Lack of host specificity of Colletotrichum spp. isolates associated with anthracnose symptoms on mango in Brazil

The aim of the present study was to analyse the genetic and pathogenic variability of Colletotrichum spp. isolates from various organs and cultivars of mango with anthracnose symptoms, collected from different municipalities of São Paulo State, Brazil. Colletotrichum gloeosporioides isolates from symptomless citrus leaves and C. acutatum isolates from citrus flowers with post‐bloom fruit drop symptoms were included as controls. Sequencing of the ITS region allowed the identification of 183 C. gloeosporioides isolates from mango; only one isolate was identified as C. acutatum. amova analysis of ITS sequences showed larger genetic variability among isolates from the same municipality than among those from different populations. fAFLP markers indicated high levels of genetic variability among the C. gloeosporioides isolates from mango and no correlation between genetic variability and isolate source. Only one C. gloeosporioides mango isolate had the same genotype as the C. gloeosporioides isolates from citrus leaves, as determined by ITS sequencing and fAFLP analysis. Pathogenicity tests revealed that C. gloeosporioides and C. acutatum isolates from either mango or citrus can cause anthracnose symptoms on leaves of mango cvs Palmer and Tommy Atkins and blossom blight symptoms in citrus flowers. These outcomes indicate a lack of host specificity of the Colletotrichum species and suggest the possibility of host migration.     

Species from the Colletotrichum acutatum,Colletotrichum boninense and Colletotrichum gloeosporioides species complexes associated with tree tomato and mango crops in Colombia

Colletotrichum species cause typical anthracnose symptoms in tree tomato and mango. To characterize species of Colletotrichum in these two crops in Colombia, 91 isolates were collected from several localities. Phylogenetic analyses using nuclear gene sequencing of the ITS region and the glyceraldehyde 3‐phosphate dehydrogenase (GAPDH) gene allowed the identification of three groups: acutatum, gloeosporioides and boninense. These three groups were further confirmed using two additional genomic regions (chitin synthase 1 and actin) for 30 isolates representative of the three previously identified complexes and one genomic region (ApMat) for the Colletotrichum gloeosporioides complex strains. The entire approach permitted a robust strain identification that allowed phylogenetic species recognition (PSR) based on the identification of well‐supported monophyletic clades and concordance between individual and multilocus phylogenies. Morphological and physiological assays were also conducted. Isolates that were morphologically identified as C. gloeosporioides showed high phenotypic diversity. Pathogenicity data revealed a considerable degree of host preference.     

Colletotrichum species associated with chili anthracnose in Australia

Phylogenetic relationships were determined for 45 Colletotrichum isolates causing anthracnose disease of chili in Queensland, Australia. Initial screening based on morphology, ITS and TUB2 genes resulted in a subset of 21 isolates being chosen for further taxonomic study. Isolates in the C. acutatum complex were analysed using partial sequences of six gene regions (ITS, GAPDH, ACT, CHS‐1, TUB2 and HIS3), and in the C. gloeosporioides complex were analysed using four gene regions (ITS, TUB2, ApMat and GS). Phylogenetic analysis delineated four Colletotrichum species including C. siamense, C. simmondsii, C. queenslandicum, C. truncatum and a new Colletotrichum species, described here as C. cairnsense sp. nov. This is the first reported association of C. queenslandicum, C. simmondsii and C. siamense with chili anthracnose in Australia; these species were previously associated with anthracnose on papaya and avocado. Furthermore, the dominant species causing anthracnose of chili in Southeast Asia, C. scovillei, was not detected in Australia. Inoculations on chili fruit confirmed the pathogenicity of C. cairnsense and the other four species in the development of chili anthracnose in Australia.     

Phylogenetic and morphological identification of Colletotrichum boninense: a novel causal agent of anthracnose in avocado

In recent years, anthracnose has become a significant disease affecting avocado fruit in the state of Michoacan, Mexico, where it significantly reduces fruit quality and commercial yield. Anthracnose has been assumed to involve Colletotrichum gloeosporioides and C. acutatum as causal agents. However, because of the increasing incidence of anthracnose, a more precise identification of the Colletotrichum spp. involved in this disease has become desirable. During the years 2004–2007, avocado fruits of different sizes exhibiting brown‐black and reddish spots on the pericarp and soft rot in the mesocarp, were gathered from orchards in nine counties. Fungal isolates were cultured on potato dextrose agar, and among these, 31 were selected for molecular, morphological and pathogenicity analyses. The molecular approaches used sequence typing of the internal transcribed spacer region and the partial nuclear large ribosomal subunit, allowing the unequivocal identification of C. gloeosporioides (71%), C. acutatum (16%) and C. boninense (13%). This last species has not been previously reported as being associated with anthracnose symptoms in avocado fruits anywhere in the world. Various morphological characteristics such as the size and shape of conidia were determined, as well as the conidial mass colour. Pathogenicity tests performed with all three species were conducted by inoculating healthy fruits. In each case, identical symptoms developed within 3 days of inoculation. Knowledge of the Colletotrichum populations in the Michoacan state, including the newly encountered avocado pathogen C. boninense, will facilitate further studies addressing the relationships between these Colletotrichum spp. and their avocado host.     

Morphological and molecular diversity of Colletotrichum spp. causing pepper spot and anthracnose of lychee (Litchi chinensis) in Australia

Since the 1980s a new disease has been affecting Australian lychee. Pepper spot appears as small, black superficial lesions on fruit, leaves, petioles and pedicels and is caused by Colletotrichum gloeosporioides, the same fungus that causes postharvest anthracnose of lychee fruit. The aim of this study was to determine if a new genotype of C. gloeosporioides is responsible for the pepper spot symptom. Morphological assessments, arbitrarily‐primed PCR (ap‐PCR) and DNA sequencing studies did not differentiate isolates of C. gloeosporioides from anthracnose and pepper spot lesions. The ap‐PCR identified 21 different genotypes of C. gloeosporioides, three of which were predominant. A specific genotype identified using ap‐PCR was associated with the production of the teleomorph in culture. Analysis of sequence data of ITS and β‐tubulin regions of representative isolates did not group the lychee isolates into a monophyletic clade; however, given the majority of the isolates were from one of three genotypes found using ap‐PCR, the possibility of a lychee specific group of C. gloeosporioides is discussed.     

Characterization and epidemiology of Colletotrichum acutatum sensu lato (C. chrysanthemi) causing Carthamus tinctorius anthracnose

In 2012, Colletotrichum isolates were collected from field‐grown safflower (Carthamus tinctorius) crops in central Italy from plants exhibiting typical anthracnose symptoms. Colletotrichum isolates were also collected from seed surfaces and from within seeds. The genetic variability of these isolates was assessed by a multilocus sequencing approach and compared with those from Colletotrichum chrysanthemi and Colletotrichum carthami isolates from different geographic areas and other Colletotrichum acutatum sensu lato‐related isolates. Phylogenetic analysis revealed that all of the strains isolated from C. tinctorius belonged to the species described as C. chrysanthemi, whereas all of the strains belonging to C. carthami had been isolated from Calendula officinalis. Phenotypic characterization of isolates was performed by assessing growth rates at different temperatures, morphology of colonies on potato dextrose agar (PDA) and the size of conidia. All C. chrysanthemi isolates from safflower had similar growth rates at different temperatures, comparable colony morphologies when grown on PDA and conidial sizes consistent with previously described C. chrysanthemi isolates. Pathogenicity tests were performed by artificially inoculating both seeds and plants and confirmed the seedborne nature of this pathogen. When inoculated on plants, C. chrysanthemi caused the typical symptoms of anthracnose on leaves. This is the first record of this pathogen on C. tinctorius in Italy, and it presents an updated characterization of Colletotrichum isolates pathogenic to safflowers in Europe.     

Aetiology of anthracnose on grapevine shoots in Brazil

Anthracnose is an important disease in vineyards in south and southeast Brazil, the main grape‐producing regions in the country. This study aimed to identify the causal agents of grapevine anthracnose in Brazil through multilocus phylogenetic analyses, morphological characterization and pathogenicity tests. Thirty‐nine Elsinoë ampelina and 13 Colletotrichum spp. isolates were obtained from leaves, stems and berries with anthracnose symptoms collected in 38 vineyards in southern and southeastern Brazil. For E. ampelina isolates, the internal transcribed spacer (ITS), histone H3 (HIS3) and elongation factor 1‐α (TEF) sequences were analysed. HIS3 was the most informative region with 55 polymorphic sites including deletions and substitutions of bases, enabling the grouping of isolates into five haplotypes. Colonies of E. ampelina showed slow growth, variable colouration and a wrinkled texture on potato dextrose agar. Conidia were cylindrical to oblong with rounded ends, hyaline, aseptate, (3.57–) 5.64 (?6.95) μm long and (2.03–) 2.65 (?3.40) μm wide. Seven species of Colletotrichum were identified: C. siamense, C. gloeosporioides, C. fructicola, C. viniferum, C. nymphaeae, C. truncatum and C. cliviae, with a wide variation in colony and conidium morphology. Only E. ampelina caused anthracnose symptoms on leaves, tendrils and stems of Vitis vinifera and V. labrusca. High disease severity and a negative correlation between disease severity and shoot dry weight were observed only when relative humidity was above 95%. In this study, only E. ampelina caused anthracnose symptoms on grapevine shoots in Brazil.     

Phenotypic and genetic characterization of Colletotrichum isolates from Belgian strawberry fields

Colletotrichum isolates (457) were collected from strawberry plant tissues with and without typical anthracnose symptoms and from symptomless weeds in 19 Belgian strawberry fields. The isolates were characterized based on genetic, morphological and pathological features. Isolates were classified according to rDNA‐ITS sequencing: 97% of 211 representative isolates were C. acutatum, 2%C. gloeosporioides and 1%C. coccodes. The C. acutatum isolates belonged to the intraspecific groups A2 (33%), A3 (5%), A4 (50%), A5 (3%) and A7 (6%). Differences in spore morphology, growth rate and colony colour of a selection of 146 isolates confirmed the genetic grouping. Multiple Colletotrichum genotypes were detected in the same field. There was no association between the most common genotypes and geographic origin, presence or absence of symptoms, nor plant species or plant part. Representative Belgian Colletotrichum isolates were used in pathogenicity tests, together with European and American reference isolates. The C. acutatum A2 and the Belgian C. gloeosporioides isolates were the most aggressive on fruits, followed by C. acutatum A3, A4, A5, A7 and C. coccodes isolates. When inoculated into crowns, C. acutatum A2, A5 and American C. gloeosporioides isolates were the most aggressive, followed by C. acutatum A3 isolates. The A4 and A7 isolates and all European C. gloeosporioides isolates were non‐pathogenic on crowns. These data indicate that an unusually diverse Colletotrichum population is present in Belgium. The traditional differentiation between C. acutatum and C. gloeosporioides as causal agents of fruit and crown rot, respectively, proved not to be valid in Belgian strawberry fields.     

Characterization of Colletotrichum truncatum from papaya,pepper and physic nut based on phylogeny,morphology and pathogenicity

Colletotrichum truncatum (syn. C. capsici) has been identified as the causal agent of anthracnose on various hosts, predominantly pepper (Capsicum spp.) plants. The aim of this study was to determine whether C. truncatum isolates infecting papaya, pepper and physic nut in southeastern Mexico are morphologically, genetically and pathogenically different, in order to improve disease management strategies. A total of 113 C. truncatum isolates collected from five producer states were subjected to phenotypic characterization and divided into six different morphological groups. These morphological traits and the location of the isolates were used to select a subset of 20 isolates for further studies. Differences in the pathogenicity of the isolates were tested with a cross‐inoculation assay using pepper, papaya and physic nut. The pathogenicity tests revealed that all isolates could infect the three hosts and produce typical anthracnose symptoms, indicating a lack of host specificity for this species and therefore its pathogenic potential on other plants. Phylogenetic analysis using internal transcribed spacer (ITS) and glyceraldehyde 3‐phosphate dehydrogenase (GAPDH) sequences of the C.   truncatum isolates from this study and reference strains was performed, grouping the isolates into a monophyletic clade. This study reports for the first time the characterization of C. truncatum causing anthracnose disease on three different hosts in Mexico.     

Factors influencing biological traits and aggressiveness of Colletotrichum species associated with cashew anthracnose in Brazil

Anthracnose is the main fungal disease on cashew orchards in Brazil, occurring on both vegetative and reproductive organs of cultivated and noncultivated host plants. Understanding the effect of physical and chemical exogenous factors on the biological traits of Colletotrichum spp. and determining their host range are key to developing appropriate anthracnose control measures. The present study aimed to estimate the optimum temperatures for mycelial growth, sporulation, and conidial germination of seven Colletotrichum species (C. chrysophilum, C. fragariae, C. fructicola, C. gloeosporioides, C. queenslandicum, C. siamense, and C. tropicale) associated with cashew anthracnose in Brazil. Their aggressiveness on cashew leaves and six alternative host fruits, and their sensitivity to three fungicides were also investigated. The optimum temperatures for mycelial growth, sporulation, and conidial germination ranged from about 25 to about 33°C. All Colletotrichum species induced anthracnose symptoms on wounded cashew leaves, but none of them caused lesions on intact leaf surfaces. The Colletotrichum species, except for C. fragariae and C. fructicola, were pathogenic to wounded fruits of avocado, banana, guava, mango, and papaya, and some isolates also produced lesions on nonwounded fruit tissues. No symptoms were observed on passion fruits, regardless of the inoculation method. Mycelial growth, sporulation, conidial germination, and/or appressorial formation of the seven Colletotrichum species were inhibited by azoxystrobin, difenoconazole, and thiophanate-methyl to varying degrees. The present study will contribute to the development of forecasting models based on prevailing weather of cashew cropping zones and improve cashew anthracnose management in Brazil.     

Isolation and pathogenicity of Colletotrichum spp. causing anthracnose of Indian mulberry (Morinda citrifolia) in tropical islands of Andaman and Nicobar, India

This study identifies the Colletotrichum spp. which cause Morinda anthracnose in the Andaman and Nicobar Islands, India. A survey of Indian mulberry plantations was carried out to ascertain the identity of isolates associated with Morinda anthracnose. These were identified as C. gloeosporioides based on morphology and sequencing analysis of internal transcribed spacer (ITS) region, demonstrating that this species is currently the causal agent of Morinda anthracnose in a tropical ecosystem. The pathogenicity test of C. gloeosporioides isolates on Morinda seedlings for 1?month revealed that the five isolates MC1, MC8, MC2 (1), MC4 and MC12 were pathogenic to the respective host with different levels of anthracnose lesion.     

Chili anthracnose: Colletotrichum taxonomy and pathogenicity

Chili anthracnose is caused by Colletotrichum species mostly associated with the acutatum, truncatum and gloeosporioides complexes. Since 2009 the Colletotrichum taxonomy has been extensively revised based on multigene phylogenetics, which has had a large impact on the number of species known to cause anthracnose disease of chili. This review discusses (i) the taxonomy of Colletotrichum spp. infecting chili, and (ii) the impact of Colletotrichum pathotypes on breeding for resistance to anthracnose. To date, 24 Colletotrichum species have been identified as pathogens of chili anthracnose, with the three main pathogens being C. scovillei, C. truncatum and C. siamense. Identification of several pathotypes within these three Colletotrichum species, particularly pathotypes that can overcome resistance in the related Capsicum species, Ca. chinense and Ca. baccatum, will be of major concern to plant breeders as they develop resistant chili genotypes from the transfer of resistance genes from these Capsicum species into Ca. annuum. Accurate identification of the Colletotrichum species causing anthracnose and improved understanding of the biology of the Colletotrichum species and their interaction with the host will enable the application of improved integrated disease management techniques.     

Colletotrichum siamense is the main aetiological agent of anthracnose of avocado in south-eastern Brazil

Anthracnose caused by species of Colletotrichum is considered one of the main postharvest diseases for avocado. In this study, Colletotrichum isolates associated with avocado anthracnose, collected in different states of Brazil, were evaluated through phylogenetic analysis, morphological characterization, and pathogenicity assays. Moreover, the events during pathogen infection of avocados were examined by scanning electron microscopy. To assess the genetic diversity of 54 Colletotrichum isolates, partial sequence analysis of the gene gapdh was performed. According to the generated groupings and the geographical origins of isolates, a subset of 14 strains was selected for performing multilocus phylogeny analysis (using sequences of gapdh, act, tub2, and ApMat). Two species previously described were identified: C. siamense belonging to the C. gloeosporioides species complex and Colletotrichum karstii belonging to the C. boninense species complex. All Colletotrichum strains evaluated caused typical symptoms of anthracnose in avocado fruits. Conidia of the most virulent strain germinated between 6 and 12 hr after inoculation (hai). Penetration through wounds occurred 48 hai, tissue colonization occurred between 144 and 240 hai, and sporulation took place at 240 hai via the production of an acervulus, conidiophores, and conidia. The findings shed light on the aetiology of avocado anthracnose in Brazil and provide a better understanding of the infection process of this pathogen, which may assist in the development of disease management strategies.     

Species of the Colletotrichum gloeosporioides and C. boninense complexes associated with olive anthracnose

The taxonomic status of Colletotrichum gloeosporioides sensu lato (s.l.) associated with olive anthracnose is still undetermined and the pathogenic ability of this species complex is controversial. In the present study, isolates obtained from olive and provisionally identified as C. gloeosporioides s.l. on the basis of morphological and cultural features were reclassified using ITS and TUB2 as DNA barcode markers and referred to seven distinct species, recently separated within C. gloeosporioides (C. aenigma, C. gloeosporioides sensu stricto (s.s.), C. kahawae, C. queenslandicum, C. siamense and C. theobromicola) and C. boninense (C. karstii) species complexes. Furthermore, isolates of C. kahawae were ascribed to the subspecies ciggaro by analysing the GS gene. A single isolate, not in either of these two species complexes, was not identified at the species level. In pathogenicity tests on detached olive drupes some of these species, including C. aenigma, C. kahawae subsp. ciggaro, C. queenslandicum, C. siamense and C. karstii, were shown to be weakly pathogenic. Moreover, they were found very sporadically on olive. In contrast, some isolates of C. gloeosporioides s.s. and isolates of C. theobromicola proved to be virulent on both green and ripening olives. This study gives a better insight into both the aetiology and the epidemiology of olive anthracnose and might have implications for biosecurity and quarantine because C. theobromicola has never been reported in major European olive‐producing countries.     

Multilocus identification and thiophanate-methyl sensitivity of Colletotrichum gloeosporioides species complex associated with fruit with symptoms and symptomless leaves of mango

Colletotrichum spp. are known causal agents of anthracnose in a broad host range, causing severe losses. Currently, the most effective way to reduce disease is by fungicide application, which could give rise to resistant populations. This study aimed to determine the Colletotrichum species present in conventional and organic mango orchards and to evaluate their pathogenicity and sensitivity to the benzimidazole fungicide thiophanate-methyl. Seventy-one isolates from fruit with symptoms and symptomless leaves were obtained. From these, 20 representative morphotypes were analysed based on glyceraldehyde-3-phosphate dehydrogenase partial gene sequencing. A subset of 10 isolates based on different species, isolation source, and fungicide sensitivity was used for morphological and multilocus phylogenetic analysis. Colletotrichum queenslandicum was only identified in conventional production systems, C. chrysophilum only in organic systems, and C. asianum and C. siamense in both. Pathogenicity tests showed all species were pathogenic, and only C. asianum caused symptoms via both unwounded and wounded inoculation methods. Overall, 25.3% of isolates (n = 18) that belong to C. siamense, isolated from a conventional orchard, grew on thiophanate-methyl amended media at 1,000 µg/ml, suggesting high resistance. Resistance was not correlated with any common point mutations at positions 198 and 200 of the β-tubulin 2 protein, as commonly found in other fungal pathogens resistant to benzimidazole. The 74.7% remaining isolates (n = 53) belonging to the other species were sensitive, reaching 100% inhibition at <10 µg/ml. Even with benzimidazole application, anthracnose symptoms persist due to the emergence of pathogenic Colletotrichum subpopulations that are resistant to thiophanate-methyl.     

Identification of <Emphasis Type="Italic">Colletotrichum</Emphasis> species associated with anthracnose disease of coffee in Vietnam

Twenty-three isolates of Colletotrichum gloeosporioides, five isolates of C. acutatum, two isolates of C. capsici and six isolates of C. boninense associated with anthracnose disease on coffee (Coffea spp.) in Vietnam were identified based on morphology and DNA analysis. Phylogenetic analysis of DNA sequences from the internal transcribed spacer region of nuclear rDNA and a portion of mitochondrial small subunit rRNA were concordant and allowed good separation of the taxa. We found several Colletotrichum isolates of unknown species and their taxonomic position remains unresolved. The majority of Vietnamese isolates belonged to C. gloeosporioides and they grouped together with the coffee berry disease (CBD) fungus, C. kahawae. However, C. kahawae could be distinguished from the Vietnamese C. gloeosporioides isolates based on ammonium tartrate utilization, growth rate and pathogenicity. C. gloeosporioides isolates were more pathogenic on detached green berries than isolates of the other species, i.e. C. acutatum, C capsici and C. boninense. Some of the C. gloeosporioides isolates produced slightly sunken lesions on green berries resembling CBD symptoms but it did not destroy the bean. We did not find any evidence of the presence of C. kahawae in Vietnam.     

Identification and species status of the mango biotype of Colletotrichum gloeosporioides in Ghana

Research work was carried out to identify and ascertain the species status of the mango biotype of Colletotrichum gloeosporioides infecting mangoes in Ghana. Forty five isolates of Colletotrichum species were collected from 12 districts in Ghana while five each were obtained from mango fruits from Florida, Mexico and Puerto Rico. The entire internal transcribed spacer region, partial beta-tubulin gene and partial glyceraldehyde-3-phosphate dehydrogenase gene of isolates were sequenced and used in phylogenetic studies. The results of the sequence analysis of the first ribosomal transcribed spacer (ITS 1) region showed that 35 % of the isolates from Ghana and all the five isolates from Mexico were the mango biotype of C. gloeosporioides, while the others were not. Phylogenetic studies showed that the mango biotype of the pathogen was Colletotrichum asianum but not C. gloeosporioides as previously thought. However, the other isolates that were not the mango biotype were identified as Colletotrichum siamense and Colletotrichum species which had probably cross-infected mango from other fruit crops in the field.     

Markers to differentiate species of anthracnose fungi identify <Emphasis Type="Italic">Colletotrichum fructicola</Emphasis> as the predominant virulent species in strawberry plants in Chiba Prefecture of Japan

Colletotrichum fungi belonging to the Colletotrichum gloeosporioides species complex include a number of economically important postharvest pathogens that often cause anthracnose. Until now, different species within this group could only be distinguished from one another reliably using multigenic phylogenetic analyses. Using a comparative genomics approach, we developed a marker that can differentiate Colletotrichum fructicola, Colletotrichum aenigma and Colletotrichum siamense within the C. gloeosporioides species complex based on PCR amplicon size differences. When we used this marker to classify 115 isolates collected over 20 years from strawberries in Chiba Prefecture, Japan, the isolates were predominantly C. fructicola. To our knowledge, this is the first report characterizing different species of Colletotrichum infecting strawberries in Japan and contributes to our understanding on the diversity of anthracnose pathogens in Japan.     

Diaporthe sclerotioides exhibits no host specificity among cucurbit species

Plant–fungal specificity between cucurbitaceous crops and Diaporthe sclerotioides, the causal agent of black root rot, was studied using cucumbers (Cucumis sativa), melons (Cucumis melo), pumpkins (Cucurbita maxima), watermelons (Citrullus lanatus) and bottlegourd (Lagenaria siceraria var. gourda). Twelve D. sclerotioides isolates from these cucurbit species were cross‐inoculated. The virulence of the isolates was evaluated as the area under the disease progress curve (AUDPC). All cucurbit species were susceptible to each isolate, but AUDPCs were significantly different among the hosts, with the order of greatest to least being melon, cucumber, watermelon, bottlegourd and finally, pumpkin. The infectiveness of isolates was assessed as the quantity of D. sclerotioides DNA detected in the hypocotyls of seedlings 2 weeks after inoculation using a real‐time PCR protocol. The fungal DNA quantities varied among the species in the same order as the AUDPCs. Whilst there were statistically significant correlations between the virulence and infectiveness of D. sclerotioides isolates in cucumbers, melons and bottlegourds, their coefficients of determination were not high (r² < 0·6). Orthogonal contrasts indicated no specificity in either the fungal virulence or infectiveness between D. sclerotioides isolates and the cucurbit hosts from which these isolates originated. Thus, although the degree of host susceptibility to D. sclerotioides varies among cucurbit species, the absence of specificity to the host species in either virulence or infectiveness suggests the pathogen may spread via various cucurbit crops, irrespective of their original host species.     

Diversity and pathogenicity of Colletotrichum species isolated from soursop in Colombia

Anthracnose, caused by Colletotrichum species is a highly limiting disease for the production of the tropical fruit tree crop, soursop (Annona muricata L.). In this study, 83 single-spore isolates of Colletotrichum were obtained from diseased soursoup tissues and subjected to a species complex-specific PCR assay. The isolates were identified as C. gloeosporioides sensu lato (n?=?60), C. boninense s. lat. (n?=?22), or C. acutatum s. lat. (n?=?1). A subset of 21 selected isolates was identified to species level by means of a multi-locus phylogenetic analysis using sequences from the ITS region and partial sequences of the actin, β-tubulin-2, glyceraldehyde-3-phosphate dehydrogenase, and chitin synthase-1 genes. The multi-locus phylogenetic analysis resolved C. theobromicola, C. tropicale, C. siamense, and C. gloeosporioides sensu stricto in the C. gloeosporioides complex; C. karstii and one undetermined species in the C. boninense complex; as well as one undetermined species in the C. acutatum complex. Significant differences in anthracnose severity were observed between Colletotrichum species when tested for pathogenicity on attached twigs of soursop cv. Elita. Colletotrichum theobromicola and C. tropicale were associated with high and intermediate virulence, respectively, whereas the remaining species were associated with low virulence.