A decrease in bulk water and mannitol and accumulation of trehalose and trehalose-based oligosaccharides define a two-stage maturation process towards extreme stress resistance in ascospores of Neosartorya fischeri (Aspergillus fischeri)

The effects of inter-crop cultivation Between rows of citrus crop on spreading of Guignardia citricarpa Ascospores and in the citrus black spot occurrence

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of temperature, leaf wetness, and rainfall on the production of Guignardia citricarpa ascospores and on back spot severity on sweet orange

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Austropleospora osteospermi gen. et sp nov and its host specificity and distribution on Chrysanthemoides monilifera ssp rotundata in Australia.

Hendersonia osteospermi was found for the first time in Australia on leaf spots of the introduced invasive plant Chrysanthemoides monilifera ssp. rotundata (bitou bush) in coastal regions of New South Wales. Pathogenicity tests on species from 11 tribes in the family Asteraceae, demonstrated that H. osteospermi caused severe necrosis on leaves and stems of C. monilifera ssp. rotundata and its congener C. monilifera ssp. monilifera (boneseed). Small necrotic spots also developed on Osteospermum fruticosum and Dimorphotheca cuneata in the Calenduleae and on Helianthus annuus (sunflower) in the Heliantheae. None of the other plant species tested developed leaf spots, although H. osteospermi was re-isolated from senescent leaves of Cynara scolymus (globe artichoke) in the Cynareae and Vernonia cinerea in the Vernonieae. Single ascospores from ascomata of a Pleospora-like fungus found on diseased stems of bitou bush produced H. osteospermi in culture, which proved the anamorph/teleomorph connection. The ITS region of both a single-ascospore isolate and a single-conidium isolate were sequenced and found to be identical. The taxonomic status of H. osteospermi is re-examined and Austropleospora osteospermi gen. et sp. nov. is described as its teleomorph based on morphology, host range tests and DNA sequence analysis. The potential of A. osteospermi for the biological control of bitou bush and boneseed in Australia is discussed.

A Demonstration of the Role ofhetGenes in Heterokaryon Formation inNeurosporaunder Simulated Field Conditions

An experimental system was developed for assessing the role ofhetgenes in heterokaryon formation inNeurosporain nature. Burned sugar cane segments planted in soil were infected using a mixture of mutant ascospores of two genotypes.Neurosporaramified in the cane and erupted as distinct pustules of conidia. When ascospores carried identicalhetalleles, the (macro) conidial pustules which formed were heterokaryotic. On the other hand, when ascospores carried dissimilarhetalleles, the pustules were homokaryotic. These results showed that stable heterokaryons between compatible strains can form in nature. When two strains are growing together on a natural substrate, heterozygosity athetloci serves to maintain their individuality.

Life-history of Neurospora intermedia in a sugar cane field

The life-history of Neurospora in nature has remained largely unknown. The present study attempts to remedy this. The following conclusions are based on observation of Neurospora on fire-scorched sugar cane in agricultural fields, and reconstruction experiments using a colour mutant to inoculate sugar cane burned in the laboratory. The fungus persists in soil as heat-resistant dormant ascospores. These are activated by a chemical(s) released into soil from the burnt substrate. The chief diffusible activator of ascospores is furfural and the germinating ascospores infect the scorched substrate. An invasive mycelium grows progressively upwards inside the juicy sugar cane and produces copious macroconidia externally through fire-induced openings formed in the plant tissue, or by the mechanical rupturing of the plant epidermal tissue by the mass of mycelium. The loose conidia are dispersed by wind and/or foraged by microfauna. It is suggested that the constant production of macroconidia, and their ready dispersal, serve a physiological role: to drain the substrate of minerals and soluble sugars, thereby creating nutritional conditions which stimulate sexual reproduction by the fungus. Sexual reproduction in the sugar-depleted cellulosic substrate occurs after macroconidiation has ceased totally and is favoured by the humid conditions prevailing during the monsoon rains. Profuse microconidiophores and protoperithecia are produced simultaneously in the pockets below the loosened epidermal tissue. Presumably protoperithecia are fertilized by microconidia which are possibly transmitted by nematodes active in the dead plant tissue. Mature perithecia release ascospores in situ which are passively liberated in the soil by the disintegration of the plant material and are, apparently, distributed by rain or irrigation water.

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.

The role of seeds and airborne inoculum in the initiation of leaf blotch (Rhynchosporium secalis) epidemics in winter barley

Both airborne spores of Rhynchosporium secalis and seed infection have been implied as major sources of primary inoculum for barley leaf blotch (scald) epidemics in fields without previous history of barley cropping. However, little is known about their relative importance in the onset of disease. Results from both quantitative real-time PCR and visual assessments indicated that seed infection was the main source of inoculum in the field trial conducted in this study. Glasshouse studies established that the pathogen can be transmitted from infected seeds into roots, shoots and leaves without causing symptoms. Plants in the field trial remained symptomless for approximately four months before symptoms were observed in the crop. Covering the crop during part of the growing season was shown to prevent pathogen growth, despite the use of infected seed, indicating that changes in the physiological condition of the plant and/or environmental conditions may trigger disease development. However, once the disease appeared in the field it quickly became uniform throughout the cropping area. Only small amounts of R. secalis DNA were measured in 24 h spore-trap tape samples using PCR. Inoculum levels equivalent to spore concentrations between 30 and 60 spores per m3 of air were only detected on three occasions during the growing season. The temporal pattern and level of detection of R. secalis DNA in spore tape samples indicated that airborne inoculum was limited and most likely represented rain-splashed conidia rather than putative ascospores.

Influence of package, type of apple juice and temperature on the production of patulin by Byssochlamys nivea and Byssochlamys fulva

Although the production of patulin in apple fruits is mainly by Penicillium expansum, there is no information on the ability of heat resistant moulds that may survive pasteurization to produce this mycotoxin in juice packages during storage and distribution. In this study, the production of patulin by Byssochlamys spp (Byssochlamys nivea FRR 4421, B. nivea ATCC 24008 and Byssochlamys fulva IOC 4518) in cloudy and clarified apple juices packaged in laminated paperboard packages or in polyethylene terephthalate bottles (PET) and stored at both 21 degrees C and 30 degrees C, was investigated. The three Byssochlamys strains were able to produce patulin in both cloudy and clarified apple juices. Overall, the lower the storage temperature, the lower the patulin levels and mycelium dry weight in the apple juices (p<0.05). The greatest variations in pH and degrees Brix were observed in the juices from which the greatest mycelium dry weights were recovered. The maximum levels of patulin recovered from the juices were ca. 150 mu g/kg at 21 degrees C and 220 mu g/kg at 30 degrees C. HPLC-UV, HPCL-DAD and mass spectrometry analyses confirmed the ability of B. fulva IOC 4518 to produce patulin. Due to the heat resistance of B. nivea and B. fulva and their ability to produce patulin either in PET bottles or in laminated paperboard packages, the control of contamination and the incidence of these fungi should be a matter of concern for food safety. Control measures taken by juice industries must also focus on controlling the ascospores of heat resistant moulds. (C) 2010 Elsevier B.V. All rights reserved.

Susceptibility of fruits of the 'Valência' and 'Natal' sweet orange varieties to Guignardia citricarpa and the influence of the coexistence of healthy and symptomatic fruits

Em pomar de laranjeiras 'Valência' e 'Natal' avaliou-se a importância da presença de frutos sintomáticos da mancha preta citros (MPC) na severidade da doença nos frutos cítricos da safra subseqüente. Adicionalmente, avaliou-se o estádio de suscetibilidade dos frutos dessas variedades. Frutos foram protegidos com sacos de papel cristal a partir do estádio de 75% de pétalas caídas em outubro de 2000, até abril de 2001. Frutos foram expostos, em intervalos semanais, da 1ª à 24ª semana. Esse processo se deu tanto em plantas onde os frutos da safra remanescente foram previamente colhidos, como naquelas cujos frutos sintomáticos da safra remanescente permaneceram até a sua queda natural. Avaliou-se a severidade da doença usando uma escala de notas que variou de 0 (ausência de sintomas) a 6 (sintomas severos). Observou-se que para as duas variedades os conídios de Phyllosticta citricarpa, formados nas lesões dos frutos da safra remanescente, não provocaram incremento significativo na severidade da doença dos frutos da safra subseqüente. A proteção dos frutos até 10ª semana após a queda de pétalas não influenciou na quantidade final de lesões, indicando que as descargas de ascósporos que ocorreram a partir desse momento foram, provavelmente, responsáveis pela severidade da doença. Frutos que ficaram expostos entre a 20ª a 24ª semanas após a queda de 75% de pétalas mostraram-se sintomáticos, indicando que nesse estádio frutos encontravam-se suscetíveis ao patógeno.

Influência da temperatura e da luminosidade no desenvolvimento de Guignardia citricarpa, agente causal da mancha preta dos frutos cítricos

O presente trabalho teve como objetivo avaliar sob condições de laboratório, o efeito de temperatura (15, 18, 20, 21, 24, 25, 27, 30, 33 e 35 ± 1 ºC) nas condições escuro, luz contínua e fotoperíodo 12/12 h, na produção de pseudotécios de isolados de Guignardia citricarpa, provenientes de regiões cítricolas dos Estados de São Paulo e Rio de Janeiro. Discos de folhas de limoeiro 'Cravo' (Citrus limonia), de 12 mm de diâmetro, foram autoclavados e depositadas (parte abaxial) na superfície do meio de cultura constituído por ágar-água 2%. Foram colocados quatro discos de folhas por placa onde, de forma conjunta e intercalar aos mesmos, depositaram-se dois discos obtidos de colônias de Phyllosticta citricarpa, com 21 dias de incubação. Foi, também, estudado o efeito da temperatura e do tempo de incubação (2, 8 e 16 h) na germinação dos ascósporos. Após 21 dias de incubação, a ótima temperatura ajustada pela função beta generalizada, para produção de pseudotécios deu-se a 26 e 22,5 a 27,5 °C, sob condição de escuro e de luz, respectivamente. Observou-se também produção de pseudotécios a 27 ºC em fotoperíodo 12/12 h. em estudo complementar foi verificado que, aos 19 dias, a 27 ºC, cerca de 90% dos pseudotécios haviam alcançado a maturidade, com abundante produção de ascósporos. A maior porcentagem de ascósporos germinados foi constatada na temperatura de 24 ºC, após 16 h de incubação. Dentre as vantagens alcançadas, incluem-se a possibilidade (i) da produção massal de ascósporos em curto período de tempo, e (ii) da padronização do inóculo, tanto qualitativa, quanto quantitativa.

Isolation and mycelial growth of Diehliomyces microsporus: effect of culture medium and incubation temperature

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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.

Controle da mancha preta dos frutos cítricos mediante manejo cultural

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)