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Diplodia intermedia. a. Culture growing on PDA; b. pycnidia developing on pine needle; c. asci; d, e. ascus, ascospores and pseudoparaphyses; f–i. conidiogenous cells; j, k. conidia in two levels of focus to show finely verruculose inner surface of the conidium wall; l, m. conidia; n, o. microconidia. — Scale bars: b = 500 μm; c, d = 20 μm; e–m = 10 μm.
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Diplodia species are known as pathogens on many woody hosts, including fruit trees, worldwide. In this study a collection of Diplodia isolates obtained mostly from apple and other Rosaceae hosts were identified based on morphological characters and DNA sequence data from ITS and EF1-α loci. The results show that the diversity of species associated...
Citations
... These metabolites could also play a crucial role in the endophytic associations of the fungus with plants mediating communication, nutrient acquisition, and defence. Although Diplodia species can establish endophytic associations with plants (Slippers and Wingfield 2007;Phillips et al. 2012), this fungal genus has primarily been studied as a plant pathogen. Therefore, little is known on the occurrence and functions of secondary metabolites produced by endophytic strains of Diplodia spp. ...
Fungi of the genus Diplodia have a worldwide distribution and are typically associated with plants. Diplodia is well‐known for the diseases it causes on economically and ecologically relevant plants. In particular, this fungal genus is responsible of various symptoms of plant diseases, including shoot blights, dieback, cankers and fruit rots. In the last decades, literature concerning metabolites produced by Diplodia species has been significantly enriched by many reports dealing with the biosynthetic potential of this fungal genus. Several polyketide- and terpenoid-derived compounds have been reported, demonstrating the biosynthetic arsenal of this fungal genus. Investigations on the biological properties of compounds isolated from in vitro cultures of these fungi have proved a broad spectrum of biological functions. In particular, bioassays disclosed that antimicrobial and phytotoxic activities are the most notable bioactivities of secondary metabolites isolated from this genus. Hence, the present review is intended as reference guide to metabolites produced by fungi currently belong to the genus Diplodia, emphasizing the implication of their occurrence, absolute configuration determinations and the structure–activity relationships.
... 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). ...
... 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). This species was reported as one of the main causal agents of apple canker in Iran (Nourian et al. 2021). ...
Apple (Malus domestica Borkh) is one of the most consumed and nutritious fruits. Iran is one of the main producers of the apple in the world. Diplodia bulgarica is the major causal agent of apple tree decline in Iran. Biological control is a nature-friendly approach to plant disease management. Trichoderma zelobreve was isolated from apple trees infected with Diplodia bulgarica in West Azarbaijan province of Iran. The results showed that T. zelobreve strongly inhibited the colony growth of D. bulgarica. In vivo assay on detached branches of apple tree cv. Golden Delicious using T. zelobreve mycelial plug showed that canker length/stem length (CL/SL) and canker perimeter/stem perimeter (CP/SP) indices decreased by 76 and 69%, respectively, 21 days after inoculation. Additionally, wettable powder formulation (WPF) containing the antagonistic fungus "T. zelobreve" decreased CL and CP/SP by 75 and 67%, respectively, 6 months after inoculation. Moreover, canker progress curves and the area under the disease progress curve (AUDPC) supported these findings. The growth temperatures of the antagonist and pathogen were similar, indicating the adaptation of T. zelobreve for biocontrol of apple canker caused by D. bulgarica. The results also showed that T. zelobreve-based WPF stored at 25 °C assure excellent shelf life at least 4 months, allowing the bioproduct to be stored at room temperature, which is a great advantage and cost-effective option.
... Conidia were brown, aseptate, ovoid, and measured 27.9 to 31.3 μm x 12.1 to 14.2 μm (mean ± S.D. of 15 conidia = 29.9 ± 0.9 μm × 13.2 ± 0.6 μm), the typical morphology of a Diplodia sp. (Phillips et al. 2012). Genomic DNA was extracted from a 7-dayold culture of a representative isolate M45-28, using the Plant/Fungi DNA Isolation Kit (Norgen Biotech, Canada). ...
A dieback of apple trees (Malus domestica (Suckow) Borkh.) associated with cankers was observed in commercial orchards in southwestern Ontario, Canada, in 2019. Fifteen 2 to 10-year-old symptomatic trees were collected from three orchards exhibiting up to 37% disease incidence. Small sections of diseased wood (1 cm long) were surface sterilized with 70% ethanol for 30 sec and 1% NaClO for 20 min, rinsed thrice in sterile water, placed on 2% PDA (Difco) amended with kanamycin (50 mg liter−1), and incubated at 22°C for 5 days in the dark (Ilyukhin et al. 2023). Fungal colonies that were consistently isolated were hyphal-tipped, transferred to individual PDA plates and incubated at 22°C for 7 days in the dark. Purified isolates with same characteristics were classed into morphotypes. One morphotype was initially white and turned dark olivaceous with dense aerial mycelium. Pycnidia were produced on pine needles on PDA (Fig. S2) after incubation at 22°C for 17 days in the dark. Conidia were brown, aseptate, ovoid, and measured 27.9 to 31.3 μm x 12.1 to 14.2 μm (mean ± S.D. of 15 conidia = 29.9 ± 0.9 μm × 13.2 ± 0.6 μm), the typical morphology of a Diplodia sp. (Phillips et al. 2012). Genomic DNA was extracted from a 7-day-old culture of a representative isolate M45-28, using the Plant/Fungi DNA Isolation Kit (Norgen Biotech, Canada). The internal transcribed spacer (ITS), translation elongation factor 1-α (EF1-α) and β-tubulin gene regions were amplified and sequenced with primers ITS1/ITS4, EF1-728F/EF1-986R and Bt2a/Bt2b and deposited in GenBank with accession numbers MZ970605, MZ995430 and MZ995431, respectively. Based on the sequence, the fungus was identified as Diplodia intermedia A.J.L. Phillips et al. and matched isolates from different hosts and countries (ITS: 100%, MG220378; EF1-α: 100%, MG220385; β-tubulin: 99.24%, MT592502). The maximum likelihood-based phylogenetic analysis of ITS, EF1-α and β-tubulin concatenated sequences was performed using IQ-Tree 2.2.2.7 (Minh et al. 2020). M45-28 was clustered with high bootstrap support values with D. intermedia isolates from the Westerdijk Fungal Biodiversity Institute collection, including the ex-holotype (CBS 124462) (Fig. S1). A living culture of M45-28 was deposited in the Canadian Collection of Fungal Cultures (DAOMC 252253). Pathogenicity assay was conducted by inoculating mycelial plugs from a 7-day-old culture of M45-28 into wounds made on the trunk of 5 eight-month-old potted healthy ‘Royal Gala’ apple seedlings. Five control seedlings were inoculated with sterile plugs. Canker symptoms appeared 15 days after inoculation, spread around, up and down the main stem from the inoculation point, and by 7 weeks the upper portion of the seedling was dead (Fig. S2). Diplodia intermedia was re-isolated from all inoculated seedlings and species identity was confirmed by sequencing as described above, fulfilling Koch’s postulates. Control seedlings remained symptomless and the fungus was not isolated from the wood. Diplodia intermedia was reported to cause cankers on apple in Uruguay (Delgado-Cerrone et al. 2016), wild apple (Malus sylvestris) in Portugal (Phillips et al. 2012), grapevines in France (Comont et al. 2016) and forest trees in Iran (Kazemzadeh Chakusary et al. 2019). To the best of our knowledge, this is the first report of D. intermedia causing canker and dieback diseases on apple trees in Canada. Further studies are required to better understand the epidemiology involved in the dynamic spread of the disease in order to recommend an adequate phytosanitary program for its control.
... and D. seriata De Not. Most of these species have a limited distribution, except for D. seriata, which has been reported globally (Cloete et al., 2011;Phillips et al., 2012;Úrbez-Torres et al., 2016;Sessa et al., 2016;Díaz et al., 2019;Nabi et al., 2020). Diplodia seriata is the most frequent species of the Botryosphaeriaceae associated with cankers on apple trees in South Africa (Slippers et al., 2007;Cloete et al., 2011;Havenga et al., 2019), Chile (Díaz et al., 2022) and northern Italy (Martino et al., 2023). ...
... Fig. 4 (A)-(F) illustrates the pycnidia and conidia of D. seriata observed on the wood pieces. The conidia illustrated in Fig. 4E indicate septation although Phillips et al. (2007Phillips et al. ( , 2012 described D. seriata as aseptate. This contrasting finding could be due to septa only developing later as conidia mature. ...
... Pathogenic taxa within Botryosphaeriaceae cause diseases that are difficult to treat and control in many economically important crops, including Eucalyptus and Pinus spp. (Burgess et al., 2006;Slippers and Wingfield, 2007), grapes (Vitis vinifera; Urbez-Torres, 2011), walnut (Juglans regia L.; Li et al., 2016), almond (Prunus dulcis; Olmo et al., 2016), apple (Malus; Phillips et al., 2012), mango trees (Mangifera indica L.; El-Komy et al., 2023), and olives (Olea europaea) (Urbez-Torres et al., 2013) among many others. With over 7.12 million ha cultivated, grapevines (V. ...
Fungal pathogens are responsible for 30% of emerging infectious diseases (EIDs) in plants. The risk of a pathogen emerging on a new host is strongly tied to its host breadth; however, the determinants of host range are still poorly understood. Here, we explore the factors that shape host breadth of plant pathogens within Botryosphaeriaceae, a fungal family associated with several devastating diseases in economically important crops. While most host plants are associated with just one or a few fungal species, some hosts appear to be susceptible to infection by multiple fungi. However, the variation in the number of fungal taxa recorded across hosts is not easily explained by heritable plant traits. Nevertheless, we reveal strong evolutionary conservatism in host breadth, with most fungi infecting closely related host plants, but with some notable exceptions that seem to have escaped phylogenetic constraints on host range. Recent anthropogenic movement of plants, including widespread planting of crops, has provided new opportunities for pathogen spillover. We suggest that constraints to pathogen distributions will likely be further disrupted by climate change, and we may see future emergence events in regions where hosts are present but current climate is unfavorable.
... Species in Diplodia are characterized by hyaline, aseptate and thick-walled conidia that may become pigmented and 1-septate either after or before discharge from the pycnidia . They have a worldwide distribution and are known as pathogens, endophytes and saprophytes on a wide range of mainly woody hosts (Damm et al. 2007, Lazzizera et al. 2008, Laveau et al. 2009, Pérez et al. 2010, Phillips et al. 2012, Linaldeddu et al. 2013, Abdollahzadeh 2015. Some Diplodia species are important pathogens causing cankers, dieback, wilt, root diseases, leaf spots and shoot/tip blight on a variety of horticultural crops, such as D. corticola on oaks, D. sapinea on pines and D. mutila and D. seriata on apples (Alves et al. 2004, Trapman et al. 2008, Stanosz et al. 2009, Phillips et al. 2012, Úrbez-Torres et al. 2016, Ferreira et al. 2021. ...
... They have a worldwide distribution and are known as pathogens, endophytes and saprophytes on a wide range of mainly woody hosts (Damm et al. 2007, Lazzizera et al. 2008, Laveau et al. 2009, Pérez et al. 2010, Phillips et al. 2012, Linaldeddu et al. 2013, Abdollahzadeh 2015. Some Diplodia species are important pathogens causing cankers, dieback, wilt, root diseases, leaf spots and shoot/tip blight on a variety of horticultural crops, such as D. corticola on oaks, D. sapinea on pines and D. mutila and D. seriata on apples (Alves et al. 2004, Trapman et al. 2008, Stanosz et al. 2009, Phillips et al. 2012, Úrbez-Torres et al. 2016, Ferreira et al. 2021. Although more than 1000 species epithets are listed in MycoBank and similar databases , DNA sequence data are available for a limited number of species. ...
... In this sense, the narrow host range of Neodeightonia species might be related with their host preference towards bamboos and, particularly, palms, since most of them have been found associated with Arecaceae members. Moreover, host preference has already been shown in many species of Diplodia , Damm et al. 2007, Lazzizera et al. 2008, Úrbez-Torres et al. 2010, Phillips et al. 2012, so it would not be surprising to find a similar pattern of host preference in the closely related genus Neodeightonia. ...
The Botryosphaeriaceae is the largest family in Botryosphaeriales and currently comprises 22 genera of important endophytes, saprobes and plant pathogens. Most botryosphaeriaceous species have a cosmopolitan distribution, occurring on a wide range of woody hosts. Nonetheless, in many hosts, including palms (Arecaceae), the complex of associated Botryosphaeriaceae taxa is as yet unknown. The present study aimed to identify the botryosphaeriaceous species associated with foliar lesions of ornamental palms in Lisbon, Portugal. Twenty-nine Botryosphaeriaceae taxa were isolated from seven different palm species and identified based on both morphological and phylogenetic analyses. Six genera were detected: Botryosphaeria, Diplodia, Dothiorella, Neodeightonia, Neofusicoccum and Sardiniella. A new species of Neodeightonia, N. chamaeropicola, is introduced. Three botryosphaeriaceous species are reduced to synonymy. Thirteen new plant host-fungus associations are reported, while four new geographical records are noted for Portugal. A synopsis of accepted and phylogenetically validated Botryosphaeriaceae taxa reported from palms worldwide is presented. A total of 31 botryosphaeriaceous species have been currently reported from Arecaceae hosts, and many of them are associated with disease symptoms. This illustrates that more systematic studies are needed to examine the complex of Botryosphaeriaceae taxa associated with palms and determine their potential pathogenicity.
... pistaciicola) in Sichuan province. Diplodia species mainly occur on woody hosts, causing rots, cankers, shoot and tip blight [11,[56][57][58][59]. Thus, the discovery and in-depth research of this genus are conducive to the protection of woody plants and the maintenance of greater economic benefits. ...
Species in the Botryosphaeriaceae are common plant pathogens, endophytes, and saprobes found on a variety of mainly woody hosts. Botryosphaeriaceae is a high-profile fungal family whose genera have been subjected to continuous revisions in recent years. Surveys conducted during 2019 and 2020 on several decaying woody hosts (from dead arial twigs, branches, stems, bark, and seed pods) in China and Thailand revealed a high diversity of Botryosphaeriaceae fungi. Identification of 16 Botryosphaeriaceae isolates was carried out based on both morphological characteristics and phylo-genetic analyses of combined ITS, LSU, tef1-α, and tub2 sequence data. Four novel species (Dothiorella ovata, Do. rosacearum, Do. septata, and Lasiodiplodia delonicis) and seven previously known species (Botryosphaeria fujianensis, Diplodia mutila, Di. seriata, L. crassispora, L. mahajangana, Macrophomina euphorbiicola and Sphaeropsis eucalypticola) were identified while new hosts and geographical records were reported. This study indicates that the fungal family Botryosphaeriaceae seems to be common and widespread on a broad range of hosts in China and Thailand.
... BLAST searches in GenBank revealed that the isolates TN.19 and TN.86 showed 99 % identity with Diplodia seriata De Not. including the ex-epitype strain CBS112555 (Phillips et al., 2012). The isolate TN.20 exhibited 100 % with Pestalotiopsis biciliata ex-epitype CBS200.65 (Maharachchikumbura et al., 2014). ...
Since 2012, a severe and widespread decline and mortality affecting several of Mediterranean forest species has been occurred throughout Tunisian forests. In 2017, field surveys showed a progressive dieback of Quercus coccifera L. (kermes oak), including shoot dieback, brown lesions on branches and innumerable pycnidia on infected tissues. For its ecological interest, preservation of Q. coccifera as valuable evergreen forest tree must be of great interest. However, information regarding kermes oak disease is still scarce. Hence, the aims of this study were to characterize the fungal pathogens associated with Q. coccifera branch canker in two Tunisian forests and evaluate their virulence. Twenty-eight isolates were obtained from symptomatic kermes oak branches. Based on morphological features and phylogenetic analysis of the internal transcribed spacer (ITS) region of the nuclear ribosomal DNA and partial sequence of the translation elongation factor 1-alpha gene (tef1-α) two fungal species namely Diplodia seriata and Pestalotiopsis biciliata were identified. Moreover, D. seriata species were the main fungi consistently obtained from symptomatic Q. coccifera branches. Pathogenicity tests carried out on asymptomatic detached branches confirmed the virulence of the two species. These findings confirm D. seriata and P. biciliata as fungal pathogens associated with kermes oak branch canker and dieback in Tunisia. To the best of our knowledge, these two plant pathogenic fungi are identified for the first time on kermes oak in Tunisia.
... Based on literature reports, these species have a cosmopolitan distribution and a wide host range (Gure et al. 2005;Slippers et al. 2007;Abdollahzadeh et al. 2010). In previous studies, D. seriata has been reported as a pathogen in woody plants such as peach (Endes et al. 2016), apricot (Smith and Stanosz 2006;Damm et al. 2007;Liu et al. 2015), plum (Phillips et al. 2012;Endes and Kayım 2022a), pear Kurbetli and Demirci (2014) and vineyard (Akgül et al. 2015), almond (Olmo et al. 2016;Gharbi et al. 2017;Holland et al. 2021). In this study, D. seriata was determined as the dominant species. ...
Türkiye is among the top-producing almond countries in the world. Almond is a precious nut in modern diets due to its bioactive compounds associated with health and disease treatment. However, the plant's yield is affected by factors, including pathogens. This study aimed to diagnose and identify biotic factors associated with decline, dieback, and gummosis in almond trees in Yozgat province with a temperate climate. Seven almond 10-year orchards were investigated and trees with symptoms were sampled. Isolated species were cultured in PDA medium to identify the morphological and cultural characterization. The results obtained with cultural and morphological characteristics along with Phylogenetic analysis of ITS, β-tubulin, and EF-1α sequence data, confirmed the presence of Diplodia seriata , Lasiodiplodia theobromae , Neofusicoccum parvum, Botryosphaeria dothidea. Among these species, Botryosphaeria dothidea was first reported from almond trees in Türkiye. Pathogenicity testing showed that although all species were virulent , L. theobromae and N. parvum isolates caused more gumming in the inoculation areas than D. seriata and B . dothidea isolates. These results confirm that multiple Botryosphaeriaceae species are associated with branch decline and dieback on almond in Türkiye, which agrees with similar studies on woody crops. Accurate diagnosis of fungal pathogens in almonds is vital for developing disease management strategies and may help improve horticultural practices in maintaining urban stands.
... Bobev is a plant pathogenic fungus of the family Botryospheriaceae. The pathogen was first described in 2012 on Malus sylvestris in Bulgaria and on Malus domestica in Iran (Phillips et al., 2012). Identity of the pest (Section 3.1) Is the identity of the pest clearly defined, or has it been shown to produce consistent symptoms and to be transmissible? ...
... Diplodia is an important genus of the family Botryosphaeriaceae, with more than 1000 species being pathogenic, endophytic or saprophytic on a wide range of mainly woody plants (Phillips et al., 2012). As pathogens, they have been reported to cause different disease symptoms on their hosts, such as canker, twig blight, gummosis, pre-and post-harvest fruit rot, dieback and tree decline (Crous et al., 2006;Phillips et al., 2007Phillips et al., , 2012Lazzizera et al., 2008;Abdollahzadeh, 2015;Hanifeh et al., 2017). ...
... Diplodia is an important genus of the family Botryosphaeriaceae, with more than 1000 species being pathogenic, endophytic or saprophytic on a wide range of mainly woody plants (Phillips et al., 2012). As pathogens, they have been reported to cause different disease symptoms on their hosts, such as canker, twig blight, gummosis, pre-and post-harvest fruit rot, dieback and tree decline (Crous et al., 2006;Phillips et al., 2007Phillips et al., , 2012Lazzizera et al., 2008;Abdollahzadeh, 2015;Hanifeh et al., 2017). Diplodia bulgarica is one of the several Diplodia species (e.g. ...
The EFSA Plant Health Panel performed a pest categorisation of Diplodia bulgarica, a clearly defined plant pathogenic fungus of the family Botryosphaeriaceae. The pathogen affects Malus domestica, M. sylvestris and Pyrus communis causing various symptoms such as canker, twig blight, gummosis, pre- and post-harvest fruit rot, dieback and tree decline. The pathogen is present in Asia (India, Iran, Türkiye) and in non-EU Europe (Serbia). Concerning the EU, the pathogen is present in Bulgaria and widespread in Germany. There is a key uncertainty on the geographical distribution of D. bulgarica worldwide and in the EU, because in the past, when molecular tools were not available, the pathogen might have been misidentified as other Diplodia species (e.g. D. intermedia, D. malorum, D. mutila, D. seriata) or other members of the Botryosphaeriaceae family affecting apple and pear based only on morphology and pathogenicity tests. Diplodia bulgarica is not included in Commission Implementing Regulation (EU) 2019/2072. Plants for planting, other than seeds, fresh fruits, and bark and wood of host plants as well as soil and other plant-growing media carrying plant debris are the main pathways for the further entry of the pathogen into the EU. Host availability and climate suitability factors are favourable for the further establishment of the pathogen in the EU. In the areas of its present distribution, including Germany, the pathogen has a direct impact on cultivated hosts. Phytosanitary measures are available to prevent the further introduction and spread of the pathogen into the EU. Diplodia bulgarica satisfies the criteria that are within the remit of EFSA to assess for this species to be regarded as potential Union quarantine pest.
