• Korean Journal Plant Pathology
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• v.14 no.5
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• pp.393-396
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• 1998

The characteristic Ramularia type conidia of Mycosphaerella nawae were formed on naturally infected leaves. Artificial inoculation with the conidia induced typical symptom on leaves, which was not distinguishable from those of ascospore infection, which has been considered as a primary inoculum source in nature. Also the morphology of the conidia produced on PDA was not different from those formed on artificially inoculated leaves or on naturally infected leaves at later stage of symptom development. Accordingly, we report the role of the conidia as a secondary inoculum of the circular leaf spot pathogen of persimmon for the first time.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.6
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• pp.528-534
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• 2005

Hairy root cultures have demonstrated great promise in terms of their biosynthetic capability toward the production of secondary metabolites, but continue to constitute a major challenge with regard to large-scale cultures. In order to assess the possibility of conducting mass production of biomass, and the extraction of useful metabolites from Panax ginseng. P. ginseng hairy roots, transformed by Rhizobium rhizogenes KCTC 2744, were used in bioreactors of different types and sizes. The most effective mass production of hairy roots was achieved in several differently Sized air bubble bioreactors compared to all other bioreactor types. Hairy root growth was enhanced by aeration, and the production increased with increasing aeration rate in a 1 L bioreactor culture. It was determined that the hairy root growth rate could be substantially enhanced by increases in the aeration rate upto 0.5vvm, but at aeration rates above 0.5vvm, only slight promotions in growth rates were observed. In 20 L air bubble bioreactors, with a variety of inoculum sizes, the hairy roots exhibited the most robust growth rates with an inoculum size of 0.1% (w/v), within the range 0.1 to 0.7% (w/v). The specific growth rates of the hairy root decreased with increases in the inoculum size.

• Research in Plant Disease
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• v.10 no.4
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• pp.209-216
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• 2004

The circular leaf spot of persimmon is occurred almost every place where persimmon is cultivated, especially the disease outbreak severely in southern part of Korea. The disease reveals unusually long incubation period after pathogen invade into leaf tissue and no practical control measure is available once the symptom has appeared. Most of the farmers just follow the suggested spray schedules calculated on the basis of weather condition of ordinary years. Therefore the damages due to circular leaf spot greatly differ year after year. In this article, we tried to describe and summarized the investigation on the circular leaf spot pathogen, Mycosphaerella nawae, related to disease outbreak such as overwintering of pathogen, inoculum formation and spread, incubation period after infection, and secondary inoculum. With the summary of these results, we suggest the disease cycle of circular leaf spot of persimmon. The pathogen overwinters in diseased leaves as mycelial form or pseudoperithecial premodium. The pseudoperitheria become matured in spring as the temperature raise and forms asci and ascospores. The maturation of pseudoperithecia are closely related to the temperatures during March and early April. The ascospores completely mature in early May and the ascospores released when the pseudoperithecia absorbed enough moisture after rainfall. The release of ascospores are diverse greatly with the variation of maturity of pseudoperithecia. Generally the spore start to release from middle of May to early of July. Duration of ascospore release is depend on the weather condition of particular year, especially amount and number of precipitation. The ascospores produced from pseudoperithecia is known to the only inoculum for circular leaf spot disease. But according to the results obtained from our investigations, the conidia formed on the lesions which incited by natural infection. This conidia are infectious to persimmon leaves and formed identical symptom as natural infection. The time of producing secondary inoculum of circular leaf spot of persimmon is considered too late to develop new disease. Generally the importance of secondary inoculum is low but the conidia produced in early September are competent to develop new disease and new infection also significantly affect to harvest of persimmon. The importance of circular leaf spot disease is recognized well to farmers. The approaches to control of the disease should be initiated on the basis of the knowledges of inoculum dynamics and ecology of disease development. The forecasting system for circular leaf spot is need to be developed.

• Korean Journal Plant Pathology
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• v.14 no.5
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• pp.408-412
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• 1998

In order to illustrate the role of conidia of Mycosphaerella nawae as a secondary inoculum in nature, pseudothecial development on persimmon leaves was investigated microscopically. The fungal ascospores have been believed as the primary or only inoculum source in nature, however, pseudothecia were readily formed on persimmon leaves infected naturally and artificially by conidia. The pseudothecia of M. nawae were found to form in the tissues of infected leaves while the leaves were still hanging on the trees. The size of pseudothecia were approximately 51.0~122.4$\times$51.0~112.2 ${\mu}{\textrm}{m}$ (82.8 $\times$72.5 ${\mu}{\textrm}{m}$in average), the shapes were spherical, ovoid or occidental pear type. The sizes of asci were approximately 30.6~61.2$\times$8.2~10.2 ${\mu}{\textrm}{m}$(46.6$\times$9.4 ${\mu}{\textrm}{m}$ in average) and the shapes were cylinder or banana. The ascospores were mostly spindle type, and the sizes were 10.2~12.2$\times$3.1~4.1 ${\mu}{\textrm}{m}$ (11.4$\times$3.2 ${\mu}{\textrm}{m}$ in average)-like. The pseudothecial formation was initiated before defoliation and morphological characteristics of the pseudothecia, ascus and ascospores on the infected leaves were fully illustrated in this study. Results indicated that conidia of M. nawae induce circular leaf spot of persimmon as much as ascospores, and might play an important role of the disease epidemics in nature.

• Korean Journal Plant Pathology
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• v.14 no.5
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• pp.397-401
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• 1998

Asexual spores of Mycosphaerella nawae were profusely produced on PDA after a prolonged incubation at $25^{\circ}C$ for 90 days. When persimmon trees were artificially inoculated by the conidial suspension, typical symptoms of circular leaf spot of persimmon appeared on the leaves two month later. The imperfect stag of the fungus was identified as Ramularia sp. based on following morphological characteristics examined under a light microscope and a scanning electron microscope. Conidia were mostly ellipsoid, but occasionally cylindrical, elongated oval, taro, peanut or gourd shapes and measured as 12.2~32.6$\times$6.1~10.2 ${\mu}{\textrm}{m}$. erect, hyaline, colorless-light brown. Conidia were formed solitarily or in chains on a medium and infected leaves. Conidiophore was erect, hyaline, colorless-light brown. and the size was 20.4~102.0$\times$3.1~10.2 ${\mu}{\textrm}{m}$, respectively. In this paper, we firstly demonstratrated that asexual spores of M. nawae induced persimmon circular leaf spot in nature as well as sexual spores of the fungus. Therefore, it is hypothesized that the imperfect stage of the fungus plays an important role in nature for epidemics as secondary inoculum.

• Research in Plant Disease
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• v.7 no.2
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• pp.107-111
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• 2001

Deterioration of pod and seed quality by pod and stem blight and purple blotch is a serious problem in the production of vegetable soybean. Major inoculum sources for the causal pathogens, Phomopsis spp. and Cercospora kikuchii, are soybean straw and debris. Phomopsis spp. have been known to be either limited for hyphal growth or latent in immature soybean tissues. In this experiment, cultural practices using these ecological traits of the pathogens were applied as a control measure. In plastic mulching, seed infection was remark ably reduced by drip irrigation as compared to overhead sprinkling, but not reduced in no mulching cultivation. Control value of plastic mulching was 28.0% for pod and stem blight and 29.3% for purple blotch, which was lower than that of benomyl application. By the first week in June, 78% of overwintered soybean straw examined contained matured alpha spores in pycnidia, acting as primary inoculum. Secondary inoculum of phomopsix spp. was observed abundantly throughout from mid June to September on fallen cotyledons from current crops and subsequently was found on petiole debris. Consequently, both plastic mulching and drip irrigation were effective to some extent for controlling pod and stem blight and purple blotch without significant reduction in soybean yield, and the cultural practices in combination with field sanitation resulted in removing more the secondary inoculum.

• 한국생물공학회:학술대회논문집
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• 2005.10a
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• pp.417-420
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• 2005

Plants generally produce secondary metabolites in nature as a defense mechanism against pathogenic and insect attack. In this study, we applied several abiotic elicitors in order to enhance growth and ginseng saponin biosynthesis in the hairy roots of P. ginseng. Generally, elicitor treatments were found to inhibit the growth of the hairy roots, although simultaneously enhancing ginseng saponin biosynthesis. The addition of selenium at inoculum time did not significantly affect ginseng saponin biosynthesis. However, when 0.5 mM selenium was added as an elicitor after 21 days of culture, ginseng saponin content and productivity increased to about 1.31 and 1.33 times control levels, respectively. These results suggest that processing time for the generation of ginseng saponin in a hairy root culture can be reduced via the application of an elicitor.

• Journal of Industrial Technology
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• v.21 no.A
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• pp.343-349
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• 2001

This study characterizes the growth of white rot fungi Phanerochaete chrysosporium IFO 31249) and lignin peroxidase(LiP) activity in different submerged culture media. P. chrysosporium was grown in the form of pellet of various sizes from a spore inoculum under shaking liquid culture condition. While the growth of mycelia was higher under the nitrogen-sufficient culture than under the nitrogen-limited culture, ligninase activity was relatively lower. The lignin peroxidase appeared in nitrogen-limited culture and was suppressed by excess nitrogen. High level(40U/l) of lignin peroxidase activity was obtained in the growth medium containing 1.5mM veratryl alcohol, a secondary metabolite of P. chrysosporium. Lignin peroxidase production was not observed under conditions of nitrogen sufficiency or in balanced media, suggesting that control parameters could increase the activity by manipulating the secondary metabolism.

• The Plant Pathology Journal
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• v.15 no.3
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• pp.162-167
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• 1999

Integration of microbial antagonists with fungicides was tried to control the gray mold caused by Botrytis cinerea on pepper in greenhouse conditions and to reduce fungicide uses. All of the selected bacterial antagonists, Bacillus amyloliquefaciens BL3, Paenibacillus polymyxa BL4, and Pseudomonas putida Cha94, completely inhibited the conidial germination of B. cinerea until 30 days after treatment. However, bacterial colonization of pepper phylloplane was poor in BL4, while the other bacterial isolates and the fungal antagonist Trichoderma harzianum TM colonized well on the phylloplane, maintaining the population density of 104-105 cfu/g until 15 days after microbial treatments. Out of 13 kinds of selected fungicides used for gray mold diseases, polyoxin B and BKF 1995 showed the most discriminatory activity on the fungal growth between B. cinerea and TM. TM grew readily on the media containing those fungicides, while B. cinerea showed poor or no mycelial growth on them. The selected fungicides and antagonists alone reduced incidence of gray mold on pepper, showing disease indices of about 2.4 to 3.0, while its was increased up to 4.2 in the untreated control. Alternate treatments with the antagonists and 2-fold diluted fungicides inhibited the disease incidence as much as the antagonists or fungicides alone, and reduced the secondary inoculum more than the single treatments. This suggests that integration of antagonists and fungicides may be an efficient way to reduce fungicide sprays with reliable control efficacy of the disease. However, there was not much difference in the early and mid-term disease progress among the treatments and the untreated control, probably due to extremely favorable environmental conditions for the disease development in this experiment.

• Horticultural Science & Technology
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• v.33 no.2
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• pp.251-258
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• 2015

Eurycoma longifolia is an important rare medicinal plant that contains valuable bioactive compounds. In the present study, cell suspension culture of E. longifolia was established for the production of biomass and phenolic compounds. Various medium parameters, such as concentration of auxin, salt strength of the medium, and sucrose and nitrogen concentrations, were optimized for the production of biomass at the flask-scale level. Full strength Murashige and Skoog (MS) medium supplemented with $3.0mg{\cdot}L^{-1}$ naphthaleneacetic acid (NAA), 3% (w/v) sucrose, 0:60 $NH{_4}^+:NO{_3}^-$ was found suitable for biomass accumulation. Based on the optimized flask-scale parameters, cell suspension cultures were established in balloon-type bubble bioreactors, and bioprocess parameters such as inoculum density and aeration rate were optimized. Inoculum density of $50g{\cdot}L^{-1}$ and increasing aeration rate from 0.05 to 0.3 vvm, with increases every 7 days, were suitable for the accumulation of both biomass and phenolic compounds. With the optimized conditions, $14.70g{\cdot}L^{-1}$ dry biomass, $10.33mg{\cdot}g^{-1}$ DW of phenolics and $3.89mg{\cdot}g^{-1}$ DW of flavonoids could be achieved. Phenolics isolated from the cell biomass showed optimal free radical scavenging activity.