Genetic Diversity and Pathogenicity of Botryosphaeriaceae Species Associated with Symptomatic Citrus Plants in Europe
by
, , and
Jadson Diogo Pereira Bezerra
1,2
,
Pedro Wilhelm Crous
2,
Dalia Aiello
3,
Maria Lodovica Gullino
4,
Giancarlo Polizzi
3 and
Vladimiro Guarnaccia
2,4,5,* 1
Setor de Micologia, Departamento de Biociências e Tecnologia, Instituto de Patologia Tropical e Saúde Pública (IPTSP), Universidade Federal de Goiás (UFG), Rua 235, s/n, Setor Universitário, Goiânia 74605-050, Brazil
2
Westerdijk Fungal Biodiversity Institute, Uppsalalaan 8, 3584 CT Utrecht, The Netherlands
3
Dipartimento di Agricoltura, Alimentazione e Ambiente, sez. Patologia Vegetale, University of Catania, Via S. Sofia 100, 95123 Catania, Italy
4
Centre for Innovation in the Agro-Environmental Sector, AGROINNOVA, University of Torino, Largo Braccini 2, 10095 Grugliasco, Italy
5
Department of Agricultural, Forest and Food Sciences (DISAFA), University of Torino, Largo Braccini 2, 10095 Grugliasco, Italy
*
Author to whom correspondence should be addressed.
Plants 2021, 10(3), 492; https://doi.org/10.3390/plants10030492
Submission received: 11 February 2021
/
Revised: 22 February 2021
/
Accepted: 2 March 2021
/
Published: 5 March 2021
(This article belongs to the Special Issue Citrus Fungal and Oomycete Diseases)
Abstract
:This study represents the first survey studying the occurrence, genetic diversity, and pathogenicity of Botryosphaeriaceae species associated with symptomatic citrus species in citrus-production areas in five European countries. Based on morphological features and phylogenetic analyses of internal transcribed spacer (ITS) of nuclear ribosomal DNA (nrDNA), translation elongation factor 1-alpha (TEF1) and β-tubulin (TUB2) genes, nine species were identified as belonging to the genera Diplodia, Dothiorella, Lasiodiplodia, and Neofusicoccum. Isolates of Neofusicoccum parvum and Diplodia pseudoseriata were the most frequently detected, while Dothiorella viticola had the widest distribution, occurring in four of the five countries sampled. Representative isolates of the nine Botryosphaeriaceae species used in the pathogenicity tests caused similar symptoms to those observed in nature. Isolates assayed were all re-isolated, thereby fulfilling Koch’s postulates. Isolates of Diplodia pseudoseriata and Diplodia olivarum are recorded for the first time on citrus and all species found in our study, except N. parvum, are reported for the first time on citrus in Europe.
1. Introduction
Citrus production represents one of the most important fruit industries worldwide in terms of total yield. Greece, Italy, Portugal, and Spain are the most important European producers of citrus fruit [1]. In 2019, nearly 11 million tons of citrus was produced in Europe on approximately 515,000 ha [2]. Most canker diseases of citrus, as well as further fruit-tree crops, are caused by a broad range of fungal species that infect the wood mainly through winter pruning wounds and a subsequent colonization of vascular tissues [3]. Several abiotic and biotic factors are considered responsible for rots and gumming on the trunk and main branches in citrus. Frost damage, sunscald, or water distribution can promote the infection of numerous ascomycetes and basidiomycetes [4]. Several fungal infections involving twigs, branches and trunks of citrus caused by Colletotrichum and Diaporthe species were reported in different continents [5,6,7,8,9]. Guarnaccia and Crous [10] reported serious cankers developing in woody tissues of lemon trees caused by Diaporthe spp., often with a gummose exudate, causing serious blight and dieback. Canker diseases of citrus are also caused by other fungal genera such as Fusarium and Neocosmospora [11], Peroneutypa [12,13], and Phaeoacremonium [14]. Recently, significant attention has been dedicated to revising species and genera of Botryosphaeriaceae, which encompass species with a cosmopolitan distribution that are able to cause diseases of numerous plant species worldwide [15,16].
Botryosphaeriaceae (Botryosphaeriales) include several species reported as endophytes, latent, and woody plant pathogens on a broad range of host [15,16,17]. This family has undergone significant revision after the adoption of molecular tools to resolve its taxonomy [15,16,18,19,20,21,22,23]. Recently, the taxonomy of Botryosphaeriaceae (and other families in Botryosphaeriales) has been reviewed by Phillips et al. [23] based on morphology of the sexual morphs, phylogenetic relationships on internal transcribed spacer (ITS) and 28S large subunit (LSU) of nuclear ribosomal DNA (nrDNA) sequences and evolutionary divergence times. The authors highlighted the main findings made by Yang et al. [16] who included new families, genera, and species in Botryosphaeriales based on morphology and multi-marker phylogenetic analyses of a large collection of isolates. Currently, six families are accepted in Botryosphaeriales and 22 genera have been included in Botryosphaeriaceae [23,24,25].
The most common symptoms observed in association with species of Botryosphaeriaceae are twig, branch and trunk cankers, die-back, collar rot, root cankers, gummosis, decline and, in severe cases, plant death [15,17]. Plant infections mainly occur through natural openings or wounds, but these fungal species could also survive in latency. This ability could lead to their spread worldwide through asymptomatic plant material, seedlings and fruit, frequently circumventing the adopted quarantine measures [22]. Moreover, stress and non-optimal plant growth conditions consistently induce the expression of diseases associated with Botryosphaeriaceae species. Thus, global warming could increase plant stress and induce favourable conditions for the development of Botryosphaeriaceae diseases [17,26,27]. Species within the Botryosphaeriaceae represent a serious threat to different crops including major fruit, berry fruit and nut crops cultivated in sub-tropical, tropical, or temperate areas [22,28,29,30].
Several species of Diplodia (Di.), Dothiorella (Do.), Lasiodiplodia, Neofusicoccum, and Neoscytalidium (Ne.) have been previously reported to affect Citrus species [13,31,32,33]. For example, Ne. dimidiatum has been reported causing citrus branch canker in California [13] and Italy [32]; Do. viticola, L. citricola, L. theobromae, and Ne. dimidiatum have been described in association with branch and trunk dieback of citrus trees in Iran [14,34] and Dothiorella spp. have been detected as causal agents of citrus gummosis in Tunisia [35]. Moreover, Di. seriata, Di. mutila, Do. viticola, L. mediterranea and L. mitidjana, have been recovered from symptomatic citrus trees in Algeria [33].
Considering the important economic value of Citrus spp., a large survey of Botryosphaeriaceae affecting plants cultivated in the major citrus production areas of Europe was considered imperative. Identification in light of modern taxonomic concepts via morphological characterization and multi-marker DNA sequence data was necessary to adopt efficient control strategies against the pathogens that could affect these crops. Thus, several surveys have been conducted in Greece, Italy, Portugal, Spain, and Malta during 2015 and 2016. In particular, the aims of this study were to (1) conduct extensive surveys for sampling symptomatic plant materials; (2) obtain a broad collection of Botryosphaeriaceae isolates; (3) subject those isolates to DNA multi-marker sequence analyses combined with morphological characterization, and (4) evaluate the pathogenicity of the isolated species to citrus plants.
2. Results
2.1. Field Sampling and Fungal Isolation
In this study, the sampling focused on symptomatic plants of Citrus limon, C. reticulata, C. sinensis, C. sinensis × Poncirus trifoliata, and Microcitrus australasica. Samples were collected in 19 orchards (Table 1). Citrus trees showed various external disease symptoms, including partial or complete yellowing, wilting leaves and twigs, and dieback of branch tips, but also defoliation and branch decline. Canker and cracking of the bark associated with gummose exudate occurred on trunks and branches. Internal observation of infected branches revealed black to brown wood discoloration in cross-sections, wedge-shaped necrosis or irregular wood discoloration. Twigs were wilted and occasionally presenting sporocarps (Figure 1). Symptoms were detected in all the orchards and regions investigated. A total of 63 fungal isolates were collected and were found to be characterized by dark green to grey, fast-growing mycelium on MEA. Moreover, the isolates produced pycnidia on pine needles within 40 days, containing pigmented or hyaline conidia. According to these characteristics, the fungal isolates were classified as Botryosphaeriaceae spp. based on comparison with the previous generic descriptions [15]. Among the collected isolates, 18 were obtained from trunk cankers, 10 were associated with branch infections, and 35 from twig dieback (Table 2).
2.2. Phylogenetic Analyses
A combined multi-marker (ITS, TEF1, and TUB2) phylogenetic tree was inferred for each genus (Diplodia, Dothiorella, Lasiodiplodia, and Neofusicoccum) obtained in this study (Figure 2, Figure 3, Figure 4 and Figure 5). The best nucleotide models for the Bayesian Inference analysis of each dataset were as follows: SYM (symmetrical model) + I (proportion of invariable sites) + G (gamma distribution) (Diplodia, Dothiorella, Lasiodiplodia, and Neofusicoccum) for ITS; GTR (generalized time-reversible model) + G (Diplodia, Dothiorella and Neofusicoccum) and HKY (Hasegawa–Kishino–Yano) + I + G (Lasiodiplodia) for TEF1 and GTR + G (Diplodia, Lasiodiplodia and Neofusicoccum) and GTR + I + G (Dothiorella) for TUB2. The Diplodia phylogenetic analysis revealed the isolates as belonging to Di. pseudoseriata (15 isolates, BPP = 1 and ML-BS = 100), Di. seriata (9 isolates, BPP = 1 and ML-BS = 95), Di. olivarum (2 isolates, Bayesian posterior probabilities (BPP) = 1 and maximum likelihood bootstrapped (ML-BS) = 99), and Di. mutila (1 isolate, BPP = 0.99 and ML-BS = 87) (Figure 2). The Dothiorella phylogeny (Figure 3) grouped the isolates together within Do. viticola (9 isolates, BPP = 1 and ML-BS = 99). The Lasiodiplodia phylogenetic analysis placed five isolates as L. theobromae (BPP = 1 and ML-BS = 98) (Figure 4). The Neofusicoccum phylogeny (Figure 5) grouped sequences from our isolates as belonging to N. luteum (2 isolates, BPP = 1 and ML-BS = 94), N. parvum (16 isolates) and N. mediterraneum (4 isolates, BPP = 1 and ML-BS = 98).
2.3. Occurrence of Botryosphaeriaceae among Countries and Citrus Species
Among countries, Do. viticola was found in Greece, Italy, Portugal, and Spain; N. parvum in Italy and Malta, and Di. pseudoseriata in Portugal and Spain. In addition, Di. mutila and Di. seriata were exclusively isolated in Greece and Spain, respectively; L. theobromae and Di. olivarum were only found in Malta, and N. luteum and N. mediterraneum were exclusively found in Portugal. Based on the citrus species, N. parvum (25.4%) and Di. pseudoseriata (23.8%) were the most frequently detected Botryosphaeriaceae spp. on C. sinensis × P. trifoliata, C. limon, C. reticulata, C. sinensis, and/or M. australasica; Di. seriata (on C. reticulata and C. sinensis); and Do. viticola (on C. aurantium and C. sinensis) had an equal percentage of frequency (14.3%); Di. mutila (exclusively found on C. sinensis), N. luteum and N. mediterraneum (only found on C. limon) and Di. olivarum and L. theobromae (exclusively found on C. sinensis) had low frequency values varying from 1.6% to 7.9%.
2.4. Pathogenicity Tests
All isolates caused lesions on wood of inoculated plants 60 d after inoculation (Figure 6) and the fungi were successfully re-isolated. No lesions were observed on control plants. The frequency of re-isolation was between 90% and 95%. The identities of the respective inoculated and re-isolated species were confirmed using culture and molecular analysis as described above, fulfilling Koch’s postulates. Lesions and internal discolouration were observed in correspondence to the inoculation points (Figure 7). The inoculated species that showed high aggressiveness on C. sinensis, C. limon, and C. reticulata were Di. seriata, Di. olivarum, L. theobromae, N. mediterraneum, N. luteum, and N. parvum (with mean lesion length (MLL) ranged from 5.25 to 6.96 cm). Weak symptoms were caused by Di. pseudoseriata, Di. mutila, and Do. viticola on the same species (with MLL ranged from 0.17 to 0.58 cm).
For each tested host species, the pairwise comparison, obtained from the Kruskal–Wallis test, showed significant differences (p < 0.05) between the species Di. seriata, Di. olivarum, L. theobromae, N. mediterraneum, N. luteum, and N. parvum and the remaining pathogens Di. pseudoseriata, Di. mutila and Do. viticola (Supplementary Tables S1–S3). No significant differences were observed within the group composed by Di. seriata, Di. olivarum, L. theobromae, N. mediterraneum, N. luteum, and N. parvum. Moreover, N. parvum revealed to be highly aggressive on M. australasica and C. sinensis x P. trifoliata with similar level of aggressiveness (Figure 8). The tested strain developed a MLL = 7.83 cm on M. australasica and a MLL = 7.45 cm on C. sinensis × P. trifoliata.
3. Discussion
Several Botryosphaeriaceae spp. have been detected in association with citrus cankers worldwide. Diplodia seriata, Di. mutila, Do. iberica, Do. viticola, L. parva, N. australe, N. luteum, N. mediterraneum, N. parvum, and Ne. dimidiatum have been recovered from necrotic tissues of branch canker and rootstock citrus samples in California [13,31,36]. Recently, Di. citricarpa was described for a fungus on twigs of Citrus sp. in Iran [16] and L. mitidjana was introduced for a fungus causing branch canker and dieback of C. sinensis in Algeria [33]. Botryosphaeriaceae spp. causing disease on citrus are known in European countries, where N. parvum and Ne. dimidiatum were reported on C. reticulata in Greece and on C. sinensis in Italy, respectively [32,37].
This study represents the first large survey aimed at studying the occurrence, genetic diversity, and pathogenicity of Botryosphaeriaceae species associated with symptomatic citrus species of citrus-producing areas in Greece, Italy, Portugal, Malta, and Spain [10,38]. Results obtained during our study have added new information about the pathogenicity of Botryosphaeriaceae spp. in citrus-producing areas of these European countries. Symptomatic plants were observed during fieldwork in all the citrus orchards and regions investigated and all isolates used in the pathogenicity test caused lesions on wood of inoculated citrus plants. Phylogenetic multi-marker analyses recognized botryosphaeriaceous isolates in four Diplodia species, with Di. pseudoseriata (15 isolates) being the most common; followed by three Neofusicoccum species, with N. parvum (16 isolates) as dominant species, Do. viticola (9 isolates), and L. theobromae (5 isolates). All species found in this study, except Di. pseudoseriata and Di. olivarum, which are reported for the first time on Citrus spp., have been found in citrus-producing areas of California (USA) [13,31,36].
Diplodia and Neofusicoccum species were dominant in this study. Different species of Neofusicoccum and Diplodia were the most frequently detected pathogens causing gummosis on citrus in California [36] and Di. citricarpa was a new species isolated from Citrus sp. in Iran [16]. Species of Diplodia, Dothiorella, Lasiodiplodia, and Neofusicoccum detected in our study are widely reported as pathogens of other host plants in Algeria and Tunisia [39,40], Australia [41], Brazil [42], China [43,44], Chile [45], Italy, Portugal [39,46,47,48], South Africa [49], and the USA [13,31,36]. The results obtained in our study provide valuable information related to the richness, occurrence, and pathogenicity of Botryosphaeriaceae species in association with citrus species. This study is also the first major survey for Botryosphaeriaceae species associated with symptomatic citrus species in citrus-producing areas of five European countries, providing essential information for future monitoring. Moreover, while previous reports of canker diseases of citrus were based exclusively on morphological observations, the current study aimed to investigate the fungi affecting the major citrus production areas in Europe by large-scale sampling, morphology, and DNA phylogeny. The information achieved with this study about Botryosphaeriaceae population and citrus canker etiology provide fundamental knowledge to start further studies aimed to improve the disease management.
4. Materials and Methods
4.1. Field Sampling and Fungal Isolation
During 2015 and 2016 more than 90 sites in the most important citrus-producing areas of Europe were investigated. The surveys were conducted in Andalusia, Valencia, and the Balearic Islands (Spain); Apulia, Calabria, Sicily, and Aeolian Islands (Italy); Algarve (Portugal); Arta, Crete, Missolonghi, and Nafplio (Greece); Malta and Gozo (Malta) [10,38]. Twig, branch and trunk portions showing cankers and dieback were collected. Investigated species of Citrus and allied genera of the Rutaceae family such as Microcitrus included: C. limon, C. reticulata, C. sinensis, M. australasica, and C. sinensis × P. trifoliata.
Wood fragments (5 × 5 mm) were collected from the margin between necrotic and healthy tissues. Then, each fragment was disinfected by immersion in 70% ethanol for 5 s, 4% sodium hypochlorite for 90 s, sterilised distilled water for 60 s and then dried on sterile filter paper. The fragments were placed into Petri dishes containing malt extract agar (MEA) [50] supplemented with penicillin (100 μg/mL) and streptomycin (100 μg/mL) (MEA-PS) and incubated at 25 °C until characteristic Botryosphaeriaceae colonies were observed. A second procedure was used with plant material incubated in moist chambers at 20 ± 3 °C for up to 10 d and inspected daily for fungal sporulation. The conidia obtained through both procedures were collected and crushed in a drop of sterile water and then spread over the surface of MEA-PS plates. After 24 h, germinating spores were individually transferred onto MEA plates. The isolates used in this study are maintained in the working collection of Pedro Crous (CPC), housed at the Westerdijk Fungal Biodiversity Institute (CBS), Utrecht, The Netherlands.
The occurrence of botryosphaeriaceous fungi among countries and citrus species was evaluated as the number of isolates from each fungal species against the total number of isolates and expressed as a percentage.
4.2. DNA Extraction, Polymerase Chain Reaction (PCR) Amplification and Sequencing
Colonies grown on MEA for 7 days were used to perform total DNA extraction using the Wizard®® Genomic DNA Purification Kit (Promega, Madison, WI, USA) standard protocol. The primer pair ITS4/ITS5 [51] was used to amplify the ITS. The primer sets EF1-728F/EF2 [52,53] and Bt2a/Bt2b [54] were used to amplify partial fragments of the TEF1 and TUB2 genes, respectively. Amplification by PCR was conducted as described by Yang et al. [16]. The PCR products were sequenced in both directions using the BigDye®® Terminator v. 3.1 Cycle Sequencing Kit (Applied Biosystems Life Technologies, Carlsbad, CA, USA), after which amplicons were purified through Sephadex G-50 Fine columns (GE Healthcare, Freiburg, Germany) in MultiScreen HV plates (Millipore, Billerica, MA, USA). Purified sequence reactions were analyzed on an Applied Biosystems 3730xl DNA Analyzer (Life Technologies, Carlsbad, CA, USA). The DNA sequences generated were analyzed and consensus sequences were computed using SeqMan Pro (DNASTAR, Madison, WI, USA). Sequences obtained in this study were deposited in GenBank https://academic.oup.com/nar/article/49/D1/D92/5983623 (accessed on 30 January 2021) (Table 2).
4.3. Phylogenetic Analyses
The phylogenetic analyses included DNA sequences generated in this study along with DNA sequences retrieved from GenBank (Table 2) and represent 124 Botryosphaeriaceae species (Diplodia = 23; Dothiorella = 31; Lasiodiplodia = 31; Neofusicoccum = 39) following recent studies [16,23,25]. Alignments were first made using MAFFT v. 7 [55] and manually checked and edited using MEGA v.7 [56]. Maximum Likelihood (ML) and Bayesian Inference (BI) analyses were conducted using RAxML-HPC BlackBox v.8.2.8 [57] and MrBayes v.3.2.7a on XSEDE, respectively, at the CIPRES Science Gateway. The best nucleotide models for the BI analysis were calculated using MrModelTest v.2.3 [58] while GTR + I + G was used for ML analysis. Clade stability of the ML phylogeny was assessed with 1000 bootstrap replicates. The BI analysis lasted for one million generations, a burning value of 25% and chains were sampled every 1000 generations. Values of ML bootstrap (ML-BS) and BI posterior probability (BPP) equal or greater than 70% and 0.95, respectively, were considered significant. Individual gene phylogenies were visually inspected and compared for topological incongruences before combining into a multi-marker sequence alignment. The combined alignments used to perform the phylogenetic inferences were deposited in TreeBASE (study ID S27709).
4.4. Pathogenicity Tests
Pathogenicity tests with nine Botryosphaeriaceae species isolated from the European citrus samples were performed to satisfy Koch’s postulates.
One isolate of Di. pseudoseriata (CPC 28084), Di. seriata (CPC 28091), Di. olivarum (CPC 27855), Di. mutila (CPC 26977), Do. viticola (CPC 27125), L. theobromae (CPC 27881), N. mediterraneum (CPC 27931), N. luteum (CPC 27961), and N. parvum (CPC 28175) were respectively inoculated onto potted 2-y-old healthy plants of lemon (C. limon), mandarin (C. reticulata) and sweet orange (C. sinensis). The strain of N. parvum was also inoculated onto potted 2-y-old healthy plants of Australasian lime (M. australasica) and Carrizo citrange (C. sinensis × P. trifoliata).
Three plants for each isolate were inoculated, each having five wounds on twigs made using a sterile blade. Mycelial plugs (5 mm diam.), taken from the margin of actively growing colonies on MEA, were placed on the wound sites on each plant. An equivalent number of plants and inoculation sites were inoculated with sterile MEA plugs and served as controls. The inoculation sites were covered with Parafilm®® (American National Can, Chicago, IL, USA). The inoculated plants were incubated with a 16 h photoperiod in a growth chamber at 100% relative humidity and 25 ± 1 °C. After 2 months external symptoms were assessed. Twigs were cut and the bark peeled off to check for any internal discolouration and the total, upward and downward lesion length was taken to evaluate the MLL. Small sections (0.5 cm) of symptomatic tissue from the edge of twig lesions were placed on MEA to re-isolate the fungal species and were identified based on TEF1 sequencing to fulfil Koch’s postulates. The experiment was conducted twice and each trial was considered a replicate. Because no normal distribution was observed in the lesion dimension data, the Kruskal–Wallis non-parametric test (at P = 0.05) was performed to determine significant differences among isolates. The data analysis was conducted using SPSS software 26 (IBM Corporate).
Supplementary Materials
The following are available online at https://www.mdpi.com/2223-7747/10/3/492/s1, Tables S1–S3. Kruskal-Wallis test results with multiple comparisons for disease severity between different Botryosphaeriaceae spp. on artificially inoculated twigs of C. sinensis (Table S1), C. limon (Table S2) and C. reticulata (Table S3).
Author Contributions
Conceptualization, V.G., G.P. and P.W.C.; methodology, J.D.P.B., D.A. and V.G.; software, J.D.P.B., V.G. and D.A.; validation, G.P., J.D.P.B. and V.G.; formal analysis, J.D.P.B., D.A. and V.G.; investigation, V.G. and D.A.; resources, P.W.C.; data curation, J.D.P.B. and V.G.; writing—original draft preparation, J.D.P.B. and V.G.; writing—review and editing, G.P., M.L.G. and P.W.C.; supervision, G.P., M.L.G. and P.W.C.; project administration, P.W.C.; funding acquisition, P.W.C. All authors have read and agreed to the published version of the manuscript.
Funding
This research received no external funding.
Institutional Review Board Statement
Not applicable.
Informed Consent Statement
Not applicable.
Acknowledgments
The authors thank G. Gilardi (AGROINNOVA—University of Torino) and A. Azzaro for technical support.
Conflicts of Interest
The authors declare no conflict of interest.
References
- FAOSTAT. Food and Agriculture Organization of the United Nations. 2019. Available online: http://www.fao.org/faostat/en/#home (accessed on 26 February 2020).
- Eurostat. Citrus Fruit Statistics; Eurostat: Luxembourg, 2020. [Google Scholar]
- Crous, P.W.; Wingfield, M.J. Fungi infecting woody plants: Emerging frontiers. Persoonia 2018, 40, i–iii. [Google Scholar] [CrossRef] [Green Version]
- Fawcett, H.S. Citrus Diseases and Their Control; McGraw-Hill: New York, NY, USA, 1936. [Google Scholar]
- Huang, F.; Chen, G.Q.; Hou, X.; Fu, Y.S.; Cai, L.; Hyde, K.D.; Li, H.Y. Colletotrichum species associated with cultivated citrus in China. Fungal Divers. 2013, 61, 61–74. [Google Scholar] [CrossRef]
- Huang, F.; Hou, X.; Dewdney, M.M.; Fu, Y.S.; Chen, G.Q.; Hyde, K.D.; Li, H. Diaporthe species occurring on citrus in China. Fungal Divers. 2013, 61, 237–250. [Google Scholar] [CrossRef]
- Mahadevakumar, S.; Yadav, V.; Tejaswini, G.S.; Sandeep, S.N.; Janardhana, G.R. First report of Phomopsis citri associated with dieback of Citrus lemon in India. Plant Dis. 2014, 98, 1281. [Google Scholar] [CrossRef]
- Guarnaccia, V.; Crous, P.W. Species of Diaporthe on Camellia and Citrus in the Azores Islands. Phytopathol. Mediterr. 2018, 57, 307–319. [Google Scholar]
- Mayorquin, J.S.; Nouri, M.T.; Peacock, B.B.; Trouillas, F.P.; Douhan, G.W.; Kallsen, C.; Eskalen, A. Identification, Pathogenicity, and Spore Trapping of Colletotrichum karstii Associated with Twig and Shoot Dieback in California. Plant Dis. 2019, 103, 1464–1473. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Guarnaccia, V.; Crous, P.W. Emerging citrus diseases in Europe caused by species of Diaporthe. IMA Fungus 2017, 8, 317–334. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Sandoval-Denis, M.; Guarnaccia, V.; Polizzi, G.; Crous, P.W. Symptomatic Citrus trees reveal a new pathogenic lineage in Fusarium and two new Neocosmospora species. Persoonia 2018, 40, 1–25. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Timmer, L.W.; Garnsey, S.M.; Graham, J.H. Compendium of Citrus Diseases, 2nd ed.; American Phytopathological Society: Saint Paul, MN, USA, 2000. [Google Scholar]
- Mayorquin, J.S.; Wang, D.H.; Twizeyimana, M.; Eskalen, A. Identification, distribution, and pathogenicity of Diatrypaceae and Botryosphaeriaceae associated with Citrus branch canker in the southern California desert. Plant Dis. 2016, 100, 2402–2413. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Espargham, N.; Mohammadi, H.; Gramaje, D.A. Survey of Trunk Disease Pathogens within Citrus Trees in Iran. Plants 2020, 9, 754. [Google Scholar] [CrossRef]
- Phillips, A.J.L.; Alves, A.; Abdollahzadeh, J.; Slippers, B.; Wingfield, M.J.; Groenewald, J.Z.; Crous, P.W. The Botryosphaeriaceae: Genera and species known from culture. Stud. Mycol. 2013, 76, 51–167. [Google Scholar] [CrossRef] [Green Version]
- Yang, T.; Groenewald, J.Z.; Cheewangkoon, R.; Jami, F.; Abdollahzadeh, J.; Lombard, L.; Crous, P.W. Families, genera, and species of Botryosphaeriales. Fungal Biol. 2017, 121, 322–346. [Google Scholar] [CrossRef] [PubMed]
- Slippers, B.; Wingfield, M.J. Botryosphaeriaceae as endophytes and latent pathogens of woody plants: Diversity, ecology and impact. Fungal Biol. Rev. 2007, 21, 90–106. [Google Scholar] [CrossRef]
- Crous, P.W.; Slippers, B.; Wingfield, M.J.; Rheeder, J.; Marasas, W.F.O.; Philips, A.J.L.; Alves, A.; Burgess, T.; Barber, P.; Groenewald, J.Z. Phylogenetic lineages in the Botryosphaeriaceae. Stud. Mycol. 2006, 55, 235–253. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Crous, P.W.; Giraldo, A.; Hawksworth, D.L.; Robert, V.; Kirk, P.M.; Guarro, J.; Robbertse, B.; Schoch, C.L.; Damm, U.; Trakunyingcharoen, T.; et al. The genera of fungi: Fixing the application of type species of generic names. IMA Fungus 2014, 5, 141–160. [Google Scholar] [CrossRef] [PubMed]
- Crous, P.W.; Muller, M.M.; Sanchez, R.M.; Giordano, L.; Bianchinotti, M.V.; Anderson, F.E.; Groenewald, J.Z. Resolving Tiarosporella spp. allied to Botryosphaeriaceae and Phacidiaceae. Phytotaxa 2015, 202, 73–93. [Google Scholar] [CrossRef]
- Schoch, C.L.; Shoemaker, R.A.; Seifert, K.A.; Hambleton, S.; Spatafora, J.W.; Crous, P.W. A multigene phylogeny of the Dothideomycetes using four nuclear loci. Mycologia 2006, 98, 1041–1052. [Google Scholar] [CrossRef]
- Slippers, B.; Boissin, E.; Phillips, A.J.L.; Groenewald, J.Z.; Lombard, L.; Wingfield, M.J.; Postma, A.; Burgess, T.; Crous, P.W. Phylogenetic lineages in the Botryosphaeriales: A systematic and evolutionary framework. Stud. Mycol. 2013, 76, 31–49. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Phillips, A.J.L.; Hyde, K.D.; Alves, A.; Liu, J. Families in Botryosphaeriales: A phylogenetic, morphological and evolutionary perspective. Fungal Divers. 2019, 94, 1–22. [Google Scholar] [CrossRef]
- Wijayawardene, N.N.; Hyde, K.D.; Al-Ani, L.K.T.; Tedersoo, L.; Haelewaters, D.; Rajeshkumar, K.C.; Zhao, R.L.; Aptroot, A.; Leontyev, D.V.; Saxena, R.K.; et al. Outline of Fungi and fungus-like taxa. Mycosphere 2020, 11, 1060–1456. [Google Scholar] [CrossRef]
- Zhang, W.; Groenewald, J.Z.; Lombard, L.; Schumacher, R.K.; Phillips, A.J.L.; Crous, P.W. Evaluating species in Botryosphaeriales. Persoonia 2021, 46, 63–115. [Google Scholar]
- Mehl, J.W.M.; Slippers, B.; Roux, J.; Wingfield, M.J. Cankers and other diseases caused by the Botryosphaeriaceae. In Infectious Forest Diseases; Gonthier, P., Nicolotti, G., Eds.; CAB International: Boston, MA, USA, 2013; pp. 298–317. [Google Scholar]
- Pour, F.N.; Ferreira, V.; Félix, C.; Serôdio, J.; Alves, A.; Duarte, A.S.; Esteves, A.C. Effect of temperature on the phytotoxicity and cytotoxicity of Botryosphaeriaceae fungi. Fungal Biol. 2020, 124, 571–578. [Google Scholar] [CrossRef]
- Guarnaccia, V.; Martino, I.; Tabone, G.; Brondino, L.; Gullino, M.L. Fungal pathogens associated with stem blight and dieback of blueberry in northern Italy. Phytopathol. Mediterr. 2020, 59, 229–245. [Google Scholar]
- Marsberg, A.; Kemler, M.; Jami, F.; Nagel, J.H.; Postma-Smidt, A.; Naidoo, S.; Wingfield, M.J.; Crous, P.W.; Spatafora, J.W.; Hesse, C.N.; et al. Botryosphaeria dothidea: A latent pathogen of global importance to woody plant health. Mol. Plant Pathol. 2017, 18, 477–488. [Google Scholar] [CrossRef] [PubMed]
- Aiello, D.; Gusella, G.; Fiorenza, A.; Guarnaccia, V.; Polizzi, G. Identification of Neofusicoccum parvum causing canker and twig blight on Ficus carica in Italy. Phytopathol. Mediterr. 2020, 59, 213–218. [Google Scholar]
- Adesemoye, A.O.; Eskalen, A. First Report of Spencermartinsia viticola, Neofusicoccum australe, and N. parvum causing branch canker of citrus in California. Plant Dis. 2011, 95, 770. [Google Scholar] [CrossRef] [PubMed]
- Polizzi, G.; Aiello, D.; Vitale, A.; Giuffrida, F.; Groenewald, J.; Crous, P.W. First report of shoot blight, canker, and gummosis caused by Neoscytalidium dimidiatum on citrus in Italy. Plant Dis. 2009, 93, 1215. [Google Scholar] [CrossRef]
- Berraf-Tebbal, A.; Mahamedi, A.E.; Aigoun-Mouhous, W.; Špetík, M.; Čechová, J.; Pokluda, R.; Baránek, M.; Eichmeier, A.; Alves, A. Lasiodiplodia mitidjana sp. nov. and other Botryosphaeriaceae species causing branch canker and dieback of Citrus sinensis in Algeria. PLoS ONE 2020, 15, e0232448. [Google Scholar] [CrossRef]
- Abdollahzadeh, J.; Javadi, A.; Goltapeh, E.M.; Zare, R.; Phillips, A.J. Phylogeny and morphology of four new species of Lasiodiplodia from Iran. Persoonia 2010, 25, 1–10. [Google Scholar] [CrossRef] [Green Version]
- Hamrouni, N.; Nouri, M.; Trouillas, F.; Said, A.; Sadfi-Zouaoui, N.; Hajlaoui, M. Dothiorella gummosis caused by Dothiorella viticola, first record from citrus in Tunisia. New Dis. Rep. 2018, 38, 10. [Google Scholar] [CrossRef] [Green Version]
- Adesemoye, A.O.; Mayorquin, J.S.; Wang, D.H.; Twizeyimana, M.; Lynch, S.C.; Eskalen, A. Identification of species of Botryosphaeriaceae causing bot gummosis in citrus in California. Plant Dis. 2014, 98, 55–61. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Vakalounakis, D.J.; Ntougias, S.; Kavroulakis, N.; Protopapadakis, E. Neofusicoccum parvum and Diaporthe foeniculina associated with twig and shoot blight and branch canker of citrus in Greece. J. Phytopathol. 2019, 167, 527–537. [Google Scholar] [CrossRef]
- Guarnaccia, V.; Groenewald, J.Z.; Li, H.; Glienke, C.; Carstens, E.; Hattingh, V.; Crous, P.W. First report of Phyllosticta citricarpa and description of two new species, P. paracapitalensis and P. paracitricarpa, from citrus in Europe. Stud. Mycol. 2017, 87, 161–185. [Google Scholar] [CrossRef]
- Linaldeddu, B.T.; Deidda, A.; Scanu, B.; Franceschini, A.; Serra, S.; Berraf-Tebbal, A.; Boutiti, M.Z.; Jamâa, M.L.B.; Phillips, A.J.L. Diversity of Botryosphaeriaceae species associated with grapevine and other woody hosts in Italy, Algeria and Tunisia, with descriptions of Lasiodiplodia exigua and Lasiodiplodia mediterranea sp. nov. Fungal Divers. 2015, 71, 201–214. [Google Scholar] [CrossRef]
- Mahamedi, A.E.; Phillips, A.J.L.; Lopes, A.; Djellid, Y.; Arkam, M.; Eichmeier, A.; Zitouni, A.; Alves, A.; Berraf-Tebbal, A. Diversity, distribution and host association of Botryosphaeriaceae species causing oak decline across different forest ecosystems in Algeria. Eur. J. Plant Pathol. 2020, 158, 745–765. [Google Scholar] [CrossRef]
- Burgess, T.I.; Tan, Y.P.; Garnas, J.; Edwards, J.; Scarlett, K.A.; Shuttleworth, L.A.; Daniel, R.; Dann, E.K.; Parkinson, L.E.; Dinh, Q. Current status of the Botryosphaeriaceae in Australia. Australas. Plant Pathol. 2018, 48, 35–44. [Google Scholar] [CrossRef]
- Machado, A.R.; Custódio, F.A.; Cabral, P.G.C.; Capucho, A.S.; Pereira, O.L. Botryosphaeriaceae species causing dieback on Annonaceae in Brazil. Plant Pathol. 2019, 68, 1394–1406. [Google Scholar] [CrossRef]
- Xu, C.; Zhang, H.; Zhou, Z.; Hu, T.; Wang, S.; Wang, Y.; Cao, K. Identification and distribution of Botryosphaeriaceae species associated with blueberry stem blight in China. Eur. J. Plant Pathol. 2015, 143, 737–752. [Google Scholar] [CrossRef]
- Li, G.; Slippers, B.; Wingfield, M.J.; Chen, S. Variation in Botryosphaeriaceae from Eucalyptus plantations in YunNan Province in southwestern China across a climatic gradient. IMA Fungus 2020, 11, 22. [Google Scholar] [CrossRef]
- Valencia, A.L.; Pilar, M.; Gil, B.A.; Latorre, I.; Rosales, M. Characterization and Pathogenicity of Botryosphaeriaceae Species Obtained from Avocado Trees with Branch Canker and Dieback and from Avocado Fruit with Stem End Rot in Chile. Plant Dis. 2019, 103, 996–1005. [Google Scholar] [CrossRef]
- Gusella, G.; Aiello, D.; Polizzi, G. First report of leaf and twig blight of Indian hawthorn (Rhaphiolepis indica) caused by Neofusicoccum parvum in Italy. J. Plant Pathol. 2020, 102, 275. [Google Scholar] [CrossRef] [Green Version]
- Alves, A.; Linaldeddu, B.T.; Deidda, A.; Scanu, B.; Phillips, A.J.L. The complex of Diplodia species associated with Fraxinus and some other woody hosts in Italy and Portugal. Fungal Divers. 2014, 67, 143–156. [Google Scholar] [CrossRef]
- Dissanayake, A.J.; Camporesi, E.; Hyde, K.D.; Yan, J.Y.; Li, X.H. Saprobic Botryosphaeriaceae, including Dothiorella italica sp nov., associated with urban and forest trees in Italy. Mycosphere 2017, 8, 1157–1176. [Google Scholar] [CrossRef]
- Jami, F.; Slippers, B.; Wingfield, M.J.; Loots, M.T.; Gryzenhout, M. Temporal and spatial variation of Botryosphaeriaceae associated with Acacia karroo in South Africa. Fungal Ecol. 2015, 15, 51–62. [Google Scholar] [CrossRef] [Green Version]
- Crous, P.W.; Verkley, G.J.M.; Groenewald, J.Z.; Samson, R.A. (Eds.) Fungal Biodiversity; CBS Laboratory Manual Series 1; Centraalbureau Voor Schimmelcultures: Utrecht, The Netherlands, 2009; pp. 1–269. [Google Scholar]
- White, T.J.; Bruns, T.; Lee, S.; Taylor, J. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In PCR Protocols: A Guide to Methods and Applications; Innis, A.M., Gelfard, D.H., Snindky, J.J., White, T.J., Eds.; Academic Press: San Diego, CA, USA, 1990; pp. 315–322. [Google Scholar]
- Carbone, I.; Kohn, L.M. A method for designing primer sets for speciation studies in filamentous ascomycetes. Mycologia 1999, 91, 553–556. [Google Scholar] [CrossRef]
- O’Donnell, K.; Kistler, H.C.; Cigelnik, E.; Ploetz, R.C. Multiple evolutionary origins of the fungus causing Panama disease of banana: Concordant evidence from nuclear and mitochondrial gene genealogies. Proc. Natl. Acad. Sci. USA 1998, 95, 2044–2049. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Glass, N.L.; Donaldson, G.C. Development of primer sets designed for use with the PCR to amplify conserved genes from filamentous ascomycetes. Appl. Environ. Microbiol. 1995, 61, 1323–1330. [Google Scholar] [CrossRef] [Green Version]
- Katoh, K.; Standley, D.M. MAFFT multiple sequence alignment software version 7: Improvements in performance and usability. Mol. Biol. Evol. 2013, 30, 772–780. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Kumar, S.; Stecher, G.; Tamura, K. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Mol. Biol. Evol. 2016, 33, 1870–1874. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Stamatakis, A. RAxML version 8: A tool for phylogenetic analysis and post-analysis of large phylogenies. Bioinformatics 2014, 30, 1312–1313. [Google Scholar] [CrossRef]
- Nylander, J. MrModeltest v. 2 Program Distributed by the Author; Evolutionary Biology Centre, Uppsala University: Uppsala, Sweden, 2004. [Google Scholar]
Figure 1.
Symptoms on citrus tissues with associated Botryosphaeriacae species. (A) Branch decline in commercial lemon orchard. (B) Trunk canker and bark cracking of C. sinensis. (C,D) Trunk and branch canker with gummosis of C. sinensis plants. (E,F) External cracking with gummosis and internal wood discoloration of the same affected branch of C. reticulata plant. (G,H) Internal wood discoloration and branch blight of C. limon. (I) Twig dieback of young C. sinensis × P. trifoliata and M. australasica (J) plants.
Figure 1.
Symptoms on citrus tissues with associated Botryosphaeriacae species. (A) Branch decline in commercial lemon orchard. (B) Trunk canker and bark cracking of C. sinensis. (C,D) Trunk and branch canker with gummosis of C. sinensis plants. (E,F) External cracking with gummosis and internal wood discoloration of the same affected branch of C. reticulata plant. (G,H) Internal wood discoloration and branch blight of C. limon. (I) Twig dieback of young C. sinensis × P. trifoliata and M. australasica (J) plants.
Figure 2.
Bayesian inference analysis of Diplodia species using ITS rDNA, TEF1 and TUB2 sequences. Isolates obtained in this study are in bold and blue. Bayesian posterior probability (BPP) and maximum likelihood-bootstrap (ML-BS) values equal or greater than 0.95 and 70%, respectively, are shown near nodes. Thickened branches represent clades with ML-BS = 100% and a BPP = 1.0. The tree was rooted to L. theobormae (CBS 111530, CBS 164.96 and CBS 124.13).
Figure 2.
Bayesian inference analysis of Diplodia species using ITS rDNA, TEF1 and TUB2 sequences. Isolates obtained in this study are in bold and blue. Bayesian posterior probability (BPP) and maximum likelihood-bootstrap (ML-BS) values equal or greater than 0.95 and 70%, respectively, are shown near nodes. Thickened branches represent clades with ML-BS = 100% and a BPP = 1.0. The tree was rooted to L. theobormae (CBS 111530, CBS 164.96 and CBS 124.13).
Figure 3.
Bayesian inference analysis of Dothiorella species using ITS rDNA, TEF1, and TUB2 sequences. Isolates obtained in this study are in bold and blue. Bayesian posterior probability (BPP) and ML bootstrap (ML-BS) values equal or greater than 0.95 and 70%, respectively, are shown near nodes. Thickened branches represent clades with ML-BS = 100% and a BPP = 1.0. The tree was rooted to N. luteum (CBS 110299 and CBS 110497).
Figure 3.
Bayesian inference analysis of Dothiorella species using ITS rDNA, TEF1, and TUB2 sequences. Isolates obtained in this study are in bold and blue. Bayesian posterior probability (BPP) and ML bootstrap (ML-BS) values equal or greater than 0.95 and 70%, respectively, are shown near nodes. Thickened branches represent clades with ML-BS = 100% and a BPP = 1.0. The tree was rooted to N. luteum (CBS 110299 and CBS 110497).
Figure 4.
Bayesian inference analysis of Lasiodiplodia species using ITS rDNA, TEF1, and TUB2 sequences. Isolates obtained in this study are in bold and blue. Bayesian posterior probability (BPP) and ML bootstrap (ML-BS) values equal or greater than 0.95 and 70%, respectively, are shown near nodes. Thickened branches represent clades with ML-BS = 100% and a BPP = 1.0. The tree was rooted to Do. viticola (CBS 117009).
Figure 4.
Bayesian inference analysis of Lasiodiplodia species using ITS rDNA, TEF1, and TUB2 sequences. Isolates obtained in this study are in bold and blue. Bayesian posterior probability (BPP) and ML bootstrap (ML-BS) values equal or greater than 0.95 and 70%, respectively, are shown near nodes. Thickened branches represent clades with ML-BS = 100% and a BPP = 1.0. The tree was rooted to Do. viticola (CBS 117009).
Figure 5.
Bayesian inference analysis of species Neofusicoccum using ITS rDNA, TEF1, and TUB2 sequences. Isolates obtained in this study are in bold and blue. Bayesian posterior probability (BPP) and ML bootstrap (ML-BS) values equal or greater than 0.95 and 70%, respectively, are shown near nodes. Thickened branches represent clades with ML-BS = 100% and a BPP = 1.0. The tree was rooted to B. dothidea (CBS 115476).
Figure 5.
Bayesian inference analysis of species Neofusicoccum using ITS rDNA, TEF1, and TUB2 sequences. Isolates obtained in this study are in bold and blue. Bayesian posterior probability (BPP) and ML bootstrap (ML-BS) values equal or greater than 0.95 and 70%, respectively, are shown near nodes. Thickened branches represent clades with ML-BS = 100% and a BPP = 1.0. The tree was rooted to B. dothidea (CBS 115476).
Figure 6.
Pathogenicity tests of selected Botryosphaeriacae isolates on citrus plants 60 d after inoculation. (A,B) Shoot blight of C. reticulata and C. sinensis plants inoculated with N. mediterraneum. (C) Internal lesion with abundant gummosis of C. sinensis plant caused by N. parvum. (D,E) Internal discoloration of C. sinensis and C. reticulata twigs inoculated with L. theobromae.
Figure 6.
Pathogenicity tests of selected Botryosphaeriacae isolates on citrus plants 60 d after inoculation. (A,B) Shoot blight of C. reticulata and C. sinensis plants inoculated with N. mediterraneum. (C) Internal lesion with abundant gummosis of C. sinensis plant caused by N. parvum. (D,E) Internal discoloration of C. sinensis and C. reticulata twigs inoculated with L. theobromae.
Figure 7.
Box plot showing the results of the pathogenicity tests. Boxes represent the interquartile range, while the horizontal line within each box indicates the average value. The Kruskal–Wallis test was carried out to compare the mean lesion lengths (cm) from inoculation with nine Botryosphaeriaceae representative isolates on C. sinensis (A), C. limon (B) and C. reticulata (C). p < 0.05 was taken to indicate a significant difference. °: Outliers. *: Extreme values.
Figure 7.
Box plot showing the results of the pathogenicity tests. Boxes represent the interquartile range, while the horizontal line within each box indicates the average value. The Kruskal–Wallis test was carried out to compare the mean lesion lengths (cm) from inoculation with nine Botryosphaeriaceae representative isolates on C. sinensis (A), C. limon (B) and C. reticulata (C). p < 0.05 was taken to indicate a significant difference. °: Outliers. *: Extreme values.
Figure 8.
The Kruskal–Wallis test was carried out to compare the mean lesion lengths (cm) from inoculation with one N. parvum representative isolate on C. sinensis × P. trifoliata and M. australasica. Significant difference was accepted for p < 0.05.
Figure 8.
The Kruskal–Wallis test was carried out to compare the mean lesion lengths (cm) from inoculation with one N. parvum representative isolate on C. sinensis × P. trifoliata and M. australasica. Significant difference was accepted for p < 0.05.
Table 1.
Geographical sites investigated and sampled.
| Site | Locality | GPS Coordinates |
|---|---|---|
| 1 | Algemesi (Spain) | 39°11′48.8″ N, 0°28′15.0″ W |
| 2 | Alginet (Spain) | 39°15′36.3″ N, 0°27′28.9″ W |
| 3 | Alhaurin El Grande (Spain) | 36°38′43.4″ N, 4°40′37.5″ W |
| 4 | Alzira (Spain) | 39°09′25.1″ N, 0°29′26.6″ W |
| 5 | Castellò (Spain) | 39°54′14.1″ N, 0°05′10.3″ W |
| 6 | Estellencs (Spain) | 39°39′12.6″ N, 2°28′54.8″ E |
| 7 | Faro (Portugal) | 37°03′45.5″ N, 7°55′02.8″ W |
| 8 | Gozo (Malta) | 36°02′15.1″ N, 14°15′36.4″ E |
| 9 | Gozo (Malta) | 36°03′18.5″ N, 14°15′35.7″ E |
| 10 | Malaga (Spain) | 36°45′42.3″ N, 4°25′37.4″ W |
| 11 | Mascali (Italy) | 37°46′05.7″ N, 15°11′40.7″ E |
| 12 | Massafra (Italy) | 40°32′41.1″ N, 17°08′38.8″ E |
| 13 | Mastro (Greece) | 38°25′49.0″ N, 21°16′49.9″ E |
| 14 | Mesquita (Portugal) | 37°12′16.3″ N, 8°17′52.1″ W |
| 15 | Moncada (Spain) | 39°35′18.8″ N, 0°23′40.5″ W |
| 16 | Nafplio (Greece) | 37°34′56.3″ N, 22°41′48.5″ E |
| 17 | Rocca Imperiale (Italy) | 40°06′30.2″ N, 16°37′04.6″ E |
| 18 | Scordia (Italy) | 37°16′53.5″ N, 14°52′08.9″ E |
| 19 | Silves (Portugal) | 37°09′50.7″ N, 8°23′21.7″ W |
Table 2.
GenBank accession numbers of sequences of Diplodia, Dothiorella, Lasiodiplodia, and Neofusicoccum species used in the phylogenetic analyses. Isolates and sequences obtained in this study are given in bold.
Table 2.
GenBank accession numbers of sequences of Diplodia, Dothiorella, Lasiodiplodia, and Neofusicoccum species used in the phylogenetic analyses. Isolates and sequences obtained in this study are given in bold.
| Species | Strains 1 | Host/Substrate | Country | GenBank Numbers 2 | ||
|---|---|---|---|---|---|---|
| ITS | TEF1 | TUB2 | ||||
| Botryosphaeria dothidea | CBS 115476 = CMW 8000, ex-epitype | Prunus sp. | Switzerland | AY236949 | AY236898 | AY236927 |
| Diplodia africana | CBS 120835 = CPC 5908, ex-type | Prunus persica, stem canker | South Africa | EF445343 | EF445382 | KF766129 |
| Di. agrifolia | CBS 132777 = UCR732, ex-type | Quercus agrifolia, cankered branch | USA: California | JN693507 | JQ517317 | JQ411459 |
| Di. allocellula | CBS 130408 = CMW 36468, ex-type | Acacia karroo, healthy branches | South Africa | JQ239397 | JQ239384 | JQ239378 |
| Di. bulgarica | CBS 124254 = CAP332, ex-type | Malus sylvestris | Bulgaria | GQ923853 | GQ923821 | – |
| Di. citricarpa | CBS 124715 = CJA 131 = IRAN 1578C, ex-type | Citrus sp., twigs | Iran | KF890207 | KF890189 | KX464784 |
| Di. corticola | CBS 112549 = CAP 134, ex-type | Quercus suber | Portugal | AY259100 | AY573227 | DQ458853 |
| Di. crataegicola | MFLU 15-13112, ex-type | Crataegus sp. | Italy | KT290244 | KT290248 | KT290246 |
| Di. cupressi | CBS 168.87, ex-type | Cupressus sempervirens, canker | Israel | DQ458893 | DQ458878 | DQ458861 |
| Di. eriobotryicola | CBS 140851 = BN-21, ex-type | Eriobotrya japonica | Spain | KT240355 | KT240193 | MG015806 |
| Di. estuarina | CMW 41231 | Avicennia marina | South Africa | KP860831 | KP860676 | KP860754 |
| Di. fraxini | CBS 136010 = CAD001, ex-type | Fraxinus angustifolia | Portugal | KF307700 | KF318747 | MG015807 |
| Di. gallae | CBS 211.25 | Quercus sp., fruit | – | KX464090 | KX464564 | KX464795 |
| Di. gallae | CBS 212.25 | Quercus sp., gall | – | KX464091 | KX464565 | KX464796 |
| Di. gallae | CBS 213.25 | Quercus sp., gall | – | KX464092 | KX464566 | KX464797 |
| Di. malorum | CBS 124130 = CAP271, ex-type | Malus sylvestris | Portugal | GQ923865 | GQ923833 | – |
| Di. mutila | CPC 26977 | Citrus sinensis, twig | Greece | MW413831 | MW419149 | MW419212 |
| Di. mutila | CBS 112553 = CAP 062 | Vitis vinifera | Portugal | AY259093 | AY573219 | DQ458850 |
| Di. mutila | CBS 121862 = PD 03708098, ex-type of Di. pyri | Pyrus sp. | The Netherlands | KX464093 | KX464567 | KX464799 |
| Di. neojuniperi | CPC 22753 = B0031, ex-type | Juniperus chinensis | Thailand | KM006431 | KM006462 | – |
| Di. olivarum | CPC 27855 | Citrus sinensis,branch | Malta | MW413832 | MW419150 | MW419213 |
| Di. olivarum | CPC 27856 | Citrus sinensis,branch | Malta | MW413833 | MW419151 | MW419214 |
| Di. olivarum | CBS 121886 | Olea europaea | Italy | EU392301 | EU392278 | – |
| Di. olivarum | CBS 121887 = CAP 254, ex-type | Olea europaea, rotting drupes | Italy | EU392302 | EU392279 | HQ660079 |
| Di. pseudoseriata | CBS 124906, ex-type | Blepharocalyx salicifolius | Uruguay | EU080927 | EU863181 | MG015820 |
| Di. pseudoseriata | CPC 27963 | Citrus sinensis, twig | Portugal | MW413834 | MW419152 | MW419215 |
| Di. pseudoseriata | CPC 27964 | Citrus sinensis, twig | Portugal | MW413835 | MW419153 | MW419216 |
| Di. pseudoseriata | CPC 27965 | Citrus sinensis, twig | Portugal | MW413836 | MW419154 | MW419217 |
| Di. pseudoseriata | CPC 27966 | Citrus sinensis, twig | Portugal | MW413837 | MW419155 | MW419218 |
| Di. pseudoseriata | CPC 27967 | Citrus sinensis, twig | Portugal | MW413838 | MW419156 | MW419219 |
| Di. pseudoseriata | CPC 28084 | Citrus reticulata, twig | Spain | MW413839 | MW419157 | MW419220 |
| Di. pseudoseriata | CPC 28086 | Citrus reticulata, twig | Spain | MW413840 | MW419158 | MW419221 |
| Di. pseudoseriata | CPC 28087 | Citrus reticulata, twig | Spain | MW413841 | MW419159 | MW419222 |
| Di. pseudoseriata | CPC 28092 | Citrus limon, twig | Spain | MW413842 | MW419160 | MW419223 |
| Di. pseudoseriata | CPC 28093 | Citrus limon, twig | Spain | MW413843 | MW419161 | MW419224 |
| Di. pseudoseriata | CPC 28094 | Citrus limon, twig | Spain | MW413844 | MW419162 | MW419225 |
| Di. pseudoseriata | CPC 28095 | Citrus limon, twig | Spain | MW413845 | MW419163 | MW419226 |
| Di. pseudoseriata | CPC 28099 | Citrus reticulata, twig | Spain | MW413846 | MW419164 | MW419227 |
| Di. pseudoseriata | CPC 28100 | Citrus reticulata, twig | Spain | MW413847 | MW419165 | MW419228 |
| Di. pseudoseriata | CPC 28102 | Citrus reticulata, twig | Spain | MW413848 | MW419166 | MW419229 |
| Di. pseudoseriata | BL132 | Fraxinus angustifolia | Italy | KF307720 | KF318767 | MG015810 |
| Di. pseudoseriata | CBS 140350, ex-type of Di. insularis | Pistacia lentiscus | Italy | KX833072 | KX833073 | MG015809 |
| Di. pseudoseriata | CBS 124931 = CMW22627, ex-type of Di. alatafructa | Pterocarpus angolensis, bark wound | South Africa | FJ888460 | FJ888444 | MG015799 |
| Di. quercivora | CBS 133852 = BL8, ex-type | Quercus canariensis | Tunisia | JX894205 | JX894229 | MG015821 |
| Di. rosulata | CBS 116470, ex-type | Prunus africana | Ethiopia | EU430265 | EU430267 | EU673132 |
| Di. sapinea | CBS 393.84, ex-epitype | Pinus nigra, cones | Netherlands | DQ458895 | DQ458880 | DQ458863 |
| Di. sapinea | CBS 124462 = CAP273, ex-type of Di. intermedia | Malus sylvestris | Portugal | GQ923858 | GQ923826 | – |
| Di. sapinea | CBS 141915 = NB7, ex-type of Di. rosacearum | Eriobotrya japonica | Italy | KT956270 | KU378605 | MG015823 |
| Di. scrobiculata | CBS 118110 = CMW 189 = BOT 1195, ex-type | Pinus banksiana | USA: Wisconsin | AY253292 | AY624253 | AY624258 |
| Di. seriata | CBS 112555 = HAP 052 = CAP 063, ex-epitype | Vitis vinifera, dead stems | Portugal | AY259094 | AY573220 | DQ458856 |
| Di. seriata | CPC 28088 | Citrus reticulata,twig | Spain | MW413849 | MW419167 | MW419230 |
| Di. seriata | CPC 28089 | Citrus reticulata,twig | Spain | MW413850 | MW419168 | MW419231 |
| Di. seriata | CPC 28090 | Citrus reticulata,twig | Spain | MW413851 | MW419169 | MW419232 |
| Di. seriata | CPC 28091 | Citrus reticulata,twig | Spain | MW413852 | MW419170 | MW419233 |
| Di. seriata | CPC 28096 | Citrus sinensis,twig | Spain | MW413853 | MW419171 | MW419234 |
| Di. seriata | CPC 28097 | Citrus sinensis,twig | Spain | MW413854 | MW419172 | MW419235 |
| Di. seriata | CPC 28098 | Citrus sinensis,twig | Spain | MW413855 | MW419173 | MW419236 |
| Di. seriata | CPC 28101 | Citrus reticulata,twig | Spain | MW413856 | MW419174 | MW419237 |
| Di. seriata | CPC 28103 | Citrus reticulata,twig | Spain | MW413857 | MW419175 | MW419238 |
| Di. seriata | CBS 119049 | Vitis sp. | Italy | DQ458889 | DQ458874 | DQ458857 |
| Di. subglobosa | CBS 124133 = JL453, ex-type | Lonicera nigra | Spain | GQ923856 | GQ923824 | – |
| Di. tsugae | CBS 418.64 = IMI 197143, ex-isotype | Tsuga heterophylla | Canada | DQ458888 | DQ458873 | DQ458855 |
| Dothiorella alpina | CGMCC 3-18001, ex-type | Platycladus orientalis | China | KX499645 | KX499651 | – |
| Do. americana | CBS 128309, ex-type | Wedge-shape canker of grapevine cv. Vignoles (complex hybrid of North America Vitis species and Vitis vinifera) | USA: Missouri | HQ288218 | HQ288262 | HQ288297 |
| Do. brevicollis | CBS 130411 = CMW 36463, ex-type | Acacia karroo, healthy branches | South Africa | JQ239403 | JQ239390 | JQ239371 |
| Do. capri-amissi | CBS 121763 = CMW 25403 = CAMS 1158, ex-paratype | Acacia erioloba | South Africa | EU101323 | EU101368 | KX464850 |
| Do. capri-amissi | CBS 121878 = CMW 25404 = CAMS 1159, ex-type | Acacia erioloba | South Africa | EU101324 | EU101369 | KX464851 |
| Do. casuarinae | CBS 120688 = CMW 4855, ex-type | Casuarina sp. | Australia: Australian Capital Territory | DQ846773 | DQ875331 | DQ875340 |
| Do. casuarinae | CBS 120689 = CMW 4856, ex-paratype | Casuarina sp. | Australia: Australian Capital Territory | DQ846772 | DQ875332 | DQ875339 |
| Do. casuarinae | CBS 120690 = CMW 4857, ex-paratype | Casuarina sp. | Australia: Australian Capital Territory | DQ846774 | DQ875333 | DQ875341 |
| Do. citricola | CBS 124728 = ICMP 16827 | Citrus sinensis | New Zealand | EU673322 | EU673289 | KX464852 |
| Do. citricola | CBS 124729 = ICMP 16828, ex-type | Citrus sinensis, twigs | New Zealand | EU673323 | EU673290 | KX464853 |
| Do. dulcispinae | CBS 121764 = CMW 25406 = CAMS 1159, ex-paratype of Dothiorella oblonga | Acacia mellifera | Namibia | EU101299 | EU101344 | KX464854 |
| Do. dulcispinae | CBS 130413 = CMW 36460, ex-type | Acacia karroo, dieback branches | South Africa | JQ239400 | JQ239387 | JQ239373 |
| Do. iberica | CBS 113188 = DA-1 | Quercus suber, branch canker | Spain | AY573198 | EU673278 | EU673097 |
| Do. iberica | CBS 113189 = DE-14 | Quercus ilex, branch canker | Spain | AY573199 | AY573230 | KX464855 |
| Do. iberica | CBS 115041 = CAP 145, ex-type | Quercus ilex, dead twigs | Spain | AY573202 | AY573222 | EU673096 |
| Do. iranica | CBS 124722 = CJA 153 = IRAN 1587C | Olea sp., twigs | Iran | KC898231 | KC898214 | KX464856 |
| Do. longicollis | CBS 122066 = CMW 26164 | Terminalia sp. | Australia: Western Australia | EU144052 | EU144067 | KX464857 |
| Do. longicollis | CBS 122067 = CMW 26165 | Lysiphyllum cunninghamii | Australia: Western Australia | EU144053 | EU144068 | KX464858 |
| Do. longicollis | CBS 122068 = CMW 26166, ex-type | Lysiphyllum cunninghamii | Australia: Western Australia | EU144054 | EU144069 | KF766130 |
| Do. mangifericola | CBS 124727 = IRAN 1584C = CJA 261, ex-type | Mangifera indica, twigs | Iran | KC898221 | KX464614 | – |
| Do. moneti | WAC 13154 = MUCC 505, ex-type | Acacia rostellifera, healthy stem | Australia: Western Australia | EF591920 | EF591971 | EF591954 |
| Do. neclivorem | DAR 80992, ex-type | Vitis vinifera cv. Chardonnay, berries | Australia: New South Wales | KJ573643 | KJ573640 | KJ577551 |
| Do. oblonga | CBS 121765 = CMW 25407 = CAMS 1162, ex-type | Acacia mellifera | South Africa | EU101300 | EU101345 | KX464862 |
| Do. oblonga | CBS 121766 = CMW 25408 = CAMS 1163, ex-paratype | Acacia mellifera | South Africa | EU101301 | EU101346 | KX464863 |
| Do. omnivora | CBS 124717 = CJA 214 = IRAN 1570C | Juglans regia, twigs | Iran | KC898233 | KC898216 | KX464865 |
| Do. omnivora | CBS 392.80 | – | France | KX464133 | KX464626 | KX464897 |
| Do. omnivora | CBS 124716 = CJA 241 = IRAN 1573C | Juglans regia, twigs | Iran | KC898232 | KC898215 | KX464864 |
| Do. omnivora | CBS 242.51 | – | Italy | EU673317 | EU673284 | EU673105 |
| Do. omnivora | CBS 188.87 | Juglans regia | France | EU673316 | EU673283 | EU673119 |
| Do. parva | CBS 124720 = CJA 27 = IRAN 1579C, ex-type | Corylus sp., twigs | Iran | KC898234 | KC898217 | KX464866 |
| Do. parva | CBS 124721 = CJA 35 | Corylus sp., twigs | Iran | KX464123 | KX464615 | KX464867 |
| Do. parva | CBS 125580 | Corylus avellana, branches | Austria | KX464124 | KX464616 | KX464868 |
| Do. plurivora | CBS 124724 = CJA 254 = IRAN 1557C, ex-type | Citrus sp., twigs | Iran | KC898225 | KC898208 | KX464874 |
| Do. pretoriensis | CBS 130404 = CMW 36480, ex-type | Acacia karroo, branches with dieback | South Africa | JQ239405 | JQ239392 | JQ239376 |
| Do. prunicola | CBS 124723 = CAP 187 = IRAN 1541C, ex-type | Prunus dulcis, necrotic twigs | Portugal | EU673313 | EU673280 | EU673100 |
| Do. rosulata | CBS 121760 = CMW 25389 = CAMS 1444, ex-type | Acacia karroo | Namibia | KF766227 | EU101335 | KX464877 |
| Do. rosulata | CBS 121761 = CMW 25392 = CAMS 1147, ex-paratype | Acacia mellifera | South Africa | EU101293 | EU101338 | KX464878 |
| Do. rosulata | CBS 121762 = CMW 25395 = CAMS 1150 | Acacia mellifera | South Africa | EU101319 | EU101364 | KX464879 |
| Do. rosulata | CBS 500.72 | Medicago sativa, stubble | South Africa | EU673318 | EU673285 | EU673118 |
| Do. santali | WAC 13155 = MUCC 509, ex-type | Santalum acuminatum, healthy stem | Australia: Western Australia | EF591924 | EF591975 | EF591958 |
| Do. sarmentorum | IMI 63581b, ex-type of Bot. sarmentorum | Ulmus sp. | UK: England | AY573212 | AY573235 | EU673102 |
| Do. sempervirentis | IRAN 1581C = CBS 124719 | Cupressus sempervirens | Iran | KC898237 | KC898220 | KX464885 |
| Do. sempervirentis | IRAN 1583C = CBS 124718 = CJA 264, ex-type | Cupressus sempervirens, twigs | Iran | KC898236 | KC898219 | KX464884 |
| Do. striata | CBS 124730 = ICMP 16819 | Citrus sinensis, twigs | New Zealand | EU673320 | EU673287 | EU673142 |
| Do. striata | CBS 124731 = ICMP 16824, ex-type | Citrus sinensis, twigs | New Zealand | EU673321 | EU673288 | EU673143 |
| Do. thailandica | CBS 133991 = CPC 21557 = MFLUCC 11-0438, ex-type of Auerswaldia dothiorella | Dead bamboo culm | Thailand | JX646796 | JX646861 | JX646844 |
| Do. thripsita | CBS 125445 = BRIP 51876a, ex-type | Acacia harpophylla, dead branches, petioles & leaves | Australia: Queensland | KJ573642 | KJ573639 | KJ577550 |
| Do. uruguayensis | CBS 124908 = CMW 26763 = UY672, ex-type | Hexachlamis edulis | Uruguay | EU080923 | EU863180 | KX464886 |
| Do. vidmadera | CBS 621.74 | Pyrus communis, leaf | Switzerland | KX464129 | KX464621 | KX464887 |
| Do. vidmadera | CBS 725.79 | Pyrus malus, dead wood and buds | Switzerland | KX464130 | KX464622 | KX464888 |
| Do. vinea-gemmae | DAR 81012, ex-type | Vitis vinifera cv. Chardonnay, dormant buds | Australia: New South Wales | KJ573644 | KJ573641 | KJ577552 |
| Do. viticola | CBS 117009, ex-type | Vitis vinifera cv. Garnatxa negra, pruned canes | Spain | AY905554 | AY905559 | EU673104 |
| Do. viticola | DAR 80529, ex-type of D. westralis | Vitis vinifera cv. Cabernet Sauvignon, discarded canes | Australia: Western Australia | HM009376 | HM800511 | HM800519 |
| Do. viticola | CPC 26174 | Citrus sinensis, twig | Italy | MW413858 | MW419176 | MW419239 |
| Do. viticola | CPC 26917 | Citrus sinensis, branch | Greece | MW413859 | MW419177 | MW419240 |
| Do. viticola | CPC 27081 | Citrus sinensis, twig | Italy | MW413860 | MW419178 | MW419241 |
| Do. viticola | CPC 27106 | Citrus aurantium, twig | Spain | MW413861 | MW419179 | MW419242 |
| Do. viticola | CPC 27123 | Citrus sinensis, branch | Italy | MW413862 | MW419180 | MW419243 |
| Do. viticola | CPC 27125 | Citrus sinensis, branch | Italy | MW413863 | MW419181 | MW419244 |
| Do. viticola | CPC 27703 | Citrus sinensis, branch | Spain | MW413864 | MW419182 | MW419245 |
| Do. viticola | CPC 27707 | Citrus sinensis, branch | Greece | MW413865 | MW419183 | MW419246 |
| Do. viticola | CPC 27968 | Citrus sinensis, twig | Portugal | MW413866 | MW419184 | MW419247 |
| Do. yunnana | CGMCC 3-17999, ex-type | Camellia sp. | China | KX499643 | KX499649 | – |
| Do. yunnana | CGMCC 3-18000 | Camellia sp. | China | KX499644 | KX499650 | – |
| Dothiorella sp. | CBS 121783 = CMW 25432 = CAMS 1187 | Acacia mearnsii | South Africa | EU101333 | EU101378 | KX464859 |
| Dothiorella sp. | CBS 121784 = CMW 25430 = CAMS 1185 | Acacia mearnsii | South Africa | EU101331 | EU101376 | KX464860 |
| Dothiorella sp. | CBS 121785 = CMW 25433 = CAMS 1188 | Acacia mearnsii | South Africa | EU101334 | EU101379 | KX464861 |
| ‘Lasiodiplodia americana’ | CERC 1961 = CFCC 50065, ex-type | Pistacia vera cv. Kerman, twigs | USA: Arizona | KP217059 | KP217067 | KP217075 |
| L. avicenniae | CMW 41467 = CBS 139670, ex-type | Avicennia marina | South Africa | KP860835 | KP860680 | KP860758 |
| L. brasiliense | CMM 4015 = URM 7118, ex-type | Mangifera indica, stems | Brazil | JX464063 | JX464049 | – |
| L. bruguierae | CMW 41470 = CBS 139669, ex-type | Bruguiera gymnorrhiza | South Africa | NR_147358 | KP860678 | KP860756 |
| L. citricola | CBS 124707 = IRAN 1522C = CJA 72, ex-type | Citrus sp., twigs | Iran | GU945354 | GU945340 | KP872405 |
| L. crassispora | CBS 118741 = WAC 12533 = CMW 14691, ex-type | Santalum album | Australia: Western Australia | DQ103550 | EU673303 | EU673133 |
| L. crassispora | CBS 121770 = CMW 25414 = CAMS 1169, ex-type of L. pyriformis | Acacia mellifera | Namibia | EU101307 | EU101352 | – |
| L. endophytica | MFLUCC 18-1121 = KUMCC 17-0233, ex-type | Magnolia candolii, fresh leaves | China | MK501838 | MK584572 | MK550606 |
| L. egyptiacae | CBS 130992 = BOT-10, ex-type | Mangifera indica, leaf | Egypt | JN814397 | JN814424 | – |
| L. euphorbicola | CMM 3609, ex-type of L. euphorbicola | Jatropha curcas, collar and root rot | Brazil | KF234543 | KF226689 | KF254926 |
| L. gilanensis | CBS 124704 = IRAN 1523C, ex-type | Citrus sp., fallen twigs | Iran | GU945351 | GU945342 | KP872411 |
| L. gilanensis | CBS 128311 = UCD 2193MO, ex-type of L. missouriana | Wedge-shape canker of grapevine cv. Catawba (complex hybrid of North America Vitis species and Vitis vinifera) | USA: Missouri | HQ288225 | HQ288267 | – |
| L. gonubiensis | CBS 115812 = CMW 14077, ex-type | Syzygium cordatum, twigs and leaves | South Africa | AY639595 | DQ103566 | DQ458860 |
| L. gravistriata | CMM 4564, ex-type | Anacardium humile | Brazil | KT250949 | KT250950 | – |
| L. hormozganensis | CBS 124709 = IRAN 1500C, ex-type | Olea sp., twigs | Iran | GU945355 | GU945343 | KP872413 |
| L. iraniensis | CBS 124710 = IRAN 1520C, ex-type | Salvadora persica, twigs | Iran | GU945346 | GU945334 | KP872415 |
| L. iraniensis | CMM 3610, ex-type of L. jatrophicola | Jatropha curcas, collar and root rot | Brazil | KF234544 | KF226690 | KF254927 |
| L. laeliocattleyae | CBS 167.28, ex-type of Diplodia laeliocattleyae | Laeliocattleya | Italy | KU507487 | KU507454 | – |
| L. lignicola | MFLUCC 11-0435 = CBS 134112, ex-type | On dead wood | Thailand | JX646797 | KU887003 | JX646845 |
| L. lignicola | CBS 342.78, ex-type of L. sterculiae | Sterculia oblonga | Germany | KX464140 | KX464634 | KX464908 |
| L. macrospora | CMM 3833, ex-type | Jatropha curcas, collar and root rot | Brazil | KF234557 | KF226718 | KF254941 |
| ‘L. magnoliae’ | MFLUCC 18-0948 = KUMCC 17-0198, ex-type | Magnolia candolii, dead leaves | China | MK499387 | MK568537 | MK521587 |
| L. mahajangana | CBS 124927 = CMW27801, ex-type | Terminalia catappa, healthy branches | Madagascar | FJ900595 | FJ900641 | FJ900630 |
| L. mahajangana | CMM 1325, ex-type of L. caatinguensis | Citrus sinensis | Brazil | KT154760 | KT008006 | KT154767 |
| L. mahajangana | CBS 137785 = BL104, ex-type of L. exigua | Retama raetam, branch canker | Tunisia | KJ638317 | KJ638336 | – |
| L. margaritacea | CBS 122519 = CMW 26162 = MOZ 11A, ex-type | Adansonia gibbosa | Australia: Western Australia | EU144050 | EU144065 | KX464903 |
| L. mediterranea | CBS 137783 = BL1, ex-type | Quercus ilex, branch canker | Italy | KJ638312 | KJ638331 | – |
| L. mitidjana | MUM 19.90 = ALG111, ex-type | Citrus sinensis, branch canker | Algeria: Mitidja | MN104115 | MN159114 | – |
| L. parva | CBS 456.78, ex-type | Cassava-field soil | Colombia | EF622083 | EF622063 | KP872419 |
| L. plurivora | CBS 120832 = CPC 5803, ex-type | Prunus salicina, wood canker | South Africa | EF445362 | EF445395 | KP872421 |
| L. pontae | CMM 1277, ex-type | Spondias purpurea | Brazil | KT151794 | KT151791 | KT151797 |
| L. pseudotheobromae | CBS 116459, ex-type | Gmelina arborea | Costa Rica | EF622077 | EF622057 | EU673111 |
| L. rubropurpurea | CBS 118740 = WAC 12535 = CMW 14700, ex-type | Eucalyptus grandis, canker | Australia | DQ103553 | EU673304 | EU673136 |
| L. subglobosa | CMM 3872, ex-type | Jatropha curcas, collar and root rot | Brazil | KF234558 | KF226721 | KF254942 |
| L. thailandica | CBS 138760 = CPC 22795, ex-type | Mangifera indica, twigs | Thailand | KJ193637 | KJ193681 | – |
| L. theobromae | CBS 111530 = CPC 2095 = JT 695 | Leucospermum sp. | USA: Hawaii | EF622074 | EF622054 | – |
| L. theobromae | CPC 27881 | Citrus sinensis, trunk | Malta | MW413867 | MW419185 | MW419248 |
| L. theobromae | CPC 27882 | Citrus sinensis, trunk | Malta | MW413868 | MW419186 | MW419249 |
| L. theobromae | CPC 27883 | Citrus sinensis, trunk | Malta | MW413869 | MW419187 | MW419250 |
| L. theobromae | CPC 27884 | Citrus sinensis, trunk | Malta | MW413870 | MW419188 | MW419251 |
| L. theobromae | CPC 27885 | Citrus sinensis, trunk | Malta | MW413871 | MW419189 | MW419252 |
| L. theobromae | CBS 124.13 | – | USA | DQ458890 | DQ458875 | DQ458858 |
| L. theobromae | CBS 164.96, ex-neotype | Fruit along coral reef coast | Papua New Guinea | AY640255 | AY640258 | EU673110 |
| L. venezuelensis | CBS 118739 = WAC 12539 = CMW 13511, ex-type | Acacia mangium, wood | Venezuela | DQ103547 | EU673305 | EU673129 |
| L. viticola | CBS 128313 = UCD 2553AR, ex-type | Wedge-shape canker of grapevine cv. Vignoles (complex hybrid of North America Vitis species and Vitis vinifera) | USA: Arkansas | HQ288227 | HQ288269 | HQ288306 |
| L. vitis | CBS 124060, ex-type | Vitis vinifera | KX464148 | KX464642 | KX464917 | |
| Neofusicoccum arbuti | CBS 117453 = CMW 13455, ex-type of N. andinum | Eucalyptus sp. | Venezuela | AY693976 | AY693977 | KX464923 |
| N. arbuti | CBS 116131 = AR 4014, ex-type | Arbutus menziesii, canker | USA: Washington | AY819720 | KF531792 | KF531793 |
| N. australe | CBS 139662 = CMW 6837, ex-type | Acacia sp. | Australia: Victoria | AY339262 | AY339270 | AY339254 |
| N. australe | CMW 6853 | Sequoiadendron | Australia | AY339263 | AY339271 | AY339255 |
| N. brasiliense | CMM 1338, ex-type | Mangifera indica | Brazil | JX513630 | JX513610 | KC794030 |
| N. buxi | CBS 116.75 | Buxus sempervirens, leaf | France | KX464165 | KX464678 | – |
| N. cordaticola | CBS 123634 = CMW 13992, ex-type | Syzygium cordatum | South Africa | EU821898 | EU821868 | EU821838 |
| N. cryptoaustrale | CBS 122813 = CMW 23785, ex-type | Eucalyptus sp., living branches and leaves | South Africa | FJ752742 | FJ752713 | FJ752756 |
| N. dianense | CSF6075 = CGMCC3.20082, ex-type | Eucalyptus urophylla × E. grandis tree, twigs | China | MT028605 | MT028771 | MT028937 |
| N. eucalypticola | CBS 115679 = CMW 6539, ex-type | Eucalyptus grandis | Australia | AY615141 | AY615133 | AY615125 |
| N. eucalyptorum | CBS 115791 = CMW 10125 = BOT 24 | Eucalyptus grandis | South Africa | AF283686 | AY236891 | AY236920 |
| N. grevilleae | CBS 129518, ex-type | Grevillea aurea | Australia | JF951137 | – | – |
| N. hellenicum | CERC 1947 = CFCC 50067, ex-type | Pistacia vera cultivar Aegina | Greece | KP217053 | KP217061 | KP217069 |
| N. hongkongense | CERC2973 = CGMCC3.18749, ex-type | Araucaria cunninghamii | China | KX278052 | KX278157 | KX278261 |
| N. illicii | CGMCC3.18310, ex-type | Illicium verum | China | KY350149 | – | KY350155 |
| N. kwambonambiense | CBS 123639 = CMW 14023, ex-type | Syzygium cordatum | South Africa | EU821900 | EU821870 | EU821840 |
| N. lumnitzerae | CBS 139674 = CMW 41469, ex-type | Lumnitzera racemosa | South Africa | KP860881 | KP860724 | KP860801 |
| N. luteum | CPC 27961 | Citrus limon, twig | Portugal | MW413872 | MW419190 | MW419253 |
| N. luteum | CPC 27962 | Citrus limon, twig | Portugal | MW413873 | MW419191 | MW419254 |
| N. luteum | CBS 110497 = CPC 4594 = CAP 037 | Vitis vinifera, grape | Portugal | EU673311 | EU673277 | EU673092 |
| N. luteum | CBS 110299 = LM 926 = CAP 002, ex-type | Vitis vinifera, cane | Portugal | AY259091 | KX464688 | DQ458848 |
| N. luteum | CBS 140738 = CMW 41365, ex-type of N. mangroviorum | Avicennia marina | South Africa | KP860859 | KP860702 | KP860779 |
| N. macroclavatum | CBS 118223 = CMW 15955 = WAC 12444, ex-type | Eucalyptus globulus, wood | Australia: Western Australia | DQ093196 | DQ093217 | DQ093206 |
| N. magniconidium | CSF5876 = CGMCC3.20077, ex-type | Eucalyptus urophylla × E. grandis tree, twigs | China | MT028612 | MT028778 | MT028944 |
| N. mangiferae | CBS 118531 = CMW 7024 | Mangifera indica | Australia | AY615185 | DQ093221 | AY615173 |
| N. mediterraneum | CBS 121718 = CPC 13137, ex-type | Eucalyptus sp., branches and leaves | Greece | GU251176 | – | – |
| N. mediterraneum | CBS 113083 = CPC 5263, ex-type of N. pistaciarum | Pistacia vera | USA: California | KX464186 | KX464712 | KX464998 |
| N. mediterraneum | CBS 113089 = CPC 5274, ex-type of N. pistaciicola | Pistacia vera | USA: California | KX464199 | KX464727 | KX465014 |
| N. mediterraneum | CPC 27931 | Citrus limon, twig | Portugal | MW413874 | MW419192 | MW419255 |
| N. mediterraneum | CPC 27932 | Citrus limon, twig | Portugal | MW413875 | MW419193 | MW419256 |
| N. mediterraneum | CPC 27935 | Citrus limon, twig | Portugal | MW413876 | MW419194 | MW419257 |
| N. mediterraneum | CPC 27936 | Citrus limon, twig | Portugal | MW413877 | MW419195 | MW419258 |
| N. microconidium | CERC3497 = CGMCC3.18750, ex-type | Eucalyptus urophylla × E. grandis tree | China | KX278053 | KX278158 | KX278262 |
| N. nonquaesitum | CBS 126655 = L3IE1 = PD484, ex-type | Umbellularia californica, cankered branch | USA: California | GU251163 | GU251295 | GU251823 |
| N. ningerense | CSF6028 = CGMCC3.20078, ex-type | Eucalyptus urophylla × E. grandis tree, twigs | China | MT028613 | MT028779 | MT028945 |
| N. occulatum | CBS 128008 = MUCC 227, ex-type | Eucalyptus grandis hybrid | Australia: Queensland | EU301030 | EU339509 | EU339472 |
| N. pandanicola | MFLUCC 17-2270 = KUMCC 17-0184, ex-type | Pandanus sp. | China | MH275072 | – | – |
| N. parviconidium | CSF5667 = CGMCC3.20074, ex-type | Eucalyptus tree, twigs | China | MT028615 | MT028781 | MT028947 |
| N. parvum | CBS 138823 = ICMP 8003 = CMW 9081 = BOT2487 = ATCC 58191, ex-type | Populus nigra, bark of dead twig | New Zealand | AY236943 | AY236888 | AY236917 |
| N. parvum | CPC 26119 | Citrus sinensisx Poncirus trifoliata, trunk | Italy | MW413878 | MW419196 | MW419259 |
| N. parvum | CPC 26120 | Citrus sinensisx Poncirus trifoliata, trunk | Italy | MW413879 | MW419197 | MW419260 |
| N. parvum | CPC 26121 | Citrus sinensisx Poncirus trifoliata, trunk | Italy | MW413880 | MW419198 | MW419261 |
| N. parvum | CPC 26122 | Citrus sinensisx Poncirus trifoliata, trunk | Italy | MW413881 | MW419199 | MW419262 |
| N. parvum | CPC 26124 | Citrus sinensisx Poncirus trifoliata, trunk | Italy | MW413882 | MW419200 | MW419263 |
| N. parvum | CPC 26126 | Citrus sinensisx Poncirus trifoliata, trunk | Italy | MW413883 | MW419201 | MW419264 |
| N. parvum | CPC 26127 | Citrus sinensisx Poncirus trifoliata, trunk | Italy | MW413884 | MW419202 | MW419265 |
| N. parvum | CPC 26128 | Citrus sinensisx Poncirus trifoliata, trunk | Italy | MW413885 | MW419203 | MW419266 |
| N. parvum | CPC 26129 | Citrus sinensisx Poncirus trifoliata, trunk | Italy | MW413886 | MW419204 | MW419267 |
| N. parvum | CPC 26130 | Citrus sinensisx Poncirus trifoliata, trunk | Italy | MW413887 | MW419205 | MW419268 |
| N. parvum | CPC 27866 | Citrus limon, branch | Malta | MW413888 | MW419206 | MW419269 |
| N. parvum | CPC 27867 | Citrus limon, branch | Malta | MW413889 | MW419207 | MW419270 |
| N. parvum | CPC 27868 | Citrus limon, branch | Malta | MW413890 | MW419208 | MW419271 |
| N. parvum | CPC 28173 | Microcitrus australasica, twig | Italy | MW413891 | MW419209 | MW419272 |
| N. parvum | CPC 28175 | Microcitrus australasica, twig | Italy | MW413892 | MW419210 | MW419273 |
| N. parvum | CPC 28177 | Microcitrus australasica, twig | Italy | MW413893 | MW419211 | MW419274 |
| N. parvum | CBS 110301 = CAP 074 | Vitis vinifera | Portugal | AY259098 | AY573221 | EU673095 |
| N. parvum | MFLUCC 15-09002, ex-type of N. italicum | Vitis vinifera | Italy | KY856755 | KY856754 | – |
| N. parvum | CBS 137504 = ALG1, ex-type of N. algeriense | Vitis vinifera, branches | Algeria | KJ657702 | KJ657721 | – |
| N. pennatisporum | WAC 13153 = MUCC 510, ex-type | Allocasuarina fraseriana, healthy stem | Australia: Western Australia | EF591925 | EF591976 | EF591959 |
| N. pistaciae | CBS 595.76, ex-isotype of Camarosporium pistaciae | Pistacia vera, fruits | Greece | KX464163 | KX464676 | KX464953 |
| N. protearum | CBS 114176 = CPC 1775 = JT 189, ex-type | Leucadendron salignum × L. laureolum cv. Silvan Red, stems | South Africa | AF452539 | KX464720 | KX465006 |
| N. ribis | CBS 115475 = CMW 7772, ex-type | Ribes vulgare | USA | AY236935 | AY236877 | AY236906 |
| N. ribis | CBS 124924 = CMW 28363, ex-type of N. batangarum | Terminalia catappa, healthy branches | Cameroon | FJ900607 | FJ900653 | FJ900634 |
| N. ribis | CBS 123645 = CMW 14058, ex-type of N. umdonicola | Syzygium cordatum | South Africa | EU821904 | EU821874 | EU821844 |
| N. sinense | CGMCC3.18315, ex-type | Unknown woody plant | China | KY350148 | KY817755 | KY350154 |
| N. sinoeucalypti | CERC2005 = CGMCC3.18752, ex-type | Eucalyptus urophylla × E. grandis tree | China | KX278061 | KX278166 | KX278270 |
| N. stellenboschiana | CBS 110864 = STE-U 4598 = CPC 4598, ex-type | Vitis vinifera | South Africa | AY343407 | AY343348 | KX465047 |
| N. terminaliae | CBS 125264 = CMW 26683 | Terminalia sericea | South Africa | GQ471804 | GQ471782 | KX465053 |
| N. ursorum | CBS 122811 = CMW 24480, ex-type | Eucalyptus sp. | South Africa | FJ752746 | FJ752709 | KX465056 |
| N. variabile | CMW 37739, ex-type | Mimusops caffra | South Africa | MH558608 | – | MH569153 |
| N. viticlavatum | CBS 112878 = CPC 5044 = JM 86, ex-type | Vitis vinifera | South Africa | AY343381 | AY343342 | KX465058 |
| N. vitifusiforme | CBS 110887 = CPC 5252 = JM5, ex-type | Vitis vinifera | South Africa | AY343383 | AY343343 | KX465061 |
| N. vitifusiforme | CBS 120081 = CPC 12925, ex-type of N. corticosae | Eucalyptus corticosa, leaves | Australia: New South Wales | DQ923533 | KX464682 | KX464958 |
| N. vitifusiforme | CBS 121112 = CPC 5912, ex-type of N. pruni | Prunus salicina | South Africa | EF445349 | EF445391 | KX465016 |
| N. yunnanense | CSF6142 = CGMCC3.20083, ex-type | Eucalyptus globulus, twigs | China | MT028667 | MT028833 | MT028999 |
1 ATCC: American Type Culture Collection, Virginia, USA; BRIP: Queensland Plant Pathology Herbarium, Brisbane, Australia; CBS: Westerdijk Fungal Biodiversity Institute, Utrecht, the Netherlands; CERC: China Eucalypt Research Centre (CERC), Chinese Academy of Forestry (CAF), China; CGMCC: China General Microbiological Culture Collection Center, Beijing, China; CMM: Culture collection of Phytopathogenic Fungi “Prof. Maria Menezes”, Universidade Federal Rural de Pernambuco, Recife, Brazil; CMW: Tree Pathology Co-operative Program, Forestry and Agricultural Biotechnology Institute, University of Pretoria, South Africa; CPC: Culture collection of Pedro Crous, housed at Westerdijk Fungal Biodiversity Institute; IMI: International Mycological Institute, Kew, U.K.; MFLUCC: Mae Fah Luang University Culture Collection, Chiang Ria, Thailand; MUCC: Murdoch University, Perth, Western Australia; URM: Culture collection Prof. Maria Auxiliadora Cavalcanti, Recife, Brazil. For other codes see the GenBank accession numbers. 2 ITS: internal transcribed spacers and intervening 5.8S nrDNA; TEF1: partial translation elongation factor 1-alpha gene; TUB2: partial β-tubulin gene.
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
MDPI and ACS Style
Bezerra, J.D.P.; Crous, P.W.; Aiello, D.; Gullino, M.L.; Polizzi, G.; Guarnaccia, V. Genetic Diversity and Pathogenicity of Botryosphaeriaceae Species Associated with Symptomatic Citrus Plants in Europe. Plants 2021, 10, 492. https://doi.org/10.3390/plants10030492
AMA Style
Bezerra JDP, Crous PW, Aiello D, Gullino ML, Polizzi G, Guarnaccia V. Genetic Diversity and Pathogenicity of Botryosphaeriaceae Species Associated with Symptomatic Citrus Plants in Europe. Plants. 2021; 10(3):492. https://doi.org/10.3390/plants10030492
Chicago/Turabian StyleBezerra, Jadson Diogo Pereira, Pedro Wilhelm Crous, Dalia Aiello, Maria Lodovica Gullino, Giancarlo Polizzi, and Vladimiro Guarnaccia. 2021. "Genetic Diversity and Pathogenicity of Botryosphaeriaceae Species Associated with Symptomatic Citrus Plants in Europe" Plants 10, no. 3: 492. https://doi.org/10.3390/plants10030492
Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.
