• The Plant Pathology Journal
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• v.33 no.3
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• pp.307-317
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• 2017

Satsuma dwarf virus (SDV) or Citrus mosaic sadwavirus (CiMV) were not consistently detected in RTPCR assay with the primer sets based on gene of Japan isolates. SDV and CiMV isolates were distinctively divided into two groups based on phylogenetic analysis of PP2 gene cloned from 22 Korean isolates, and the Korean CiMV and SDV isolates shared 95.5-96.2% and 97.1-97.7% sequence identity with Japanese isolate, respectively. We developed PP2-1 primer set based on the PP2 gene sequence of Korean isolates to simultaneously and effectively detect SDV and CiMV. And CTLV-2013 and CTV-po primer sets were newly designed for detection of Citrus tatter leaf virus (CTLV) and Citrus tristeza virus (CTV), respectively. Using these primer sets, a new multiplex PCR assay was developed as a means to simultaneously detect 4 citrus viruses, CTV, CTLV, SDV, and CiMV. The degree of detection by the multiplex PCR were consistent with those of uniplex RT-PCR for detection of each of the viruses. Therefore, the new multiplex PCR provides an efficient method for detecting 4 citrus viruses, which will help diagnose many citrus plants at the same time. We verified that 35.2% and 72.1% of 775 trees in 155 orchards were infected with SDV or CiMV (SDV/CiMV) and CTV by the multiplex-PCR assay, respectively, and CTLV was not detected in any of the trees tested.

• The Plant Pathology Journal
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• v.37 no.3
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• pp.268-279
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• 2021

Citrus scab, caused by the fungal pathogen Elsinoë fawcettii, is one of the most important fungal diseases affecting Citrus spp. Citrus scab affects young tissues, including the leaves, twigs, and fruits, and produces severe fruit blemishes that reduce the market value of fresh fruits. To study the molecular responses of satsuma mandarin (C. unshiu) to E. fawcettii, plant hormone-related gene expression was analyzed in response to host-compatible (SM16-1) and host-incompatible (DAR70024) isolates. In the early phase of infection by E. fawcettii, jasmonic acid- and salicylic acid-related gene expression was induced in response to infection with the compatible isolate. However, as symptoms advanced during the late phase of the infection, the jasmonic acid- and salicylic acid-related gene expression was downregulated. The gene expression patterns were compared between compatible and incompatible interactions. As scabs were accompanied by altered tissue growth surrounding the infection site, we conducted gibberellic acid- and abscisic acid-related gene expression analysis and assessed the content of these acids during scab symptom development. Our results showed that gibberellic and abscisic acid-related gene expression and hormonal changes were reduced and induced in response to the infection, respectively. Accordingly, we propose that jasmonic and salicylic acids play a role in the early response to citrus scab, whereas gibberellic and abscisic acids participate in symptom development.

• The Plant Pathology Journal
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• v.39 no.4
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• pp.309-318
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• 2023

Huanglongbing (HLB) is one of the most destructive diseases in citrus, which imperils the sustainability of citriculture worldwide. The presumed causal agent of HLB, 'Candidatus Liberibacter asiaticus' (CLas) is a non-culturable phloem-limited α-proteobacterium transmitted by Asian citrus psyllids (ACP, Diaphorina citri Kuwayama). A widely adopted method for HLB diagnosis is based on quantitative real-time polymerase chain reaction (qPCR). Although HLB diagnostic qPCR provides high sensitivity and good reproducibility, it is limited by time-consuming DNA preparation from plant tissue or ACP and the requirement of proper lab instruments including a thermal cycler to conduct qPCR. In an attempt to develop a quick assay that can be deployed in the field for CLas detection, we developed a real-time loop-mediated isothermal amplification (rt-LAMP) assay by targeting the CLas five copy nrdB gene. The rt-LAMP assay using various plant sample types and psyllids successfully detected the nrdB target as low as ~2.6 Log10 copies. Although the rt-LAMP assay was less sensitive than laboratory-based qPCR (detection limit ~10 copies), the data obtained with citrus leaf and bark and ACP showed that the rt-LAMP assay has >96% CLas detection rate, compared to that of laboratory-based qPCR. However, the CLas detection rate in fibrous roots was significantly decreased compared to qPCR due to low CLas titer in some root DNA sample. We also demonstrated that the rt-LAMP assay can be used with a crude leaf DNA extract which is fully deployable in the field for quick and reliable HLB screening.

• Mycobiology
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• v.49 no.3
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• pp.267-274
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• 2021

Two Diaporthe species isolated from fruit of Citrus sinensis in China were characterized based on morphology and multilocus phylogeny of ITS, tef1, and tub2 gene sequences. The phylogeny indicated that the two species match Diaporthe taoicola and D. siamensis. A critical examination of phenotypic characteristics confirmed the phylogenetic results. Diaporthe taoicola was morphologically characterized by producing Alpha conidia with tapering toward both ends. Meanwhile, D. siamensis produced cylindrical or ellipsoidal Alpha conidia with two oil drops. Pathogenicity tests revealed that both species were pathogenic to fruit of C. sinensis. To our knowledge, the two species were firstly reported on Citrus sinensis in China.

• The Plant Pathology Journal
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• v.34 no.6
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• pp.499-505
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• 2018

Huanglongbing (HLB, Citrus greening disease) is one of the most devastating diseases that threaten citrus production worldwide. Although HLB presents systemically, low titer and uneven distribution of these bacteria within infected plants can make reliable detection difficult. It was known loop-mediated isothermal amplification (LAMP) method has the advantages of being highly specific, rapid, efficient, and laborsaving for detection of plant pathogens. We developed a new LAMP method targeting gene contained tandem repeat for more rapid and sensitive detection of Candidatus Liberibacter asiaticus (CLas), putative causal agent of the citrus huanglongbing. This new LAMP method was 10 folds more sensitive than conventional PCR in detecting the HLB pathogen and similar to that of real-time PCR in visual detection assay by adding SYBR Green I to mixture and 1% agarose gel electrophoresis. Positive reactions were achieved in reaction temperature 57, 60 and $62^{\circ}C$ but not $65^{\circ}C$. Although this LAMP method was not more sensitive than real-time PCR, it does not require a thermocycler for amplification or agarose gel electrophoresis for resolution. Thus, we expect that this LAMP method shows strong promise as a reliable, rapid, and cost-effective method of detecting the CLas in citrus and can be applied for rapid diagnosis is needed.

• Research in Plant Disease
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• v.11 no.2
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• pp.115-121
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• 2005

This studies were carried out to elucidate the control effect of copper fungicides against some citrus diseases, assess copper spray injury on leaf and fruit of citrus by spraying of copper fungicides according to growth stages and spray seasons and by tank mixing with paraffin oil in field experiments for 3 years (2003-2005 season). As results, the control value was approximately $75\%$ against scab disease in all tested copper fungicides, $85-88\%$ to canker and $75-86\%$ to melanose. In case of leaf, the copper injury was the severest in early stage of shooting, and the injury was gradually weakened according to growth of leaf. In fruit, the copper injury was more severe in midsummer (late of July and August) than early of summer and spring season (June and ealy of July). Among copper fungicides, the bordeaux mixture which made by level of farm induced more injury compared to other copper fungicides commercialized. Less copper spray injury was observed on treatments in which copper fungicide was sprayed as tank mixtures with paraffin oil when compared to treatment which sprayed with only copper fungicide.

• 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

• Journal of Korean Clinical Health Science
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• v.2 no.1
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• pp.73-78
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• 2014

Purpose. Citrus aurantium L.(familyRutaceae), alsoknownasbitter orange, have been used as traditional herbal medicine in many Asian countries since ancient times. Hence, the purpose of the study was to briefly discuss the new findings about anti-inflammatory and anti-cancer activities of Citrus aurantium L in-vitro. Methods. The articles for this study were collected from pubmed and scopus electronic resources. Results. Citrus aurantium L contains an abundant Flavonoids, including hesperidin, naringin and nobiletin. These Flavonoids has reported to have various medicinal benefits that include antioxidant, antimicrobial, anti-inflammatory, anticancer, anti-diabetic activities, and also used to treat cardiovascular diseases. Conclusion. Based on the above evidence, we propose that Flavonoids from Korea Citrus aurantium L would be a therapeutic potential for cancer treatment and pharmacological benefit for inhibiting the inflammatory effect.

• 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.

• Horticultural Science & Technology
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• v.30 no.2
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• pp.214-219
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• 2012

This study aimed to develop a new type of functional citrus beverage (Citurs-F) containing flavonoids extracted from the young fruits of satsuma mandarin ($Citrus$ $unshiu$) and matured fruits of Jeju native dangyooja ($C.$ $grandis$). We made beverages that contained 30% of satsuma mandarin extract with different percentages of concentrated dangyooja-derived flavonoid extracts. In sensory evalution, the highest response indices of color, taste and aroma were from the beverages based on the 30% young fruit extracts plus 15% (Citrus-F-15) and 20% (Citrus-F-20) flavonoids extract from the dangyooja using the KILO prep. In the changes of body weight after oral administration of the Citrus-F, the rat group with HF diet plus the Citrus-F decreased the body weight compared to the rat group fed only HF diet. This effect was to be continued for 9 weeks until the end of experiment. In the lipid content in blood, the rat group with oral administration of citrus extractions merely tended to resolve it in serum test. However, all the 0.1% Citrus-F-15 and Citrus-F-20 treated rat groups from the beginning or after 5 weeks appeared the lowest lipid contents in the blood. In the cholesterol contents, the rat group feeding the KILO-prep's extraction from the beginning weren't significantly recognized them in the group but the rat group feeding 0.1% Citrus-F-15 acted to reduce in the cholesterol contents from 5 weeks. The results indicated that the Citrus-F-15 with rich flavonoids might be main source alleviating the vascular diseases and obesity in human diet.