• The Plant Pathology Journal
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• v.37 no.6
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• pp.681-686
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• 2021

It is difficult to distinguish melanose and melanoses-like symptoms with the naked eye because they appear similar. To accurately detect melanose symptoms caused by Diaporthe citri from melanose-like symptoms, we developed PCR-based specific primers Dcitri by aligning the internal transcribed spacer (ITS) region of D. citri with the ITS of Diaporthe cytosporella, Diaporthe foeniculina, Colletotrichum gloeosporioides, Botrytis cinerea, Alternaria citri, and Fusarium oxysporum found on citrus peel. PCR results showed that the specific product was amplified in D. citri but not in other isolates including, C. gloeosporioides, B. cinerea, A. citri, F. oxysporum. In addition, specific products were observed in melanose symptoms caused by D. citri but not in melanose-like symptoms, such as copper-injury, sunscald, damages by yellow tea thrips, and pink citrus rust mite. Using the Dcitri primers developed in this study, it is expected that melanose caused by D. citri could be accurately distinguished from melanose-like symptoms.

• The Plant Pathology Journal
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• v.35 no.5
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• pp.417-424
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• 2019

Melanose, caused by Diaporthe citri, is one of severe diseases in citrus, a major economic resource in Jeju island. To reduce the usage amount of organic synthetic fungicide, bio-sulfur was tested as an alternative chemical to control citrus melanose in the present study. Direct antifungal activity of bio-sulfur against D. citri was determined through in vitro experiment using artificial nutrient media. Disease severity of melanose on bio-sulfur pretreated citrus leaves was lower than that on untreated ones. To illustrate the mechanism of disease suppression by bio-sulfur, infection structures were observed with a fluorescent microscope and a scanning electron microscope. In fluorescent microscopic observation, most conidia rarely germinated. In addition, hyphal growth on leaves pretreated with bio-sulfur was inhibited compared to that on untreated ones. In scanning electron microscope images of bio-sulfur pretreated leaves, surfaces of most conidia were shrunk while hyphae were morphologically changed and frequently branched. Such microscopic observations were also found for leaves pretreated with a commercial fungicide Dithianon. These results suggest that bio-sulfur may be used to control citrus melanose as an environment friendly alternative to organic synthetic fungicides

• Research in Plant Disease
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• v.18 no.4
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• pp.331-337
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• 2012

Citrus melanose is one of important disease in citrus cultivation, reducing quality of citrus fruits and resulting in economic loss. Like other diseases in citrus, melanose was mainly protected by chemical fungicide in the field. Recently, alternative method of disease control is highly required due to the side effect of the chemicals. In this study four rhizobacterial strains TRH423-3, MRL408-3, THJ609-3, and TRH415-2 are selected by dual-culture testing its antifungal activity against Diaporthe citri causing citrus melanose. To investigate the protection efficacy of the selected rhizobacterial strains to citrus melanose, the bacteria were pre-treated on citrus leaves following inoculation with melanose pathogen. Pre-treatment with all selected rhizobacterial strains showed disease suppression in which the levels of protection rates were different by the rhizobacterial strains. Additional treatment with the rhizobacterial strains after the pathogen inoculation enhanced protection rates in all cases. The strain MRL408-3 and TRH423-3 were identified as Burkholderia gladioli, TRH415-2 as Pseudomons fluorescens and THJ609-3 as Pseudomonas pudia as a result of analyzing the internal transcript spaces of the rhizobacterial strains rDNA. The selected rhizobacterial strains may be valuable as biological control agents in the environment-friendly citrus farm in which chemical application is limited.

• Journal of Environmental Science International
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• v.29 no.10
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• pp.997-1002
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• 2020

In this study, we investigated the effects of the eco-friendly chemical bio-sulfur, on the citrus melanose-causing pathogen, Diaporthe citri, and on the pest, Panonychus citri. In an open field experiment with a plot-scale application of the chemicals: Mancozeb, lime sulfur, lime sulfur + machine oil, and bio-sulfur, the control group showed 70.6% disease severity compared with 10.3% for the Mancozeb-treated group. Among the eco-friendly treatments, disease severity was the lowest for the group treated with lime sulfur + machine oil (32.2%) and was 53.9%, 58.8%, and 58.1% following treatment with lime sulfur, and bio-sulfur diluted 500 and 1000 times, respectively. The proportion of diseased fruit showed similar results, suggesting that bio-sulfur is an effective alternative to lime sulfur. Three days after treatment acaricidal effects on P. citri showed a 197.6% control survival rate whereas the machine oil, and bio-sulfur diluted 500 and 1000 times treatments showed rates of 2.9%, 5.8%, and 9.0%, respectively. After three days, the control value for bio-sulfur diluted 1000 times was 73.2% compared with the values for the machine oil (96.4%) and bio-sulfur diluted 500 times (94.6%) treatments. Therefore, we suggest that additional research is needed on the combined application of bio-sulfur and oils to enhance the additive control effect on citrus melanose and Panonychus citri.

• The Korean Journal of Pesticide Science
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• v.13 no.4
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• pp.332-335
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• 2009

Rhizo-bacteria were isolated from organic-farming soils to select antagonistic agent for controlling citrus melanose disease. Among several antagonistic bacteria, KB-401 effectively inhibited mycelial growth of several plant fungal pathogens, including the pathogen of citrus melanose, Diaporthe citri. KB-401 also inhibited spore germination of the fungal pathogen. The tip of germ tube was swollen when conidia of D. citri were co-culture with KB-401 in PD broth amended 1% glucose. KB-401 was identified as Bacillus subtilis through the investigation for physiological characters and the analysis of nucleotide sequences of 16S rDNA.

• Research in Plant Disease
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• v.20 no.3
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• pp.196-200
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• 2014

This study was carried out to determine the effect of mixing with paraffin oil on rainfastness of mancozeb on citrus fruits and assay the improvement of control effect of mancozeb against citrus melanose by mixing with paraffin oil. In artificial rainfall condition (7.2 mm/hr), the attached contents of mancozeb on detached fruits were the most in treatment of mancozeb 0.2% + paraffin oil 0.1% as the contents was $7.43{\mu}g/cm^2$ after treatment of rainfall for 10 hr. In open field condition, the contents of attached mancozeb on fruits were significantly more in treatment of mancozeb 0.2% + paraffin oil 0.1% or 0.25% than the other treatments 1, 15 and 25 days after treatment 2009 and 2010 seasons. The disease incidence was significantly lower in treatment of mancozeb 0.2% + paraffin oil 0.1% or 0.25% than treatment of mancozeb 0.2% only 2009, 2010 and 2011 seasons. Based on this study, it was suggested that the control effect of mancozeb against citrus melanose can be improved by mixing with paraffin oil.

• Research in Plant Disease
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• v.29 no.2
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• pp.130-136
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• 2023

Citrus melanoses caused by Diaporthe citri, has been one of the serious diseases in many citrus orchards of Jeju Island. To protect melanose in citrus farms, a fast and exact diagnosis method is necessary. In this study, diseased leaves and dieback twigs were collected from a total of 49 farms within March to April in 2022. A total of 465 fungal isolates were obtained from a total of 358 isolated plant samples. Among these fungal isolates, 40 representatives of D. citri isolates which were isolated from 22 twigs and 18 leaves on 23 farms were found based on cultural characteristics on potato dextrose agar and conidial morphology. Additionally, the molecular assay was carried out and compared with those by morphological diagnosis. All isolates were identified as D. citri by analyzing the sequences at the internal transcribed spacer (ITS) rDNA region using primers of ITS1/ITS4 or at β-tubulin using primer Btdcitri-F/R. Therefore, based on the present study, where the results of morphological identification of conidial type were consistent with DNA sequence analysis of certain gene, choosing a suitable method for a fast diagnosis of citrus melanose was suggested.

• The Plant Pathology Journal
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• v.39 no.4
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• pp.351-360
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• 2023

Citrus melanose, caused by Diaporthe citri, has been one of the serious diseases, and chemical fungicides were used for protection in many citrus orchards of Jeju Island. Establishing a disinfectant resistance management system and reducing pesticide usage would be important for contributing to safe agricultural production. In this study, monitoring of chemical resistance was performed with 40 representative D. citri isolates from many citrus orchards in Jeju Island. Four different fungicides, kresoxim-methyl, benomyl, fluazinam, and prochloraz manganese, with seven different concentrations were tested in vitro by growing the mycelium of the fungal isolates on the artificial medium potato dextrose agar. Among the 40 fungal isolates, 12 isolates were investigated as resistant to kresoxim-methyl which could not inhibit the mycelium growth to more than 50%. Especially isolate NEL21-2 was also resistant against benomyl, whose hyphae grew well even on the highest chemical concentration. However, any chemical resistance of fungal isolates was found against neither fluazinam nor prochloraz manganese. On the other hand, in vivo bio-testing of some resistant isolates was performed against both kresoxim-methyl and benomyl on young citrus leaves. Typical melanose symptoms developed on the citrus leaves pre-treated with both agrochemicals after inoculation with the resistant isolates. However, no or less symptoms were observed when the susceptible isolates were inoculated. Based on these results, it was suggested that some resistant isolates of D. citri occurred against both systemic fungicides, which may be valuable to build a strategy for protecting citrus disease.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2002.11a
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• pp.70.3-71
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• 2002

• Research in Plant Disease
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• v.9 no.3
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• pp.153-158
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• 2003

Mycological characteristics of Phomopsis citri isolates obtained from blighted twigs of citrus cultivation areas in Jeju island were all identified as Phomopsis citri known as the anamorph stage of Diaporthe citri. Pycnidia the isolates sized from 112.2 to 614.4${\mu}m$ (av. 328.8${\mu}m$) and shaped conical to lenticulata in black. Two types of phialospores, ${\alpha}$-and ${\beta}$-spores were observed from pycnidia. The ${\alpha}$-spores were hyaline, unicellula, fusiform to ellipsoidal and sized 4.7~8.7 1.9 ${\times}$ 3.5${\mu}m$ (av. 6.7 ${\times}$ 2.3 ${\mu}m$). The ${\beta}$-spores were hyaline, unicellula, filiform, curved and often strongly hooked and sized 13.2~27.1 ${\times}$ 0.8~1.6 ${\mu}m$ (av. 22.1 ${\times}$ 1.0 ${\mu}m$). All isolates grew well and produced abundant pycnidia and spore horns on PDA. In addition, all isolates showed strong pathogenicity to citrus fruit inducing melanose symptoms when artificially inoculated with the pycnidial spores.