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Symptoms of Cytospora canker on apple trees in the Semirom region. (A and B) Dieback of a 25-yr-old Golden Delicious cultivar of apple tree following the entire girdling of the trunk by a canker. (C) Diffuse resinous twig cankers on Golden Delicious cultivar (D & E) Conidiomata of a Cytospora species on Red Delicious cultivar. 

Symptoms of Cytospora canker on apple trees in the Semirom region. (A and B) Dieback of a 25-yr-old Golden Delicious cultivar of apple tree following the entire girdling of the trunk by a canker. (C) Diffuse resinous twig cankers on Golden Delicious cultivar (D & E) Conidiomata of a Cytospora species on Red Delicious cultivar. 

Contexts in source publication

Context 1
... trees ( Malus pumila Mill.) are widely planted in the Semirom region of Isfahan Province in Iran. Unfortunately, decline of mature apple trees in this region is generally caused by fungal disease, especially Cytospora canker. Cytospora canker is characterized by diffuse resinous branch cankers, with fruiting bodies of the causal fungi usually forming on infected parts. Cankers first occur on the lower branches, and then affect limbs progressively higher up the tree ( Fig. 1). The damage caused by this disease has attracted attention from orchardists as well as plant pathologists. Species of the genus Cytospora Ehrenb., and their related teleomorphs, are among the major causal agents of annual and perennial canker diseases and sudden death of many hardwood and coniferous trees, rarely herbaceous plants and monocots (Adams et al. , 2005). Members of Cytospora are associated with cankers and dieback on a wide range of trees and shrubs (Farr et al. , 1989; Speilman, 1985). Cytospora canker, caused by species of the genus Cytospora Ehrenb., is a worldwide problem and affects more than 70 species of woody shrubs and trees. The genus Cytospora was first described by Ehrenberg (1818). Cytospora refers to the anamorphic (asexual) stage of the causal fungi commonly found forming cankers. Numerous species of Cytospora are listed as causal agents of Cytospora cankers. However, species identification is difficult, because Cytospora fruiting and vegetative structures, as well as spore size, vary greatly even in the isolates of the same species (Spielman, 1985). The species of Cytospora is the anamorphs of Valsa Fr., Leucostoma (Nitschke) Höhn., Valsella Fuckel and Valseutypella Höhn. (Adams et al ., 2005). Fruiting bodies consist of stromata that usually contain either labyrinthine chambers, having filamentous conidiophores and allantoid hyaline conidia. In moist conditions, the conidia exude from the fruiting bodies in gelatinous matrices, usually as yellow, orange, red or pallid droplets or tendrils (Adams et al ., 2006; Sutton, 1980). Kobayashi (1970) in Japan, Gvritishvili (1982) and Vasilyeva (1994) in the former Union of Soviet Socialist Republics (USSR), Spielman (1983, 1985) for North American species on hardwoods and Adams et al . (2005, 2006) in South Africa, described the numerous species of genus of Cytospora . The first report on species of genus Cytospora in Iran was published by Fragozo (1918) to introduce Cytospora silenes Gonz. Frag. on Silene boryi Boiss. Thereafter, other researchers such as Petrak and Esfandiari (1941), Esfandiari (1946, 1948), Steyeart (1953), Ershad (1995), Ashkan (1979), Fotouhifar (2007) and Fotouhifar et al. (2007, 2008) introduced many other species of the genus Cytospora in Iran. These taxa have however, not been extensively investigated. Thus, awareness of existing species is essential for effective disease management. The main objective of the present study was to identify Cytospora species associated with canker diseases on apple trees in Semirom region of Iran. The apple orchards with dieback and canker symptoms were obesreved during spring and autumn of year 2007 that cankered twigs showed fruiting bodies of Cytospora spp. and their related teleomorphs were collected. A total of 170 infected twigs were collected from apple orchards almost all regions of the Semirom. The geographic origins of the collected isolates are listed in Table 1. Of these, 114 fungal specimens belonging to different anamorphic and teleomorphic species of this group of fungi were selected using morphological characteristics of sexual and asexual fruiting bodies (Fig. 1). Single spore isolation of fungi from infected host tissues with fruiting bodies was carried out by removing a mass of conidia from the surface of the specimens and spreading the suspension on the surface of agar plates. Single germinating spores were picked off and placed on fresh plates. Alternatively, plant tissues were cut in almost 5 mm diameter discs and placed in 70% ethanol for 1min and then surface sterilized by brief flaming. Masses of conidia from fruting bodies were discharged by placing the treated tissues in a moist chamber for 1hr at 25oC. Small amount of conidial mass from a single fruiting body was streaked on water agar medium using sterile needle and then the plates were incubated at 25oC for 24hr. If perithecia were presented in infected tissues, the contents of a single perithecium were similarly transferred to an agar plate. Single-spore isolates were obtained by transferring 3-10 germinated conidia or ascospores to PDA plates separately (Fotouhifar, 2007). Colors of the surface and reverse parts of the pure fungal colonies were compared using a color chart of (Rayner, 1970). Identification of species were based on morphological features of the sexual and asexual fruiting bodies produced on infected plant tissues. For this purpose, thin cross sections were prepared by hand from fruiting bodies. Morphological characteristics of the fruiting bodies including; size and arrangement of stromata, presence or absence of conceptacle in stromata, number and diameter of ostioles per disc, color, shape and size of discs, arrangement type of locules and perythecia, size and number of perithecia, size and shape of conidiophores and asci, size of conidia and ascospores were determined using a dissecting microscope. Microscopic measurements were made with a calibrated microscope. For each isolate, over 30 conidiomata were sectioned and diameters of 50 conidia were measured under an Olympus light microscope (SZH10) . If the perithecia were presented, over 50 perithecia were sectioned and then over 100 ascospores were measured under an Olympus microscope. For other morphological features, at least 50 specimens were measured. Representative isolates and samples were deposited at the Iranian Research Institute of Plant Protection (IRAN, Tehran, Iran). Isolates were placed in three genera based on their morphological features including; Cytospora , Valsa and Leucostoma . In the genus Cytospora , the following species were identified; Cytospora cincta, C. schulzeri, C. leucostoma and C. chrysosperma . In the genus Valsa only Valsa malicola was identified and in the genus Leucostoma also a single species, Leucostoma cinctum was identified. Out of 107 isolates studied, 50 isolates belonged to Valsa malicola (anamorph: Cytospora schulzeri ), 51 isolates belonged to Leucostoma cinctum (anamorph ; Cytospora cincta ), three isolates belonged to C. leucostoma , and finally, three isolates belonged to C. chrysosperma . The distinctive features of the identified species are shown as ...
Context 2
... trees ( Malus pumila Mill.) are widely planted in the Semirom region of Isfahan Province in Iran. Unfortunately, decline of mature apple trees in this region is generally caused by fungal disease, especially Cytospora canker. Cytospora canker is characterized by diffuse resinous branch cankers, with fruiting bodies of the causal fungi usually forming on infected parts. Cankers first occur on the lower branches, and then affect limbs progressively higher up the tree ( Fig. 1). The damage caused by this disease has attracted attention from orchardists as well as plant pathologists. Species of the genus Cytospora Ehrenb., and their related teleomorphs, are among the major causal agents of annual and perennial canker diseases and sudden death of many hardwood and coniferous trees, rarely herbaceous plants and monocots (Adams et al. , 2005). Members of Cytospora are associated with cankers and dieback on a wide range of trees and shrubs (Farr et al. , 1989; Speilman, 1985). Cytospora canker, caused by species of the genus Cytospora Ehrenb., is a worldwide problem and affects more than 70 species of woody shrubs and trees. The genus Cytospora was first described by Ehrenberg (1818). Cytospora refers to the anamorphic (asexual) stage of the causal fungi commonly found forming cankers. Numerous species of Cytospora are listed as causal agents of Cytospora cankers. However, species identification is difficult, because Cytospora fruiting and vegetative structures, as well as spore size, vary greatly even in the isolates of the same species (Spielman, 1985). The species of Cytospora is the anamorphs of Valsa Fr., Leucostoma (Nitschke) Höhn., Valsella Fuckel and Valseutypella Höhn. (Adams et al ., 2005). Fruiting bodies consist of stromata that usually contain either labyrinthine chambers, having filamentous conidiophores and allantoid hyaline conidia. In moist conditions, the conidia exude from the fruiting bodies in gelatinous matrices, usually as yellow, orange, red or pallid droplets or tendrils (Adams et al ., 2006; Sutton, 1980). Kobayashi (1970) in Japan, Gvritishvili (1982) and Vasilyeva (1994) in the former Union of Soviet Socialist Republics (USSR), Spielman (1983, 1985) for North American species on hardwoods and Adams et al . (2005, 2006) in South Africa, described the numerous species of genus of Cytospora . The first report on species of genus Cytospora in Iran was published by Fragozo (1918) to introduce Cytospora silenes Gonz. Frag. on Silene boryi Boiss. Thereafter, other researchers such as Petrak and Esfandiari (1941), Esfandiari (1946, 1948), Steyeart (1953), Ershad (1995), Ashkan (1979), Fotouhifar (2007) and Fotouhifar et al. (2007, 2008) introduced many other species of the genus Cytospora in Iran. These taxa have however, not been extensively investigated. Thus, awareness of existing species is essential for effective disease management. The main objective of the present study was to identify Cytospora species associated with canker diseases on apple trees in Semirom region of Iran. The apple orchards with dieback and canker symptoms were obesreved during spring and autumn of year 2007 that cankered twigs showed fruiting bodies of Cytospora spp. and their related teleomorphs were collected. A total of 170 infected twigs were collected from apple orchards almost all regions of the Semirom. The geographic origins of the collected isolates are listed in Table 1. Of these, 114 fungal specimens belonging to different anamorphic and teleomorphic species of this group of fungi were selected using morphological characteristics of sexual and asexual fruiting bodies (Fig. 1). Single spore isolation of fungi from infected host tissues with fruiting bodies was carried out by removing a mass of conidia from the surface of the specimens and spreading the suspension on the surface of agar plates. Single germinating spores were picked off and placed on fresh plates. Alternatively, plant tissues were cut in almost 5 mm diameter discs and placed in 70% ethanol for 1min and then surface sterilized by brief flaming. Masses of conidia from fruting bodies were discharged by placing the treated tissues in a moist chamber for 1hr at 25oC. Small amount of conidial mass from a single fruiting body was streaked on water agar medium using sterile needle and then the plates were incubated at 25oC for 24hr. If perithecia were presented in infected tissues, the contents of a single perithecium were similarly transferred to an agar plate. Single-spore isolates were obtained by transferring 3-10 germinated conidia or ascospores to PDA plates separately (Fotouhifar, 2007). Colors of the surface and reverse parts of the pure fungal colonies were compared using a color chart of (Rayner, 1970). Identification of species were based on morphological features of the sexual and asexual fruiting bodies produced on infected plant tissues. For this purpose, thin cross sections were prepared by hand from fruiting bodies. Morphological characteristics of the fruiting bodies including; size and arrangement of stromata, presence or absence of conceptacle in stromata, number and diameter of ostioles per disc, color, shape and size of discs, arrangement type of locules and perythecia, size and number of perithecia, size and shape of conidiophores and asci, size of conidia and ascospores were determined using a dissecting microscope. Microscopic measurements were made with a calibrated microscope. For each isolate, over 30 conidiomata were sectioned and diameters of 50 conidia were measured under an Olympus light microscope (SZH10) . If the perithecia were presented, over 50 perithecia were sectioned and then over 100 ascospores were measured under an Olympus microscope. For other morphological features, at least 50 specimens were measured. Representative isolates and samples were deposited at the Iranian Research Institute of Plant Protection (IRAN, Tehran, Iran). Isolates were placed in three genera based on their morphological features including; Cytospora , Valsa and Leucostoma . In the genus Cytospora , the following species were identified; Cytospora cincta, C. schulzeri, C. leucostoma and C. chrysosperma . In the genus Valsa only Valsa malicola was identified and in the genus Leucostoma also a single species, Leucostoma cinctum was identified. Out of 107 isolates studied, 50 isolates belonged to Valsa malicola (anamorph: Cytospora schulzeri ), 51 isolates belonged to Leucostoma cinctum (anamorph ; Cytospora cincta ), three isolates belonged to C. leucostoma , and finally, three isolates belonged to C. chrysosperma . The distinctive features of the identified species are shown as ...

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... Apple canker disease is caused usually by the species in Botryosphaeriaceae and Valsaceae families in Iran (Ashkan 1994;Mehrabi et al. 2011;Arzanlou and Bakhshi 2012;Nourian et al. 2021) and in other regions of the world (Brown and Britton 1986;Proffer and Jones 1989;Adams et al. 2006;Phillips et al. 2012;Nabi et al. 2020). D. bulgarica was first reported from apple trees displaying canker symptoms in Bulgaria (Phillips et al. 2012), and thereafter, in Iran (Arzanlou and Bakhshi 2012;Nourian et al. 2021). ...
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... However, recent studies employing molecular phylogeny revealed many cryptic species with similar morphology on the same host of known species of this genus [11][12][13][14][15]. For example, up to 28 Cytospora species were discovered from Eucalyptus spp. in South Africa with the help of DNA sequence evidence [2], eight from willow (Salix spp.) trees in China [16], six from Castanea mollissima in China [17], six from Populus hosts in China [18] and six from apple trees in Iran [19]. The taxonomy of Cytospora is currently more dependent on combined evidence of DNA sequence data, morphological features and ecology than species morphology and host associations [1,20]. ...
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