Hyekyung Shim;Lee, Hyunjeong;Seungbeom Hong;Park, Dae-Sup;DaeRobert A Samson;Hyeongjin Jee;Lee, Sukchan
Proceedings of the Korean Society of Plant Pathology Conference
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2003.10a
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pp.122-123
/
2003
Apple stem grooving virus (ASGV) belongs to Capillovirus and infects pome fruits. Transmission mode of ASGV is known by grafting and mechanical inoculation into susceptible hosts, not by any other natural vectors. But we have observed the spread of ASGV in the field without mechanical inoculation or grafting. Transmission seems to be occurred from tree-to-tree and tree-to-susceptible herbaceous plants along but not across ditches in the field. In order to ascertain this possibility, various fungi were isolated and cultured from ASGV-infected plants and 69 isolates were characterized. By means of RNA dot-blot hybridization and PCR analysis, 3 isolates were sorted out for further studies. The isolates were identified to Tataromyces sp. and belonged to Phenicillium by morphological characteristics and molecular markers. As an experimental host, 10 kidney beans (Phaseolus vulgaris) were screened and Kyunggi-5 was selected for virus amplification and symptom development. Kyunggj-5 infected by fungi which seemed to carry ASGV showed the typical disease symptoms and viral coat protein genes were detected from all tested plants. To confirm the Koch's rule, fungi cultured from inoculation origins of kidney bean were grown on PDA media and re-inoculated to hosts. The fungi isolated from inoculation origins induced the typical disease symptoms on hosts. However virus free fungi did not induce any symptom on the experimental hosts. This bioassay showed that these typical symptoms were caused by virus, not fungi.
This study was carried out to investigate the management characteristics and growth performance of L. edodes from the cooling stage to incubation. Bags of different heights and weights are available for bagging. When the medium size of $17{\times}13cm$ was used and the size of the inoculation hole was changed from 1/3 to 2/3, the browning period was shortened to 30 days. Mycelial growth was evaluated according to the cooling temperature after sterilization. It was observed to be the highest at 122 mm/15 days at $10^{\circ}C$ and 114 mm/15 days and 117 mm/15 days at $15^{\circ}C$ and $20^{\circ}C$, respectively. The contamination rate of the sawdust media before inoculation was measured as 0, $4.5{\times}10$, $1.3{\times}10^2$, $4.0{\times}10^3cfu$ at $5^{\circ}C$, $10^{\circ}C$, $15^{\circ}C$, and $24^{\circ}C$ respectively. The average of $1.6{\times}10^8$ colony forming units (cfu) of microorganisms was observed in the sawdust that had been piled for six months outdoors. In summer, the sawdust has to be used immediately after mixing. The sterilized medium had an average of $4{\times}10^3cfu$ of microorganisms at $24^{\circ}C$ and $1.3{\times}10^2cfu$ at $15^{\circ}C$. After 15 days of inoculation in vitro, the growth conditions of the sawdust was the best at 132 mm, followed by grain and liquid. When inoculated with liquid spawn, the moisture content of the substrate should be adjusted between 50% and 55% in advance.
Journal of Physiology & Pathology in Korean Medicine
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v.18
no.4
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pp.1061-1070
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2004
Chronic oral aphthae, recurrent ulcer and uveitis are the three main festations of Behcet's disease(BD). The aetiopathogenesis of Behcet's disease is still obscure, but herpes simplex virus is one of the possible casual factors. Gamchosasim-tang (Gancaoxiexin-tang), Banhasasim-tang(Banxiaxiexin-tang) and Saenggangsasim-tang( Shengjiangxiexin-tang) are traditional medication in Oriental medicine, that has been used to treat inflammatory disease. Especially, Gamchosasim-tang used to treat Behcet's disease like symptoms. ICR mice were used for this study. The earlobe of the mice were scratched with a needle, then inoculation with 1.0×10/sup 6/ plaque forming units/㎖ of HSV type I. Virus inoculation was performed twice with 10 day interval, followed by 16 weeks of observation. Using the HSV-induced Behcet's disease mouse model, kinds of Sasim-tang were administered variously before and after inoculation. In order to. classify the symptomatic mice as having Behcet's disease like symptoms. We followed the revised Japanese classification with minor modifications. Ulceration of the mice were monitored. In addition, spleen cytokine expression were measured by polymerase chain reaction, ELISA. HSV DNA was detected in HSV inoculation mice. HSV-induced mice treated with kinds of Sasim-tang showed improvement in symptom. In RT-PCR results, IFN-γ was expressed for all groups, IL-2 was expressed for the treated groups, and IL-10 was also expressed. IL-4 was expressed nothing. In ELISA, IL-2 was increased for GSST 2, BSST 2, GSST 2, GSST3 and INF-γ was increased for GSST 2, BSST 2, SSST 2, SSST 3. This model suggest the possible role of immune response to viral infection in the development and activation of Behcet's disease.
Kim, Ki-Yoon;Hwang, Seong-Woong;Saravanan, Venkatakrishnan Sivaraj;Sa, Tong-Min
Korean Journal of Soil Science and Fertilizer
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v.45
no.1
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pp.51-58
/
2012
Salinity is one of the most relevant abiotic factor limiting crop yield and its net primary productivity. In addition, salinity induces an increased stress ethylene synthesis in plants which, in turn, exacerbate the responses to the stressor. Bacterial single or co-inoculation effect was tested using previously characterized plant growth promoting (PGP) bacteria Brevibacterium iodinum RS16 and Methylobacterium oryzae CBMB20 on maize and sorghum-sudan grass hybrid under different concentrations of NaCl. Non-inoculated maize and sorghum-sudangrass hybrid showed 33.4% and 20.0% reduction in seed germination under highest NaCl (150 mM) level tested. However, under the same NaCl concentration, co-inoculation with B. iodinum RS16 and M. oryzae CBMB20 PGP strains increased the seed germination in maize (16.7%) and sorghum-sudangrass hybrid (4.4%). In Gnotobiotic growth pouch experiments conducted for maize and sorghum-sudangrass hybrid, co-inoculation of PGP B. iodinum RS16 and M. oryzae CBMB20 mitigated the salinity stress and promoted root length by 22.9% and 29.7%, respectively. Thus the results of this study could help in development of potential bioinoculants that may be suitable for crop production under saline conditions.
Barley mild mosaic virus(BaMMV) is a soilborne Bymovirus vectored by root-infecting fungus, Polymyxa graminis. Mechanism of cultivar's resistance to BaMMV in field tests are difficult to assess since resistance could be either due to the virus or to P. graminis, or both. Whereas, available mechanical inoculation methods for BaMMV and other related viruses are labor intensive, give inconsistent results and generally result in low infection rates. Inoculation method using stick with gauze(SWG) was developed for BaMMV. The improved method proved to be simple, efficient, and reliable. The infected leaf tissues were preserved by drying in a frozen state under high vaccum(freeze dried barley infected leaves) to circumvent reduction of virus infectivity during storage. Five Korean barley cultivars were mechanically inoculated with BaMMV-infected sap by the improved method. Infection rates obtained were compared with natural infection. Cultivar Naehanssalbori showed resistance to BaMMV in the field trials but was found highly susceptible in the greenhouse tests by mechanical inoculation, indicating that the field resistance may be possibly due to resistance to P. graminis.
In our preliminary study to find antiviral or antitumor agents from Korean natural products, we found that the Shope fibroma virus (SFV) induced fibromas reaching maximum size at $5{\sim}6$ days with spontaneous disappearance at $15{\sim}20$ days after SFV intracutaneous inoculation into Korean domestic rabbits. However, the sizes of fibromas of rabbits at day 5 after virus inoculation were significantly different individually. Assuming that the variation of tumor size was due to either susceptibility or the preexisting antibodies against SFV in the Korean domestic rabbits, the rabbits were checked for the antibodies against SFV by IFAT using SFV infected RK13 cells. The antibody positive rate of normal Korean domestic rabbits was 32.8% and the sizes of the fibromas of the positive rabbits were significantly smaller than those of negative rabbits (p<0.0001). The fibroma sizes were dependent on the antibody titers of rabbits to SFV. The sizes of fibromas after inoculation of SFV into immunized rabbits were about one tenth of those by the first inoculation into normal rabbits. This is the first report on the antibody prevalence against SFV among normal Korean domestic rabbits and it suggest the existence of a wild fibroma virus or related virus in Korea.
Journal of The Korean Society of Grassland and Forage Science
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v.15
no.4
/
pp.245-252
/
1995
To select a carrier as Rhizobiwn inoculants for alfalfa, severeal Rhizobium inoculants were produced by adding or not sub-materials($CaCO_3$ and sucrose) to main materials(organic or inorganic materials). The root nodule bacteria, Rhizobium meliloti 1061 distributed from KAIST, and Vemal alfalfa was used in this experiment. The Rhizobium populations and inoculation effects of the Rhizobium inoculants produced in several materials were scrutinized at laboratory and greenhouse in Livestock Experiment Station, RDA from 1993 to 1994. Moisture contents of the caniers were varied from 32 to 50% on dry weight basis according to material characteristics and the pH ranges of these were varied from pH 4.56 to 10.06 according to raw material characteristics and preparations. Initial Rhizobium numbers of the carriers were higher in organic material-inoculants than in inorganic, and among the inoculants, the inoculant made of Bentonite+Vermiculite(l:3 w/w) was excellent because of high rhizobium population($7.8~8.3\times10^8/g$ inoculant) and high rhizobium reappearance of inoculant in severed different production time. The root nodules of the alfalfa inoculated with different inoculants were fast formed in the fermented sawdust with cattle dung (FSC) inoculant, and bentonite(B)+vermiculite(V) than others. Plant length of alfalfa was differentiated on 15 days after inoculation but was not nearly different between higher inoculants than rhizobium number $10^7/g$ inoculant. Total dry matter of alfalfa was yielded by 20.65, 20.34mg per pot in FSC + sucrose 0.5% and B + V + sucrose 1% inoculants respectively that were higher inoculation effect by 17 times compared with non-inoculation, 1.2 mg per pot.
Citrus canker disease is caused by bacteria Xanthomonas axonopodis .pv. Citri. Shiranuhi cultivar, a hybrid of Kiyomi tangor and Nakano No.3 ponkan was evaluated for resistance to citrus canker based on initiation of disease, percent area of lesion infected and growth rate of bacteria in the leaf under growth chamber condition. Significant differences between susceptible plant and resistant plants were observed in these assays. Resistant plants showed delayed disease symptoms compared to the susceptible plants after spray inoculation of the pathogen. The resistant verities, satsuma, yuzu, and Shiranuhi showed symptoms after six days where as susceptible, mexican lime showed the symptoms just after three days of inoculation. 18 days after inoculation, percent area of lesions developed on leaf and disease severity differed significantly in susceptible and resistant plants, and were ranked as follows: mexican lime > early satsuma =Shiranuhi =yuzu (P <, 0.01). However, 30 days after inoculation, percent area of lesion was further differentiated into resistant and highly resistant plants. That was ranked as follows: sweet orange> early satsuma =Shiranuhi =Kiyomi > yuzu (P < 0.01). These results indicate that host reaction to the bacterial was more distinct when the disease developed for a longer period. Growth rates of a citrus canker bacterium during 16 40 h also were distinct after infiltration into leaves of susceptible and resistant plants, and were ranked as follows: sweet orange> early satsuma =Shiranuhi =Kiyomi =yuzu (P < 0.01). Based on these results, we concluded that Shiranuhi is resistant to citrus canker as compared to Kiyomi, early satsuma, and yuzu.
Infection structures were observed using a fluorescence microscope at the penetration sites on the leaves of potato plants pre-inoculated with the bacterial strains Pseudomonas putida TRL2-3, Micrococcus luteus TRK2-2, and Flexibacteraceae bacterium MRL412, which mediated an induced systemic resistance on potato plants against late blight disease caused by Phytophthora infestans. In order to compare the infection structures on the leaves expressing systemic acquired resistance, the leaves of potato plants pre-treated with DL-3-amino butyric acid (BABA) were also observed after challenge inoculation with the same pathogen. The infection structures were investigated. The total number of germination and appressorium formation of P. infestans were counted. Furthermore, the frequencies of fluorescent epidermal cells at the penetration sites, which indicate a defense response of plant cell, were estimated. There were no differences on the germination rates of the fungal cysts among the untreated control, BABA pre-treated, and bacterial strains pre-inoculated plants. However, appressorium formation was slightly decreased on the leaves of BABA pre-treated plants compared to those of untreated as well as bacterial strains pre-inoculated plants. Furthermore, the frequencies of fluorescent cells of BABA pre-treated and bacterial strains pre-inoculated were higher than that of untreated plants, indicating an active defense reaction of the host cells against the fungal attack. On the other hand, the pre-treatment with BABA caused a stronger fluorescent of epidermal cells at the penetration sites compared to the pre-inoculation with the bacterial strains. Interestingly, the frequency of fluorescent cells by BABA, however, was lower than that by the bacterial strains. Based on the results it is suggested that the infection structures showing resistance reaction on the leaves of potato plants were different between by pre-inoculation with bacterial strains and by pre-treatment with BABA against the late blight pathogen.
The present study was carried out to investigate the incidence of C jejuni and C coli in chicken. Also were examined the pathogenicity of the isolates in chick by experimental inoculation. Thermophilic Campylobacter were isolated from 34(45.9%) of the 74 specimens, and classified as 21.6% C jejuni, and 24.3% C coli. In the biotyping of 16 stranis of C jejuni isolates, 37.5% of the strains were grouped as biotype I, 62.5% as biotype II. In the case of 18 strains of C coli isolates, 49.9% of isolates were grouped as biotype I, 55.6% as biotype II. n oral inoculation with $10^4cfu$ of Campylobacter isolates into infant chicks(1 to 3 days-old), 17 days-old and 34 days-old chicks, 32.5% of the chicks developed diarrhea on day 1, 52.5% on day 3, 70.0% on day 5, and 27.5% on day 7, and the peak incidence of diarrhea was reached on day 5. The organisms were found to be discharged in feces one day afterwards. C jejuni and C coli strains were detected from the feces in 87.5% of the chicks on day 5. The organisms were multiplied from $10^4$ to $10^6cfu/gm$ in feces 5 to 7 days after inoculation. C jejuni and C coli recovered from 100% of the cecum, 64.3% of the duodenum, 50.0% of the spleen, 42.9% of the livers, and from 21.4% of gallbladders 7 days after inoculation.
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