Background : During 2016 to 2017, Bacterial Rot symptom has been observed on Atractylodes macrocephala Koidz. in Yeongju-si, Mungyeong-si, Jecheon-si and Eumseong-gun. This experiment was carried out to identify pathogenic bacteria that has not been reported up to now from A. macrocephala and to test pathogenicity of isolated bacteria against A. macrocephala. Methods and Results : Nine types of representative bacteria strains depending on colony size and color were isolated from surface disinfested symptomatic tissue that was macerated and streaked onto lysogeny broth (LB) medium with agar. Fungi were not recovered from any tissue that was surface disinfested and placed into acidified potato dextrose agar. Only one strain cause dark brown leaf rot symptom on A. macrocephala leaves soaked in bacterial suspensions. Potted A. macrocephala plants were used to test for pathogenicity. Inoculum was prepared by suspending the bacteria in sterile distilled water (SDW) for a final concentration of approximately 105 CFU/㎖. Suspensions were sprayed until runoff onto three replicate plants. Control plants were sprayed with SDW until runoff. Plants maintained in a dew chamber with 100% relative humidity at 30℃. After 3 days, leaf rot lesions developed on all inoculated plants; lesions later turned dark brown and appeared similar to symptoms observed in the field. Plants treated with water developed no symptoms. Same bacteria re-isolated onto LB from symptomatic tissues. Conclusion : On the basis of 16S rRNA sequence analysis, the strain isolated from A. macrocephala was identified as Pseudomonas viridiflava. Biological assay method using Potted plants confirmed the pathogenicity of Pseudomonas viridiflava. This is the first report of bacterial rot caused by Pseudomonas viridiflava on A. macrocephala.

Background : Ginseng root rot is a devastating disease caused by the fungus, Ilyonectria mors-panacis that generally attacks younger roots (-2 years), leading to defects in root quality, ginsenoside accumulation and also life cycle of the plant. Hence, there is an indispensable need to develop strategies resulting in tolerance against ginseng root rot. The protective role of silicon during pathogen infestation is well documented in other plant systems and a previous study demonstrated that silica nanoparticles are absorbed and accumulated more than the bulk silica in maize. However, the role of silica in ginseng-root rot pathosystem is unknown. Methods and Results : In the present study, we evaluated the effect of silica nanoparticles (N-SiO2) in Panax ginseng during I. mors-panacis infection. Long term analysis (30 dpi) revealed a striking 50% reduction in disease severity index upon 1 mM and 2 mM treatment of N-SiO2. However, N-SiO2 did not have any direct antifungal activity against I. mors-panacis. Candidate genes and metabolites based approach revealed jasmonic acid (JA) mediated sterol accumulation and incresed ginsenside biosyntesis as the key transcriptional reprogramming events orchestrated by N-SiO2 during the fungal infection. Conclusion : In a nut shell, N-SiO2 administration induces transcriptional reprogramming in ginseng roots, leading to increased phytosterol and ginsenosides synthesis resulting in enhanced tolerance against I. mors-panacis.

Background : Replant failure of ginseng is caused by soil-born pathogens causing root rot such as Cylindrocarpon destructans and Fusarium solani. Dazomet are widely used as soil fumigant to solve soilborne problems, and the degradation intermediates are toxic to nematodes, fungi, bacteria, insects and weeds. Methods and Results : The effects of cultivation of green manure crop, maize before and after soil fumigation on the control of ginseng root rot disease were compared using soil of field where 6-years-old ginseng was harvested. Fumigant (dazomet) were used for soil fumigation in May and September, respectively. Maize was grown for soil management before and after soil fumigation. Maize cultivation after May fumigation was delayed the sowing day by 15 days, and the fresh and dry weight decreased significantly. Maize cultivation after May fumigation increased pH but decreased EC, NO3, P2O5, and K significantly. Maize cultivation after May fumigation decreased fungi population and ratio of fungi and bacteria. Growth of 2-years-old ginseng was improved and the incidence of ginseng root rot was significantly decreased by maize cultivation after May fumigation. After harvesting 2-years-old ginseng, the population of Cylindrocarpon destructans was not different among treatments, but Fusarium solani showed a significant increase in September fumigation after maize cultivation. Conclusion : Maize cultivation after soil fumigation was effective in inhibiting ginseng root rot by improvement of mineral composition and microorganism in soils.

Pectobacterium carotovorum subsp. carotovorum was found to be highly virulent to various vegetables, including Chinese cabbage. The antibacterial isolate CAB12243-2 was tested in a field bioassay for suppressing soft rot disease. The nucleotide sequencing of the 16S rRNA gene identified, the CAB12243-2 strain used in this study as Bacillus toyonensis. B. toyonensis CAB12243-2 inhibited the pectate lyase process by soft rot pathogens, and used trehalose and glucose as carbon sources. In field tests, the antibacterial isolate B. toyonensis CAB12243-2 suppressed soft rot disease with 73.0% control efficacy on the spring cultivar “Norangbom” and with 68.9% efficacy on the fall cultivar “Bulam 3”. These results suggest that B. toyonensis CAB12243-2 can be used as a biological control agent for the control of soft rot diseases on vegetables.

Background: To investigate the possibility of using Asarum sieboldii as an environment-friendly fumigant for protecting organic cultural heritages, the inhibitory effect of A. sieboldii extract against wood rot fungi on Hanji was examined. Methods and Results: The physical, optical, and morphological properties of Hanji inoculated with Trametes versicolor and Tyromyces palustris, and exposed to the n-hexane fraction of A. sieboldii extract, were measured. The physical properties were expressed as weight loss, zero-span tensile strength and viscosity and the optical properties were depicted by luminance and chromaticity (L*, a*, and b*). The results showed that, the n-hexane fraction of A. sieboldii extract inhibited the growth of fungi on Hanji, and preserved its condition. At a concentration of 25 ㎎, the n-hexane fraction of A. sieboldii extract maintained zero-span tensile strength, increased viscosity, and restricted discoloration of Hanji. It also was confirmed that the weight of fungi infested Hanji exposed to the extract did not decrease. Scanning electron microscopic images revealed that the spores and hyphae of T. versicolor and T. palustris were not present on Hanji during treatment with > 25 ㎎ of the n-hexane fraction of A. sieboldii extract. Conclusions: These results indicate that the n-hexane fraction of A. sieboldii extract by virtue of its antifungal effectiveness may help in preserving Korean paper cultural heritages, including Hanji.

Background: The application of crop rotation systems may reduce the occurrence of soil-borne diseases by releasing allelochemicals and by subsequent microbial decomposition. Methods and Results: For reduction of ginseng root rot by the crop rotation system, after harvesting 6-year-old ginseng, fresh ginseng was grown along with continuous cultivation of sweet potato, peanut, and bellflower. Growth of 2-year-old ginseng was significantly inhibited in the continuous cultivation than in the first cultivation. Sweet potato, peanut and bellflower cultivations assisted in obtaining normal yields of ginseng in the first year after the harvest of 6-year-old ginseng. Salt concentration, potassium and sodium contents were gradually decreased, and, organic matter was gradually increased through cirp rotation. Phosphate, calcium and magnesium contents were not altered. The density of the root rot fungus was gradually decreased by the increase in crop rotation; however it was decreased distinctly in the first year compared to the second and third year. The severity of root rot disease tended to decrease gradually by the increase of crop rotation. Conclusions: Short-term crop rotation for three years promoted the growth of ginseng, however root rot infection was not inhibited significantly, although it was somewhat effective in lowering the density of the root rot pathogen.

Spent coffee grounds, activated sludge, chicken manure, and agricultural waste-derived biochar were used to manufacture eco-friendly and functional compost via the bioaugmentation of white rot fungus and plant growth that promotes beneficial microorganisms. Six lab-scale composting reactors were established to perform composting. After composting was completed over 45 days, the composts were analyzed for major elements, physico-chemical characteristics, compost maturity, and compost effectiveness on crop growth and quality. Concentrations of T-N and PO4 3− significantly increased in the composts that had been amended with biochar and/or white rot fungus compared to the control, while those of NO3 −-N, TOC and TOC/T-N had significantly decreased, indicating the occurrence of effective composting. Besides, the germination indices of these composts were also generally higher than the control by 10-34%, indicating that the composts were mature. The four composts amended with biochar and/or white rot fungus (TR-3, TR- 4, TR-5, TR-6) also appeared to stimulate more growth in lettuce compared to commercial organic fertilizers (by 36- 104%). Besides, composts TR-3 and TR-4 respectively enhanced DPPH scavenging activity in lettuce leaves by 58% and 49%, while TR-4 and TR-5 respectively enhanced the total phenolic content (TPC) by 44% and 37%. This implies that the amendment of biochar and the bioaugmentation of white rot fungus could facilitate the composting process for the production of quality functional compost that is able to enhance the antioxidant content in crops. Quality composts could better compete with the commercially available fertilizers in the market, leading to the eco-friendly recycling of organic wastes such as spent coffee grounds, sludge, chicken manure, and agricultural waste.

Background : Ginseng (Panax ginseng C.A. Meyer) is one of the most important medicinal plants in Korea, but its yields are often reduced by a variety of root pathogens. The root rot of ginseng is a destructive soil-borne disease caused by Cylindrocarpon destructans (teleomorph: Ilyonectria radicicola). Methods and Results : Ilyonectria radicicola and its Cylindrocarpon-like anamorph represent a species complex that is commonly associated with root rot disease symptoms on virulence. During the course of this study, several species could be distinguished from I. radicicola sensu stricto based on morphological and culture characteristics. As results of investigated culture characteristics, optimal temperature for mycelial growth of isolates were 20℃, and colony pattern and color were slightly different on PDA. Isolates of I. radicicola were analyzed for their genetic relatedness based on several genes and microsatellite region. I. radicicola group was divided into two small groups. Conclusion : Therefore, we were able to confirm pathogenicity and genetic difference between the isolates in each of the groups of the pathogen. Among these isolates, 21.5% were classified as highly virulent and 78.5% were weakly virulent. *(Corresponding

Background : Korean ginseng is an important cash crop in Asian countries. However, plant yield is reduced by pathogens. Among the Ilyonectria radicicola-species complex, I. mors-panacis is responsible for root-rot and replant failure of ginseng in Asia. It is essential to find a way to reveal the existence of the pathogen before starting cultivation. Therefore, qRT-PCR method is developed to detect and quantify the pathogen in ginseng soils. Methods and Results : In this study, species specific Histone H3 primer set is developed for the quantification of I. mors-panacis. The primer set was applied on DNA of other microbes to evaluate its sensitivity and selectivity on I. mors-panacis DNA. Sterilized soil samples artificially infected by the pathogen in different concentrations were used to evaluate the ability of the primer set to detect the pathogen population in the soil DNA. Finally, the pathogen was quantified in many natural soil samples. The designed primer set was found to be sensitive and selective to I. mors-panacis DNA. In artificially infected sterilized soil samples, the estimated template using qRT-PCR was positively correlated with the pathogen concentration in soil samples (R2=0.94), disease severity index (R2=0.99), and colony forming unit (R2=0.87). In the natural soils, the pathogen was recorded in the most of fields produce bad yields with the range of 5.82 ± 2.35 to 892.34 ± 103.70 pg/g of soil. Conclusion : According to the presented results, the proposed primer set is applicable for estimating soils quality before ginseng cultivation. This will help in the disease management and crop protection in the future.

Background : The fungus Cylindrocarpon destructans (Zins) Scholten is the cause of root rot in many ginseng production areas. Root exudate composition and quantity vary in relation to plant nutritional status, but the impact of the differences on rhizosphere microbial communities is not known. Methods and Results : Five kinds of rotation crops, sudan grass soybean peanut sweet potato, perilla were grown for one year in ginseng garden harvested 6-year-old ginseng. The ratio of gram-negative bacteria, fungi, bacteria, total microbial biomass, aerobic/anaerobic microbes were increased by rotational crop cultivation, while the ratio of actinomycetes and the ratio of saturated to unsaturated fatty acids were decreased. The increase in the fungal density or the increase in the proportion of fungi to the bacteria tended to increase the incidence of root rot, but there was no significant difference. The yield of ginseng root showed a highly significant negative correlation with actinomycetes. The correlation between the soil chemical properties and the incidence of root rot was analyzed by cultivating 23 kinds of green manure crops for one year in field where cultivated ginseng continuously. The survival rate of ginseng showed a highly significant positive correlation with soil acidity and a highly significant negative correlation with nitrate nitrogen, and a significant negative correlation with soil salt concentration. Conclusion : Rotation crops improved soil microbial communities, lowered the rate of fungi and increased the proportion of bacteria, the survival rate of ginseng was significantly correlated with soil acidity, nitrate nitrogen and soil salinity.

Background: Some plants have harmful effects on fungi and bacteria as well as plants. Incorporating into soil as green manures are effective in reducing population densities of soil pathogens. Methods and Results: Twenty-three species of green manure crops were cultivated after the harvest of 6-year-old ginseng and then incorporated green manure into the soil at the flowering stage. The following year, the root rot ratio of 2-year-old ginseng and soil chemical properties were investigated. Sorghum sudanense, Helianthus annuus, and Helianthus tuberosus were relatively high in dry matter production. Without incorporating green manure into soil, NO3, EC (electric conductivity) and K were decreased by 95%, 79% and 65%, respectively. When green manure was incorporated to soil, P2O5 and NO3, were reduced by 41% and 25%, respectively. The survived root ratio of 2-year-old ginseng were significantly increased by 56.2%, 47.5%, and 47.3% in Sorghum sudanense, Ricinus communis and Helianthus tuberosus, respectively, In addition, there was a significant increase in Secale cereale, Chrysanthemum morifolium, Atractylodes macrocephala, and Smallanthus sonchifolius. The survived root ratio of ginseng showed a significant positive correlation with soil pH and a negative correlation with NO3, and EC. Conclusion: Cultivation of Chrysanthemum family mainly using rhizome and root as green manure was effective for root rot disease of ginseng.

Background: Some plants have harmful effects on fungi and bacteria as well as other plants. Incorporating such plant into soil as green manure is effective in reducing population densities of soil pathogens. Methods and Results: Twenty-three species of green manure crops were cultivated after the harvest of 6-year-old ginseng and then incorporated into the soil at the flowering stage. The following year, the root rot ratio of 2-year-old ginseng and soil chemical properties were investigated. In the absence of green manure addition, the NO3 content, electric conductivity (EC), and K content decreased by 95%, 79% and 65%, respectively. In the presence of green manure addition, P2O5 and NO3 contents reduced by 41% and 25%, respectively. The “survived root ratio” of 2-year-old ginseng significantly increased by 56.2%, 47.5%, and 47.3%, in the Sorghum sudanense, Ricinus communis and Helianthus tuberosus treatment, respectively. In addition, there was a significant increase in the “survived root ratio” in the Secale cereale, Chrysanthemum morifolium, Atractylodes macrocephala, and Smallanthus sonchifolius treatments. The “survived root ratio” of ginseng showed a significant positive correlation with the soil pH and a negative correlation with the NO3 contents, and EC. Conclusions: Cultivation of plant form the Chrysanthemum family as green manure, using mainly the rhizomes was effective for the control of root rot disease of ginseng.

Background : Ginseng is a perennial, and damage occurs due to various diseases and pests. As a result, yield decreases and quality deteriorates. In particular, it is impossible to be repeatedly cultivated due to root rot, soil-borne disease. For this, in order to control root rot and repeatedly cultivate ginseng by using soil fumigation from virgin soil, this experiment was conducted. Methods and Results : This experiment was performed in ginseng farm field, Geumsan-gun, Chungnam province. And total area was 3ha. And the experiment was conducted in soil where apple trees had been grown. Apple trees were rooted up in 2006. And soil was fumigated in May 2007. With regard to soil fumigant, soil was treated with Basamid of 40kg/10a. After soil fumigation, 5 ton/10a of cattle manure fermented for 1 year was put in the soil in June, and then the soil was cared for by being plowed and rotary-tilled as occasion. Ginseng seeds were sowed in the soil cared for in October 2007. And 4-year-old ginseng was harvested in 2011. The soil from which ginseng was harvested was fumigated by the same method in May 2012, and then ginseng seeds were sowed in 1.5ha in October 2012 and in 1.5ha in October 2013. In October 2013, 3-year-old and 4-year-old ginseng was harvested. And ginseng growth characteristics and root rot incidence was examined. It was shown that 4-year-old ginseng yield after the fumigation of virgin soil was nearly 2 times as high as that of conventional virgin soil cultivation. And root rot didn't occur. After that time, the soil was fumigated again. And when ginseng was cultivated, 3-year-old and 4-year-old ginseng yield was 650kg and 960kg per 10a. And it was shown that root rot incidence was 1.3% and 15.3% in 3-year-old and 4-year-old ginseng respectively. Conclusion : This study showed the results where ginseng can be cultivated repeatedly if soil was fumigated again after ginseng was harvested following the soil fumigation from virgin soil so as to control the ginseng root rot and cultivate ginseng repeatedly.

Background : Ginseng (Panax ginseng C.A. Meyer) is one of the most important medicinal plants in Korea, but its yields are often reduced by a variety of root pathogens. The root rot of ginseng is a destructive soil-borne disease caused by Cylindrocarpon destructans (teleomorph: Ilyonectria radicicola). To monitor contamination with C. destructans in ginseng harvested in 2015 were sampled from 57 different growing fields. The spore number of C. destructans was quantified by use of a specific primers and selective media (radicicol) in soils of ginseng fields. Methods and Results : The ginseng samples were surface-sterilized and placed on potato dextrose agar plates for 7 day incubation at 20℃. Emerging fungal colonies were counted primarily based on colony and conidia morphology. Further species level identification was confirmed by ITS rDNA sequencing. For quantification of the soil-borne C. destructans, the genomic DNA was extracted from the soil using a NucleoSpin soil kit (MN, Germany). Density of C. destructans was determined by species specific real time PCR (qPCR). The qPCR was completed by running a melting curve analysis. Conclusion : The C. destructans associated with root rot disease of ginseng were detected in more than 60% in pyeongtaek-1, pochenon-1, jecheon-1, chungju-1 and jinan-4. As results of the study, the correlation between pathogen density and identification clearly clarified in the soil.