Effect of Metalaxyl Resistance and Cultivar Resistance on Control of Phytophthora nicotianae in Tobacco

Phytophthora nicotianae is a devastating root and stem pathogen of tobacco (Nicotiana tabacum) in South Africa. Growers strive to control the resulting disease, known as black shank, with metalaxyl treatments and resistant cultivars. The aim of this study was to consider whether development of metalaxyl resistance in P. nicotianae has contributed to poor disease control and if recently developed cultivars with high levels of resistance require metalaxyl for effective control. One hundred and thirty-two isolates of P. nicotianae were screened for sensitivity to metalaxyl. P. nicotianae isolates from most tobacco farms were metalaxyl sensitive. Growth of most isolates was inhibited completely at 1.0 μg a.i./ml. However, isolates from the MKTV tobacco producing area showed EC50 values ranging from 1.02 μg a.i./ml to 3.57 μg a.i./ml. Twenty-one tobacco cultivars were planted and treated with and without metalaxyl in two different growing seasons to evaluate their resistance to P. nicotianae and the value of using metalaxyl. Hicks was the most susceptible cultivar. Vuma/3/46, LK30/40/60-1, and LK33/60 exhibited the greatest resistance to P. nicotianae. Use of metalaxyl in combination with moderately resistant cultivars such as NC60 × TL33 and LK10/80/60 effectively reduced black shank in the field. Resistant cultivars were healthy and no significant difference between metalaxyl treated and untreated plants was observed.

Polyphasic identification of a Rhizopus species and a putative novel Gongronella which both degrade the fungicide metalaxyl

Apresentação efetuada na MicroBiotec’15

Enhanced degradation of metalaxyl in Gley Humic and Dark Red Latosol

Enhanced degradation of the fungicide metalaxyl was investigated in two soils: a gley humic (GH) and a Dark Red Latosol (LE), collected at sites never exposed to the fungicide. The soil samples were treated with successive applications of metalaxyl as a commercial formulation and 14C-metalaxyl in laboratory. Metalaxyl biodegradation was analyzed during 63 days by means of radiometric techniques to verify biomineralization and degradation product formation from the applied 14C-metalaxyl. Although biomineralization (maximum of 14 and 8% in the GH and LE soils, respectively), and partial degradation (about 32 and 48%, respectively) were detected in both soils, enhanced degradation was verified only in the GH soil. Results proved that metalaxyl behaves differently in soils.

Enhanced degradation of metalaxyl in agricultural soils of São Paulo State, Brazil

This work investigated the effect of repeated applications on enhanced degradation of metalaxyl in two different agricultural soils used for cultivation of orange and lemon from Casa Branca and Itapetininga districts of São Paulo State, Brazil. Soil samples were collected from areas repeatedly treated with commercial ridomil 50GR for six successive years, and from other areas never exposed to this fungicide. At the laboratory, soil samples received a 14C-metalaxyl solution and its degradation was studied through radiometric techniques to measure biomineralization and recovery of extractable- and soil-bound products. Enhanced degradation was verified only in one soil, although partial degradation and mineralization of the fungicide were detected in both soils. The different rates and patterns of metalaxyl degradation in the soils were probably due to their different physical, chemical, and biological characteristics.

Toxicity of triclosan, penconazole and metalaxyl on Caulobacter crescentus and a freshwater microbial community as assessed by flow cytometry.

Biocides are widely used for domestic hygiene, agricultural and industrial applications. Their widespread use has resulted in their introduction into the environment and raised concerns about potential deleterious effects on aquatic ecosystems. In this study, the toxicity of the biocides triclosan, penconazole and metalaxyl were evaluated with the freshwater bacterium Caulobacter crescentus and with a freshwater microbial community using a combination of single- and double-stain flow cytometric assays. Growth of C.  crescentus and the freshwater community were repressed by triclosan but not by penconazole or metalaxyl at concentrations up to 250 μM. The repressive effect of triclosan was dependent on culture conditions. Caulobacter crescentus was more sensitive to triclosan when grown with high glucose at high cell density than when grown directly in sterilized lake water at low cell density. This suggests that the use of conventional growth conditions may overestimate biocide toxicity. Additional experiments showed that the freshwater community was more sensitive to triclosan than C.  crescentus, with 10 nM of triclosan being sufficient to repress growth and change the phylogenetic composition of the community. These results demonstrate that isolate-based assays may underestimate biocide toxicity and highlight the importance of assessing toxicity directly on natural microbial communities. Because 10 nM of triclosan is within the range of concentrations observed in freshwater systems, these results also raise concerns about the risk of introducing triclosan into the environment.

Mineralisation of the fungicide 14C-labelled metalaxyl fungicide in Brazilian soils.

Laboratory incubation experiments were carried out to estimate the mineralisation of metalaxyl 14C {N-(2-6 dimethyphenyl)-N-(methoxyacetyl) alanine methyl ester} in four Brazilian soils with different physico-chemical properties, at 3 and 30 ?g a.i. g-1. In the Petrolina sandy soil the mineralisation presented higher 14CO2 production rates, at two essayed concentrations, after 70 days. Microbiological studies were done to determine the numbers of bacteria, actinobacteria and fungi (CFU g-1 soil). In relation with other microbial community, bacterial population demonstrated to be a major component of the cultivable heterotrophic community after the application.of the compound. No detectable metabolites were found in this study. The results suggest that soil properties and application history may have a strong influence on the fungicide behavior in these soil samples.

Mineralisation of 14c-labelled metalaxyl fungicide in Brazilian soils.

2008

A global marker database for Phytophthora infestans

A marker database was compiled for isolates of the potato and tomato late blight pathogen, Phytophthora infestans, originating from 41 locations which include 31 countries plus 10 regions within Mexico. Presently, the database contains information on 1,776 isolates for one or more of the following markers: restriction fragment length polymorphism (RFLP) fingerprint consisting of 23 bands; mating type; dilocus allozyme genotype; mitochondrial DNA haplotype; sensitivity to the fungicide metalaxyl; and virulence. In the database, 305 entries have unique RFLP fingerprints and 258 entries have unique multilocus genotypes based on RFLP fingerprint, dilocus allozyme genotype, and mating type. A nomenclature is described for naming multilocus genotypes based on the International Organization for Standardization (ISO) two-letter country code and a unique number, Forty-two previously published multilocus genotypes are represented in the database with references to publications. As a result of compilation of the database, seven new genotypes were identified and named. Cluster analysis of genotypes from clonally propagated populations worldwide generally confirmed a previously published classification of old and new genotypes. Genotypes from geographically distant countries were frequently clustered, and several old and new genotypes were found in two or more distant countries. The cluster analysis also demonstrated that A2 genotypes from Argentina differed from all others. The database is available via the Internet, and thus can serve as a resource for Phytophthora workers worldwide.

Identifying pesticide use patterns among flower growers to assess occupational exposure to mixtures

Objectives Exposure assessment to a single pesticide does not capture the complexity of the occupational exposure. Recently, pesticide use patterns analysis has emerged as an alternative to study these exposures. The aim of this study is to identify the pesticide use pattern among flower growers in Mexico participating in the study on the endocrine and reproductive effects associated with pesticide exposure. Methods A cross-sectional study was carried out to gather retrospective information on pesticide use applying a questionnaire to the person in charge of the participating flower growing farms. Information about seasonal frequency of pesticide use (rainy and dry) for the years 2004 and 2005 was obtained. Principal components analysis was performed. Results Complete information was obtained for 88 farms and 23 pesticides were included in the analysis. Six principal components were selected, which explained more than 70% of the data variability. The identified pesticide use patterns during both years were: 1. fungicides benomyl, carbendazim, thiophanate and metalaxyl (both seasons), including triadimephon during the rainy season, chlorotalonyl and insecticide permethrin during the dry season; 2. insecticides oxamyl, biphenthrin and fungicide iprodione (both seasons), including insecticide methomyl during the dry season; 3. fungicide mancozeb and herbicide glyphosate (only during the rainy season); 4. insecticides metamidophos and parathion (both seasons); 5. insecticides omethoate and methomyl (only rainy season); and 6. insecticides abamectin and carbofuran (only dry season). Some pesticides do not show a clear pattern of seasonal use during the studied years. Conclusions The principal component analysis is useful to summarise a large set of exposure variables into smaller groups of exposure patterns, identifying the mixtures of pesticides in the occupational environment that may have an interactive effect on a particular health effect.

Efeito do míldio na concentração de sólidos solúveis totais em frutos do meloeiro

O míldio causado por Pseudoperonospora cubensis se constitui em uma das mais importantes doenças do meloeiro (Cucumis melo) no Nordeste brasileiro, região que concentra mais de 80% da produção nacional de melão. Epidemias de míldio são referidas, constantemente, causando severas perdas na produção de frutos. O efeito da doença no teor de sólidos solúveis totais (SST), importante fator de aferição qualitativa do fruto, foi determinado em meloeiro do tipo amarelo inodoro sob condições naturais. Intensidades variáveis da doença foram obtidas, variando-se o período de cultivo e a aplicação de fungicidas recomendados para o controle (tiofanato metílico + chlorotalonil, metalaxyl + mancozeb e oxicloreto de cobre). Os resultados revelaram que o aumento da severidade da doença resulta numa acentuada redução no teor de SST, proporcionando decréscimos de até 49%. Entre os parâmetros usados para se estimar os efeitos relacionados à doença, a área abaixo da curva de progresso da doença foi o mais eficiente, principalmente quando a pressão da doença aumentou.

Monitoramento da população de Phytophthora infestans na região da Zona da Mata de Minas Gerais de 1998 a 2000

Foram caracterizados 212 isolados de Phytophthora infestans obtidos de 51 lavouras de tomate (Lycopersicon esculentum)e batata (Solanum tuberosum), em sete municípios da Zona da Mata, MG. Todos os isolados tiveram o grupo de compatibilidade determinado; 96 isolados foram caracterizados para a isoenzima glucose 6-fosfato-isomerase (Gpi); 71 isolados foram analisados quanto à resistência ao metalaxyl; e determinou-se o espectro de virulência de 46 isolados. Todos os 212 isolados testados foram classificados como do grupo A1 de compatibilidade. A maioria dos isolados testados para Gpi (95) apresentou o fenótipo 86/100, típico da linhagem clonal US-1. Apenas um isolado apresentou o fenótipo 100/100 para Gpi. Quanto à resistência ao metalaxyl, em 1998 a freqüência de isolados sensíveis, intermediários e resistentes foi de 40%, 40% e 20%, respectivamente, e em 2000 de 3,2%; 61,3% e 35,5%, respectivamente. Quanto ao espectro de virulência, todos os 46 isolados analisados foram virulentos sobre a cultivar de tomate 'Kada'. A maioria foi virulenta em plantas de tomate com os genes Ph1 (91%) ou Ph2 (95 %). Todos os isolados foram virulentos em batata 'Bintje'. Houve pequena variação do espectro de virulência sobre clones de batata, quando esses foram inoculados com isolados coletados em diferentes anos. Há evidências que a população de P. infestans da Zona da Mata, MG é constituída de isolados da linhagem clonal US-1, de várias raças e baixa sensibilidade ao fungicida metalaxyl.

Combinação dos tratamentos biológico e químico de semente de milho

A integração de métodos de proteção de semente de milho (Zea mays) poderá reduzir consideravelmente o uso de pesticidas nessa cultura, melhorando a eficiência de controle e diminuindo a poluição do ambiente. O objetivo do presente trabalho foi determinar o efeito da combinação dos controles biológico e químico no tratamento de semente de milho. Como controle biológico, foi usado o bioprotetor Embr. 144 e, como controle químico, foi empregado fludioxonil + metalaxyl M. Dois experimentos de laboratório e dois de campo foram conduzidos na município de Passo Fundo, RS. Os tratamentos que continham o bioprotetor Paenibacillus macerans (Embr. 144) ou o fungicida fludioxonil + metalaxyl-M, ou ambos, reduziram significativamente o nível de infecção dos patógenos nas sementes, destacando-se os tratamentos que continham ambos, os quais eliminaram por completo todos os patógenos da semente de milho, demonstrando que a interação entre o bioprotetor e o fungicida apresentou benefícios. Mesmo quando as doses do bioprotetor e do fungicida no tratamento integrado foram reduzidas à metade, o nível de controle dos patógenos de milho foi superior ao nível de controle do bioprotetor e do fungicida isoladamente, com as doses normais. Todos os tratamentos aumentaram significativamente a germinação de sementes e o rendimento de grãos de milho em relação à testemunha, entretanto, os efeitos foram superiores quando houve a combinação dos tratamentos do bioprotetor com o fungicida. A combinação do bioagente P. macerans com o produto fludioxonil + metalaxyl-M poderá ter importante impacto na redução do uso de fungicidas, para alcançar a agricultura sustentável e proteger o ambiente.

Efeito do início de ocorrência do míldio na produção de frutos do meloeiro

O míldio, causado por Pseudoperonospora cubensis, é a principal doença fúngica da cultura do meloeiro (Cucumis melo) no período chuvoso nas regiões produtoras do Nordeste brasileiro, e para o seu controle são realizadas até dez aplicações de fungicidas. O estabelecimento de sistemas de previsão de danos causados por esta doença requer uma estimativa sob várias situações epidêmicas. Os danos causados pelo míldio à produção de frutos do meloeiro, levando-se em consideração o início do aparecimento dos sintomas e o estádio de desenvolvimento da planta, foram determinados em três experimentos de campo. Utilizou-se a aplicação de uma das misturas fungicidas tiofanato metílico + clorotalonil ou metalaxyl + mancozeb no estabelecimento de menores intensidades de míldio. Houve uma significativa redução do rendimento de frutos quando a doença teve início aos 24 e 36 dias, porém, quando a doença iniciou-se aos 47 dias, nenhum efeito na produção foi observado. Concluiu-se que os aspectos relacionados com a duração e, notadamente, com o início da epidemia, em relação ao ciclo do hospedeiro, devem ser considerados no estabelecimento de modelos de prognósticos de danos à produção.