bônus II' melon plants were resistant to the fungus, whereas ungrafted ones were susceptible. The adopted potassium levels did not influence the stem lesion size or the survival of plants.

Fungicides, seed dresser adjuvants and storage time in the control of Drechslera teres in barley seeds

In experiments conducted in laboratory, the effect of fungicides, seed dresser adjuvants and storage time in the control of Drechslera teres in seeds of barley cultivar BRS Elis, with 58% incidence, was quantified. Fungicides indicated by barley research (carboxin + thiram, difenoconazole and triadimenol) compared with the mixture carbendazim + iprodione were tested. As seed dresser adjuvants, water (500m mL/100 Kg) and a polymer (150 mL/100 Kg) were used. Treated seeds were stored in paper bags and kept in the refrigerator at 5ºC. At 30-day intervals during six months, seeds were plated on semi-selective Reis's medium (1983). The best control was obtained by the mixtures carboxin + thiram and carbendazim + iprodione and the polymer as seed dresser. The control efficiency was improved by the storage time without negatively affecting seed germination. Due to the transmission efficiency, the fungus eradication in seeds should be pursued.

Progress of Asian soybean rust and airborne urediniospores of Phakopsora pachyrhizi in southern Brazil

Asian soybean rust, caused by the fungus Phakopsora pachyrhizi, was reported at epidemic levels in 2003/2004 and is the main soybean disease in Brazil. The aim of this study was to investigate the spread of Asian soybean rust and to quantify airborne urediniospores in the region of Campo Mourão, Paraná State, Brazil. Three experiments were conducted under field conditions during the 2007/08 and 2008/09 crop seasons. Using the disease gradient method, provided by the application of increasing levels of the fungicide tebuconazole, four Asian soybean rust epidemics at different intensities were obtained in each experiment. To quantify the urediniospores, weathercock-type spore collectors were installed during and between the two crop seasons. Disease progress curves were plotted for each epidemic, and maximum severity was estimated. The curves were fit to the logistic model, which provided higher coefficients of determination and more randomly distributed residuals plotted over time. Analyses of the area under the disease progress curve showed that the largest epidemics occurred in the 2007/2008 crop season and that the progress rates were higher for severity, even among plants protected with the fungicide. The number of urediniospores collected in the air was related to the presence of soybean plants in the cultivated crops. The quantity of urediniospores was also positively correlated to the disease severity and incidence, as well as to cumulative rainfall and favorable days for P. Pachyrhizi infection.

Meloidogyne javanica control by Pochonia chlamydosporia, Gracilibacillus dipsosauri and soil conditioner in tomato

Organic matter plays a fundamental role in the antagonistic activity of microorganisms against phytonematode populations on the soil. In this study, the compatibility between the fungus Pochonia chlamydosporia (Pc-12) and the rhizobacterium Gracilibacillus dipsosauri (MIC 14) was evaluated in vitro, as well as the effect of the fungus at the concentration of 5,000 chlamydospores per gram of soil, rhizobacterium at 4.65 x 10(9) cells/g of soil, and the soil conditioner Ribumin® at 10 g/pot, either alone or in combination, against Meloidogyne javanica population in tomato plants (3,000 eggs/pot). A suspension of water or Ribumin® alone was applied on the soil as negative control, while a suspension of nematode eggs was applied as positive control. The reduction in the number of galls in roots per plant was 48 and 41% for the treatments Ribumin + MIC 14 + Pc-12 and MIC 14 + Pc-12, respectively. Regarding to the number of eggs per plant, MIC 14 and Pc-12 + Ribumin led to a reduction by 26 and 21%, respectively, compared to the control treatment. Interaction between the nematophagous fungus and the rhizobacterium was positive for the nematode control, even though G. dipsosauri inhibited P. chlamydosporia growth by up to 30% in in vitro tests.

Diagrammatic scale for the assessment of blast on wheat spikes

The correct quantification of blast caused by the fungus Magnaporthe oryzae on wheat (Triticum aestivum) spikes is an important component to understand the development of this disease aimed at its control. Visual quantification based on a diagrammatic scale can be a practical and efficient strategy that has already proven to be useful against several plant pathosystems, including diseases affecting wheat spikes like glume blotch and fusarium head blight. Spikes showing different disease severity values were collected from a wheat field with the aim of elaborating a diagrammatic scale to quantify blast severity on wheat spikes. The spikes were photographed and blast severity was determined by using resources of the software ImageJ. A diagrammatic scale was developed with the following disease severity values: 3.7, 7.5, 21.4, 30.5, 43.8, 57.3, 68.1, 86.0, and 100.0%. An asymptomatic spike was added to the scale. Scale validation was performed by eight people who estimated blast severity by using digitalized images of 40 wheat spikes. The precision and the accuracy of the evaluations varied according to the rater (0.82

Orange rust in sugarcane: molecular identification in Rio de Janeiro State

Sugarcane (Saccharum spp.) cultivation is one of the major agricultural activities in the Brazilian states. This study aimed to molecularly identify the pathogen associated with rust in sugarcane cultivars in the state of Rio de Janeiro and to suggest a control strategy. Among the 14 PCR-tested cultivars, Puccinia kuehnii infection was identified for RB947520, RB92606, RB835486, RB72454, SP89-11I5, SP83-2847, both from infected leaf sample and from urediniospores. Puccinia kuehnii was not detected by PCR for the cultivars RB955971, RB951541, RB92579, RB867515, RB855536, SP91-1049, SP80-3280, SP80-1816. This is the first molecular detection of this fungus in the state of Rio de Janeiro for six of the 14 analyzed cultivars.

Sensitivity reduction in Blumeria graminis f. sp. hordei to triadimenol fungicide applied as barley seed treatment

Experiments were carried out in a growth chamber with controlled temperature and photoperiod to test two populations of Blumeria graminis f. sp. hordei from Guarapuava, Paraná State, and Passo Fundo, Rio Grande do Sul State, Brazil. Treatments consisted in application of the fungicide triadimenol (Baytan 150 SC®) at three rates of its commercial formulation: 150, 250, 350 mL/100 Kg barley seeds. The experiments were conducted separately in a growth chamber for each population, adopting the same temperature and photoperiod. For inoculation, pots containing barley seedlings colonized by the fungus were placed among the plots. After emergence of the first symptoms, the disease severity was assessed at two-day intervals. The experiments were repeated twice for each fungus population. Data were expressed as area under the disease progress curve and as powdery mildew control by comparing the severity after the fungicide treatments to that of control. Data were subjected to analysis of variance and regression analysis; the area under the disease progress curve was also calculated. Comparing the data obtained in the present study with those reported in the literature and the control, the maximum value of 26.1% is considered insufficient to prevent the damages caused by the disease. The control response to the fungicide rate was significant. We can conclude that there was a reduction in the sensitivity of both B. graminis f.sp. hordei populations to the fungicide triadimenol, which explains the control failure observed in barley farms.

Semi-selective culture medium for Exserohilum turcicum isolation from corn seeds

Northern corn leaf blight, caused by Exserohilum turcicum (Et), is a disease of widespread occurrence in regions where corn, sweetcorn and popcorn are grown. This disease has great potential to cause damage and has been studied for years, but the association of its causal agent with seeds remains unconfirmed. Thus, the availability of a sensitive method to detect and quantify the inoculum in seeds, even at low incidence, is essential. The aim of this study was to develop a method to detect and quantify the presence of the fungus infecting and infesting corn and popcorn seeds. Artificially and naturally infected seeds were employed to develop the medium. The semi-selective medium was composed of carbendazim (active ingredient) (60 mg/L), captan (30 mg/L), streptomycin sulfate (500 mg/L) and neomycin sulfate (600 mg/L) aggregated to the medium lactose casein hydrolysate agar medium. By using this, Et was detected in naturally infected corn seeds, showing 0.124% incidence, in four out of ten analyzed samples. In addition, 1.04 conidia were detected per infested seed. By means of isolation, pathogenicity test, morphological characterization and comparison with descriptions of the species in the literature, the fungus isolated from the seeds was confirmed to be Et. Both infection and infestation were considered low; thus, for studies of Et detection in corn seeds, the use of semi-selective medium and more than 1,200 seeds/sample is suggested.

Etiology of phomopsis root rot in soybean

In a survey of damages caused by soybean root rot to crops in the south of Brazil for several years, a root rot caused by Phomopsis sp has been found with increasing frequency. The primary symptoms are seen when the main root is cut longitudinally, including the death of the wood which shows white coloration and well-defined black lines that do not have a defined format. Thus, based on similarity, it has been called geographic root rot due to its aspect resembling irregular lines that separate regions on a map. In isolations, colonies and alpha spores of Phomopsis have prevailed. Pathogenicity test was done by means of inoculation in the crown of plants cultivated in a growth chamber. The geographic symptoms were reproduced in plants and the fungus Phomopsis sp. was reisolated. In soybean stems naturally infected with pod and stem blight, geographic symptoms caused by Phomopsis phaseoli are found. To the known symptoms on stems, pods and grains, that of root rot caused by P. phaseoli is now added.