• The Plant Pathology Journal
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• v.32 no.3
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• pp.251-259
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• 2016

The present study investigated the suppression of the disease development of anthracnose caused by Colletotrichum gloeosporioides and C. acutatum in harvested apples using an antagonistic rhizobacterium Paenibacillus polymyxa APEC128 (APEC128). Out of 30 bacterial isolates from apple rhizosphere screened for antagonistic activity, the most effective strain was APEC128 as inferred from the size of the inhibition zone. This strain showed a greater growth in brain-heart infusion (BHI) broth compared to other growth media. There was a reduction in anthracnose symptoms caused by the two fungal pathogens in harvested apples after their treatment with APEC128 in comparison with non-treated control. This effect is explained by the increased production of protease and amylase by APEC128, which might have inhibited mycelial growth. In apples treated with different APEC128 suspensions, the disease caused by C. gloeosporioides and C. acutatum was greatly suppressed (by 83.6% and 79%, respectively) in treatments with the concentration of $1{\times}10^8$ colony forming units (cfu)/ml compared to other lower dosages, suggesting that the suppression of anthracnose development on harvested apples is dose-dependent. These results indicated that APEC128 is one of the promising agents in the biocontrol of apple anthracnose, which might help to increase the shelf-life of apple fruit during the post-harvest period.

• Research in Plant Disease
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• v.29 no.4
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• pp.345-362
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• 2023

Anthracnose, caused by the Colletotrichum genus, comprises a significant number of plant pathogens and poses a major threat to fruit production worldwide, including South Korea. Colletotrichum species were identified associated with anthracnose in fruits such as apple, persimmon, plum, peach, jujube, walnut, and grape. A polyphasic approach, including morphology, multigene phylogenetics, and pathogenicity testing, was used. Additionally, the in-vitro sensitivity of identified Colletotrichum species to common fungicides was also evaluated. A total of nine Colletotrichum species within two complexes, namely gloeosporioides and acutatum, have been identified as the causal agents of anthracnose in common fruits in South Korea. In the gloeosporioides complex, we found Colletotrichumaenigma, C. fructicola, C. gloeosporioides, C. horii, C. siamense, and C. viniferum. Meanwhile, in the acutatum complex, C. fioriniae, C. nymphaeae, and C. orientalis were identified. Notably, C. fructicola, C. siamense, C. fioriniae, and C. nymphaeae were reported for the first time from apple, C. siamense, C. fioriniae and C. nymphaeae from plum, C. siamense, C. fructicola, and C. fioriniae frompeach, C. siamense and C. horii from persimmon, C. fioriniae from Omija (Schisandra), C. orientalis from walnut, C. nymphaeae from jujube, and C. aenigma, C. fructicola, and C. siamense fromgrape. Fungicide sensitivity tests revealed significant variation in the EC₅₀ values among specific Colletotrichum species when exposed to different fungicides. Moreover, the same Colletotrichum species isolated from different host plants displayed varying sensitivity to the same fungicide.

• Korean Journal of Environmental Agriculture
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• v.36 no.4
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• pp.299-305
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• 2017

BACKGROUND: Although the filamentous fungal pathogen Colletotrichum species causing anthracnose disease on various fruits including peach, apple, persimmon and grape, there is no report on Japanese plum in Korea. METHODS AND RESULTS: In 2016, diseased fruits showing typical anthracnose symptoms of Japanese plum were collected in market and ochards. Diseased tissue was cut off and disinfected subsequently with 70% ethanol for 1 min, and in 1% sodium hypochloride solution for 1 min, followed by three washes with sterile distilled water. The disinfected tissues were placed onto potato dextrose agar (PDA), and incubated at $25^{\circ}C$ in the dark for 5 to 7 days. For single-spore isolation, conidia were scraped off the plate using a loop, and suspended with 10 mL sterile distilled water. One hundred microliter of the conidial suspension was spread on PDA plates and incubated at $25^{\circ}C$. Finally, one germinated conidium was transferred onto PDA plates. Morphological and cultural characteries of colonies and spores of isolated Colletotrichum were observed after 7 to 10 days incubation on PDA. Molecular identification of isolates were analyzed by comparing rDNA-ITS gene sequences with NCBI GeneBank. CONCLUSION: Of eleven isolates of Colletotrichum isolated from anthracnose diseased Japanese plum fruits, six were identified as C. acutatum, and five as C. gloeosporioides based on diagnostic characteristics such as colony growth rate, shape and size of conidia, and rDNA-ITS sequences. This is the first report of Colletotrichum causing the anthracnose on Japanese plum in Korea.

• Journal of Microbiology and Biotechnology
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• v.16 no.2
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• pp.280-285
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• 2006

Methanol extract of the rhizomes of turmeric, Curcuma longa L., effectively controlled the development of red pepper anthracnose caused by Colletotrichum coccodes. In addition three antifungal substances were identified from the methanol extract of C. longa rhizomes as curcumin, demethoxycurcumin, and bisdemethoxycurcumin using mass and $^{1}H-NMR$ spectral analyses. The curcuminoids in a range $0.4-100\;{\mu}g/ml$ effectively inhibited the mycelial growth of three red pepper anthracnose pathogens, C. coccodes, C. gloeosporioides, and C. acutatum. The three curcuminoids inhibited mycelial growth of C. coccodes and C. gloeosporioides to an extent similar to the synthetic fungicide dithianon did, but the synthetic agent was a little more effective against C. acutatum. The curcuminoids also effectively inhibited spore germination of C. coccodes, and bisdemethoxycurcumin was the most active. Among the three curcuminoids, only demethoxycurcumin was effective in a greenhouse test in suppressing red pepper anthracnose caused by C. coccodes.

• Research in Plant Disease
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• v.26 no.1
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• pp.8-18
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• 2020

Red pepper, one of the major economic crops in Korea, is being affected by anthracnose disease caused by Colletotrichum acutatum. To control this disease, an antagonistic bacterial strain, Bacillus subtilis YGB36 identified by 16S rDNA sequencing, physiological and biochemical analyses is used as a biological control agent. In vitro screening revealed that the strain YGB36 possess strong antifungal activity against the pathogen Cylindrocarpon destructans. The strain exhibited cellulase, protease, amylase, siderophore production and phosphate solubility. In vitro conidial germination of C. acutatum was most drastically inhibited by YGB36 cell suspensions (10⁶ cfu/ml) or culture filtrate. Development of anthracnose symptoms was reduced on detached immature green pepper fruits by treatment with cell suspensions, and its control value was recorded as 65.7%. The YGB36 bacterial suspension treatment enhanced the germination rate of red pepper seeds and promoted root development and growth under greenhouse conditions. The in vitro screening of fungicide and insecticide sensitivity test against YGB36 revealed that the bacterial growth was not affected by any of the insecticides, and 11 fungicides out of 21 used. Collectively, our results clearly suggest that the strain YGB36 is considered as one of the potential biocontrol agents against anthracnose disease in red pepper.

• The Korean Journal of Mycology
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• v.46 no.4
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• pp.479-490
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• 2018

Red pepper is seriously damaged by thrips (Thrips palmi) and anthracnose caused by Colletotrichum acutatum throughout its development. Because of biotic constraints, producers often depend on chemicals that are expensive and have adverse effects on the environment, operator, and beneficial insects. In addition, resistance is developed because of the repeated use of chemicals. In recent decades, the use of microorganisms in crop protection has become a credible alternative because it is eco-friendly. In this study, we aimed to select isolates with insecticidal and fungicidal activities against the pathogens that cause anthracnose and thrips. We treated T. palmi adults and juveniles with 13 strains of entomopathogenic fungi (isolated from the soil by using the insect-bait method), and 6 strains showed excellent insecticidal activity (70-100%) 5 days after the treatment. The selected isolates were cultured with C. acutatum to screen for the strain with excellent anti-fungal activities, among which an isolate FT333 showed more than 95% control efficacy against C. acutatum in vitro. The isolate was identified as Isaria javanica through its morphological characteristics and phylogenetic analysis of the ITS and ${\beta}-tubulin$ nucleotide sequences. The Isaria javanica FT333 isolate could be used effectively for dual bio-control of thrips and anthracnose during red pepper cultivation.

• The Plant Pathology Journal
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• v.24 no.4
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• pp.392-399
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• 2008

Detached chili pepper fruits were inoculated with the conidial suspension of Colletotrichum acutatum JC-24 simultaneously (simultaneous inoculation, SI) and at delayed time (delayed inoculation, DI) after wounding with (delayed wound inoculation, DWI) or without additional wounding (delayed non-wound inoculation, DNI) at the inoculation time. Disease severity was significantly lowered by DNI, compared to SI. By DNI, the disease reduction rates were proportional with the length of delayed time, and greater at the high temperature range (18, 23 and $28^{\circ}$) than at the low temperature ($13^{\circ}$) tested. DWI was also effective in reducing the disease severity especially at 18oC; however, its effectiveness was lower than for DNI. In light microscopy, parenchyma cells at the wounding sites were modified structurally, initially forming new cell walls crossing cytoplasm, enlarged with multiple periclinal cell divisions, and finally layered like wound periderms. In DWI, the above structural modifications occurred, showing the restriction of the fungal invasion by the cell walls in enlarged modified cells, while no definite cellular modifications were found with proliferation of fungal hyphae in SI. Sclerenchyma-like cells with thickened cell walls were proliferated around the wounding sites, which were partially dissolved by DWI, probably leading to some disease development. All of these results suggest that the decline of the anthracnose disease in pepper fruit by the delayed inoculations may be derived from the structural modifications related to the healing processes of the previous wound inflicted on the tissues.

• Korean journal of applied entomology
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• v.53 no.2
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• pp.157-169
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• 2014

Between 2010 and 2012, diseases and insect pests of sweet persimmon were surveyed at sweet persimmon export complexes and non-export orchards in Suncheon, Jeonnam Province; Jinju, Changwon (Dongeup and Bukmyeon), and Gimhae, Gyeongnam Province; and Ulzu, Ulsan. The following diseases were found in the sweet persimmon orchards: angular leaf spot (Cercospora kaki), anthracnose (Colletotrichum gloeosporioides and Colletotrichum acutatum), circular leaf spot (Mycosphaerella nawae), powdery mildew (Phyllactinia kakicola), and gray mold (Botrytis cinerea). Circular leaf spot was the most frequent and serious disease, and C. gloeosporioides and C. acutatum were found on fruits. Thirty-three insect pest species that belonged to 32 genera of 20 families in 5 orders were found in the sweet persimmon orchards; the two-spotted spider mite, Tetranychus urticae, was also found in the surveyed orchards. Apolygus spinolae, Pseudaulacaspis cockerelli, and Adoxophyes orana were widely found in the surveyed orchards; Spodoptera litura and Homona magnanima were also recorded. Damage by insect pests was low, and the quarantine insect pests peach pyralid moth (Dichocrocis punctiferalis) and persimmon fruit moth (Stathmopoda masinissa) were rarely or not found in the sweet persimmon export complexes. In addition, other quarantine insect pests, such as persimmon false spider mite (Tenuipalpus zhizhilashviliae) and Japanese mealybug (Planococcus kraunhiae), were not detected. These quarantine insect pests were also not found in the sorting places, storage houses, and fruits for export; however, scale insects and two-spotted spider mites were found at a low rate. Although anthracnose (C. acutatum) infested fruit was found in the storage houses, only one in Jinju and Gimhae.

• The Plant Pathology Journal
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• v.35 no.6
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• pp.575-584
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• 2019

Colletotrichum acutatum is a species complex responsible for anthracnose disease in a wide range of host plants. Strain C. acutatum KC05, which was previously isolated from an infected pepper in Gangwon Province of South Korea, was reidentified as C. scovillei using combined sequence analyses of multiple genes. As a prerequisite for understanding the pathogenic development of the pepper anthracnose pathogen, we optimized the transformation system of C. scovillei KC05. Protoplast generation from young hyphae of KC05 was optimal in an enzymatic digestion using a combined treatment of 2% lysing enzyme and 0.8% driselase in 1 M NH₄Cl for 3 h incubation. Prolonged incubation for more than 3 h decreased protoplast yields. Protoplast growth of KC05 was completely inhibited for 4 days on regeneration media containing 200 ㎍/ml hygromycin B, indicating the viability of this antibiotic as a selection marker. To evaluate transformation efficiency, we tested polyethylene glycol-mediated protoplast transformation of KC05 using 19 different loci found throughout 10 (of 27) scaffolds, covering approximately 84.1% of the entire genome. PCR screening showed that the average transformation efficiency was about 17.1% per 100 colonies. Southern blot analyses revealed that at least one transformant per locus had single copy integration of PCR-screened positive transformants. Our results provide valuable information for a functional genomics approach to the pepper anthracnose pathogen C. scovillei.

• The Plant Pathology Journal
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• v.38 no.6
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• pp.629-636
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• 2022

Colletotrichum species is known as the major causal pathogen of red pepper anthracnose in Korea and various groups of fungicides are registered for the management of the disease. However, the consistent use of fungicides has resulted in the development of resistance in many red pepper-growing areas of Korea. Effective management of the occurrence of fungicide resistance depends on constant monitoring and early detection. Thus, in this study, various methods such as agar dilution method (ADM), gene sequencing, allele-specific polymerase chain reaction (PCR), and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) were applied for the detection of benzimidazole resistance among 24 isolates of Colletotrichum acutatum s. lat. and Colletotrichum gloeosporioides s. lat. The result of the ADM showed that C. gloeosporioides s. lat. was classified into sensitive and resistant isolates to benomyl while C. acutatum s. lat. was insensitive at ≥1 ㎍/ml of benomyl. The sequence analysis of the β-tubulin gene showed the presence of a single nucleotide mutation at the 198th amino acid position of five isolates (16CACY14, 16CAYY19, 15HN5, 15KJ1, and 16CAYY7) of C. gloeosporioides s. lat. Allele-specific PCR and PCR-RFLP were used to detect point mutation at 198th amino acid position and this was done within a day unlike ADM which usually takes more than one week and thus saving time and resources that are essential in the fungicide resistance management in the field. Therefore, the molecular techniques established in this study can warrant early detection of benzimidazole fungicide resistance for the adoption of management strategies that can prevent yield losses among farmers.