Differentiation of lineages within “Colletotrichum gloeosporioides s.l.” associated with cassava anthracnose disease by BOX‐ and ERIC‐PCRs

Several molecular techniques have been used to differentiate species or genetic lineages of microorganisms prior to sequencing. Among them, BOX‐ and ERIC‐PCRs may provide specific banding patterns for different species, allowing its differentiation. Therefore, the objective of this study was to evaluate these techniques as a tool for differentiation of phylogenetic lineages belonging to the Colletotrichum gloeosporioides species complex associated with cassava anthracnose disease. Sets of BOX‐ and ERIC‐PCR primers were used to assess the differentiation of lineages belonging to the complex with 81 C. gloeosporioides sensu lato (s.l.) isolates from different cassava producing regions. Some were identified by sequencing, such as Colletotrichum fructicola, Colletotrichum tropicale, C. gloeosporioides s.s, Colletotrichum theobromicola, Colletotrichum siamense, Colletotrichum brevisporum and Colletotrichum sichuanensis. The primers were able to amplify DNA fragments from all isolates. The ERIC‐PCR presented a wider range of banding patterns in comparison to BOX‐PCR, providing better differentiation of the individuals, as well as a higher correlation with the phylogenetic data was obtained by ERIC‐PCR and the combined data set for “BOX‐/ERIC‐PCRs,” inferred by Mantel test. However, the use of concatenated data (BOX‐/ERIC‐PCRs) reduced the discriminatory capacity presented by ERIC‐PCR alone, probably due to the lowest resolution of BOX‐PCR. Therefore, ERIC‐PCR technique enabled efficient differentiation of isolates belonging to the C. gloeosporioides complex and can be used to analyse multiple isolates in a collection and also being an important tool as a guide in the decision‐making process prior to sequencing. Based on this methodology, it was possible to identify two new species associated with cassava anthracnose disease, C. brevisporum and C. sichuanensis, being the first report of these two species associated with cassava anthracnose disease in Brazil.     

Identification and Characterization of Colletotrichum spp. affecting Fruit after Harvest in Brazil

Colletotrichum spp. cause anthracnose in various fruits post‐harvest and are a particularly important problem in tropical and subtropical fruits. The disease in fruits of avocado, guava, papaya, mango and passion fruit has been reported to be caused by C. gloeosporioides, and in banana by C. musae. In subtropical and temperate crops such apple, grape, peach and kiwi, the disease is caused by C. acutatum. The variation in pathogenic, morphological, cultural and molecular characteristics of Brazilian isolates of Colletotrichum acutatum Simmonds and isolates from post‐harvest decays of avocado, banana, guava, papaya, mango and passion fruit was evaluated. The fruits were inoculated with mycelium of C. acutatum, Colletotrichum spp. and C. musae on a disc of potato dextrose agar. The morphological, cultural and molecular characteristics studied were conidia morphology, colony growth at different temperatures, colony coloration and PCR with primers CaInt2 and ITS4 for C. acutatum and CgInt and ITS4 for C. gloeosporioides. C. acutatum was pathogenic to avocado, guava, papaya, mango and passion fruit, but it was not pathogenic to banana. The morphological, cultural and molecular studies indicated that the avocado, papaya, mango and passion fruit isolates were C. gloeosporioides. The natural guava isolate was identified as C. acutatum, which had not been found previously to produce anthracnose symptoms on guava in Brazil.     

Intraspecific Differentiation of Colletotrichum gloeosporioides sensu lato Based on In Silico Multilocus PCR-RFLP Fingerprinting

Colletotrichum gloeosporioides sensu lato is one of the most common and widely distributed plant pathogens in the world. Understanding fungal biodiversity is hinged on accurate identification and delimitation at the inter- and intraspecific levels. Sequences of the ITS1-5.8S-ITS2 region (ITS), β-tubulin (TUB), actin (ACT), and glyceraldehyde-3-phosphate dehydrogenase (GPDH) genes of 30 C. gloeosporioides sensu lato isolates, collected from anthracnose infected papaya fruits grown in the main production areas in Trinidad, were analyzed by in silico PCR-RFLP analysis with the aim of identifying which gene region(s) had the highest level of intraspecific polymorphism. Restriction site polymorphisms generated from 13 restriction enzymes enabled the identification of specific enzymes that were successful at intraspecific discrimination of the C. gloeosporioides isolates. Genetic distance values were reflective of the level of polymorphisms obtained for the four different gene regions. In both cases (calculated genetic distance and percentage of polymorphic loci from RFLP profiles), ACT and ITS gene regions had the highest level of restriction site polymorphisms and genetic diversity, GPDH and TUB had the lowest. Cluster analysis based on PCR-RFLP genetic distance data revealed sub-specific placement of the isolates which appeared to be gene-dependent. The implications of these findings are discussed relative to biodiversity monitoring and the need for multilocus, polyphasic investigations which must take into account the possibility of exaggerated estimates of genetic diversity.     

PCR-based detection and characterization of the fungal pathogens Colletotrichum gloeosporioides and Colletotrichum capsici causing anthracnose in papaya (Carica papaya l.) in the Yucatan peninsula

Colletotrichum gloeosporioides is the common causal agent of anthracnose in papaya (Carica papaya L.) fruits, and infection by this fungal pathogen results in severe post-harvest losses. In the Yucatán peninsula (Mexico) a different Colletotrichum species was isolated from papaya fruits with atypical anthracnose lesions. The DNAs from a variety of Colletotrichum isolates producing typical and atypical lesions, respectively, were amplified by PCR with C.gloeosporioides-specific primers. All isolates from typical anthracnose lesions yielded a 450 bp PCR product, but DNAs from isolates with atypical lesions failed to produce an amplification product. For further characterization, the rDNA 5.8S-ITS region was amplified by PCR and processed for sequencing and RFLP analysis, respectively, to verify the identity of the papaya anthracnose pathogens. The results revealed unequivocally the existence of two Colletotrichum species causing anthracnose lesions on papaya fruits: C. gloeosporioides and C. capsici. PCR-RFLP using the restriction endonuclease MspI reliably reproduced restriction patterns specific for C. capsici or C. gloeosporioides. The generation of RFLP patterns by MspI (or AluI or RsaI) is a rapid, accurate, and unequivocal method for the detection and differentiation of these two Colletotrichum species.     

Diversity of <Emphasis Type="Italic">Colletotrichum</Emphasis> spp. isolated from chili pepper fruit exhibiting symptoms of anthracnose in Thailand

Colletotrichum spp. are causal agents of anthracnose disease in chili fruits and other tropical crops. The disease is increasing in chili fruits in Thailand and significantly reduces fruit quality and fruit production. Forty-eight isolates of Colletotrichum spp. associated with chili anthracnose were collected from different areas of Thailand during 2010–2015. Based on morphological characteristic identification, 10 isolates were shown to belong to the C. gloeosporioides species complex, 24 isolates belong to the C. acutatum species complex and 14 isolates to C. capsici. For molecular identification, two primer sets, ITS1/ITS4 and ACT528/ACT738, were used for amplification of the internal transcribed spacer of rRNA gene (ITS1–5.8S–ITS2) and partial region actin gene (ACT), respectively. The phylogenetic analysis of individual and combined ITS region and actin nucleotide sequences identified the collected isolates into 4 species: C. gloeosporioides, C. siamense, C. acutatum and C. capsici. The pathogenicity test demonstrated that all four species were pathogenic on intact unwounded and healthy fruits. These results indicated that C. capsici, C. acutatum, C. gloeosporioides and C. siamense were the causal agents of chili anthracnose disease.     

Antifungal effects of dimethyl trisulfide against Colletotrichum gloeosporioides infection on mango

Colletotrichum gloeosporioides, one of the main agents of mango anthracnose, causes latent infections in unripe mango and can lead to huge losses during fruit storage and transport. Dimethyl trisulfide (DMTS) is an antifungal agent produced by several microorganisms or plants, but its effects on the infection process of C. gloeosporioides have not been well characterized. A histological investigation demonstrated that DMTS exhibits strong inhibitory effects on the infection process of C. gloeosporioides in planta by inhibiting the germination of conidia and formation of appressoria, damaging cytoplasm to cause cells to vacuolate and contributing to deformation of appressoria prior to penetration. This is the first study to demonstrate antifungal activity of DMTS against C. gloeosporioides on mango by suppression of the infection process, thus providing a novel postharvest biorational control for mango anthracnose.     

Elucidating the Colletotrichum spp. diversity responsible for papaya anthracnose in Brazil

Papaya (Carica papaya L.) is among the most important tropical fruits produced in Brazil and is grown in nearly every state. However, several diseases can affect papaya production. Anthracnose stands out among these diseases due to high postharvest yield losses. Previous studies identified Colletotrichum magna (invalid name) and Colletotrichum gloeosporioides causing anthracnose of papaya in Brazil, but species identification was inadequate due to reliance on nuclear ribosomal internal transcribed space (nrITS) and glutamine synthetase (GS) sequences. Thus, the diversity of Colletotrichum spp. causing papaya anthracnose in Brazil may be underestimated. The present study aims to identify the Colletotrichum species associated with papaya anthracnose in Brazil based on broad geographical sampling and multilocus phylogenetic analysis, as well as to assess the prevalence and aggressiveness of the species found. Here, we report C. chrysophilum, C. fructicola, C. gloeosporioides, C. karsti, C. okinawense, C. plurivorum, C. queenslandicum, C. siamense, C. theobromicola, Colletotrichum truncatum causing papaya anthracnose in Brazil. We are also synonymizing Colletotrichum corchorum-capsularis under C. truncatum. Colletotrichum okinawense was the most prevalent species in general and in most sampled locations, and with C. truncatum represents the most aggressive species.     

Colletotrichum fructicola and C. siamense are involved in chilli anthracnose in India

Chilli anthracnose is a major problem in India and worldwide. In this study, we investigated the phylogenetic relationships of 52 fungal isolates associated with chilli anthracnose in southern India. All the 52 isolates were sequenced for partial ITS/5.8S rRNA and glyceraldehyde-3-phosphate dehydrogenase (gapdh) genes and showed affinities with Colletotrichum siamense and C. fructicola within Colletotrichum. gloeosporioides species complex. Further, a reduced subset of 17 selected isolates was made and in a maximum parsimony analysis of a multigene data-set including partial ITS/5.8S rRNA, actin (act), calmodulin (cal), chitin synthase (chs1), gapdh and β-tubulin (tub2) gene sequence data, these fungal isolates clustered with the type strain of C. fructicola, except for strain MTCC 3439 that showed phylogenetic affinities with C. siamense. The pathogenicity tests involving two representative isolates: UASB-Cg-14 and MTCC 3439, confirmed the involvement of C. fructicola and C. siamense in the development of disease symptoms on fresh chilli fruits. This is the first report of the association of C. fructicola and C. siamense in causing chilli anthracnose in India.     

Characterisation of husk rot in macadamia

The causal agent of husk rot of macadamia is often attributed to Colletotrichum gloeosporioides sensu lato. However, in recent husk rot outbreaks, the characteristic concentric ring of pycnidia of C. gloeosporioides that is associated with the disease was often absent. Due to its sporadic occurrence, the importance of husk rot is often underrated and attributed to environmental and physiological factors. In order to determine the significance, prevalence and factors that influence husk rot in macadamia, this study examined the aetiology of husk rot in Australia. The relative incidence and severity of husk rot was evaluated in several macadamia orchards over eight consecutive years. Pathogenicity assays were developed to confirm the identity of the causal agent. A range of fungi from several genera including: Diaporthe, Lasiodiplodia, Colletotrichum, Pestalotiopsis, Aspergillus, Alternaria, Nigrospora and Epicoccum were isolated from samples of macadamia pericarps with husk rot symptoms from different orchards. Fungi in the genus Diaporthe were most frequently isolated, often from symptomatic fruit. Results from pathogenicity trials showed the characteristic soft or spongy black lesions characteristic of husk rot symptoms in wounded fruits that were incubated with the diseased fruit or inoculated with a conidial suspension of Diaporthe spp. Our results suggest that injury to the macadamia fruit pericarp not only predisposes the pericarp to pathogen infection but it is a prerequisite for infection. Large variations in husk rot severity were observed over years. Husk rot severity was linked to days after anthesis and was associated with mean weekly relative humidity and minimum temperatures. This study confirmed that Diaporthe species cause husk rot in macadamia, hence, a rationale for adopting Phomopsis husk rot as the name of the disease is discussed.     

Cytochrome b Gene-Based Assay for Monitoring the Resistance of Colletotrichum spp. to Pyraclostrobin

Resistance to pyraclostrobin due to a single nucleotide polymorphism at 143rd amino acid position on the cytochrome b gene has been a major source of concern in red pepper field infected by anthracnose in Korea. Therefore, this study investigated the response of 24 isolates of C. acutatum and C. gloeosporioides isolated from anthracnose infected red pepper fruits using agar dilution method and other molecular techniques such as cytochrome b gene sequencing, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), and allele-specific polymerase chain reaction (PCR). The result showed that four isolates were resistant to pyraclostrobin on agar dilution method and possessed GCT (alanine) codon at 143rd amino acid position, whereas the sensitive isolates possessed GGT (glycine). Furthermore, this study illustrated the difference in the cytochrome b gene structure of C. acutatum and C. gloeosporioides. The use of cDNA in this study suggested that the primer Cacytb-P2 can amplify the cytochrome b gene of both C. acutatum and C. gloeosporioides despite the presence of various introns in the cytochrome b gene structure of C. gloeosporioides. The use of allele-specific PCR and PCR-RFLP provided clear difference between the resistant and sensitive isolates. The application of molecular technique in the evaluation of the resistance status of anthracnose pathogen in red pepper provided rapid, reliable, and accurate results that can be helpful in the early adoption of fungicide-resistant management strategies for the strobilurins in the field.     

Morphological and molecular identification of Colletotrichum species associated with cassava anthracnose in Thailand

The objective of this study was to identify the causal agent of anthracnose disease of cassava in Thailand. The study was carried out by collecting cassava samples with anthracnose symptoms from various planting areas including 10 districts of eight provinces in Thailand. One hundred and thirty‐six Colletotrichum samples were isolated from cassava anthracnose lesions on leaves, petioles and stems. Thirty‐eight single‐spore isolates were subsequently obtained and cultured on half potato dextrose agar for morphological and molecular characterizations. All 38 isolates were pathogenic with varying degrees of virulence when tested on detached leaves of Kasetsart 50, a susceptible cassava cultivar. Based on their growth habit, colony morphology, conidial morphology and the internal transcribed spacer sequences similarity to that of Colletotrichum accessions in the GenBank, one isolate was identified as C. capsici, one as C. lindemuthianum, two as C. aeschynomene, four as C. boninense and 28 C. gloeosporioides species complex. Geographically, the cosmopolitan C. gloeosporioides species complex was found in all regions, but other species were found only in particular regions. This is, so far, the first report of Colletotrichum complex species associated with cassava anthracnose in Thailand.     

Genetic diversity of Colletotrichum gloeosporioides species complex associated with Citrus wither‐tip of twigs in Tunisia using microsatellite markers

Anthracnose Citrus disease has been associated with several symptoms worldwide and it is recently compromising Citrus production in the Mediterranean area. Four species complexes are mainly involved: Colletotrichum boninense, C. acutatum, C. gloeosporioides and C. truncatum. In this study, we investigated the genetic diversity of Colletotrichum spp. in Tunisia associated with wither‐tip of twigs on Citrus. Specific primers ITS4‐CgInt allowed the identification of C. gloeosporioides species complex in all the 54 isolates, sampled from three regions and four Citrus species. Overall, our genotypic analysis using 10 SSR markers showed a moderate diversity level in Tunisian C. gloeosporioides population and highlighted that C. gloeosporioides reproduce mainly clonally. In addition, heterothallic isolates were present in our population, suggesting that the pathogen population may undergo parasexual recombinations. The highest genetic diversity in C. gloeosporioides was recorded in Nabeul and on orange, which likely constitutes the area and the host of origin for the Citrus anthracnose disease in Tunisia. In addition, no population subdivision was detected at the geographic, host species or cultivars’ origin levels. However, our study identified two genetic subpopulations and indicated a rapid C. gloeosporioides population change at temporal scale that should be further examined over several consecutive growing seasons in order to understand its population dynamics.     

Effect of 2-deoxy-D-glucose on the Respiration of Tropical Anthracnose Fungi

This study begins an investigation of the chemical activities of one of the most destructive groups of spoilage organisms of tropical foods, the anthracnose fungi. The present paper treats of an introductory investigation on carbohydrate metabolism. Two fungi isolated from rotted banana tissue, Gloeosporium musarum and Colletotrichum gloeosporioides, were grown in the presence of 2-deoxy-D-glucose, a metabolic blocking agent, in shake culture. The mycelia produced were studied in the Warburg respirometer in the presence of glucose, 2-deoxyglucose, and 2,4-dinitrophenol. Gloeosporium musarum was found to be incapable of growth in the presence of 2-deoxyglucose as the sole carbon source, but did grow when glucose was added. Colletotrichum gloeosporioides grew normally, but at a slower rate on 2-deoxyglucose than on glucose. Glucose grown G. musarum and C. gloeosporioides both exhibited respiratory inhibition in the presence of 2-deoxyglucose, whereas 2-deoxyglucose adapted C. gloeosporioides did not. The uncoupling agent 2, 4-dinitrophenol induced respiratory stimulation except in the presence of 2-deoxyglucose in glucose grown C. gloeosporioides where it caused a drop in oxygen uptake.     

Antifungal activity of plant extracts against Colletotrichum gloeosporioides (Penz.) the causative agent of yam anthracnose disease

Yam anthracnose disease is a major constraint to yam production world-wide. The hazardous effects of synthetic fungicides on both humans and the environment have necessitated the use of alternative environmentally friendly fungicides for the control of the disease. This study tested the efficacy of aqueous and ethanol extracts of Azadiratcha indica, Balanites aegyptiaca, Jatropha curcas, and Khaya senegalensis seeds, Icacina oliviformis leaves and Capsicum annuum (Legon 18 variety) fruit against Colletotrichum gloeosporioides (Penz.), the causative agent of yam anthracnose. The antifungal activity of each plant extract was assessed in vitro on potato dextrose agar using the food poison technique. Each extract inhibited significantly (p?≤?.05) the mycelia growth and spore germination of C. gloeosporioides. Qualitative phytochemical tests detected alkaloids, anthraquinones, cardiac glycosides, flavonoids, phlobatinnins, saponins, steroids, tannins and terpenoids. The potential antifungal activity exhibited by these plant material makes them suitable candidates for the control of anthracnose disease of yam.     

Pathogenic and genetic variability among Colletotrichum gloeosporioides isolates from different yam hosts in the agroecological zones in Nigeria

Anthracnose, caused by Colletotrichum gloeosporioides Penz., is the most severe foliar disease of water yam (Dioscorea alata) worldwide. Population genetic analyses can yield useful insights into the evolutionary potential of C. gloeosporioides and thus lead to the development of appropriate disease management strategies. The genetic structure of C. gloeosporioides populations from yam and non‐yam hosts in three agroecological zones of Nigeria was investigated. Microsatellite‐primed polymerase chain reaction (MP‐PCR), virulence phenotyping using five putative D. alata differentials, cross‐inoculation tests, and the presence/absence of a Glomerella teleomorph in yam fields were used to infer the evolutionary potential of C. gloeosporioides on yam. We observed high genotypic diversity (GD = 0.99 to 1.00) for populations from all hosts and agroecological zones, with multiple pathogen genotypes in individual anthracnose lesions. Genetic differentiation was low among pathogen populations from different hosts (G_ST = 0.10, θ = 0.034), and agroecological zones (G_ST = 0.04, θ = 0.018), indicating limited host differentiation and significant gene flow. No evidence was found for the existence of C. gloeosporioides f. sp. alatae reported in previous studies. The fungus was recovered from several non‐yam host species commonly found in yam fields but non‐yam isolates caused only mild to moderate symptoms on yam. Eighteen C. gloeosporioides virulence phenotypes were identified among 217 isolates but there was a weak correlation (r = 0.02, P = 0.40) between virulence phenotype and MP‐PCR haplotype. Consistent with the above findings, we observed for the first time the Glomerella teleomorph on anthracnose‐infected yam plants in Nigeria, indicating that sexual recombination might play an important role in anthracnose epidemics on yam. The implications of these findings for C. gloeosporioides evolutionary potential and anthracnose resistance breeding are discussed.     

Marine yeasts and bacteria as biological control agents against anthracnose on mango

The potential of four yeasts (Debaryomyces hansenii, Rhodotorula minuta, Cryptococcus laurentii and Cryptococcus diffluens) and three bacteria (Bacillus amyloliquefaciens, Bacillus subtilis and Stenotrophomonas rhizophila) with antagonistic capacity against anthracnose caused by Colletotrichum gloeosporioides in mango cv. Ataulfo fruit was investigated. Germination of C. gloeosporioides spores was significantly inhibited by all marine yeasts and bacteria strains of an in vitro test. When yeasts and bacteria were tested on mango fruit, the marine yeast D. hansenii 1R11CB strain and marine bacteria S. rhizophilaKM02 strain were the best antagonists to anthracnose (C. gloeosporioides), which significantly decreased disease incidence by 56% and 89%, respectively, and reduced lesion diameter by 91% and 92%, respectively. All the isolated strains of the phytopathogen, yeasts and bacteria were molecularly identified. Our results from in vitro and in vivo experiments suggest that marine yeasts and bacteria strains can be used as some effective biological control agents for anthracnose in mango.     

Characterization and Pathogenicity of Colletotrichum gloeosporioides and C. karstii Causing Preharvest Disease on Citrus sinensis in Italy

During the period from 2010 to 2013 preharvest symptoms were detected on different cultivars of sweet orange in six orchards in Catania, Siracusa and Enna provinces, Southern Italy. A total of 56 monosporic fungal isolates were obtained, and among these, 44 were identified as Colletotrichum gloeosporioides and 12 as C. karstii through morphological and molecular analysis. PCR with primers ITS1 and ITS4, primers TubGF1 and TubGR specific for β‐tubulin gene, primers GDF‐GDR, specific for Glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) gene, were used to confirm the identification of Colletotrichum isolates from citrus. The ITS1‐5.8S‐ITS2 region, a portion of approximately 500 bp of β‐tubulin gene and a fragment of 220 bp of GAPDH gene of the isolates were sequenced and analysed with the BLASTn program. Koch's postulates were fulfilled by pathogenicity tests carried out on fruit of ‘Tarocco Scirè’ and ‘Tarocco Nucellare’ with representative isolates of C. gloeosporioides and C. karstii. Field surveys and pathogenicity tests revealed significant differences in fruit susceptibility between ‘Tarocco Scirè’ and ‘Tarocco Nucellare’ and in virulence between the fungal species. To our knowledge, this is the first report on the emergence of Colletotrichum spp. causing anthracnose in preharvest conditions.     

Genetic differentiation of Colletotrichum gloeosporioides and C. truncatum associated with Anthracnose disease of papaya (Carica papaya L.) and bell pepper (Capsium annuum L.) based on ITS PCR-RFLP fingerprinting

Members of the genus Colletotrichum include some of the most economically important fungal pathogens in the world. Accurate diagnosis is critical to devising disease management strategies. Two species, Colletotrichum gloeosporioides and C. truncatum, are responsible for anthracnose disease in papaya (Carica papaya L.) and bell pepper (Capsicum annuum L.) in Trinidad. The ITS1–5.8S–ITS2 region of 48 Colletotrichum isolates was sequenced, and the ITS PCR products were analyzed by PCR-RFLP analysis. Restriction site polymorphisms generated from 11 restriction enzymes enabled the identification of specific enzymes that were successful in distinguishing between C. gloeosporioides and C. truncatum isolates. Species-specific restriction fragment length polymorphisms generated by the enzymes AluI, HaeIII, PvuII, RsaI, and Sau3A were used to consistently resolve C. gloeosporioides and C. truncatum isolates from papaya. AluI, ApaI, PvuII, RsaI, and SmaI reliably separated isolates of C. gloeosporioides and C. truncatum from bell pepper. PvuII, RsaI, and Sau3A were also capable of distinguishing among the C. gloeosporioides isolates from papaya based on the different restriction patterns that were obtained as a result of intra-specific variation in restriction enzyme recognition sites in the ITS1–5.8S–ITS2 rDNA region. Of all the isolates tested, C. gloeosporioides from papaya also had the highest number of PCR-RFLP haplotypes. Cluster analysis of sequence and PCR-RFLP data demonstrated that all C. gloeosporioides and C. truncatum isolates clustered separately into species-specific clades regardless of host species. Phylograms also revealed consistent topologies which suggested that the genetic distances for PCR-RFLP-generated data were comparable to that of ITS sequence data. ITS PCR-RFLP fingerprinting is a rapid and reliable method to identify and differentiate between Colletotrichum species.     

The antagonistic effect and mechanisms of Bacillus amyloliquefaciens DGA14 against anthracnose in mango cv. ‘Carabao’

Bacillus amyloliquefaciens strain DGA14 was tested for in vitro antagonism towards Colletotrichum gloeosporioides, a causal pathogen of anthracnose in mango cv. ‘Carabao’. DGA14 produced extracellular metabolites in solid and liquid media that suppressed the growth of C. gloeosporioides. The cells of DGA14 were often observed adjacent to the pathogen so affecting its spore germination and mycelium development. DGA14 colonised mango fruit 48 h after artificial inoculation and persisted 14 days after storage at 18–20°C. On fruit surfaces, DGA14 attached and produced dents to spores of C. gloeosporioides. Dipping mangoes in aqueous cell suspension (10⁸ mL L^?1) of DGA14 significantly decreased the incidence of anthracnose as compared to untreated fruit.