In vitro experimental environments lacking or containing soil disparately affect competition experiments of Aspergillus flavus and co-occurring fungi in maize grains

In vitro experimental environments are used to study interactions between microorganisms, and predict dynamics in natural ecosystems. This study highlights that experimental in vitro environments should be selected to closely match the natural environment of interest during in vitro studies to strengthen extrapolations about aflatoxin production by Aspergillus and competing organisms. Fungal competition and aflatoxin accumulation was studied in soil, cotton wool or tube (water-only) environments, for Aspergillus flavus competition with Penicillium purpurogenum, Fusarium oxysporum or Sarocladium zeae within maize grains. Inoculated grains were incubated in each environment at two temperature regimes (25oC and 30oC). Competition experiments showed interaction between main effects of aflatoxin accumulation and environment at 25oC, but not so at 30oC. However, competition experiments showed fungal populations were always interacting with their environments. Fungal survival differed after the 72-hour incubation in different experimental environments. Whereas, all fungi incubated within the soil environment survived; in the cotton-wool environment, none of the competitors of A. flavus survived at 30 oC. With aflatoxin accumulation, F. oxysporum was the only fungus able to interdict aflatoxin production at both temperatures. This occurred only in the soil environment and fumonisins accumulated instead. Smallholder farmers in developing countries face serious mycotoxin contamination of their grains, and soil is a natural reservoir for the associated fungal propagules, and a drying and storage surface for grains on these farms. Studying fungal dynamics in the soil environment and other environments in vitro can provide insights into aflatoxin accumulation post harvest.

What influences fungal communities in cotton soils

Soilborne diseases such as Fusarium wilt, Black root rot and Verticillium wilt have significant impact on cotton production. Fungi are an important component of soil biota with capacity to affect pathogen inoculum levels and their disease causing potential. Very little is known about the soil fungal community structure and management effects in Australian cotton soils. We analysed surface soils from ongoing field experiments monitoring cotton performance and disease incidence in three cotton growing regions, collected prior to 2013 planting, for the genetic diversity and abundance as influenced by soil type, environment and management practices and link it with disease incidence and suppression. Results from the 28S LSU rRNA sequencing based analysis indicated a total of 370 fungal genera in all the cotton soils and the top 25 genera in abundance accounted for the major portion of total fungal community. There were significant differences in the composition and genetic diversity of soil fungi between the different field sites from the three cotton growing regions. Results for diversity indices showed significantly greater diversity in the long-term crop rotation experiment at Narrabri (F6E) and experiments at Cowan and Goondiwindi compared to the Biofumigation and D1 field experiments at ACRI, Narrabri. Diversity was lowest in the soils under brassica crop rotation in Biofumigation experiment. Overall, the diversity and abundance of soil fungal community varied significantly in the three cotton growing regions indicating soil type and environmental effects. These results suggest that changes in soil fungal community may play a notable role in soilborne disease incidence in cotton.

Fumonisina B1 em arroz: validação de método e efeito de tratamento térmico nos níveis da micotoxina

O arroz (Oryza sativa, L.), como todos os cereais, pode ser contaminado por fungos responsáveis por danos tecnológicos, nutricionais e toxicológicos, dentre eles a produção de micotoxinas. Diversas toxinas fúngicas produzidas pelo gênero Fusarium tem sido relatadas em arroz, no entanto a fumonisina B1 (FB1) é pouco estudada neste grão. As principais características da FB1 é a alta solubilidade em solventes polares, estabilidade a altas temperaturas além de efeitos neurotóxicos e carcinogênicos. Assim o objetivo deste trabalho foi avaliar o efeito do tratamento térmico e hidrotérmico nos teores de fumonisina B1 e nas características químicas de arroz comercial. Na primeira etapa do trabalho foi adaptado um método para detecção e quantificação de FB1 em arroz cru e após cocção, por HPLC-FL. O método foi avaliado quanto aos indicativos de eficiência destacando-se o LOD (30 µg.kg-1) e a recuperação ( 90% para arroz cru e 86% pra arroz cozido). Na segunda etapa realizou-se o levantamento de ocorrência de FB1 em 05 diferentes amostras comerciais de arroz integral, branco e parboilizado da cidade de Rio Grande, RS, totalizando 9 amostras. Foi detectada a presença de FB1 em 7 das 9 amostras, sendo que os maiores índices foram encontrados em amostras de arroz parboilizado e integral apresentando níveis de contaminação entre 30 e 170 µg.kg-1. A terceira etapa do trabalho consistiu no estudo do efeito de tratamentos térmicos sobre os níveis de FB1 em amostras após aplicação de calor. Foram testados tratamento hidrotérmico com evaporação, tratamento hidrotérmico com autoclavagem e tratamento térmico seco. O maior nível de redução dos teores iniciais de FB1 foi 82,8% quando se empregou tratamento térmico seco a 125 °C/3 min. Ainda foram avaliados os efeitos do t ratamento hidrotérmico com evaporação de água na composição química e na digestibilidade protéica. Esta característica proporcionou aumento de até 100% na digestibilidade in vitro das proteínas e reduziu em média 73% do teor de contaminação com FB1.

Collecting banana diversity in eastern Indonesia

Major diseases, including Fusarium wilt tropical race 4, threaten banana production systems worldwide. New sources of genetic resistance are considered necessary in the fight against such diseases. The triangular region of Indonesia taking in Sulawesi, the Maluku Islands and Lesser Sunda Islands was prioritized by the Global Musa Genetic Resources Network, MusaNet for exploration and collecting. It is just east of the Wallace Line, which is recognized as a transition zone for flora in southeast Asia, and had been little explored. Bioversity International funded a team of scientists from Indonesia and Australia to make collecting missions in the triangle in October 2012 and February 2013. Suckers and seeds of 35 promising new accessions were collected. About 90% of these are either wild species or diploid cultivars of more direct use to breeding programs. These were morphologically characterized during the collecting missions and included a set of photographs recommended by Bioversitys Taxonomic Advisory Group. Cigar leaf samples were also collected and sent as fresh samples to the International Banana Genotyping Centre in the Czech Republic. Ploidy and DNA (SSR) genotyping determinations from these samples have been invaluable in quickly interpreting and better appreciating what has been discovered. The new accessions have been grown on at Solok field collection, West Sumatra and will be made available by Indonesia to the international community, including breeding programs, for evaluation and utilization. Information on wild Eumusa prompts a rethinking of the phytogeography of Musa acuminata. The variation within the Australimusa species M. lolodensis highlights the need for broader study of this Musa section. French Plantain-like edible AAs and prospects for the generation of African plantains in the region were identified. The mission indicated existence of local edible ABs in eastern Indonesia in association with balbisiana hybrids origins in the region. Further explorations in the region should add to Musa diversity knowledge.

Research into crown rot and wilt diseases of strawberries in Queensland

Plant losses due to fungal diseases in strawberry (Fragaria × ananassa Duch.) can potentially cause total loss of production. Three fungal pathogens, Fusarium oxysporum f. sp. fragariae, Colletotrichum gloeosporioides and Macrophomina phaseolina, cause similar crown rot and wilt symptoms in strawberries in Queensland. Since the withdrawal of methyl bromide in 2005, no effective chemical control of any of the three pathogens has been available. This study aims at identifying sources of plant genetic resistance that can be used in the breeding program to develop resistant cultivars for use as part of an integrated disease management plan in commercial strawberry production. Results from glasshouse pathogenicity and screening trials on the three pathogens showed that when breeding for resistance against a pathogen, resistance to other pathogens also needs to be considered, e.g., cultivar 'Festival' is resistant to F. oxysporum f. sp. fragariae, but is highly susceptible to C. gloeosporioides. The cultivars 'Earlisweet', 'Kabarla' and 'Phenomenal', two seedling clones and four DAFF breeding lines were resistant to C. gloeosporioides. Cultivar 'Suncoast Delight' showed the most promising level of resistance to M. phaseolina. These cultivars, breeding lines and seedling selections have been made available for incorporation into the crossing program to support the Queensland strawberry breeding program.

Analyses of the community compositions of root rot pathogenic fungi in the soybean rhizosphere soil

Soybean ( Glycine max [L.] Merr.) root rot is an important disease of soybean under continuous cropping, and root rot is widely distributed throughout the world. This disease is extremely harmful, and it is difficult to prevent and control. The study aimed to elucidate the composition of root rot pathogenic fungal communities in the continuous cropping of soybean. In this study, we employed PCRDGGE technology to analyze the communities of root rot pathogenic fungi in soybean rhizosphere soil subjected to continuous cropping during a season with a high incidence of root rot in Heilongjiang province, China, the main soybean producing area in China. The results of 13 DGGE bands were analyzed by phylogenetic revealed that the predominant root rot pathogenic fungi in rhizosphere soil in the test area were Pythium ultimum and Fusarium species. The results of cluster analysis showed that the duration of continuous cropping, the soybean variety and the plant growth stage all had significant effects on the diversity of root rot pathogenic fungi in rhizosphere soil.

Chemical profile and antifungal potential of essential oils from leaves and flowers of Salvia algeriensis (Desf.): A comparative study

Salvia is a plant genus widely used in folk medicine in the Mediterranean area since antiquity. A large number of Salvia essential oils have been reported against diverse microorganisms. In the current study, chemical composition of essential oils from leaves and flowers of Salvia algeriensis (Desf.) was determined using gas chromatography-electron impact mass spectrometry (GC-EIMS) as well as their antifungal activity against phytopathogenic fungi Alternaria solani and Fusarium oxysporum exploring disk method. The GC-EIMS analysis identified 59 compounds (84.8%) in the essential oil obtained from leaves of S. algeriensis. Its major constituents were benzaldehyde (9.7%), eugenol (8.7%) and phenylethyl alcohol (8.4%). In flowers oil, 34 compounds (92.8%) were detected. The main ones were viridiflorol (71.1%) and globulol (8.6%). The essential oil obtained from leaves exhibited the highest antifungal activity, where the effective dose inhibiting 50% of mycelial fungal (ED50) against A. solani was 0.90 μL mL-1 with minimum inhibitory concentration (MIC) equal to 2 μL mL-1, whereas the ED50 and MIC in F. oxysporum culture was 1.84 μL mL-1 and 3 μL mL-1 respectively. The mycelial inhibition by flowers oil varies from 1.77 μL mL-1 (ED50) with A. solani culture (MIC 6.5 μL mL-1) to the lowest effect recorded (ED50 3.00 μL mL-1 and MIC 9.33 μL mL-1) against F. oxysporum. To our best knowledge, this is the first report on S. algeriensis, their leaves oil can constitute an alternative biocontrol against phytopathogenic fungi commonly controlled by chemical fungicides.

EFFECT OF SORGHUM SEED TREATMENT IN BURKINA FASO VARIES WITH BASELINE CROP PERFORMANCE AND GEOGRAPHICAL LOCATION

Sorghum [ Sorghum bicolor (L.) Moench] is a major subsistence crop throughout the region of Sahel. With the exception of seeds and labour, no agricultural inputs are in general used in sorghum production since the grain is of a relatively low commercial value and the risk of losing the crop to drought, flooding, etc. is substantial. A meta-analysis of 118 field experiments was carried out to identify conditions in which two protective seed treatments could support a yield increase of sorghum in Burkina Faso. The two treatments were: i) treatment with the pesticide Calthio C (thiram and chlorpyrifos) and ii) treatment with an aqueous extract from the plant Eclipta alba . Both treatments were found to produce a yield increase (Medians: Calthio C +199 kg ha-1, P<2x10-9; E. alba +90.5 kg ha-1 P<4x10-4). A strong relative effect of Calthio C on yield (+36%) was found for field experiments with a low baseline yield. A strong relative effect of E. alba extract on yield (+22%) was found for experiments with a low baseline of emergence. ANOVA of the 118 field tests showed that baseline crop performance (yield and emergence) and the effect of seed treatments were strongly linked to geographical location (twelve different villages included). Roots from sorghum in the village showing the strongest effect of both seed treatments (>40% yield increase) were found to carry a comparatively high load of the infectious ascomycetes: Fusarium equiseti , Macrophomina phaseolina and Curvularia lunata .

Optimização das condições de cultura da Saccharomyces Cerevisiae e de outros fungos produtores de oligossacáridos

Este estudo envolve o controlo e a optimização das condições de culturas dos microrganismos: Saccharomyces cerevisiae CCMI 396, S. cerevisiae v. lab., Aspergillus oryzae CCMI 125, Aspergillus japonicus CCMI 443, Fusarium oxysporum CCMI 866, Aspergillus niger CCMI 296 com vista à produção de oligossacáridos. Determinaram-se os parâmetros característicos das culturas de duas diferentes estirpes de Saccharomyces com diferentes fontes de carbono e em diferentes condições ambientais. O perfil de crescimento da S. cerevisiae CCMI 396 foi semelhante nos diferentes meios de cultura estudados, sendo a velocidade específica de crescimento mais elevada no meio com glucose a pH 5 e a 30°C (0,36h-1). A S. cerevisiae v. lab. Teve velocidade específica de crescimento idêntica nas mesmas condições da outra estirpe, no entanto, o perfil de crescimento foi diferente nos outros meios de cultura. Estudou-se o efeito da adição de sumo de laranja ou de tomate ao meio de cultura com sacarose e avaliou-se a evolução glucídica no meio de cultura durante o ensaio por HPLC com detector RI. Determinou-se a frutosiltransferase no sobrenadante e na fracção intracelular e determinou-se a evolução dos oligossacáridos. Numa segunda parte deste trabalho efectuaram-se culturas dos quatro fungos filamentosos com vista a avaliar a capacidade de produção, nomeadamente, de fruto­oligassacáridos. Os resultados mostraram que a espécie Aspergillus japonicus CCMI 443 originou, nas mesmas condições de cultura, valores superiores, sendo a percentagem de produção FOStotais/GluCtotais de 61% para as enzimas intracelulares e 40% para as enzimas no sobrenadante. ABSTRACT; This study involves control and optimization of the cultures of microorganisms: Saccharomyces cerevisiae CCMI 396, S. cerevisiae v. lab., Aspergillus oryzae CCMI 125, Aspergillus japonicus CCMI 443, Fusarium oxysporum CCMI 866, Aspergillus níger CCMI 296 for oligosaccharides production. Were determined the parameters characteristic of the cultures of two different strains of Saccharomyces with different sources of carbon and in different environmental conditions. The growth profile of S. cerevisiae CCMI 396 was similar in different cultures media, but the highest specific growth was obtained in a medium with glucose, pH 5, at 30°C (0.36h-1). S. cerevisiae v. lab. had similar growth profile in a medium with glucose but with others culture media was different. We studied the effect of adding orange juice or tomato to the culture medium with sucrose and evaluated the evolution glucidic in the culture medium during the test by HPLC with RI detector. Fructosyltransferase was determined in the extracellular and the intracellular fractions and determined the evolution of oligosaccharides. ln the second part of this work were carried out cultures of four filamentous fungi in order to assess production capacity, in particular, fructoligosaccharides. The results showed that the specie Aspergillus japonicus CCMI 443 originated in the same culture conditions, higher values and the percentage of production FOStotal/Guctotal of 61% for intracellular enzymes and 40% for extracellular enzymes.

Vicilinas de sementes de leguminosas selvagens:purificação, caracterização, efeito de deletério e mecanismo de ação para bruquídeos e para fungos fitopatogênicos

Globulins fractions of legume seeds of Crotalaria pallida, Erytrina veluntina and Enterolobium contortisiliquum were isolated and submitted to assays against serine, cysteine and aspartic proteinases, as also amylase present in midgut of C. maculatus and Z. subfasciatus. Hemagglutination assays indicated presence of a lectin in E. veluntina globulin fractions. This lectin had affinity to human erythrocytes type A, B and O. Vicilins were purified by chromatography on Sephacryl S-300 followed of a chromatography on Sephacryl S-200, which was calibrated using protein markers. Vicilins from C. pallida (CpV) and E. veluntina (EvV) seeds had a molecular mass of 124.6 kDa and E. contortisiliquum a molecular mass of 151kDa. Eletrophoresis in presence of SDS showed that CpV was constituted by four subunities with apparent molecular mass of 66, 63, 57 and 45 kDa, EvV with three subunities with apparent molecular mass of 45kDa and EcV four subunities, two with 37.1 kDa and two with 25.8 kDa. Non denaturantig eletrophoresis displayed single bands with high homogeneity, where CpV had lower acidic behavior. All vicilins are glycoproteins with carbohydrate contents at 1 to1.5%. Bioassays were done to detect deleterious effects of vicilins against C. maculatus and Z. subfasciatus larvae. CpV, EvV and EcV exhibited a WD50 of 0.28, 0.19 and 1.03%; LD50 0.2, 0.26, and 1.11% respectively to C. maculatus. The dose responses of CpV, EvV and EcV to Z. subfasciatus were: WD50 of 0.12, 0.14, 0.65% and LD50 of 0.09, 0.1, and 0.43% respectively. The mechanism of action of these proteins to bruchids should be based on their properties of bind to chitin present in mid gut of larvae associated with the low digestibility of vicilin. In assays against phytopatogenous fungus, only EcV was capable of inhibit F. solani growth at concentrations of 10 and 20 µg and its action mechanism should be also based in the affinity of EcV to chitin present in the fungi wall

Incidência de fungos em pós-colheita de banana 'Prata anã'(Musa AAB)

Este trabalho teve por objetivo determinar a ocorrência e a freqüência de fungos em banana 'Prata anã' e elucidar o agente causal das podridões em pós-colheita de frutos provenientes do norte de Minas Gerais. Dois métodos de isolamento foram adotados: diluição em placas, a partir da lavagem de frutos verdes, e direto de frutos maduros. Os fungos Colletotrichum musae, Trichoderma harzianum, Fusarium equisetii, Penicillium sp. Aspergillus parasiticus, Trichothecium roseum, Colletotrichum acutatum, Alternaria sp., Cladosporium musae e Curvularia lunata foram os mais freqüentemente associados aos frutos. A patogenicidade desses fungos foi testada pela substituição de discos da casca de frutos verdes por discos de micélio. Colletotrichum musae apresentou área média lesionada em torno do ponto de inoculação igual a 5,8 cm², enquanto para os demais fungos testados não passou de 1,50 cm². Os resultados mostraram que C. musae é o agente primário das podridões dos frutos examinados com 100 % de incidência e os demais fungos limitaram-se a necrosar os ferimentos em torno do ponto de inoculação. O modo de infecção latente, causada por C. musae, parece favorecer, primeiramente, a colonização interna dos tecidos e, posteriormente, a ação dos fungos oportunistas, que aceleram as podridões nos frutos e na coroa.

Intoxicaciones por plantas y micotoxinas en rumiantes diagnosticadas en Uruguay

Resumen Las intoxicaciones por plantas y micotoxinas ocasionan importantes pérdidas económicas en países pecuarios, principalmente por muertes de animales y mermas en la producción. Este trabajo fue realizado con el propósito de brindar al veterinario que actúa en el diagnóstico de enfermedades de animales productivos, información breve y accesible sobre las intoxicaciones por plantas y micotoxinas diagnosticadas en rumiantes de Uruguay. Para llevar a cabo el mismo se utilizaron como fuentes bibliográficas: revistas científicas arbitradas, tesis de grado, libros y comunicaciones en congresos, jornadas nacionales e internacionales. Se encontraron 37 géneros y 45 especies de plantas tóxicas, distribuidos en 20 familias botánicas. En cuanto a los hongos toxicogénicos, se hallaron 13 especies pertenecientes a 9 géneros de 6 familias diferentes. Entre las plantas diagnosticadas, la familia Asteraceae (Compositae) cuenta con la mayor cantidad de especies tóxicas, seguida por Gramineae, Solanaceae y Fabaceae (Leguminoseae). Los principales géneros de hongos toxicogénicos Aspergillus, Penicillium, Fusarium y Claviceps han sido diagnosticados en el país. Desde el punto de vista patológico, las plantas y micotoxinas hepatotóxicas resultaron ser las más numerosas. La información presentada en las tablas, permitirá a los veterinarios un fácil abordaje en esta temática

Óleos essenciais e extratos vegetais no controle de fitopatógenos de grãos de soja.

A utilização de sementes sadias é um importante método para diminuir a ocorrência de doenças desde o inicio de seu cultivo. Uma das alternativas ao uso de agrotóxicos é a utilização de óleos essenciais e extratos de plantas que possuam propriedades tóxicas aos fitopatógenos. O objetivo deste trabalho foi avaliar o efeito de óleos essenciais e extratos hidroalcoólicos de plantas medicinais na sanidade e germinação de sementes de soja. Os óleos essenciais e os extratos hidroalcoólicos foram diluídos em água destilada, na concentração de 2 mg/mL para os extratos e 20% para os óleos essenciais. Em seguida, as sementes foram pulverizadas com os tratamentos e secas a sombra durante 4 horas em temperatura ambiente (25ºC). Os tratamentos utilizados foram: T1=testemunha (água destilada); T2 = óleo essencial de gengibre; T3 = óleo essencial de limão Taiti; T4 = extrato de pariparoba e T5 = extrato de penicilina. Os testes de germinação e sanidade foram realizados segundo as Regras para Análises de Sementes (RAS). Os resultados obtidos no teste de germinação demonstraram que o percentual de germinação inicial das sementes não tratadas (4%) encontrava-se muito abaixo dos níveis recomendados pelas RAS (80%), não tendo como avaliar a interferência dos tratamentos na germinação. Em função deste resultado, foi realizado o teste de sanidade visando o tratamento de grãos. O óleo essencial de gengibre apresentou uma redução da incidência de 75% de Cladosporium sp; Rhizopus sp.; Fusarium spp. quando comparado à testemunha. Pode-se concluir que o óleo essencial de gengibre na concentração de 20% apresenta atividade positiva no controle de Cladosporium sp, Rhizopus sp e Fusarium spp em grãos de soja.

Avaliação probabilística do risco de giberela do trigo considerando a influencia do ENOS.

A giberela é uma importante doença na cultura do trigo cujas epidemias ocorrem sob a influência marcante de condições ambientais específicas durante o florescimento. Neste trabalho, a influência do fenômeno El Nino/Oscilação Sul (ENOS) sobre o risco de epidemias da giberela do trigo, em Passo Fundo, RS, estimado com modelos de simulação, foi avaliada via abordagem de anos análogos. Foi utilizada uma série local histórica (1957 a 2006) de dados meteorológicos para estimar um índice de risco de giberela (IRG) para diferentes datas de plantio (1 a 30/06) nos 50 anos da série. Para quantificar a influência do ENOS, foram comparadas medianas e funções probabilidade de excedância (FPE) do IRG (Prob(IRG>y)para três fases do ENOS (quente, fria e neutra) nos anos da série, utilizando métodos não paramétricos. Foi observado um efeito pronunciado do ENOS, principalmente para a fase fria, onde o risco estimado de epidemias moderadas ou severas foi nulo para todas as datas de plantio. Nas demais fases, o referido risco variou de 0,10 a 0,25.

Registro de Meloidogyne enterolobii em mudas de goiabeiras formadas em viveiros de Petrolina, PE.

Objetivou-se avaliar a qualidade das mudas de goiabeira no município de Petrolina, PE.