Bio-formulation of fluorescent Pseudomonas spp. induces systemic resistance against red rot disease and enhances commercial sugar yield in sugarcane

Abstract

Isolates of Pseudomonas spp. collected from the rhizosphere of sugarcane and cane stalks were screened for their antagonistic activity against Colletotrichum falcatum causing red rot disease in sugarcane. Talc formulations of the selected Pseudomonas spp. isolates improved the sugarcane vegetative sett germination and sugarcane growth under field conditions. Optimal talc formulations were assessed for their effect on induction of systemic resistance against the pathogen in the canes under artificial inoculation. All the four isolates CHAO, EP1, KKM1 and VPT4 were effective in inducing systemic resistance against C. falcatum in two seasons. In other studies, the bacterial formulations were assessed to induce resistance in sugarcane in a sick plot situation. In pathogen-infested soil the isolates KKM1 and CHAO suppressed the red rot disease development in susceptible sugarcane cultivar. Pseudomonas strains also protected sugarcane in a disease-endemic location. Pseudomonas spp treatment substantially improved the cane juice quality parameters affected by the pathogen infection. Standardization of talc formulations and application methods in the field offers potential for large-scale application of biocontrol formulations for the management of red rot disease in sugarcane growing regions.     

Suppression of red rot caused by <Emphasis Type="Italic">Colletotrichum falcatum</Emphasis> on sugarcane plants using plant growth-promoting rhizobacteria

Bacterial strains with ability to suppress Colletotrichum falcatum were isolated from the rhizosphere of sugarcane. Thirty nine candidates, chosen on the basis of in vitro antagonism, inhibited C. falcatum growth by 15–65% on test plates. Twenty two isolates causing 50% or more in vitro inhibition were screened for their root colonization ability and biocontrol activity on micropropagated sugarcane plants under greenhouse conditions. Twelve strains suppressed red rot infection in plantlets, but no significant correlation was observed between in vitro pathogen inhibition and in vivo disease suppression. However, isolates showing root colonization over 5.2 log₁₀ CFU g⁻¹ of soil showed highest suppression of C. falcatum and reduction of red rot disease. Six strains with the capability to maintain a significant population in the sugarcane rhizosphere and with a high potential to control red rot were identified by 16S rDNA as Ochrobacterum intermedium NH-5, Pseudomonas putida NH-50, Bacillus subtilis NH-100, Bacillus subtilis NH-160, Bacillus sp NH-217 and Stenotrophomonas maltophilia NH-300.     

Induction of pathogenesis-related proteins in sugarcane leaves and cell-cultures by a glycoprotein elicitor isolated from <Emphasis Type="Italic">Colletotrichum falcatum</Emphasis>

The induction of pathogenesis-related (PR) proteins in sugarcane (Saccharum officinarum L.) leaves and suspension-cultured cells in response to treatment with a glycoprotein elicitor isolated from Colletotrichum falcatum (the red rot pathogen) was investigated. Treatment of leaves and cells with the elicitor resulted in a much marked increase in the activities of chitinase and β-1,3-glucanase in red rot resistant (BO 91) than susceptible (CoC 671) sugarcane cultivar. SDS-PAGE analysis revealed that C. falcatum elicitor induced the accumulation of several proteins in suspension-cultured cells of resistant cultivar (BO 91); among them the 35 kDa protein was predominant. Whereas, a 27 kDa protein was induced predominantly in the cells of susceptible cultivar upon treatment with the elicitor. When sugarcane leaves were treated with C. falcatum elicitor, two proteins with apparent molecular masses of 25 and 27 kDa were induced both in the resistant and susceptible cultivars. However, the induction was stronger in the resistant than the susceptible cultivar. Immunoblot analysis for chitinase indicated that a protein with an apparent molecular mass of 37 kDa cross-reacting with barley chitinase antiserum was strongly induced in the suspension cultured cells of both the cultivars. The induction of 37 kDa chitinase was more in the cells of resistant cultivar than in the susceptible cultivar. Western blot analysis revealed that a 25 kDa thaumatin-like protein (TLP) cross-reacting with bean TLP antiserum was strongly induced in leaves and cultured cells of both resistant and susceptible cultivars due to elicitor treatment.     

Molecular and Pathological Characterization of Colletotrichum falcatum Infecting Subtropical Indian Sugarcane

Red rot, caused by Colletotrichum falcatum, is the most significant problem of sugarcane worldwide. Pathological studies and three different marker systems were used to characterize 25 C. falcatum isolates collected from 18 subtropical sugarcane cultivars from 15 different sugarcane‐growing regions of three north‐eastern states of India to assess pathogen diversity. Of these 25 isolates, three were new (RR2A, RR15, RR83) from cultivars Co 7717, Co J83 and Co S88230, respectively, pathologically characterized on 13 standard differential hosts. Isolates Cf 01, Cf 08 and RR15 were the most, and Cf‐07 the least virulent. Molecular characterization using random amplified polymorphic DNA, universal rice primers (URP) and inter simple sequence repeat markers amplified a total of 161 alleles of which 159 were polymorphic (98.76%). Unweighted paired group method with arithmetic averages analysis of combined data of all the DNA markers obtained by three marker systems classified 25 isolates into six clusters at 34% genetic similarity with high Mantel matrix correlation (r = 0.83). The principal component analysis (PCA) of marker data explained 68% of the variation by first three components. Molecular diversity as revealed in these isolates is very high, but non‐structured. Isolate Co Pant 84212 was found to be genetically most diverse. We demonstrated for the first time that URPs derived from weed rice could successfully assess genetic diversity in C. falcatum. Molecular characterization of the C. falcatum isolates prevalent in north‐eastern India would enable red rot management strate‐gies, selection for resistance genes and development of resistant cultivars.     

甘蔗赤腐病内生拮抗菌YC89的筛选及鉴定

【目的】筛选防治甘蔗赤腐病(sugarcane red rot)的生防菌株。【方法】实验以前期分离获得的甘蔗内生细菌为目标菌,以甘蔗赤腐病的病原真菌镰孢炭疽菌(Colletotrichum falcatum Went.)为指示菌,采用平板对峙法筛选对该病菌有较强抑制作用的菌株,然后通过琼脂扩散法测定菌株代谢产物对抑菌活性的影响,并对具有较好拮抗效果的高效菌株进行抑菌广谱性分析并对其进行鉴定。最后通过形态学、生理生化特征以及16SrDNA和gyrA序列分析对高效菌株YC89进行鉴定。【结果】经初筛筛选到抑菌带均大于1.60 cm的5株拮抗细菌,其中X22、W2、YC89抑菌带均高达1.87 cm。对初筛得到的5株内生菌进行复筛,结果所示菌株YC89、H1、X22、W2、YT93对镰孢炭疽菌的抑菌率都在75%以上,其中菌株YC89对该病菌的抑菌效果最好,其抑菌率为78%。菌株YC89的发酵液、上清液、过滤液及粗蛋白提取液对镰孢炭疽菌的生长有较强的抑制作用,且菌株YC89对玉米大斑病、甘蔗梢腐病、草莓灰霉病等7种病原菌也有较好的抑制效果。通过菌株鉴定结果,初步将YC89菌株鉴定为贝莱斯芽孢杆菌(Bacillus velezensis)。【结论】菌株YC89对镰孢炭疽菌具有较好的抑制效果,表明其对甘蔗赤腐病的生物防治具有较大的应用潜力。     

Variations in random amplified polymorphic DNA profile and toxin production among isolates of Colletotrichum falcatum Went from sugarcane in Tamil Nadu,India

Red rot, caused by Colletotrichum falcatum Went, is one of the most important diseases of sugarcane (Saccharum officinarum L.). The pathogen shows a great diversity in virulence as a number of pathotypes are known to occur in nature. In the present study, the toxin producing ability and genetic variability among isolates of C. falcatum collected from major sugarcane growing areas of Tamil Nadu, India were analysed. The C. falcatum isolates differed significantly in their ability to produce toxin in vitro. The toxin from C. falcatum isolate Cf 671a induced the maximum electrolyte leakage (300 μS) from sugarcane leaf tissues. The genetic relatedness of the isolates of C. falcatum differing in toxin production potential was investigated by using RAPD analysis. Analysis of the genetic coefficient matrix derived from the scores of RAPD profiles showed that minimum and maximum percent similarities among the tested C. falcatum isolates were in the range of 19 to 95% respectively. The phylogenetic analysis by the UPGMA identified two main clusters. Cluster A contains only one isolate (Cf 98061) and all the other isolates were placed in Cluster B confirming high genetic diversity among the isolates. No correlation was observed between clustering of the C. falcatum isolates in the dendrogram and their toxin producing abilities.     

Expression analysis on mycoparasitism related genes during antagonism of <Emphasis Type="Italic">Trichoderma</Emphasis> with <Emphasis Type="Italic">Colletotrichum falcatum</Emphasis> causing red rot in sugarcane

Present study was aimed to select a suitable Trichoderma isolate as candidate antagonist based on its efficacy in producing cell wall degrading enzymes (CWDEs), its mycoparasitism activity and expression of related genes against the red rot pathogen caused by Colletotrichum falcatum in sugarcane. For which, six different isolates of Trichoderma selected from our earlier studies (T. harzianum, T. asperullum) were evaluated based on their capability in releasing cell wall degrading enzymes individually and during antagonism with C. falcatum in dual plate. Amongst T. harzianum (T20) exhibited the greatest mycoparasitic potential against the C. falcatum, by producing higher concentration of  CWDEs viz., chitinase and β-1, 3-glucanase, slightly lower amounts of cellulase and protease with significant reduction in polygalacturonase produced by pathogen. Further microscopic observation on interaction of C. falcatum with the selected isolate of T. harzianum (T20) exhibited the mycoparasitic activity of antagonist over pathogen in dual culture and inhibition of C. falcatum pathogenesis in detached sugarcane leaves. In addition, expression pattern of eight genes coding various enzymes involved in mycoparasitism by T. harzianum over C. falcatum were analyzed using qRT-PCR in vitro and on sugarcane leaves. In in vitro interactions, five genes of  cell wall degrading enzymes viz., chitinase (chit33), endochitinase (endo42), β-1, 3-glucanase (glu), exochitinase 1 (exc1), exochitinase 2 (exc2), were upregulated during and after contact as compared to before contact, while three genes related with proteases such as alkaline proteinase (prb1), trypsin-like protease (Pra1), subtilin-like serine protease (ssp), genes were upregulated during the contact with C. falcatum and slightly down regulated after contact. In detached leaves, seven genes were potentially upregulated except subtilin-like serine protease, which was down regulated during interaction of C. falcatum and T. harzianum as compared to T. harzianum inoculation alone. All these biochemical and molecular results confirm the efficacy of T. harzianum (T20) against C. falcatum and justify the right selection of candidate antagonist for our further studies on identification of antifungal genes/proteins against C. falcatum in sugarcane.     

Why species delimitation matters for fungal ecology: Colletotrichum diversity on wild and cultivated cashew in Brazil

Anthracnose is one of the most important plant diseases globally, occurring on a wide range of cultivated and wild host species. This study aimed to identify the Colletotrichum species associated with cashew anthracnose in Brazil, determine their phylogenetic relationships and geographical distribution, and provide some insight into the factors that may be influencing community composition. Colletotrichum isolates collected from symptomatic leaves, stems, inflorescences, and fruit of cultivated and wild cashew, across four Brazilian biomes, were identified as Colletotrichum chrysophilum, Colletotrichum fragariae, Colletotrichum fructicola, Colletotrichum gloeosporioides sensu stricto, Colletotrichum queenslandicum, Colletotrichum siamense and Colletotrichum tropicale. Colletotrichum siamense was the most dominant species. The greatest species richness was associated with cultivated cashew; leaves harbored more species than the other organs; the Atlantic Forest encompassed more species than the other biomes; and Pernambuco was the most species-rich location. However, accounting for the relative abundance of Colletotrichum species and differences in sample size across strata, the interpretation of which community is most diverse depends on how species are delimited. The present study provides valuable information about the Colletotrichum/cashew pathosystem, sheds light on the causal agents identification,and highlights the impact that species delimitation can have on ecological studies of fungi.     

Assessment of genetic diversity in Colletotrichum falcatum Went accessions based on RAPD and ISSR markers

Sugarcane is susceptible to red rot disease caused by phytopathogenic fungus Colletotrichum falcatum Went which ultimately affect the economy of farmers as well as sugar based industry. One of the various ways to control this devastating disease is to develop disease resistance sugarcane cultivar and this requires the complete understanding of genetic makeup of pathogen. Although South Gujarat is well known sugarcane cultivating area, less published data can be found about PCR-based genetic diversity in prevalent C. falcatum accessions. So, present investigation aims at finding molecular variation among the ten accessions of C. falcatum using RAPD and ISSR molecular markers. A total of 35 RAPD and 39 ISSR primers were screened across 10 C. falcatum accessions, of which 15 RAPD and 21 ISSR primers have showed consistent amplification. Statistics related to genetic variation were estimated using NTSYS-PC by means of Dice’s coefficient. The results revealed 80.6% and 68.07% polymorphism and similarity coefficient ranged from 0.43 to 0.91 and 0.73 to 0.93 in RPAD and ISSR analysis respectively. The dendrogram generated using RAPD, ISSR and combined RAPD-ISSR grouped accessions into different clusters which reveal considerable level molecular variation among the C. falcatum accessions. It is also evident from PCA plots that accessions are rather dispersed with tested marker systems indicating good genetic base. So, in nut shell, we found considerable genetic variation and relatedness within C. falcatum accessions collected from different areas of south Gujarat, India using RAPD and ISSR markers.     

Detection of Colletotrichum falcatum causing red rot of sugarcane by enzyme-linked immunosorbent assay

Red rot disease of sugarcane caused by Colletotrichum falcatum Went is one of the most destructive diseases of sugarcane (Saccharum officinarum L.) worldwide. The pathogen spreads primarily through infected sugarcane setts and hence the use of disease-free setts is essential to prevent the disease. In order to develop immunological method for detection of C. falcatum, two proteins with molecular weights of 27 kDa and 45 kDa were purified from the mycelium of C. falcatum race Cf 05 and used as antigen source to raise polyclonal antibodies in NewZealand white rabbit. The developed polyclonal antibodies were tested for detection of C. falcatum by enzyme-linked immunosorbent assay (ELISA) and immunoblot analysis. The polyclonal antibodies specifically detected C. falcatum in extracts from infected plants, both in immunoblot and ELISA. The ELISA results showed that the developed polyclonal antibodies were highly specific to C.falcatum. The developed antibodies were very sensitive and could detect C.falcatum proteins even at a dilution of 1:50,000. Higher ELISA absorbance values were recorded even at an antigen dilution of 1:500. In western blot analysis, protein bands with molecular weights of 27 kDa and 45 kDa reacting to antisera raised against 27 kDa and 45 kDa mycelial proteins of C. falcatum, respectively, were detected in protein samples from red rot infected canes. The high specific reactivity and sensitivity of the antisera indicate its potential suitability for ELISA-based detection of C. falcatum.     

Pseudomonas spp. Mediate defense response in sugarcane through differential exudation of root phenolics

Pseudomonas spp., a ubiquitous biocontrol agent, protects the plants from phytopathogens by suppressing them directly by reinforcing the plant’s intrinsic defense mechanism. Root exudated phenolics play an important role in establishing the rhizobacteria population and cross the host boundaries in beneficial plant–microbe interaction. In this study, Pseudomonas spp. HU-8 & HU-9 antagonized the sugarcane red rot pathogen (C. falcatum) and showed a positive chemotactic response against different concentrations (10–30 µM) of synthetic phenolic acids like p-coumaric, vanillic, and 3,4 di-hydroxybenzoic acid. In a pot experiment, they effectively colonized the sugarcane rhizosphere and mediated defense response in sugarcane plants challenged with red rot pathogen C. falcatum by regulating the exudation of root phenolics under hydroponic conditions. They significantly induced the activity of the antioxidant enzymes CAT (1.24–1.64 fold), PO (0.78–1.61 fold), PAL (0.77–0.97 fold), and PPO (3.67–3.73 fold) over untreated plants in sugarcane. They also induced the total phenolic contents (TPC) in sugarcane in the presence (6.56–10.29 mg/g GAE) and absence (2.89–4.16 mg/g GAE) of the pathogen quantified through the Folin-Ciocalteu (FC) method. However, their effect was lower than that of the pathogen (4.34–8 mg/g GAE). The Pseudomonas spp. significantly colonized the sugarcane rhizosphere by maintaining a cell population of (1.0E + 07–1.3E + 08 CFU/mL). A significant positive Pearson’s correlation was observed between the root exudated total phenolic contents, antioxidant enzymatic activities, and rhizospheric population of inoculated bacteria. The 16S rRNA and rpoD gene analysis showed sequence conservation (C: 0.707), average number of nucleotide differences (k: 199.816), nucleotide diversity, (Pi): 0.09819), average number of informative nucleotide sites per site (Psi: 0.01275), GC content (0.57), and polymorphic sites (n = 656). These diverse Pseudomonas spp. could be an ideal bio-inoculants for a broad range of hosts especially graminaceous crops.     

Mechanism of resistance induced by plant activators against Colletotrichum falcatum in sugarcane

Abstract

A search for plant activators capable of inducing systemic resistance in sugarcane showed that plants pre-treated with synthetic signal inducers confer a high degree of resistance to Colletotrichum falcatum – the red rot pathogen. Among the various treatments, Acibenzolar S- methyl (ASM) was found to be very effective in restricting the pathogen colonization inside the inoculated cane stalk tissues. The induction of resistance was accompanied by a significant increase in peroxidases and polyphenoloxidases activities. A considerable decrease of pathogen titre in the pre-treated tissues as determined by ELISA, clearly demonstrated the restriction of pathogen colonization and proliferation in the sensitized cane stalks. Specific induction of new isoforms of peroxidases and polyphenoloxidases in C. falcatum elicitor treatment indicates the pathogen elicitor induced specific cellular response of sugarcane suspension-cultured cells.     

Colletotrichum species associated with cassava anthracnose in China

Anthracnose caused by Colletotrichum (CAD) is an economically important disease of cassava; however, research on its species diversity and geographical distribution of Colletotrichum in China remains limited. In this study, we investigated the phylogenetic diversity of Colletotrichum isolates associated with symptomatic leaf tissues of cassava from Guangxi and Yunnan provinces in China and aimed to confirm their identification using advanced techniques. Based on multi‐locus phylogenetic analyses and phenotypic characteristics, we identified four species from the isolates, comprising C. plurivorum, C. karstii, C. fructicola and C. siamense, and we found variation in degrees of virulence among the species and between cassava varieties. Our results are the first to report these species from cassava, and they provide a basis for the development of management strategies for CAD in cassava.     

Der mit der Versuchsdauer anwachsende Resistenzfaktor

Abstract

Pseudomonas spp. strains capable of inducing systemic resistance were applied to sugarcane by sett treatment followed by soil applications in the field. Later the fungal pathogen Colletotrichum falcatum causing red rot disease of sugarcane was inoculated in the treated canes and its colonization was assessed by ELISA at different nodal positions above the point of inoculation. Studies with three cvs showed a significant variation in pathogen colonization only in disease susceptible cv CoC 671 and not with cvs Co 8021 and BO 91, moderately susceptible and moderately resistant to the disease, respectively. In further studies when pathogen colonization was assessed on the entire stalks of cv CoC 671, the pathogen titre was significantly reduced from three nodes upwards in the treated canes. In the upper nodes no pathogen colonization was noticed in bacteria-treated canes, whereas in the control all the nodes recorded higher titre for pathogen infection. Incorporation of chitin in the talc formulation caused further reduction in fungal colonization in the stalks.     

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.     

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.     

Morphological and molecular identification of Colletotrichum siamense,a novel leaf pathogen associated with Sterculia lanceolata recorded in China

Sterculia lanceolata, an important tropical woody plant, has high ornamental and medicinal value. To our knowledge, only brown root disease in this plant has been reported. In Nanning, Guangxi, China, an outbreak of leaf blight disease was observed on S. lanceolata in June 2019, with the leaf infection rate ranging from 80% to 100%. The disease seriously affected the leaves of trees and caused economic loss. Eight isolates were recovered from the infected leaves of different trees, and the pathogenicity was then determined by the methods of mycelial disc and conidial suspension, fulfilling Koch's postulates. According to the morphological and molecular biological characteristics of isolates, the pathogen causing leaf blight on S. lanceolata was identified as Colletotrichum siamense. Accurate identification of the pathogen provides a reliable basis for the control of the disease.     

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.     

Identification and characterization of Colletotrichum species associated with anthracnose on persimmon in Brazil

Persimmon (Diospyros kaki) anthracnose is a major threat in production areas worldwide. Most of the studies are focused on Colletotrichum horii, but other species have been reported as well. The association of distinct Colletotrichum species present in Brazilian persimmon production regions as well as their host ranges are yet elusive. The aims of this work were to identify and characterize Colletotrichum species associated with the persimmon anthracnose. In a survey performed in four production regions of Brazil, 88.7% and 11.3% out of 231 isolates were identified as members of Colletotrichum gloeosporioides species complex (Cgc) or Colletotrichum acutatum species complex (Cac), respectively. A subset of 18 isolates were identified through multilocus phylogenetic analysis, using ITS-rDNA region and two loci, namely GAPDH and TUB2. This study revealed the presence of four species: C. horii (38.8%) and Colletotrichum fructicola (27.7%) from the Cgc and Colletotrichum nymphaeae (27.7%) and Colletotrichum melonis (5.8%), from the Cac. Additionally, 13 isolates were selected for morphological, physiological, and pathogenic analyses. Contrasting characteristics were observed among species of the Cgc and Cac complexes. The optimal temperature for mycelial growth and germination were higher for Cgc species. The percentage of appressoria melanisation also varied across complexes. All the identified species were able to cause anthracnose-like symptoms on persimmon fruit, leaves, shoots, and sepals. Colletotrichum species from persimmon were also able to infect apple and pear. The findings will support decisions to manage anthracnose of persimmon under high infection risk due to multiple host susceptibility.