Glomerella leaf spot (GLS) of apple is caused by three different Colletotrichum species complexes. This study evaluated the dispersal of Colletotrichum spores related to GLS temporal progress and defoliation. Spores were monitored by air and water runoff in different plant heights, and the temporal progress of GLS and defoliation were assessed. Spores of the pathogen were first cached in the lower part of the tree closer to the ground, confirming the importance of dead leaves on the ground as main source of primary inoculum. In plots with high primary inoculum, the disease increases exponentially during favorable weather conditions. The highest initial inoculum was found in the lower part of the tree, but the highest rate of the disease progress in the upper.

Glomerella leaf spot (GLS) and bitter rot (BR) on apples are often caused by Colletotrichum acutatum in Paraná State, Brazil. GLS control is difficult because of its rapid development, with an incubation period of only 2 days under favorable conditions. Therefore, producers use successive fungicide applications every season; however, failure to control GLS has been commonly reported. The objectives of this study were to determine the sensitivity of isolates of the C. acutatum species complex obtained from apple orchards in Brazil to mancozeb, thiophanate-methyl, and azoxystrobin fungicides. Isolates from the different parts of the plant (leaves, flowers, buds, and twigs) and cultivars (Gala and Eva) showed different levels of sensitivity to mancozeb, thiophanate-methyl, and azoxystrobin. For mancozeb, the frequencies of isolates were 25% highly resistant, 50% low-resistance, and 25% sensitive. For thiophanate-methyl, the frequencies of isolates were 72.2% highly resistant, 11.1% resistant, and 16.7% moderately resistant. For azoxystrobin, the frequencies of isolates were 11.1% highly resistant, 5.6% resistant, and 83.3% sensitive. Interestingly, no mutations in the β-tubulin and cytochrome b genes were observed in any of the isolates resistant to thiophanate-methyl and azoxystrobin fungicides.

The Glomerella leaf spot (GLS) and bitter rot (BR) are important apple diseases in Brazil, caused by species of Colletotrichum, which are usually related with quiescent infection. This study aimed to detect quiescent infections of Colletotrichum spp. in flowers and in unripe fruits from apple commercial orchard cultivars Eva and Gala. The pathogen was observed in various structures of the flower in both cultivars. In unripe fruits, the pathogen was recovered only from 'Eva'. Five isolates were obtained and were inoculated by mycelial (with and without wound) on ripe apple fruits and by conidia suspension (without wound) on ripe apple fruits and leaves of 'Gala'. On inoculated fruits by mycelial, the isolates induced symptoms of BR, and on inoculated fruits by conidia without wound the isolates induced symptoms of GLS. On inoculated leaves, isolates induced symptoms of GLS. The isolates were characterized by cultural, morphological and pathogenic aspects. Detection of Colletotrichum spp. in asymptomatic apple flowers is epidemiologically important, since the flowers can be a route for infection in unripe fruits. This is the first investigation of quiescent infection of this pathogen on apple flowers.

RESUMOColletotrichum gloeosporioides e C. acutatum são os agentes etiológicos da Mancha da Gala (MG), na cultura da macieira, no Brasil. Programas de melhoramento visando o desenvolvimento de resistência genética à doença necessitam do conhecimento pleno das características das espécies do patógeno envolvidas. Uma maneira de separar estas espécies é através do estudo da sensibilidade de isolados ao fungicida benomyl. Através do teste de sensibilidade de 39 isolados de C. acutatum e C. gloeosporioides relacionados à MG, em que foram avaliados a taxa de crescimento micelial e o índice de redução deste crescimento, observou-se que dentre os isolados de C. gloeosporioides há um grupo resistente ao fungicida e que difere de todos os isolados desta espécie relatados neste patossistema. Este grupo mostrou-se resistente às concentrações de 5 e 100 µg.mL -1 com índice de redução (IR) variando entre 0% e 8%, e entre 15 e 30%, respectivamente, enquanto o isolado padrão (CG197) apresentou IR de 100% em ambas as concentrações testadas. Os isolados de C. acutatum foram todos resistentes ao fungicida. Devido à resistência de alguns isolados de C. gloeosporioides, a separação de espécies de Colletotrichum associadas à macieira, baseada apenas na resistência ou suscetibilidade ao fungicida benomyl, não deve ser utilizada isoladamente. Palavras-chave: Malus spp., resistência a fungicida, crescimento in vitro. ABSTRACT In vitro assessment of sensitivity to benomyl of Colletotrichum spp. isolates associated with Gala leaf spot of appleIn Brazil, Colletotrichum gloeosporioides and C. acutatum are responsible for the occurrence of Gala leaf spot or Glomerella Leaf Spot (GLS) on apple. Breeding programs aiming to develop genetic resistance to disease need a complete understanding of the characteristics of involved species. A study of the sensibility of the isolates to benomyl is a way to separate those species. A sensitivity test of 39 isolates of C. acutatum and C. gloeosporioides involved with GLS, evaluated by growth rate and reduction rate of mycelium, revealed a group of C. gloeosporioides isolates that are resistant to the fungicide. This group showed resistance to concentrations of 5 and 100 µg.mL -1 with reduction rate varying between 0% and 8%, and between 15 and 30%, respectively. While the standard C. gloeosporioides isolate (CG197) showed a reduction rate of 100% at both tested concentrations, all isolates of C. acutatum were resistant to the fungicide. Due to the resistance of some isolates of C. gloeosporioides, the separation of Colletotrichum species on apple should not be based only on resistance or susceptibility to benomyl.

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