• Journal of Ginseng Research
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• v.25 no.3
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• pp.136-140
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• 2001

Effects of incubation temperature and pH on chlamydospore germination of Cylindrocarpon destrcutans (isolate CY-9802) causing root rot of Panax ginseng were studied. Germination rate of the chlamydospores on Czapek solution agar(CSA) was higher than on potato dextrose agar(PDA) at the incubation temperatures tested. The chlamydospores were able to be germinated at range of 5$\^{C}$ to 30$\^{C}$ after 48 hours incubation on CSA. Germination rate was 53.2∼6.27% at range of 15$\^{C}$ to 25$\^{C}$, and the optimum temperature was 20$\^{C}$, whereas they were very low at 30$\^{C}$ on PDA. Germination rate was 43.6% to 47.9% at range of 10$\^{C}$ to 20$\^{C}$, and the optimum temperature was 20$\^{C}$ as well. They were able to be germinated at pH of 5.2 to 8.1 on CSA and 5.2 to 7.2 on PDA. Optimum pHs for the germination on CSA and PDA were from 6.4 to 8.2 and from 5.2 to 6.0, respectively. Mycelial color of the fungus on CSA was pale brown at pH from 5.2 to 6.0 and white from pH 6.4 to 8.1, while it was typical dark brown ar range of pH 5.2 to 7.1 and brown at pH 7.2 on PDA after 21 days incubation.

• The Korean Journal of Mycology
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• v.39 no.1
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• pp.16-21
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• 2011

Ginseng root rot caused by Cylindrocarpon destructans is one of the most destructive diseases of ginseng(Panax ginseng). We analyzed the features of the species through pathogenicity test and genetic diversity analysis of C. destructans in Korea, for its application as basic data to attempt for effective control. C. destructans isolated from rotted ginseng roots exhibited a variety of colonial colors on media. It was assumed that there may exist genetic diversity in the population by the diversity of pathogenicity among isolates observed when artificially inoculated into ginseng roots. Pathogenicity tests using ex vivo wound inoculation with agar mixture inoculation on ginseng roots were performed similar results as were observed appear to be useful for rapid pathogen inspection. According to RAPD analysis results, Korean C. destructans isolates formed a single genetic group which can be distinguished readily from closely related other fungi. C. destructans group was divided into two small groups. Therefore, we were able to confirm pathogenicity and genetic difference between the isolates in each of the groups of the pathogen.

• Korean Journal of Plant Resources
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• v.16 no.1
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• pp.40-48
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• 2003

Cylindrocarpon destructans is the major pathogen inducing the root rot disease in ginseng. Up to now, there is no reliable and convenient method to analyze the spore density or population of this pathogen in ginseng-growing soil or any contaminated farmlands. Therefore, it will be very valuable to develop a new and reliable method in detecting the spore of this pathogen. In this study, a molecular biological technique using two step nested PCR method, was developed. Two universal ITS primers, ITS5F and ITS4R were used in the first round of PCR to amplify a fragment of ITS region from the genomic DNA of C. destructans. The specific prmers Nest 1 and Nest 2 were designed and used in the second round of PCR to amplify a inner fragment from the first round PCR product of C. destructans. C. destructans spore, only soil samples from the diseased ginseng farm produced the positive bands, suggesting its usefulness in detecting the C. destructans spores in soil samples. Thus it is recommended to first extract the whole genomic DNA from soil samples and use it for the PCR reaction, thereby eliminating the inhibitory activity of soil components.

• The Plant Pathology Journal
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• v.27 no.4
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• pp.333-341
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• 2011

Two antifungal bacteria were selected from forest soils during the screening of microorganisms antagonistic to Cylindrocarpon destructans, a cause of ginseng root rot. The antifungal bacteria were identified as Bacillus subtilis (I4) and B. amyloliquefaciens (yD16) based on physiological and cultural characteristics, the Biolog program, and 16S rRNA gene sequencing analyses. Antagonistic activity of both bacterial isolates to C. destructans increased with increasing temperature. More rapid starch hydrolytic activity of the bacteria was seen on starch agar at higher temperatures than at lower temperatures, and in the higher density inoculum treatment than in the lower density inoculum treatment. The bacterial isolates failed to colonize ginseng root the root tissues inoculated with the bacteria alone at an inoculum density of $1{\times}10^6$ cfu/ml, but succeeded in colonizing the root tissues co-inoculated with the bacteria and C. destructans. Scanning electron microscopy showed that the pathogen was damaged by the low-density inoculum treatment with the bacterial isolates as much as by the high-density inoculum treatment. Both bacterial isolates were more effective in reducing root rot when they were treated at a concentration of $1{\times}10^6$ cfu/ml than at $1{\times}10^8$ cfu/ml. Also, only the former treatment induced prominent wound periderm formation, related to structural defense against pathogen infection. The results suggest that the bacterial antagonists may have high potential as biocontrol agents against ginseng root rot at relatively low-inoculum concentrations.

• The Plant Pathology Journal
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• v.25 no.1
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• pp.1-5
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• 2009

Root rot caused by Cylindrocarpon destructans is one of the most important diseases of ginseng (Panax ginseng C. A. Meyer). Two types of symptoms found in ginseng root rot are black root rot and rusty root (rusty spots), in which disease severities are high and low, respectively. Symptom development and related histopathological changes were examined in an inoculation test on 2-year-old ginseng roots using virulent (Cy9801) and avirulent (Cy0001) isolates of C. destructans under different temperature conditions (13, 18, 23, and $28^{\circ}C$). Black root rot was only induced by Cy9801 in the lower temperature range (13, 18, and $23^{\circ}C$) and not at the higher temperature ($28^{\circ}C$). No black root rot, but only rusty spot symptoms, were induced by Cy0001 at all temperatures tested except $13^{\circ}C$, at which no symptoms occurred on over half of inoculation sites, suggesting disease development was influenced by pathogen virulence and temperature. Wound periderms were formed in all root tissues with rust spot symptoms at $28^{\circ}C$ caused by Cy9801 and at 18, 23, and $28^{\circ}C$ temperatures caused by Cy0001. No wound periderm was formed at $13^{\circ}C$ by either Cy9801 or Cy0001. Light microscopy revealed that the wound periderm was formed by initial cell divisions in cell wall formation and/or additional cell wall layering in parenchyma cells without obvious nuclear division, followed by layering of the divided cells adjacent to the inoculation sites, blocking the spread of the rot. These results suggest that disease development declined at lower temperatures and by the formation of a wound periderm at higher temperatures, and that ginseng rusty root may develop under conditions unfavorable for further disease development of C. destructans.

• Mycobiology
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• v.42 no.2
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• pp.174-180
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• 2014

We analyzed the genetic diversity of Cylindrocarpon destructans isolates obtained from Korean ginseng (i.e., Panax ginseng) roots by performing virulence tests and nuclear ribosomal gene internal transcribed spacer (ITS) and mitochondrial small subunit (mt SSU) rDNA sequence analysis. The phylogenetic relationship analysis performed using ITS DNA sequences and isolates from other hosts helped confirm that all the Korean C. destructans isolates belonged to Nectria/Neonectria radicicola complex. The results of in vivo and ex vivo virulence tests showed that the C. destructans isolates could be divided into two groups according to their distinctive difference in virulence and the genetic diversity. The highly virulent Korean isolates in pathogenicity group II (PG II), together with foreign isolates from P. ginseng and P. quinquefolius, formed a single group. The weakly virulent isolates in pathogenicity group I, together with the foreign isolates from other host plants, formed another group and exhibited a greater genetic diversity than the isolates of PG II, as confirmed by the mt SSU rDNA sequence analysis. In addition, as the weakly virulent Korean isolates were genetically very similar to the foreign isolates from other hosts, they were likely to originate from hosts other than the ginseng plants.

• Journal of Ginseng Research
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• v.20 no.1
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• pp.88-95
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• 1996

The effects of media, incubation time, temperature and pH on production of conidia and chlamydospore of Cylindrocarpon destructans (Zinssm.) Scholen causing root rot of Panax ginseng were studied. Microconidia of the pathogen were abundantly produced on V-8 juice agar as a solid substrate with 5.73(log conidia/mm2) and in V-8 broth as a liquid substrate with 6.65 (log conidia/ml) among media tested. No difference was observed on the length of microconidia produced from the media with a range of 9.50∼11.38 $\mu\textrm{m}$. However, tryptic soy agar produced the broadest microconidia (average 5.00 $\mu\textrm{m}$) among the media tested. All the media produced chlamydospores In a range of 1.06∼4.37 (log chlamydospores/mm2) without a significant difference in number, while V-8 juice agar produced the bigger one (18.39 $\mu\textrm{m}$ in diameter) as compared to the tested media. The fungus began to sporulate conidia after three days of incubation and reached maximum at the 8th day. It seemed to be in a stationary phase until 30 days of incubation but was decreased thereafter. Chlamydospore was produced at 4th day after incubation. Maximum production was observed at 8th day and the number seemed to be maintained during the observation period. Both conidia and chlamydospore of the pathogen were able to be spoluated at 10∼25$^{\circ}C$. However, optimum temperatures of conidia and chlamydospore formation were 15∼25$^{\circ}C$ and 10∼20$^{\circ}C$, respectively. C. destrmtans produced conida with an wide range of pH from 3.3 to 8.0 and chlamydospore from 2.8 to 8.0. Number of conidia was increased with an increase of pH up to 4.0. There was no significant difference in the number between 4.0 to 8.0. It seemed to have two optimum pH ranges, 3.3∼4.0 and 7.1∼8.0 for the chlamydospore formation.

• Journal of Ginseng Research
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• v.42 no.1
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• pp.9-15
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• 2018

Cylindrocarpon destructans/Ilyonectria radicicola is thought to cause both rusty symptom and root-rot disease of American and Korean ginseng. Root-rot disease poses a more serious threat to ginseng roots than rusty symptoms, which we argue result from the plant defense response to pathogen attack. Therefore, strains causing rotten root are characterized as more aggressive than strains causing rusty symptoms. In this review, we state 1- the molecular evidence indicating that the root-rot causing strains are genetically distinct considering them as a separate species of Ilyonectria, namely I. mors-panacis and 2- the physiological and biochemical differences between the weakly and highly aggressive species as well as those between rusty and rotten ginseng plants. Eventually, we postulated that rusty symptom occurs on ginseng roots due to incompatible interactions with the weakly aggressive species of Ilyonectria, by the established iron-phenolic compound complexes while root-rot is developed by I. morspanacis infection due to the production of high quantities of hydrolytic and oxidative fungal enzymes which destroy the plant defensive barriers, in parallel with the pathogen growth stimulation by utilizing the available iron. Furthermore, we highlight future areas for study that will help elucidate the complete mechanism of root-rot disease development.

• Journal of Ginseng Research
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• v.44 no.1
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• pp.161-167
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• 2020

Background: The ascomycete fungi Cylindrocarpon destructans (Cd) and Fusarium solani (Fs) cause ginseng root rot and significantly reduce the quality and yield of ginseng. Cd produces the secondary metabolite radicicol, which targets the molecular chaperone Hsp90. Fs is resistant to radicicol, whereas other fungal genera associated with ginseng disease are sensitive to it. Radicicol resistance mechanisms have not yet been elucidated. Methods: Transcriptome analyses of Fs and Cd mycelia treated with or without radicicol were conducted using RNA-seq. All of the differentially expressed genes (DEGs) were functionally annotated using the Fusarium graminearum transcript database. In addition, deletions of two transporter genes identified by RNA-seq were created to confirm their contributions to radicicol resistance. Results: Treatment with radicicol resulted in upregulation of chitin synthase and cell wall integrity genes in Fs and upregulation of nicotinamide adenine dinucleotide dehydrogenase and sugar transporter genes in Cd. Genes encoding an ATP-binding cassette transporter, an aflatoxin efflux pump, ammonium permease 1 (mep1), and nitrilase were differentially expressed in both Fs and Cd. Among these four genes, only the ABC transporter was upregulated in both Fs and Cd. The aflatoxin efflux pump and mep1 were upregulated in Cd, but downregulated in Fs, whereas nitrilase was downregulated in both Fs and Cd. Conclusion: The transcriptome analyses suggested radicicol resistance pathways, and deletions of the transporter genes indicated that they contribute to radicicol resistance.

• Research in Plant Disease
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• v.28 no.4
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• pp.204-208
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• 2022

Hydrogen peroxide is an eco-friendly oxidizing agent, which has exhibited a broad spectrum of antimicrobial activity without adverse environmental impact. This study was conducted to investigate the antifungal effect of hydrogen peroxide treatment against Cylindrocarpon destructans, and consequently to evaluate its control efficacy against root rot disease of 2-year-old ginseng plants. Hydrogen peroxide treatment strongly inhibited the viability of C. destructans conidia in vitro. The hydrogen peroxide at a concentration of 300 mg/l significantly reduced disease infection of the ginseng root when treated to spore suspension (10⁷ conidia/ml). Spraying with 300 mg/l of hydrogen peroxide reduced the root rot disease of the ginseng sprouts by 15% compared to the untreated control at 14 days after the inoculation. However, 300 mg/l of hydrogen peroxide delayed the emergence of ginseng plants during sprouting under aeroponic conditions. Further works need to be done to provide an acceptable control efficacy of hydrogen peroxide against the disease and its good safety to ginseng plants.