Dynamics of QoI sensitivity in Mycosphaerella fijiensis in Costa Rica during 2000 to 2003

From 1997 onward, the strobilurin fungicide azoxystrobin was widely used in the main banana-production zone in Costa Rica against Mycosphaerella fijiensis var. difformis causing black Sigatoka of banana. By 2000, isolates of M. fijiensis with resistance to the quinolene oxidase inhibitor fungicides were common on some farms in the area. The cause was a single point mutation from glycine to alanine in the fungal target protein, cytochrome b gene. An amplification refractory mutation system Scorpion quantitative polymerase chain reaction assay was developed and used to determine the frequency of G 143A allele in samples of M. fijiensis. Two hierarchical surveys of spatial variability, in 2001 and 2002,found no significant variation in frequency on spatial scales <10 in. This allowed the frequency of G143A alleles on a farm to be estimated efficiently by averaging single samples taken at two fixed locations. The frequency of G 143A allele in bulk samples from I I farms throughout Costa Rica was determined at 2-month intervals. There was no direct relationship between the number of spray applications and the frequency of G143A on individual farms. Instead, the frequency converged toward regional averages, presumably due to the large-scale mixing of ascospores dispersed by wind. Using trap plants in an area remote from the main producing area, immigration of resistant ascospores was detected as far as 6 km away both with and against the prevailing wind.

Effects of fungicide dose and mixtures on selection for triazole resistance in Mycosphaerella graminicola under field conditions

The effects of varying doses of fungicides, alone or in mixtures, on selection for triazole resistance were examined under field conditions. Two experiments were conducted using the triazole fungicide fluquinconazole with the strobilurin fungicide azoxystrobin as a mixture partner. Inoculated wheat plots with a known ratio of more sensitive to less sensitive isolates of the leaf blotch fungus Mycosphaerella graminicola were sprayed with fungicide and sampled once symptoms had appeared. Selection for fluquinconazole resistance increased in proportion to the dose, up to one-half of the full dose (the maximum tested) in both experiments. At the higher doses of fluquinconazole, the addition of azoxystrobin was associated with a decrease in selection (nonsignificant in the first experiment) for triazole resistance. Control by low doses of fluquinconazole was increased by mixture with azoxystrobin, but at higher doses mixture with azoxystrobin sometimes decreased control, so that reduced selection was obtained at the cost of some reduction in control. The effects on resistance are not necessarily general consequences of mixing fungicides, and suggest that the properties of any specific mixture may need to be demonstrated experimentally. Selection was inversely related to control in the unmixed treatments in both experiments, but the relationship was weaker in the mixtures with azoxystrobin.

Sensitivity distributions and cross-resistance patterns of Mycosphaerella graminicola to fluquinconazole, prochloraz and azoxystrobin over a period of nine years

The sensitivity of 73 isolates of Mycosphaerella graminicola collected over the period 1993–2002 from wheat fields in South England was tested in vitro against the triazole fluquinconazole, the strobilurin azoxystrobin and to the imidazole prochloraz. Over the sampling period, sensitivity of the population to fluquinconazole and prochloraz decreased by factors of approximately 10 and 2, respectively, but there was no evidence of changes in sensitivity to azoxystrobin. There was no correlation between sensitivity to fluquinconazole and prochloraz, but there was a weak negative cross-resistance between fluquinconazole and azoxystrobin.

Accounting for the economic risk, caused by variation in disease severity, in fungicide dose decisions: exemplified for Mycosphaerella graminicola on winter wheat

A method is presented to calculate economic optimum fungicide doses accounting for the risk-aversion of growers responding to variability in disease severity between crops. Simple dose-response and disease-yield loss functions are used to estimate net disease-related costs (fungicide cost, plus disease-induced yield loss) as a function of dose and untreated severity. With fairly general assumptions about the shapes of the probability distribution of disease severity and the other functions involved, we show that a choice of fungicide dose which minimises net costs on average across seasons results in occasional large net costs caused by inadequate control in high disease seasons. This may be unacceptable to a grower with limited capital. A risk-averse grower can choose to reduce the size and frequency of such losses by applying a higher dose as insurance. For example, a grower may decide to accept ‘high loss’ years one year in ten or one year in twenty (i.e. specifying a proportion of years in which disease severity and net costs will be above a specified level). Our analysis shows that taking into account disease severity variation and risk-aversion will usually increase the dose applied by an economically rational grower. The analysis is illustrated with data on septoria tritici leaf blotch of wheat caused by Mycosphaerella graminicola. Observations from untreated field plots at sites across England over three years were used to estimate the probability distribution of disease severities at mid-grain filling. In the absence of a fully reliable disease forecasting scheme, reducing the frequency of ‘high loss’ years requires substantially higher doses to be applied to all crops. Disease resistant cultivars reduce both the optimal dose at all levels of risk and the disease-related costs at all doses.

Sulphate and sulphurous acid alter the relative susceptibility of wheat to Phaeosphaeria nodorum and Mycosphaerella graminicola

The relative abundances of DNA of Mycosphaerella graminicola and Phaeosphaeria nodorum in archived wheat samples are closely correlated with UK anthropogenic emissions of oxidized sulphur over the last 160 years. To test whether this could be a causal relationship, possible modes of action of sulphur on the two fungi were examined. Mycelial growth of the two fungi in solutions of sulphurous acid was similar. Sulphurous acid at pH 4 reduced percentage germination of P. nodorum conidia more strongly than M. graminicola conidia. In spray inoculations of wheat cv. Squarehead’s Master, Cappelle Desprez and Riband with water or sulphurous acid (pH 4), the ratio of leaves infected by P. nodorum to leaves infected by M. graminicola was increased by factors of 2.5, 2.1 and 0.6, respectively at pH 4. The same three cultivars of wheat were grown in sand and vermiculite and fertilized with nutrient solution containing 2.5 or 0.5 mM sulphate. Both pathogens infected less frequently at 2.5 mM sulphate, by a factor of about 2. The severity of infection by M. graminicola was reduced on all three cultivars by a factor of about 4-5 at 2.5mM sulphate, but severity of P. nodorum was reduced only by a factor of about 2. Both elevated free sulphate concentrations in soil and sulphite in rainwater could therefore increase the prevalence of P. nodorum relative to M. graminicola, which is consistent with the historical changes in abundance

Diversity of Mycosphaerella spp. and Teratosphaeria spp. in Eucalyptus globulus plantations in Brazil

Among the most important diseases affecting Eucalyptus is Mycosphaerella Leaf Disease (MLD) caused by Mycosphaerella spp. and Teratosphaeria spp. MLD has led to significant losses in eucalypt plantations in the South and Southeast Region of Brazil, as well as in several countries such as Portugal, Spain, South Africa and Australia. Symptoms of MLD include localized necrotic spots, early defoliation in juvenile plants, stem cankers, early death of branches, and in some cases, atrophy and death. In the present study, single spore isolations from leaves of E. globulus from five locations in Brazil allowed the differentiation of species of Mycosphaerella and Teratosphaeria based on ascospore germination and growth in culture. These isolates were also subjected to sequence analysis of the ribosomal RNA internal transcribed spacer regions, which allowed their identification to species level. The results of this study showed that six species of Mycosphaerella and four species of Teratosphaeria were associated with leaves showing symptoms of MLD in E. globulus plantations in various locations of Brazil. © 2013 KNPV.

Ação de óleos essenciais no controle de sigatoka-negra (Mycosphaerella fijiensis) Morelet de bananeiras (Musa sp)

Pós-graduação em Ciências Biológicas (Botânica) - IBB

Population differentiation in the banana leaf spot pathogen Mycosphaerella musicola, examined at a global scale

Single-copy restriction fragment length polymorphism (RFLP) markers were used to determine the genetic structure of the global population of Mycosphaerella musicola, the cause of Sigatoka (yellow Sigatoka) disease of banana. The isolates of M. musicola examined were grouped into four geographic populations representing Africa, Latin America and the Caribbean, Australia and Indonesia. Moderate levels of genetic diversity were observed for most of the populations (H = 0.22-0.44). The greatest genetic diversity was found in the Indonesian population (H = 0.44). Genotypic diversity was close to 50% in all populations. Population differentiation tests showed that the geographic populations of Africa, Latin America and the Caribbean, Australia and Indonesia were genetically different populations. Using F-ST tests, very high levels of genetic differentiation were detected between all the population pairs (F-ST > 0.40), with the exception of the Africa and Latin America-Caribbean population pair. These two populations differed by only 3% (F-ST = 0.03), and were significantly different (P < 0.05) from all other population pairs. The high level of genetic diversity detected in Indonesia in comparison to the other populations provides some support for the theory that M. musicola originated in South-east Asia and that M. musicola populations in other regions were founded by isolates from the South-east Asian region. The results also suggest the migration of M. musicola between Africa and the Latin America-Caribbean region.

The genetic structure of Australian populations of Mycosphaerella musicola suggests restricted gene flow at the continental scale

Mycosphaerello musicolo causes Sigatoka disease of banana and is endemic to Australia. The population genetic structure of M. musicola in Australia was examined by applying single-copy restriction fragment length polymorphism probes to hierarchically sampled populations collected along the Australian cast coast. The 363 isolates studied were from 16 plantations at 12 sites in four different regions, and comprised 11 populations. These populations displayed moderate levels of gene diversity (H = 0.142 to 0.369) and similar levels of genotypic richness and evenness. Populations were dominated by unique genotypes, but isolates sharing the same genotype (putative clones) were detected. Genotype distribution was highly localized within each population, and the majority of putative clones were detected for isolates sampled from different sporodochia in the same lesion or different lesions on a plant. Multilocus gametic disequilibrium tests provided further evidence of a degree of clonality within the populations at the plant scale. A complex pattern of population differentiation was detected for M. musicola in Australia. Populations sampled from plantations outside the two major production areas were genetically very different to all other populations. Differentiation was much lower between populations of the two major production areas, despite their geographic separation of over 1,000 km. These results suggest low gene flow at the continental scale due to limited spore dispersal and the movement of infected plant material.

Mycosphaerella areola - the Teleomorph of Ramularia areola of cotton in Brazil, and its epidemiological significance.

2016

Sigatoka disease reduces the greenlife of bananas

Sigatoka disease (SD) of bananas is caused by the pathogenic fungus Mycosphaerella musicola Leach. This disease provokes necrotic lesions on leaves and serious infestations can lead to a substantial reduction in the leaf area of infected plants and thus to yield losses. In addition to these effects on yield, SO was found to have an impact on fruit quality, especially because exported bananas ripen prematurely. In the present work, a plantation survey and experiments have been conducted in Guadeloupe (FWI) to assess the effect of this disease on the greenlife of bananas harvested at a constant physiological age, as measured in degree-days (dd). Our results revealed that bananas harvested at 900 dd from plants with high Sigatoka disease severity had normal diameter growth, but a shorter greenlife (GL) than bananas harvested from uninfected plants. These results indicate that SD is directly responsible for the reduction of banana greenlife since the reduction of GL could not be attributed to the harvest of fruits at a more advanced physiological age (dd). Furthermore, a correlation was noted between SO severity and GL The potential physiological mechanisms involved are also discussed. (C) 2008 Elsevier Ltd. All rights reserved.

Características físico-quimicas, análise sensorial e conservação de frutos de cultivares de bananeira resistentes à sigatoca-negra

O cultivar Prata Anã é o mais plantada no Norte de Minas Gerais, o que demonstra estar a bananicultura regional baseada numa variedade suscetível à sigatoca-negra. Apesar de existirem cultivares resistentes em fase de recomendação para o cultivo na região, ainda são escassos os trabalhos relacionados a metodologias de conservação pós-colheita. Assim, o objetivo deste trabalho foi avaliar o período de armazenamento e as características físico-químicas e sensoriais dos cultivares de bananeira Fhia - 02 e Precioso, ambos resistentes à sigatoca-negra, e o efeito da atmosfera modificada associada à refrigeração. Foram realizados três experimentos, o primeiro em temperatura ambiente, o segundo a 12 ºC e o terceiro a 15 ºC. Esses experimentos foram instalados no delineamento em blocos casualizados em esquema de parcelas subdivididas no tempo, tendo nas parcelas um fatorial 2 (com e sem embalagem) x 2 (Fhia - 02 e Precioso) e nas subparcelas as avaliações em intervalo de seis e dois dias para frutos armazenados nas temperaturas de 12, 15 e 25 ºC, com quatro repetições e três frutos por unidade experimental. Após o tratamento, os frutos foram dispostos em bandejas de poliestireno expandido (sem embalagem) e embalagens de 10 e 16 µm e umidade relativa de 90%. As características avaliadas foram: tamanho, diâmetro, cor, firmeza, perda de matéria fresca, sólidos solúveis totais (SST), acidez total titulável (ATT), SST/ATT, pH e análise sensorial. O uso da embalagem associada à refrigeração permitiu armazenar os frutos por 28 dias, mantendo as características físico-químicas.

Studies on the eucalyptus leaf disease complex in Portugal

Native from south eastern Australia, Eucalyptus globulus is the main species in eucalypts plantations in Portugal. The most serious foliar disease in eucalypt plantations is linked to Mycosphaerella senso lato, which affects young trees in the juvenile phase foliage causing leaf necrosis. This disease results in reduced growth rate of the host and lower wood volume, thus causing significant productivity losses. The most common name for this disease was Mycosphaerella Leaf Disease that became inappropriate when most of the pathogens on eucalypts were re-distributed into several genera. The term "Eucalyptus Leaf Disease Complex" is now more appropriate. The overall aim of this thesis was to investigate the Eucalyptus Leaf Disease Complex in Portugal, focusing on species diversity, taxonomy and the role played by each species in the disease complex on Eucalyptus globulus. Literature on the Eucalyptus Leaf Disease Complex was reviewed and the species were distributed into several genera. A survey based on symptomatic leaves collected from several Eucalyptus globulus plantations and characterized by morphological and molecular tools provided an overview of species incidence and of the most frequent species in the disease complex. The present work reveals additional species of Mycosphaerella senso lato associated with eucalypt plantations in Portugal. Thus, five new records of Teratosphaeria and phylogenetically related species were added to the Iberian Peninsula, namely, Neodevriesia hilliana, for the first time on Myrtaceae; Quasiteratosphaeria mexicana, Teratosphaericola pseudoafricana, Teratosphaeria pluritubularis and Teratosphaeria lusitanica, a new species. Furthermore, new anamorphic structures were found and two new combinations were made. Regarding other genera, some species were observed for the first time, such as Cladosporium cladosporioides, Fusicladium eucalypti, Mycosphaerella madeirae, in the mainland. In addition to leave diseases, Teratosphaeria gauchensis was found causing a severe stem and trunk canker on Eucalyptus globulus. The aggressiveness of several species was compared to evaluate each species individually in the complex, permitting to distinguish different behaviours, from primary to secondary pathogens. Cladosporium cladosporioides, M. communis and M. lateralis, appeared to be more aggressive than Teratosphaeria nubilosa. In fact, contrary to the prevailing views on this disease complex, Teratosphaeria nubilosa is not the only species responsible for the disease, which clearly involves a complex of species acting together.

Plantio adensado não controla a sigatoka-negra da bananeira

Algumas publicações relatam que o adensamento populacional dos bananais reduz a severidade da sigatoka-negra (Mycosphaerella fijiensis). Instalou-se um ensaio com cinco tratamentos e quatro repetições. Os tratamentos 1.600, 2.000, 2.400, 2.800 e 3.200 plantas.ha-1 da cultivar D'Angola foram instalados em parcelas agrupadas de 2.000 m² cada. O tamanho das parcelas foi fixo e o número de plantas por parcela variou conforme o espaçamento adotado para cada população. A área de 2.000 m² foi dividida em quatro subáreas de 500 m², considerando-as como parcelas. Em cada subárea selecionaram-se 15 plantas centrais para serem avaliadas. Na época do florescimento registraram-se a severidade da doença na folha n.°10 e o número de folhas viáveis. Na colheita, a altura e o diâmetro do pseudocaule e o peso dos cachos, das pencas e dos frutos. A análise conjunta dos dados indica que todos os tratamentos foram semelhantes entre si e que o adensamento das plantas não controlou a sigatoka-negra.