Colletotrichum Species Research Articles

Genomic Resources of Four Colletotrichum Species (C.fioriniae, C. chrysophilum, C. noveboracense, and C.nupharicola) Threatening Commercial Apple Production in the Eastern United States.

The genus Colletotrichum includes nine major clades with 252 species and 15 major phylogenetic lineages, also known as species complexes. Colletotrichum spp. are one of the top fungal plant pathogens causing anthracnose and pre- and postharvest fruit rots worldwide. Apple orchards are imperiled by devastating losses from apple bitter rot, ranging from 24 to 98%, which is a serious disease caused by several Colletotrichum species. Bitter rot is also a major postharvest rot disease, with C. fioriniae causing from 2 to 14% of unmarketable fruit in commercial apple storages. Dominant species causing apple bitter rot in the Mid-Atlantic United States are C. fioriniae from the Colletotrichum acutatum species complex and C. chrysophilum and C. noveboracense from the C. gloeosporioides species complex (CGSC). C. fioriniae is the dominant species causing apple bitter rot in the Northeastern and Mid-Atlantic states. C. chrysophilum was first identified on banana and cashew but has been recently found as the second most dominant species causing apple bitter rot in the Mid-Atlantic. As the third most dominant pathogen, C. noveboracense MB 836581 was identified as a novel species in the CGSC, causing apple bitter rot in the Mid-Atlantic. C. nupharicola is a sister group to C. fructicola and C. noveboracense, also causing bitter rot on apple. We deliver the resources of 10 new genomes, including two isolates of C. fioriniae, three isolates of C. chrysophilum, three isolates of C. noveboracense, and two isolates of C. nupharicola collected from apple fruit, yellow waterlily, and Juglans nigra. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Two Newly Identified Colletotrichum Species Associated with Mango Anthracnose in Central Thailand

Anthracnose caused by Colletotrichum spp. is one of the major problems in mango production worldwide, including Thailand. All mango cultivars are susceptible, but Nam Dok Mai See Thong (NDMST) is the most vulnerable. Through a single spore isolation method, a total of 37 isolates of Colletotrichum spp. were obtained from NDMST showing anthracnose symptoms. Identification was performed using a combination of morphology characteristics, Koch's postulates, and phylogenetic analysis. The pathogenicity assay and Koch's postulates on leaves and fruit confirmed that all Colletotrichum spp. tested were causal agents of mango anthracnose. Multilocus analysis using DNA sequences of internal transcribed spacer (ITS) regions, β-tubulin (TUB2), actin (ACT), and chitin synthase (CHS-1) was performed for molecular identification. Two concatenated phylogenetic trees were constructed using either two-loci of ITS and TUB2, or four-loci of ITS, TUB2, ACT, and CHS-1. Both phylogenetic trees were indistinguishable and showed that these 37 isolates belong to C. acutatum, C. asianum, C. gloeosporioides, and C. siamense. Our results indicated that using at least two loci of ITS and TUB2, were sufficient to infer Colletotrichum species complexes. Of 37 isolates, C. gloeosporioides was the most dominant species (19 isolates), followed by C. asianum (10 isolates), C. acutatum (5 isolates), and C. siamense (3 isolates). In Thailand, C. gloeosporioides and C. acutatum have been reported to cause anthracnose in mango, however, this is the first report of C. asianum and C. siamense associated with mango anthracnose in central Thailand.

First Report of Leaf Blight Caused by Colletotrichum karsti on Peppermint (Mentha x piperitavar. citrata) in Mexico.

Spearmint (Mentha x piperita var. citrata (Ehrh.) Briq.: Lamiaceae) is an aromatic herb widely cultivated owing to its industrial properties. In June 2020, symptoms of leaf blight were observed on 1,500 peppermint plants in a commercial nursery located in Cuautla (18°52'18"N 98°57'58"W), Morelos, Mexico. The incidence of the disease was 89%. Symptoms were initially observed as irregular, small black necrotic spots, that grew rapidly until the leaves were blighted. Fungal isolation was done using diseased leaf tissue on potato dextrose agar (PDA) as described by Ayvar-Serna et al. (2020) and Colletotrichum-like colonies were obtained. Six isolates were purified by single spore culture and only a single morphotype was obtained. One isolate was used for pathogenicity tests, morphological characterization, and multilocus phylogenetic analysis. The isolate (accession no. UACH449) was deposited in the Culture Collection of Phytopathogenic Fungi of the Department of Agricultural Parasitology at the Chapingo Autonomous University. Colonies in PDA grow at a rate of 7.0-10.0 mm/d. After 14 days, the colony was white to orange, and conidia (n =100) were hyaline, cylindrical, and straight with rounded ends, measuring 15.0-17.0 × 4.5-6.5 μm. Appressoria were brown and bullet-shaped. In 28-day-old colonies, the formation of perithecia was observed. Asci were hyaline, unitunicate, 8-spored, fasciculate, and cylindrical to clavate. Ascospores (n =100) were hyaline, unicellular, allantoid, inaequilateral, often straight on the inner side, apices rounded, arranged biseriately within the asci, and measured 14-19 × 4.0-7.5 μm. Morphological features of the isolate placed it tentatively within the Colletotrichm boninense species complex (Damm et al. 2012). For molecular identification, genomic DNA was extracted, and the internal transcribed spacer (ITS) region (White et al. 1990), partial sequences of calmodulin (CAL), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and actin (ACT) (Damm et al. 2012) genes were amplified and sequenced. A phylogenetic tree including published ITS, CAL, GAPDH, and ACT data for Colletotrichum species was constructed and the isolate UACH449 was grouped in the clade of Colletotrichum karsti. Sequences were deposited in GenBank under the accession numbers: ITS, OL825605; CAL, OL855890; GAPDH, OL855891 and ACT, OL855889. Pathogenicity was tested by spraying a suspension of 1 × 10^5 conidia/ml, onto eight healthy peppermint plants 30-days-old var. citrata, while eight control plants were sprayed using sterile distilled water. All plants were kept at 25 +/- 2°C and 70% RH. The characteristic symptoms of the disease were observed seven days after inoculation, while control plants remained symptomless. The pathogenicity test was repeated twice. The fungus was consistently reisolated from the eight inoculated plants and was morphologically identical to that originally isolated from diseased leaves, fulfilling Koch's postulates. To date, this pathogen has not been reported on peppermint (Farr and Rossman, 2022). To our knowledge, this is the first report of Colletotrichum karsti causing foliar blight on peppermint worldwide. According to our field observations, this disease is a threat to the production of peppermint plants.

Endophytic Colletotrichum (Sordariomycetes, Glomerellaceae) species associated with Citrusgrandis cv. "Tomentosa" in China.

Colletotrichum species are well-known plant pathogens, saprobes, endophytes, human pathogens and entomopathogens. However, little is known about Colletotrichum as endophytes of plants and cultivars including Citrusgrandis cv. "Tomentosa". In the present study, 12 endophytic Colletotrichum isolates were obtained from this host in Huazhou, Guangdong Province (China) in 2019. Based on morphology and combined multigene phylogeny [nuclear ribosomal internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (gapdh), chitin synthase 1 (chs-1), histone H3 (his3) actin (act), beta-tubulin (β-tubulin) and glutamine synthetase (gs)], six Colletotrichum species were identified, including two new species, namely Colletotrichumguangdongense and C.tomentosae. Colletotrichumasianum, C.plurivorum, C.siamense and C.tainanense are identified as being the first reports on C.grandis cv. "Tomentosa" worldwide. This study is the first comprehensive study on endophytic Colletotrichum species on C.grandis cv. "Tomentosa" in China.

Colletotrichum Species on Cultivated Solanaceae Crops in Russia

Colletotrichum species are the causal agents of potato and tomato diseases, such as black dot and anthracnose. Several new species and species complexes were recently established. Thereby, a reassessment of the genus diversity is required. The study revealed two species, Colletotrichum coccodes and Colletotrichum nigrum, as Russia’s main disease agents of cultivated Solanaceae plants. Black dot and anthracnose in potato were caused exclusively by C. coccodes, whereas the same diseases in tomato, eggplant, and pepper were predominately caused by C. nigrum. However, one isolate of C. coccodes was also identified as an agent of the tomato disease. Five potentially hybrid isolates were discovered. Morphological examination and pathogenicity assessment revealed no significant differences between the two Colletotrichum species. All isolates were sensitive to the fungicides azoxystrobin, difenoconazole, and thiabendazole, which are currently used in agriculture. This is the first report of the occurrence of C. nigrum in Russia.

First Report of Colletotrichum fructicola Causing Fruit Rot and Leaf-Tip Dieback on Pineapple in Northern Thailand.

Pineapple is one of the most economically important fruits in tropical countries, particularly in Thailand. Canned pineapple is currently Thailand's main exported commodity to many countries, including the United States, Russia, Germany, Poland, and Japan. Fungal diseases are considered a permanent threat to fruits in the pre- and post-harvest stages, leading to considerable economic losses. Fungal disease is one of the primary causes of massive yield losses in pineapples around the world. Colletotrichum species are the most common fungal pathogens affecting different tropical fruits. Although there are many reports regarding Colletotrichum species associated with pineapple, they do not have molecular data to confirm species identification. However, the occurrence of Colletotrichum species on pineapple has not been reported in Thailand so far. In this study, we isolated and identified Colletotrichum fructicola on pineapple in northern Thailand and have proven its pathogenicity to the host. This is the first report of the occurrence of Colletotrichum in pineapple, based on morpho-molecular approaches.

First Report of Colletotrichum aenigma Causing Anthracnose on Mulberry Leaves in China.

Mulberry (Morus alba L.) has been grown worldwide as a crop for silkworm rearing for over five thousand years (Jiao et al. 2020). In July 2021, a leaf spot disease was observed on mulberry leaves in Wuhan city (114°33'E, 30°48'N), Hubei province, China, with approximately 40% of leaves (about 300 trees) affected. Early symptoms were light brown, with small lesions subsequently expanding to larger sometimes irregular dark brown or black spots surrounded by yellow-brown margins, with easily perforated necrotic lesions. Leaf tissues (5 mm×5 mm) were excised from the border between diseased and healthy tissues, surface sterilized with 75% ethanol solution for 30 s and 2.5% sodium hypochlorite for 2 min, washed thrice in sterile distilled water, and then placed on potato dextrose agar (PDA), and incubated at 25°C in darkness. Four isolates (C1, C9, CHS2, and CHS6) were subcultured using the single-spore method. On PDA, colonies were cottony, pale white from above, and white to grayish-green on the reverse side. Conidia were aseptate, hyaline, subcylindrical with broadly rounded ends, 8.4 to 18.3×4.1 to 7.7 μm (mean = 13.9×5.5 μm, n = 30). Appressoria were typically elliptic or irregular with a few lobes, dark brown, 5.9 to 9.6×4.2 to 8.1 μm (mean = 7.9 ×5.7 μm, n = 30). The morphological characteristics of the isolates matched the descriptions of Colletotrichum gloeosporioides species complex (Weir et al. 2012). The isolates were further identified by analysis of the ribosomal internal transcribed spacers (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), calmodulin (CAL), actin (ACT), chitin synthase (CHS-1), glutamine synthetase (GS), and β-tubulin 2 (TUB2) genes, amplified respectively with ITS1/ITS4, GDF/GDR, CL1C/CL2C, ACT-512F/ACT-783R, CHS-79F/CHS-345R, GSF1/GSR, and Bt2a/Bt2b (Glass and Donaldson 1995; Weir et al. 2012; White et al. 1990). The sequences were deposited in GenBank (ON492187-ON492214). Concatenated sequences of the seven genes in addition to Colletotrichum species sequences from GenBank were used to conduct a phylogenetic analysis using Maximum-Likelihood (ML) method in MEGA7. The four isolates were grouped into a clade with Colletotrichum aenigma supported by a high bootstrap value (89%), and hence, they were identified as C. aenigma based on the morphological and molecular analyses. To confirm Koch's postulates, wounded leaves of six healthy 2-month-old seedlings made by a sterile needle were inoculated with each isolate by spraying 10 ml of conidial suspensions (105 conidia/ml) on each plant, and the control plants were treated with sterile distilled water. All the treated plants were kept in a plastic box containing sterile water and incubated at 28°C in a 12 h/12 h light/dark cycle. The test was performed three times. After 7 days, typical anthracnose lesions appeared on all inoculated leaves, whereas control plants remained asymptotic. Furthermore, C. aenigma was only reisolated from the symptomatic leaves. Previous studies reported five Colletotrichum species (C. morifolium, C. fioriniae, C. brevisporum, C. karstii, and C. kahawae subsp. ciggaro) to cause this disease on mulberry in China (Tian, 1981; Xue et al. 2019). To our knowledge, this is the first report of C. aenigma causing anthracnose on mulberry in China. The finding will facilitate epidemiological studies and the development of effective control strategies for the disease.

Identification and Characterization of Colletotrichum Species Associated with Cherry Leaf Spot Disease in China.

Leaf spot is a common and serious disease of sweet cherry worldwide and has become a major concern in China. From 2018 to 2020, disease investigations were carried out in Beijing City, Sichuan, Shandong, and Liaoning Provinces in China, and 105 Colletotrichum isolates were obtained from diseased samples. Isolates were identified by morphological characterization coupled with multigene phylogenetic analyses based on six loci (internal transcribed spacer region, glyceraldehyde 3-phosphate dehydrogenase, calmodulin, actin, chitin synthase, and β-tubulin). A total of 13 Colletotrichum species were identified, namely Colletotrichum aenigma, C. gloeosporioides, C. fructicola, C. siamense, C. temperatum, C. conoides, C. hebeiense, C. sojae, C. plurivorum, C. karsti, C. truncatum, C. incanum, and C. dematium. Among these, C. aenigma (25.7%) was the most prominent species isolated from diseased leaves, followed by C. gloeosporioides (19.0%) and C. fructicola (12.4%). Pathogenicity was tested on detached leaves of cv. 'Tieton' and 'Summit' and young seedlings of cv. 'Brooks' under greenhouse conditions. All 13 species were pathogenic to cherry leaves, and C. aenigma, C. conoides, and C. dematium showed high levels of virulence. Seedlings inoculated with the isolates developed similar symptoms to those seen in the orchards. This study provides the first reports for 11 of the 13 Colletotrichum species on sweet cherry in the world, excluding C. aenigma and C. fructicola. This is the first comprehensive study of Colletotrichum species associated with cherry leaf spot in China, and the results will provide basic knowledge to develop sustainable control measures for cherry leaf spot.

Antifungal activity of selected lactic acid bacteria from olive drupes

In this study, 16 Lactobacilli (LABs) isolated from the drupes of olive (Olea europaea) oil varieties were identified as Lactiplantibacillus plantarum (seven isolates), Pediococcus pentosaceus (six isolates), Enterococcus faecium (two isolates) and Streptococcus salivarius (a single isolate) by peptide mass fingerprinting and sequencing of the 16S rRNA. Antifungal activity of LABs and their cell-free fermentates (CFSs) against several plant pathogenic fungi and oomycetes (fungi sensu lato), including Alternaria, Aspergillus Colletotrichum, Penicillium, Plenodomus and Phytophthora, was evaluated in vitro using the culture overlaying and the agar diffusion tests. Minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) were determined. LABs showed antifungal activity against the fungi sensu lato tested. The most noticeable inhibitory activity was shown by isolates of L. plantarum and P. pentosaceus against Fusarium oxysporum, Colletotrichum species and Penicillium nordicum. Chemical analysis revealed CFSs contained acid lactic and variable quantities of 14 diverse phenolic acids and 26 volatile organic compounds (VOCs). No obvious correlation was found between the metabolic profile of LABs and their antifungal efficacy. However, it is the first time that the potential of fermentates of LABs, recovered from drupes of olive oil varieties, as natural fungicides, was demonstrated.

Characterization of colletotrichum species associated with soybean pod blight in soybean growing districts of Chhattisgarh

Characterization of colletotrichum species associated with soybean pod blight in soybean growing districts of Chhattisgarh

Identification and characterization of Colletotrichum species causing kiwifruit anthracnose in Kagawa Prefecture, Japan

Identification and characterization of Colletotrichum species causing kiwifruit anthracnose in Kagawa Prefecture, Japan

Colletotrichum truncatum Causing Anthracnose of Tomato (Solanum lycopersicum L.) in Malaysia.

Tomato (Solanum lycopersicum L.) is a popular nutritious vegetable crop grown in Malaysia and other parts of the world. However, fungal diseases such as anthracnose pose significant threats to tomato production by reducing the fruit quality and food value of tomato, resulting in lower market prices of the crop globally. In the present study, the etiology of tomato anthracnose was investigated in commercial tomato farms in Sabah, Malaysia. A total of 22 fungal isolates were obtained from anthracnosed tomato fruits and identified as Colletotrichum species, using morphological characteristics. The phylogenetic relationships of multiple gene sequence alignments such as internal transcribed spacer (ITS), β-tubulin (tub2), glyceraldehyde 3-phosphate dehydrogenase (gapdh), actin (act), and calmodulin (cal), were adopted to accurately identify the Colletotrichum species as C. truncatum. The results of pathogenicity tests revealed that all C. truncatum isolates caused anthracnose disease symptoms on inoculated tomato fruits. To our knowledge, the present study is the first report of tomato anthracnose caused by C. truncatum in Malaysia. The findings of this study will be helpful in disease monitoring, and the development of strategies for effective control of anthracnose on tomato fruits.

Morphometric Analysis of Isolated Conidia of Various Species of Colletotrichum sp. from Avocado and Mango in Côte d'Ivoire

Morphometric Analysis of Isolated Conidia of Various Species of Colletotrichum sp. from Avocado and Mango in Côte d'Ivoire

Identification and Characterization of Colletotrichum Species Causing Sorghum Anthracnose in Kenya and Screening of Sorghum Germplasm for Resistance to Anthracnose.

Anthracnose caused by Colletotrichum species is one of the most destructive fungal diseases of sorghum with annual yield losses of up to 100%. Although the resistance to anthracnose has been identified elsewhere, the usefulness of the resistance loci differs depending on the pathogen species and pathotypes. Accurate species identification of the disease-causing fungal pathogens is essential for developing and implementing suitable management strategies. The use of host resistance is the most effective strategy of anthracnose management and therefore identification of sources for resistance against unique pathogen pathotypes is fundamental. The aims of this study were to identify and characterize Colletotrichum species associated with sorghum anthracnose and screen sorghum germplasm for resistance to anthracnose. Symptomatic sorghum leaf samples were collected from smallholder farmers in lower eastern Kenya and used for the isolation, identification and characterization of Colletotrichum species using morpho-cultural and phylogenetic analyses with the sequences of the rDNA internal transcribed spacer (ITS) region. Pathogenicity tests of the seven fungal isolates showed that there were no significant differences in the pathogenicity on host plants. The fungal isolates were variable in cultural and morphological characters such as colony type and color, colony diameter, mycelia growth and hyaline. The phenotypic characters observed were useful in the identification of the genus Colletotrichum and not the species. Based on the sequence and phylogenetic analysis of ITS, Colletotrichum sublineola was revealed to be associated with anthracnose on sorghum. Germplasm screening for resistance to anthracnose showed differential reactions of sorghum genotypes to anthracnose under greenhouse and field conditions. The results revealed four resistant genotypes and ten susceptible genotypes against Colletotrichum sublineola. Significant (p ≤ 0.05) differences were observed in grain weight, grain yield, weight of 100 seeds and harvest index among the tested sorghum genotypes. The present study indicated that the Kenyan accessions could be an important source of resistance to anthracnose. The findings from this study provide a platform towards devising efficient disease control strategies and resistance breeding.

Identification, Pathogenicity, and Sensitivity to Fungicide of Colletotrichum Species That Causes Walnut Anthracnose in Beijing

Walnuts (Juglans regia L.) are a major food crop in Beijing. Recently, walnut anthracnose has become a serious problem in walnut plantations of Beijing, and the diversity of pathogens that cause this disease is poorly understood, making targeted treatment difficult. This study investigated incidences of walnut anthracnose in seven districts of Beijing (Changping, Fangshan, Haidian, Huairou, Mentougou, Miyun and Pinggu). A total of 377 isolates of Colletotrichum spp. were obtained from walnut anthracnose infected leaves and fruits. Morphological observation and multigene phylogenetic analysis of the genes ACT, CAL, CHS-1, GAPDH, ITS and TUB2 revealed that the isolates consisted of six species, C. aenigma, C. fructicola, C. gloeosporioides, C. siamense, C. liaoningense and C. sojae. Among these, C. gloeosporioides was the dominant species, and, for the first time, C. liaoningense and C. sojae were found to cause anthracnose in walnuts. Sensitivity tests showed that prochloraz and SYP-14288 had the strongest inhibitory effect on mycelial growth. These findings have clarified the species that cause walnut anthracnose in these seven districts of Beijing, which provides a scientific basis for diagnosis and control of walnut anthracnose.

Population Structure and Genetic Diversity of Colletotrichum gloeosporioides on Citrus in China

To analyze the genetic structure and genetic diversity of Colletotrichum gloeosporioides as the dominant Colletotrichum species on Citrus, the Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genetic diversity, including 63 strains isolated and selected from 8 different sites and 5 different citrus species, was studied. A total of 19 GAPDH haplotypes were identified by genetic analysis, and the main haplotype (haplotype 5) was distributed in 28 isolates, mainly from Citrus unchiu Hort. ex Tanaka (WG) and Citrus reticulata Blanco cv. Succosa (BDZ) in Huangyan (HY), Linhai (LH), and Jiande (JD) of Zhejiang province, and Mashui tangerine (MSJ) in Mengshan of Guangxi province (GX). Using the genetic differentiation index, Fst revealed significant genetic differentiation in C. gloeosporioides populations between Jiangxi province (JXGZ) and GX, HY, LH, JD, and Chun’an (CA) of Zhejiang province, and also revealed slightly less genetic differentiation for C. gloeosporioides populations between HY, LH, JD, GX, Shaanxi province (SX), and Quzhou (QZ) of Zhejiang province. In addition, Fst revealed great genetic differentiation between the C. gloeosporioides populations obtained from MSJ and Citrus paradise Macf (PTY), and also revealed weak genetic differentiation between the C. gloeosporioides populations obtained from Citrus sinensis Osbeck (QC), WG, and BDZ. The AMOVA test showed that the levels of genetic differentiation for C. gloeosporioides were 19% and 81% among and within geographic populations, respectively. It also showed that C. gloeosporioides had levels of genetic differentiation among and within host populations of 12% and 88%, respectively. The Mantel test showed that the genetic distance was not linearly correlated with geographical distance and the haplotype phylogenetic analysis showed that C. gloeosporioides from different regions and hosts were scattered in the phylogenetic tree, implying that the genetic differentiation was independent of host variety and geographical origin. We speculated that genetic differentiation may be mainly due to gene mutation, gene recombination, or gene migration within native populations and has nothing to do with natural selection triggered by geography or host variety.

Etiology of Colletotrichum diseases on Sastuma mandarin in Croatia

During the last decade anthracnose has become a major disease of Satsuma mandarin, the most important citrus crop in Croatia. The aim of this study was to determine Colletotrichum species associated with different symptoms and to identify the origin of inoculum. From 2013 to 2016, 437 samples of plant material were collected. Colletotrichum spp. was isolated from 93% of dried twigs, 35% of dropped flowers, 89% of leaf spots, all fruit (100%) with anthracnose or calyx-end rot symptoms, 12% of fruit with post-harvest soft rot and from 40% of fruit showing spots remaining on trees after harvest. Out of 258 Colletotrichum isolates, 253 has been morphologically identified as C. gloeosporioides (Penz.) Penz. & Sacc. species complex. Twenty-seven representative isolates were selected for phylogenetic analysis. Sequencing the inter-spacer gene region of ribosomal DNA confirmed the identity of the species. Artificial inoculation of flowers led to more than 2-fold higher young fruit drop compared to control. Pathogenicity tests on green fruit induced typical anthracnose symptoms on 82% of inoculated fruit two months after inoculation. Inoculation of mature fruit caused the appearance of typical anthracnose symptoms on 87% of inoculated fruit. These results showed that C. gloeosporioides species complex is responsible for different disease types on Satsuma mandarin, and that the fungus is present throughout the year on different plant organs.

First Report of Colletotrichum fioriniae Causing Grapevine Anthracnose in New York.

Grapevine is one of the most widely-planted fruit crops in the world, and is the most economically important fruit crop in the state of New York, USA. Symptoms of anthracnose on grapevine are similarly widely-reported on grapevine fruit and foliage, and such symptoms are commonly attributed to Elsinöe ampelina (Wilcox et al., 2015). However, similar symptoms, if not identical, to those associated with E. ampelina have been sporadically attributed to various species in the genus Colletotrichum. In September 2021, a survey was conducted in three research vineyards at Cornell AgriTech in Geneva, NY. Symptoms of anthracnose werebserved on four Vitis interspecific hybrid breeding lines in a 1 ha vineyard. Leaves, fruit, and petioles showing symptoms of anthracnose, i.e., sunken necrotic lesions with grayish centers and brownish margins, were collected. Symptomatic and healthy portions of surface-sterilized tissues were placed on PDA medium and incubated at 23oC for 7 days. Several petiole samples yielded colonies of white to greyish mycelium, with some red to orange pigmentation (Fig. 1A and 1B), similar to those described by Chowdappa et al. (2009) for Colletotrichum species isolated from grapevine in India. Cultures were allowed to sporulate. Slides from cultures were prepared and examined at 400X magnification. Conidia from cultures were cylindrical with rounded ends, 13.5-15.2 μm in length and 7.6-9.0 μm in width (Fig. 1C). Koch's postulates were fulfilled by inoculating detached healthy leaves of V. vinifera 'Chardonnay' that had been surface sterilized in 10% sodium hypochlorite and triple-rinsed in sterile distilled water. Drop inoculation was used from a suspension of 105 conidia/ml from the foregoing pure cultures as five 2 µL droplets per leaf. Inoculated detached leaves were maintained on water agar in a Petri dish at 23oC. Four days after inoculation, symptoms were observed and compared with the originally collected samples. Inoculated leaves displayed symptoms typically found on the collected tissues, and the original pathogen, as confirmed by colony morphology and conidial characteristics and dimensions, was reisolated from inoculated leaves, and not from non-inoculated controls. For molecular characterization, fungal DNA was isolated by using Qiagen DNeasy kit and amplified using the following primer pairs: ITS1/ITS4, TEF (Hyun et al., 2009), E. ampelina F/R (Santos et al. 2018), TUB2, ACT, HIS3, GAPDH and CHS1 (Damm et al., 2001). PCR products were purified using ExoSAP-IT, and samples were Sanger sequenced. Sequences were analyzed using Geneious Prime software, and the resulting sequences (NCBI accessions OL720215, OL720216, OL720217, OL720218, OL853836, OM982612, OM982613, OM982614, OM982615 and OM982616) had 94 to 100% identity to Colletotrichum fioriniae NCBI accessions MN944922.1, MK646015.1, MN944922.1, MN856415.1, KU847413.1, MN520490.1, MN544294.1, KY695259.1, MN535117.1 and MN544295.1. Symptoms of grapevine anthracnose caused by Colletotrichum species have been reported from India (Chowdappa et al., 2009) and Korea (Kim et al., 2021). To our knowledge this is the first report of grapevine anthracnose caused by C. fioriniae Anthracnose and ripe rot are diseases of increasing importance, particularly as new grapevine cultivars with resistance to powdery mildew or downy mildew are adopted. Taxonomy of the causal agents (E. ampelina and Colletotrichum spp.) has undergone considerable revision. Consequently, distribution and relative prevalence of the various taxa will require further study.

Avaliação da sensibilidade micelial a fungicidas de Colletotrichum spp. associadas à atemoia no estado de Alagoas.

Anthracnose, caused by Colletotrichum spp., is an important disease of atemoya causing damage both pre and post-harvest. Symptoms of this disease range from foliar anthracnose, flower abortion, branch blight and necrotic fruit lesions. Knowing the action of certain fungicides in the fight against the etiological agent is essential to develop control techniques. The objective of this work was to evaluate the mycelial sensitivity of Colletotrichum spp., from atemoya, to the fungicides thiophanate methyl, azoxystrobin and tebuconazole. To determine the sensitivity of Colletotrichum species to fungicides, mycelial growth in synthetic potato-dextrose-agar (PDA) supplemented with fungicide was evaluated. The fungicides were dissolved in dimethylsulfoxide (DMSO) and added to the fluxing culture medium (45°C), to reach concentrations of 0.0, 0.1, 0.5, 1.0, 5.0 and 10 μg/ml-1 of a.i. The mycelial growth of each colony was measured daily to obtain the mycelial growth index (ICM). EC50 was also calculated. After calculating the EC50, the Colletotrichum species were classified into three sensitivity categories, where: EC50: <10 μg/ml-1: high sensitivity (SA); EC50: 10-100 μg/ml-1: moderate sensitivity (MS); EC50: 100-500 μg/ml-1: insensitivity (I). The species C. theobromicola, C. fruticola, C. siamense and C. karstii were highly sensitive to the fungicides tebuconazole and thiophanate methyl. The fungicide azoxystrobin proved to be efficient in the in vitro control of the species C. theobromicola, however, it proved to be inefficient for the control of the species C. siamense. The species C. fruticola and C. karstii were moderately sensitive to the fungicide azoxystrobin.

First report of anthracnose on sugar beet (Beta vulgaris) caused by Colletotrichum incanum in Michigan, USA

In 2016, sugar beet leaf samples were received at the Michigan State University plant diagnostic laboratory with distinctive oblong lesions on the petiole (Fig. 1). The lesions contained acervuli with dark setae (Fig. 2). Lesions on the blade had light and dark rings that showed similar acervuli in a 90% humidity chamber. Pure cultures from single spore isolation were grown on malt extract agar (MEA). The resulting cultures produced acervuli-like structures with dark setae distributed throughout the conidiomata. Spores were curved and had a distinct gutule (Fig. 3). Based on these characteristics, isolates were identified as a Colletotrichum sp. A representative isolate, Co16-4, was grown in liquid V8 medium with shaking for seven days. Fungal tissue was lyophilised and DNA extracted using a DNeasy Plant Mini Kit (Qiagen, Germany). Partial sequences of the internal transcribed spacer (ITS), actin (ACT), and β-tubulin (TUB2) genes were amplified according to Damm et al. (2009) and submitted to the Research Technology Support Facility (East Lansing, USA) for Sanger sequencing. Sequences were compared to data in GenBank where ITS gave the closest match to C. spaethianum (>98%), while ACT and TUB2 gave the closest match to C. incanum (99 and 96%, respectively). Sequences were submitted to GenBank as Accession Nos. OP628186 (ITS), OP627081 (ACT) and OP627082 (TUB2). Due to the BLAST search ambiguity, sequences of 19 Colletotrichum species were retrieved from GenBank as identified in Yang et al. (2014). Sequences were concatenated and manually adjusted using Geneious Prime 2021.1.1 (Biomatters Ltd., Auckland, New Zealand). Alignment and a maximum-likelihood tree were generated in Mega version 11.0.13 (Tamura et al., 2021) using the Tamura-Nei substitution model with 1000 bootstrap replicates (Fig. 4). Based on this data, isolate Co16-4 was identified as Colletotrichum incanum. Koch's postulates were fulfilled by spraying sugar beet foliage (C869) at the 10- to 12-leaf growth stage with 10ml of a spore suspension (1×104/ml with 0.1% Tween 20). After seven days, an oblong lesion was observed on the petiole and irregular lesions developed on inoculated leaves. No lesions were observed on control plants. A morphologically identical Colletotrichum sp. was isolated from lesions. Colletotrichum incanum has been reported on soybean (Yang et al., 2014), but to our knowledge, no Colletotrichum spp. have been reported causing anthracnose on beet in the United States. The symptoms and morphology of the lesions on sugar beets are consistent with beet anthracnose reported in Japan and Canada which was attributed to C. dematium (Chiko et al., 1984; Gourley, 1966). In Japan, a radish isolate, originally identified as C. dematium, was recently identified as C. incanum (Gan et al., 2016). Leaves with similar lesions were found in commercial sugarbeet fields in 2021 and 2022, indicative of continuing disease presence, but yield effects are unknown.