Cordyceps pruinosa grows upon dead pupae of Lepidoptera and produces one or $3{\sim}4$ club-shaped stromata per host. The stromata have distinct club-shaped head and long stalk. The length of stromata varies from $1{\sim}3\;cm$. Apical head consists of densely crowded semi-immersed perithecia, which are $360{\sim}400\;{\times}\;180{\sim}200\;{\mu}m$ in size. Asci are $150\;{\mu}m$ in length and $2.8{\sim}3\;{\mu}m$ in diameter. Ascospores, which are $124{\sim}141\;{\mu}m$ in length, have thin thread-like structures in the middle with part-spores attached on both sides. Each ascospore does not separate into part-spores after dispersal, but each part-spore germinates and together develops a colony. The imperfect form produces phialides of $15{\sim}24\;{\times}\;2{\sim}3\;{\mu}m$ size, with spherical or spindle shaped conidia of $4{\sim}6\;{\times}\;1.8{\sim}2.4\;{\mu}m$ size, The anamorph was identified as Mariannaea elegans Samson. YMA and SDAY agar media with pH 7 was produced abundant mycelial growth with high density. Best mycelial growth was observed when dextrin was used as a carbon source. Lactose, saccharose and sucrose also produced high mycelial growth. Peptone, yeast extract and tryptone produced abundant mycelial growth, when used as nitrogen sources. Highest mycelial growth and density was observed when C/N ratio was 1 : 1 at the concentration of 12.5 g/l each. $KH_2PO_4$ was the best mineral source for mycelial growth. Highest mycelial dry wt. was produced in YM and SDAY broths. Optimum inoculum for 100 ml of liquid broth was 6 mycelial discs. Similarly, optimum liquid culture period was 7 days.
Two attenuated Cucumber mosaic virus (CMV) isolates, Paf-CMV and Rs2-CMV that had been selected from CMV isolates associated with satellite RNA (satRNA) were tested for cross-protection effect in pepper plants. The viruses selected as attenuated strains appeared to be identical serologically and physically to the challenge virus (Mf-CMV), but they were lower in the dilution end-point of infectivity of crude sap than Mf-CMV When symptoms were observed in several indicator plants after inoculation, Paf-CMV and Rs2-CMV were symptomless or showed mild mosaic symptoms while another satRNA isolate Ap-CMV developed severe mosaic symptoms on the leaves as Mf-CMV. The nucleotide sequences of the satRNAs were determined by sequencing full-length cDNA clones. Paf-, Rs2- and Ap-satRNAs were 386, 335, and 347 nucleotides long, respectively, The sequences were then compared with the other known Y-satRNA, revealing that nucleotide sequences of the satRNAs consisted of 5'- and 3'-terminal conserved regions. However variations occurred on the middle regions of the sequences, especially those related to symptom interference, showing significant differences between Paf-satRNA and other isolates. Infectious transcripts of Paf-satRNA and Rs2-satRNA induced mild mosaic symptoms in pepper plants when supported by genomic RNAs of Mf-CMV. Under greenhouse conditions, Paf-CMV and Rs2-CMV were tested for cross-protection effect in pepper and tobacco (Nicotiana tabacum cv, Xanthi nc) plants against Mf-CMV. No symptoms were developed on the plants vaccinated with Paf-CMV until 3 weeks after inoculation with the virulent strain; however another attenuated isolate, Rs2-CMV, showed less effectiveness in cross-protection. Depending on the concentration of the challenged virus, symptoms sometimes appeared later in the upper leaves. However, in plants challenged with low concentrations (below 0.2 mg/ml) of the challenge inoculum, symptoms caused by the virulent strain did not develop on the plants vaccinated with Paf-CMV. In the field experiments, the number of pepper plants with severe mosaic symptoms in the control plots was progressively increased after transplanting and reached approximately 50% after 50 days. On the other hand, the incidence of mosaic disease appeared very low on the plants that had received the protective inoculation with Paf-CMV.
Lee, Won Jeong;Jang, Kyoung Soo;Choi, Yong Ho;Kim, Heung Tae;Kim, Jin-Cheol;Choi, Gyung Ja
Research in Plant Disease
/
v.21
no.3
/
pp.201-207
/
2015
This study was conducted to establish a simple mass-screening method for resistant melon to Fusarium wilt caused by Fusarium oxysporum f. sp. melonis (FOM). Root-dipping inoculation method has been used to investigate resistance of melon plants to Fusarium wilt. However, the inoculation method requires a lot of labor and time because of complicate procedure. To develop a simple screening method on melon Fusarium wilt, occurrence of Fusarium wilt on susceptible and resistant cultivars of melon according to inoculation method including root-dipping, soil-drenching, tip, and scalpel methods was investigated. Scalpel and tip methods showed more clear resistant and susceptible responses in the melon cultivars than root-dipping inoculation method, but tip method represented slightly variable disease severity. In contrast, in the case of soil-drenching inoculation method, disease severity of the susceptible cultivars was very low. Thus we selected scalpel method as inoculation method of a simple screening method for melon Fusarium wilt. By using the scalpel inoculation method, resistance degrees of the cultivars according to incubation temperature after inoculation (25 and $30^{\circ}C$) and inoculum concentration ($1{\times}10^6$ and $1{\times}10^7conidia/ml$) were measured. The resistance or susceptibility of the cultivars was hardly affected by all the tested conditions. To look into the effectiveness of scalpel inoculation methods, resistance of 22 commercial melon cultivars to FOM was compare with root-dipping inoculation method. When the melon cultivars were inoculated by scalpel method, resistance responses of all the tested cultivars were clearly distinguished as by root-dipping method. Taken together, we suggest that an efficient simple mass-screening method for resistant melon plant to Fusarium wilt is to sow the seeds of melon in a pot (70 ml of soil) and to grow the seedlings in a greenhouse ($25{\pm}5^{\circ}C$) for 7 days, to cut the root of seedlings with a scalpel and then pour a 10 ml-aliquot of the spore suspension of $1{\times}10^6conidia/ml$ on soil. The infected plants were cultivated in a growth room at 25 to $30^{\circ}C$ for about 3 weeks with 12-hr light a day.
Park, Myung-Soo;Jeong, Bo-Ram;Jang, Kyoung-Soo;Choi, Yong-Ho;Kim, Jin-Cheol;Choi, Gyung-Ja
Horticultural Science & Technology
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v.30
no.4
/
pp.426-431
/
2012
This study was conducted to establish an efficient screening method for resistant tomato to Fusarium wilt caused by Fusarium oxysporum f. sp. lycopersici (FOL). The resistance degrees of the six commercial cultivars of tomato to the pathogen were evaluated by dipping roots of the seedlings in spore suspension of five FOL isolates. On the basis of the results, two cultivars (Dotaerangmaster, resistant cultivar to FOL race 1; Supersunload, resistant cultivar to FOL race 2) and two isolates (KACC40043, FOL race 2; TF104, FOL race 3) were selected for system establishment. The disease development of the FOL isolates on the cultivars according to several conditions including root wounding, incubation temperature, inoculum concentration and dipping period of roots in spore suspension was investigated. The resistance of each cultivar to the disease was a race-specific response and hardly affected by the tested conditions except for incubation temperature of $20^{\circ}C$. The optimum temperature for disease development caused by FOL was 25 to $30^{\circ}C$. On the basis of the results, we suggest that an efficient screening method for resistant tomato cultivars to Fusarium wilt is to dip the non-cut roots of tomato seedlings at two-leaf stage in spore suspension of $1{\times}10^7\;conidia{\cdot}mL^{-1}$ for 0.5 hours and transplant the seedling to plastic pot with horticulture nursery media, and then to cultivate the plants in a growth room at $25^{\circ}C$ for 3 weeks with 12 hours light a day.
Escherichia coli O157:H7, Staphylococcus aureus and Salmonella enteritidis are food borne pathogens involved in food poisoning in numerous countries. This study aimed to obtain knowledges on the survival of Esc coli O157:H7, Sta aureus and Sal. enteritidis in the yoghurt added with water extract of Omija(Schizandra chinensis). The growth inhibition of Schizandra chinensis extract on the food borne pathogens were measured by total microbial count and effect of growth inhibition was correspondent to the concentration of Schizandra chinensis extract. The highest growth inhibition effect of Schizandra chinensis extract was shown on the Sta aureus followed by Sal. enteritidis and Esc. coli O157:H7. The number of surviving Esc. coli O157:H7 cell(3.55${\times}$10$\^$5/ CFU/mL) was decreased to 1.00${\times}$10$^1$∼3.00${\times}$10$^1$ CFU/mL after 24 hours incubation by the addition of 0.4∼l.0% of Schizandra chinensis extract in the yoghurt. And also the viable cell counts of surviving Sta. aureus cells (initial inoculum 1.24${\times}$10$\^$5/ CFU/mL) were decreased gradually to 4.00${\times}$10$^2$∼8.50${\times}$10$^2$ CFU/mL after 48 hours of incubation, but the viable cells of Sal. enteritidis were not detected after 24 hours of incubation. Growth of the food borne pathogens was strongly inhibited by the addition and incubation of Schizandra chinensis extract for 48 hours in the yoghurt.
Kim, Min-Jeong;Shim, Chang-Ki;Kim, Yong-Ki;Hong, Sung-Jun;Park, Jong-Ho;Han, Eun-Jung;Jee, Hyeong-Jin;Kwon, Jin-Hyeuk;Kim, Seuk-Chul
Research in Plant Disease
/
v.21
no.2
/
pp.58-66
/
2015
This study was conducted to evaluate control efficacy of a fermented food 'Cheonggukjang' against cucumber powdery mildew caused by Sphaerotheca fuliginea in greenhouse. Sterilized Daepung beans were inoculated with the rice straw as natural inoculum and then incubated for 72 hrs at $42^{\circ}C$ in the household cheonggkjang maker. After 72 hrs of cheonggukjang fermentation, white zymogens were grown on the surface of a sterile Daepung beans. The pH of the 72 hrs fermented soybean was not significantly changed and electrical conductivity was found to increase by about 2 times than before fermentation. The population density of soybean zymogen showed a peak of growth at 60 hrs after fermentation and the concentration of zymogen was $8.2{\times}10^7cfu/ml$. Soybean zymogen form of the colony was divided into three kinds of bacteria and a white and a large colony (WL) was predominant bacteria among those up to 60 hrs of fermentation. To control the cucumber powdery mildew, diluted solutions of cheonggukjang was applied from 6.0% to 30.0% on cucumber leaves and they showed injury symptoms on cucumber leaves in more than 15% of them. However, more than 6.0% diluted cheonggukjang solutions showed more than 77.8% control effect of cucumber powdery mildew at 15 days after treatment. The fermented bacteria of Chenggukjang were well established in the cucumber leaf area at 15 days after treatment. The antifungal activity of 10% diluted cheonggukjang solutions was excellent for four species of plant fungal pathogens, Colletotrichum gloeosporioides, Sclerotinia cepivorum, Rhizoctonia sloani and Phytophthora capsici in the dual culture test. Results indicated that foliar application of Cheonggukjang solution could be used for the control of powdery mildews occurring on organically cultivated cucumber.
Kim, Jung-Beom;Kim, Jae-Kwang;Kim, Hyunjung;Cho, Eun Jung;Park, Yeon-Joon;Lee, Hae Kyung
Annals of Clinical Microbiology
/
v.21
no.4
/
pp.80-85
/
2018
Background: The aim of this study was to comparatively evaluate the bactericidal effects of copper, brass (copper 78%, tin 22%), and stainless steel against methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus faecium (VREFM), and multidrug-resistant Pseudomonas aeruginosa (MRPA). Methods: The isolates (MRSA, VREFM, MRPA) used in this study were mixed wild type 3 strains isolated from patients treated at Uijeongbu St. Mary's Hospital in 2017. These strains showed patterns of multidrug resistance. The lyophilized strains were inoculated into and incubated for 24 hr in tryptic soy broth at $35^{\circ}C$. The initial bacterial inoculum concentration was adjusted to $10^5CFU/mL$. A 100-mL bacterial suspension was incubated in containers made of brass (copper 78%, tin 22%), copper (above 99% purity), and stainless steel at $35^{\circ}C$. Viable counts of bacteria strains were measured for 9 days. Results: In this study, the bactericidal effects of copper and brass on MRSA, VREFM, and MRPA were verified. The bactericidal effect of stainless steel was much weaker than those of copper and brass. The bactericidal effect was stronger on MRPA than on MRSA or VREFM. Conclusion: To prevent cross infection of multidrug resistant bacteria in hospitals, further studies of longer duration are needed for testing of copper materials on objects such as door knobs, faucets, and bed rails.
This study investigated the effectiveness of using pathogens and aqueous acids to reduce the Aspergillus ochraceus and Rhodotorula mucilaginosa contamination in livestock production environments. For this study, 1 mL of each bacterial suspension (107-108 spores/mL) was inoculated on a knife surface, dried at 37℃, and used under each treatment condition. First, to investigate the effect of organic acids, acetic, lactic, and citric acids were used. Subsequently, to select the appropriate concentration, they were prepared at concentrations of 0.5, 1, 2, 3, 4, and 5%, respectively. Accordingly, to further maximize the effect of organic acid treatment, we combined the treatment with ultraviolet light. The two strains showed a significant difference (P<0.05) compared to the initial strain, with a greater than 90% decrease in the concentrations of all organic acids. Consequently, acetic and lactic acids decreased by approximately 5 and 2 log colony forming unit (CFU)/cm2, respectively, when treated with ultraviolet light (360 mJ/cm2); however, citric acid decreased by less than 1 log CFU/cm2. However, when manufactured with 4% acetic acid, a severe malodor was emitted, making it difficult for workers to use it in a production environment. Accordingly, the optimal treatment conditions for organic acid and ultraviolet light for application were selected as follows: immersion in a 4% lactic acid solution for 1 minute and then, sterilization with ultraviolet light at 360 mJ/cm2. Finally, when a pork meat sample was cut with a knife that was finally washed with lactic acid and treated with ultraviolet light, the low level of inoculum transferred from the cleaned knife to the surface of the sample was not detected. In conclusion, using this established method can prevent cross-contamination of the surface of the meat during processing.
Attempts to search infection period, infection speed in the tissue of neck blast of rice plant, location of inoculum source and effects of several conditions about the leaf sheath of rice plants for neck blast incidence have been made. 1. The most infectious period for neck blast incidence was the booting stage just before heading date, and most of necks have been infected during the booting stage and on heading date. But $Indica{\times}Japonica$ hybrid varieties had shown always high possibility for infection after booting stage. 2. Incubation period for neck blast of rice plants under natural conditions had rather a long period ranging from 10 to 22 days. Under artificial inoculation condition incubation period in the young panicle was shorter than in the old panicle. Panicles that emerged from the sheath of flag leaf had long incubation period, with a low infection rate and they also shown slow infection speed in the tissue. 3. Considering the incubation period of neck blast of rice plant, we assumed that the most effective application periods of chemicals are 5-10 days for immediate effective chemicals and 10-15 days for slow effective chemicals before heading. 4. Infiltration of conidia into the leaf sheath of rice plant carried out by saturation effect with water through the suture of the upper three leaves. The number of conidia observed in the leaf sheath during the booting stage were higher than those in the leaf sheath during other stages. Ligule had protected to infiltrate of conidia into the leaf sheath. 5. When conidia were infiltrated into the leaf sheath, the highest number of attached conidia was observed on the panicle base and panicle axis with hairs and degenerated panicle, which seemed to promote the infection of neck blast. 6. The lowest spore concentration for neck blast incidence was variable with rice varietal groups. $Indica{\times}Japonica$ hybrid varieties were infected easily compared to the Japonica type varieties, especially. The number of spores for neck blast incidence in $Indica{\times}Japonica$ hybrid varieties was less than 100 and disease index was higher also in $Indica{\times}Japonica$ hybrid than in Japonica type varieties. 7. Nitrogen content and silicate content were related with blast incidence in necks of rice plants in the different growing stage changed during growing period. Nitrogen content increased from booting stage to heading date and then decreased gradually as time passes. Silicate content increased from booting stage after heading with time. Change of these content promoted to increase neck blast infection. 8. Conidia moved to rice plant by ascending and desending dispersal and then attached on the rice plant. Conidia transfered horizontally was found very negligible. So we presumed that infection rate of neck blast was very low after emergence of panicle base from the leaf sheath. Also ascending air current by temperature difference between upper and lower side of rice plant seemed to increase the liberation of spores. 9. Conidial number of the blast fungus collected just before and after heading date was closely related with neck blast incidence. Lesions on three leaves from the top were closely related with neck blast incidence, because they had high potential for conidia formation of rice blast fungus and they were direct inoculum sources for neck blast. 10. The condition inside the leaf sheath was very favorable for the incidence of neck blast and the neck blast incidence in the leaf sheath increased as the level of fertilizer applied increased. Therefore, the infection rate of neck blast on the all panicle parts such as panicle base, panicle branches, spikelets, nodes, and internodes inside the leaf sheath didn't show differences due to varietal resistance or fertilizers applied. 11. Except for others among dominant species of fungi in the leaf sheath, only Gerlachia oryzae appeared to promote incidence of neck blast. It was assumed that days for heading of varieties were related with neck blast incidence.
Throughout the studies the following experimental results were obtained and are summarized: 1. Multiplication of agents in primary cell cultures of both GF classical and CR-64 acute strain of Marek's disease infected chicken kidneys was accompanied by the formation of distinct transformed cell foci. This characteristic nature of cell transformation was passaged regularly by addition of dispersed cell from infected cultures to normal chicken kidney cell cultures, and also transferred was the nature of cell transformation to normal chick-embryo liver and neuroglial cell cultures. No cytopathic changes were noticed in inoculated chick-embryo fibroblast cultures. 2. The same cytopathic effects were noticed in normal kidney cell monolayers after the inoculation of whole blood and huffy coat cells derived from both forms of Marek's disease infected chickens. In these cases, however, the number of transformed cell foci appearing was far less than that of uninoculated monolayers prepared directly from the kidneys of Marek's disease infected chickens. 3. The change in cell culture IS regarded as a specific cell transformation focus induced by an oncogenic virus rather than it plaque in slowly progressing cytopathic effect by non-oncogenic viruses, and it is quite similar to RSV focus in chick-embryo fibroblasts in many respects. 4. The infective agent (cell transformable) were extremely cell-associated and could not be separated in an infective state from cells under the experimental conditions. 5. The focus assay of these agents was valid as shown by the high degree of linear correlation (r=0.97 and 0.99) between the relative infected cell concentration (in inoculum) and the transformed cell foci counted. 6. No differences were observed between the GF classical strain and the CR-64 acute strain of Marek's disease as far as cell culture behavior. 7. Characterization of the isolates by physical and chemical treatments, development of internuclear inclusions in Infected cells, and nucleic acid typing by differential stainings and cytochemical treatments indicated that the natures of these cell transformation agents closely resemble to those described fer the group B herpes viruses. 8. Susceptible chicks inoculated with infected kidney tissue culture cells developed specific lesions of Marek's disease, and in a case of prolonged observation after inoculation (5 weeks) the birds developed clinical symptoms and gross lesions of Marek's disease. Kidney cell cultures prepared from those inoculated birds and sacrificed showed a superior recovery of cell transformation property by formation of distinct foci. 9. Electron microscopic study of infected kidney culture cells (GF agent) by negative staining technique revealed virus particles furnishing the properties of herpes viruses. The particle was measured about $100m{\mu}$ and, so far, no herpes virus envelop has been seen from these preparations. 10. No relationship of both isolates to avian leukosis/sarcoma group viruses and PPLO was observed.
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