Resumo
Twenty six isolates of Fusarium graminearum from grains of maize hybrids harvested in ±west Argentina were grown on autoclaved rice grain to assess their ability to produce type B trichothecenes. Chemical analysis indicated that 38% of isolates were nivalenol (NIV) producers only, 31% were major NIV producers with high DON(deoxynivalenol)/NIV ratios, 8% were major DON producers with minor NIV production, and 23% were DON producers only. Isolates showed a high variability in their toxigenic potential which was not related to fungal biomass. The distribution of the different chemotypes as well as the high and the low trichothecene-producing Fusarium isolates could not be associated to a geographical origin. Our results confirmed for the first time that isolates of Fusarium graminearum from maize of northwest Argentina are able to produce DON and NIV. A substancial contamination with both NIV and DON is likely in maize from northwest Argentina. Their contents should be quantified in regional surveillances for mycotoxin contamination.(AU)
Assuntos
Fusarium , Tricotecenos , Ergosterol , Zea mays , FungosResumo
Ear rots caused by Fusarium spp. are among the main fungal diseases that contribute to poor quality and the contamination of maize grains with mycotoxins. This study aimed to determine the visual incidence of fungal-damaged kernels (FDKs), the incidence of two main Gibberella (a teleomorph of Fusarium) complexes (G. fujikuroi and G. zeae) associated with maize using a seed health blotter test, and the fumonisin levels, using high performance liquid chromatography, in samples of maize grains grown across 23 municipalities during the 2008/09 and 2009/10 growing seasons. Additionally, 104 strains that were representative of all of the analysed samples were identified to species using PCR assays. The mean FDK was seven per cent, and only six of the samples had levels greater than six per cent. Fusarium spp. of the G. fujikuroi complex were present in 96% of the samples, and G. zeae was present in 18% of the samples (5/27). The mean incidence of G. fujikuroi was 58%, and the incidence of G. zeae varied from 2 to 6%. FB1 was found in 58.6%, FB2 in 37.9%, and both toxins in 37.9% of the samples. The FB1 and FB2 levels were below the quantification limits for 41.3% of the samples, and the mean FB1 levels (0.66 µg/g) were higher than the mean FB2 levels (0.42 µg/g). The PCR identification separated the 104 isolates into three of the G. fujikuroi complex: F. verticillioides (76%), F. subglutinans (4%) and F. proliferatum (2%); and G. zeae (anamorph = F. graminearum) (18%). Our results confirmed the dominance of F. verticillioides, similar to other regions of Brazil, but they differed due to the relatively higher incidence of F. graminearum. Total fumonisin levels were below the maximum limit determined by current Brazilian regulations.(AU)
Assuntos
Micoses/patologia , Fumonisinas/toxicidade , Regulamentação Governamental , Fusarium/ultraestrutura , Zea maysResumo
Fusarium head blight of wheat in Brazil is caused mainly by two species of the Fusarium graminearum species complex: F. graminearum sensu stricto (Fgss) and F. meridionale (Fmer), which vary in relation to toxigenic potential. The aim of this study was to evaluate the effect of phenotypic traits and aggressiveness of these two species, associated with the tebuconazole fungicide, of the triazole group, on the reduction of wheat yield. Ten Fgss strains and Fmer strains were evaluated with regards to: total sporulation, germination rate and tebuconazole sensitivity. The strains were spray-inoculated onto BRS Guamirim cv. 'plants' at full flowering and disease severity, Fusarium-damaged kernels, and kernel weight were evaluated. The effect of tebuconazole in disease control was evaluated through spraying the fungicide and then inoculating the central-floret of the wheat head with the following inoculum treatments: a mixture of all Fgss strains, a mixture of all Fmer strains or a mixture of all isolates of both species (1:1). Strains of Fgss showed higher total sporulation and germination rates and were less sensitivity to tebuconazole. The weight of Fusarium-damaged kernels from inoculations with Fmer strains was 50% higher than those inoculated with Fgss. A fungistatic effect on the disease was found for tebuconazole application and harvested kernels showed 25% higher grain weight than the untreated kernels. It is suggested that the distinct yield loss potential by these two species, in association with the fungistatic effect of triazoles could explain the co-occurrence of different mycotoxins in the harvested kernels, which needs to be proven with field data.
No Brasil, a giberela em trigo é causada por espécies do complexo Fusarium graminearum, especialmente F. graminearum sensu stricto (Fgss) e F. meridionale (Fmer), as quais variam quanto ao potencial toxigênico. O objetivo deste trabalho foi avaliar a relação entre características fenotípicas e agressividade dessas duas espécies associadas ao uso do fungicida tebuconazole, da classe dos triazóis, com a redução de rendimento do trigo. Em dez isolados Fgss e nove Fmer, foram avaliados: esporulação total, taxa de germinação e sensibilidade ao fungicida tebuconazole. A inoculação, para cada isolado, foi feita por aspersão em espigas e as variáveis severidade da doença, incidência de grãos giberelados e o peso de grãos foram avaliados. O efeito de tebuconazole na redução da doença foi avaliado em ação protetora, seguida de inoculação na espigueta central de plantas da cultivar 'BRS Guamirim', com os seguintes tipos de inóculo: somente Fgss, somente Fmer ou a mistura de ambos (1:1). Isolados Fgss apresentaram maior esporulação total, maior taxa de germinação e foram menos sensíveis ao tebuconazole em comparação a Fmer. Grãos giberelados por isolados Fmer apresentaram 50% maior peso do que aqueles provenientes de inoculações com Fgss. O tebuconazole apresentou efeito fungistático e os grãos de espigas tratadas com o fungicida apresentaram peso 25% superior aos não tratados. Sugere-se que diferenças no potencial de dano aos grãos pelas duas espécies, assim como o efeito fungistático de triazóis, podem ajudar a explicar a co-ocorrência de diferentes micotoxinas, o que ainda necessita ser confirmado com dados de campo.
Resumo
Fusarium head blight of wheat in Brazil is caused mainly by two species of the Fusarium graminearum species complex: F. graminearum sensu stricto (Fgss) and F. meridionale (Fmer), which vary in relation to toxigenic potential. The aim of this study was to evaluate the effect of phenotypic traits and aggressiveness of these two species, associated with the tebuconazole fungicide, of the triazole group, on the reduction of wheat yield. Ten Fgss strains and Fmer strains were evaluated with regards to: total sporulation, germination rate and tebuconazole sensitivity. The strains were spray-inoculated onto BRS Guamirim cv. 'plants' at full flowering and disease severity, Fusarium-damaged kernels, and kernel weight were evaluated. The effect of tebuconazole in disease control was evaluated through spraying the fungicide and then inoculating the central-floret of the wheat head with the following inoculum treatments: a mixture of all Fgss strains, a mixture of all Fmer strains or a mixture of all isolates of both species (1:1). Strains of Fgss showed higher total sporulation and germination rates and were less sensitivity to tebuconazole. The weight of Fusarium-damaged kernels from inoculations with Fmer strains was 50% higher than those inoculated with Fgss. A fungistatic effect on the disease was found for tebuconazole application and harvested kernels showed 25% higher grain weight than the untreated kernels. It is suggested that the distinct yield loss potential by these two species, in association with the fungistatic effect of triazoles could explain the co-occurrence of different mycotoxins in the harvested kernels, which needs to be proven with field data.
No Brasil, a giberela em trigo é causada por espécies do complexo Fusarium graminearum, especialmente F. graminearum sensu stricto (Fgss) e F. meridionale (Fmer), as quais variam quanto ao potencial toxigênico. O objetivo deste trabalho foi avaliar a relação entre características fenotípicas e agressividade dessas duas espécies associadas ao uso do fungicida tebuconazole, da classe dos triazóis, com a redução de rendimento do trigo. Em dez isolados Fgss e nove Fmer, foram avaliados: esporulação total, taxa de germinação e sensibilidade ao fungicida tebuconazole. A inoculação, para cada isolado, foi feita por aspersão em espigas e as variáveis severidade da doença, incidência de grãos giberelados e o peso de grãos foram avaliados. O efeito de tebuconazole na redução da doença foi avaliado em ação protetora, seguida de inoculação na espigueta central de plantas da cultivar 'BRS Guamirim', com os seguintes tipos de inóculo: somente Fgss, somente Fmer ou a mistura de ambos (1:1). Isolados Fgss apresentaram maior esporulação total, maior taxa de germinação e foram menos sensíveis ao tebuconazole em comparação a Fmer. Grãos giberelados por isolados Fmer apresentaram 50% maior peso do que aqueles provenientes de inoculações com Fgss. O tebuconazole apresentou efeito fungistático e os grãos de espigas tratadas com o fungicida apresentaram peso 25% superior aos não tratados. Sugere-se que diferenças no potencial de dano aos grãos pelas duas espécies, assim como o efeito fungistático de triazóis, podem ajudar a explicar a co-ocorrência de diferentes micotoxinas, o que ainda necessita ser confirmado com dados de campo.
Resumo
Zearalenone, a mycotoxin produced by fungi of the genus Fusarium, including F. graminearum, triggers reproduction disorders in certain animals and hyperestrogen syndromes in humans. Current research investigates three concentrations of neem oil extract (0.1, 0.25 and 0.5%) in reducing the production of zearalenone. Neem oil extract decreased zearalenone amount in the three concentrations but highest inhibition (59.05%) occurred at 0.1%.
Resumo
The main objective of this work was to develop a PCR protocol for the identification of Fusarium graminearum, based on a pair of primers targeted to a segment of the 3' coding region of the gaoA gene that codes for the enzyme galactose oxidase (GO). This region has low homology with the same region of GO genes from other fungi. Genomic DNA from 17 strains of Fusarium spp. isolated from diseased cereals, from several other Fusarium species, and from other fungi genera was analyzed in a PCR assay using this primer set. The 17 strains of Fusarium spp. were also analyzed for the GO enzyme production in submerse fermentation in a new formulated liquid medium. All strains that were morphologically and molecularly identified as F. graminearum were able to secrete the enzyme and had a positive result in the used PCR protocol. No DNA fragment was amplified using genomic DNA from other Fusarium species and species of other fungi genera. The results suggest that the proposed PCR protocol is specific and can be considered as a new molecular tool for the identification of F. graminearum. In addition, the new formulated medium is a cheap alternative for screening for GO screening production by F. graminearum.
O principal objetivo deste trabalho foi desenvolver um novo protocolo de PCR para identificação de isolados de Fusarium graminearum, baseado no uso de um par de iniciadores direcionado para um segmento da região 3' codificadora do gene gaoA que codifica a enzima galactose oxidase (GO). Esta região possui baixa homologia com a mesma região de genes da GO de outros fungos. O DNA genômico de 17 cepas de Fusarium spp. isoladas de cereais infectados com sintomas, de vários outras espécies de Fusarium e de outros gêneros de fungos foi analisado em um protocolo de PCR utilizando os iniciadores desenhados. Os 17 isolados de Fusarium spp. também foram analisados para a produção da enzima GO em fermentação submersa em um novo meio líquido. Todas as cepas que foram morfologicamente e molecularmente identificadas como F. graminearum foram capazes de secretar a enzima e tiveram um resultado positivo no protocolo de PCR, utilizando os iniciadores direcionados para o gene gaoA. Nenhum fragmento de DNA foi amplificado quando foi utilizado o DNA genômico de várias outras espécies de Fusarium e de espécies de outros gêneros de fungos. Os resultados sugerem que o protocolo de PCR gerado é específico e pode ser considerado como uma nova ferramenta molecular para a identificação de cepas de F. graminearum. Além disso, o meio líquido formulado é uma alternativa barata para a avaliação da produção de GO por F. graminearum.
Resumo
Fusarium graminearum isolates causing Fusarium head blight in wheat were collected in Brazil and analyzed by random amplified polymorphic DNA (RAPD) markers and vegetative compatibility grouping (VCG). Nitrate non-utilizing mutants (nit) from each isolate were paired to verify heterokaryon formation. Three VCGs were identified among F. graminearum isolates: VCG1 included F-2, F-3 and F-4 isolates; VCG2 included F-1, F-6 and F-9 isolates; VCG3 included F-5, F-7 and F-8 isolates. Based on PCR amplification with eight different primers, the isolates showed great genetic similarity among themselves. Dendrogram analysis demonstrated two RAPD groups: Group A, consisting of isolates F-2 and F-9, and Group B, composed of the remaining isolates. Results suggest the clonal origin of F. graminearum isolates.
Isolados de Fusarium graminearum, obtidos de espigas de trigo com sintomas de Giberela, foram analisados pela técnica do Polimorfismo de DNA Amplificado ao Acaso (RAPD) e pelos Grupos de Compatibilidade Vegetativa (GCV). Mutantes auxotróficos (nit) de cada isolado foram pareados em todas as combinações possíveis, para a formação de heterocários. Três GCVs foram identificados: GCV1, incluindo os isolados F-2, F-3 e F-4; GCV2, incluindo os isolados F-1, F-6 e F-9; e GCV3, formado pelos isolados F-5, F-7 e F-8. Dois grupos foram identificados com base nos marcadores de RAPD: o grupo A, formado pelos isolados F-2 e F-9, e o grupo B, composto pelos demais isolados, os quais apresentaram grande similaridade entre si. Os resultados sugerem a origem clonal dos isolados de F. graminearum analisados.
Resumo
Fusarium graminearum isolates causing Fusarium head blight in wheat were collected in Brazil and analyzed by random amplified polymorphic DNA (RAPD) markers and vegetative compatibility grouping (VCG). Nitrate non-utilizing mutants (nit) from each isolate were paired to verify heterokaryon formation. Three VCGs were identified among F. graminearum isolates: VCG1 included F-2, F-3 and F-4 isolates; VCG2 included F-1, F-6 and F-9 isolates; VCG3 included F-5, F-7 and F-8 isolates. Based on PCR amplification with eight different primers, the isolates showed great genetic similarity among themselves. Dendrogram analysis demonstrated two RAPD groups: Group A, consisting of isolates F-2 and F-9, and Group B, composed of the remaining isolates. Results suggest the clonal origin of F. graminearum isolates.
Isolados de Fusarium graminearum, obtidos de espigas de trigo com sintomas de Giberela, foram analisados pela técnica do Polimorfismo de DNA Amplificado ao Acaso (RAPD) e pelos Grupos de Compatibilidade Vegetativa (GCV). Mutantes auxotróficos (nit) de cada isolado foram pareados em todas as combinações possíveis, para a formação de heterocários. Três GCVs foram identificados: GCV1, incluindo os isolados F-2, F-3 e F-4; GCV2, incluindo os isolados F-1, F-6 e F-9; e GCV3, formado pelos isolados F-5, F-7 e F-8. Dois grupos foram identificados com base nos marcadores de RAPD: o grupo A, formado pelos isolados F-2 e F-9, e o grupo B, composto pelos demais isolados, os quais apresentaram grande similaridade entre si. Os resultados sugerem a origem clonal dos isolados de F. graminearum analisados.
Resumo
Fusarium fungi are known to be pathogenic for plants and mycotoxin producers. The in vitro production of deoxynivalenol and zearalenone was qualitatively evaluated in 24 different isolates of Fusarium graminearum collected from small cereals associated with the scab disease, in southern Brazil. Isolates were cultivated in rice during 14 days at 28ºC. Cultivates were extracted with methanol:water (40:60 v/v) and analyzed by thin layer chromatography. Other trichothecenes (diacetoxyscirpenol, fusarenon-X, neosolaniol and nivalenol) and zearalenol, often produced by Fusarium, were also analyzed. In the conditions used, it was possible to detect zearalenone and deoxynivalenol in 67% and 33% of the isolates, respectively. The presence of zearalenol, diacetoxyscirpenol and fusarenone was also detected. None of the isolates was found to produce nivalenol or neosolaniol.
Fungos do gênero Fusarium são bem conhecidos como patógenos para plantas e como produtores de micotoxinas. O objetivo deste trabalho foi avaliar qualitativamente a produção "in vitro" de desoxinivalenol e de zearalenona, em 24 diferentes isolados de Fusarium graminearum coletados a partir de cereais associados à doença Giberela na Região Sul do Brasil. Os isolados foram cultivados em arroz, durante 14 dias, a 28ºC. Os cultivos foram extraídos com metanol:água (40:60, v/v) e analisados por cromatografia em camada delgada. Outros tricotecenos (diacetoxiscirpenol, fusarenona-X, neosolaniol e nivalenol) e zearalenol, freqüentemente produzidos por Fusarium, também foram avaliados. Nas condições utilizadas, foi possível determinar o perfil de produção dessas micotoxinas, sendo que 67% dos isolados produziram zearalenona e 33% dos isolados produziram desoxinivalenol. Também foram detectadas as presenças de zearalenol, diacetoxiscirpenol e fusarenona. Finalmente, em nenhum dos isolados estudados foram encontrados nivalenol e neosolaniol.