• Research in Plant Disease
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• v.20 no.2
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• pp.101-106
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• 2014

Citrus canker caused by Xanthomonas citri subsp. citri (Xcc) is one of the most important diseases on citrus. Although Satsuma mandarin cultivating mostly in Korea is moderately resistance to canker, occurrence of the disease were more frequently reported since last decade. Like other diseases in citrus, citrus canker was mainly protected by chemical fungicide in the field. Due to the side effect of the chemicals, alternative method of disease control is recently required. In this study four rhizobacterial strains TRH423-3, MRL408-3, THJ609-3 and TRH415-2 are selected by testing its antifungal activity against Xcc. Pre-inoculation with the selected rhizobacterial strains caused disease suppression on the citrus leaves after inoculation with the citrus canker pathogen. Similarly, in the field test symptoms of citrus canker were less developed in the citrus trees applied several times with the selected rhizobacterial strains compared with those of untreated trees. Therefore, it is suggested that the selected rhizobacterial strains may be valuable as an alternative method in the environment-friendly citrus farm.

• Animal Bioscience
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• v.35 no.10
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• pp.1592-1605
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• 2022

Objective: The objective of this study was to select an effective in vitro digestion-fermentation model to estimate the effect of decreasing dietary crude protein (CP) on odor emission during pig production and to suggest potential prediction markers through in vitro and in vivo experiments. Methods: In the in vitro experiment, three diet formulations with different CP contents (170 g/kg, 150 g/kg, and 130 g/kg) but containing the same standardized ileal digestible essential amino acids (SID-EAA) were assessed. Each diet was evaluated by two different in vitro gastric-intestinal phase digestion methods (flask and dialysis), combined with fresh pig feces-ferment inoculation. Eighteen growing barrows (31.9±1.6 kg) were divided into three groups: control diet (180 g CP/kg, without SID-EAA adjustment), 170 g CP/kg diet, and 150 g CP/kg diet for 4 weeks. Results: The in vitro digestion results indicated that in vitro digestibility was affected by the gastric-intestinal phase digestion method and dietary CP level. According to the gas kinetic and digestibility results, the dialysis method showed greater distinguishability for dietary CP level adjustment. Nitrogen-related odor compounds (NH₃-N, indole, p-cresol, and skatole) were highly correlated with urease and protease activity. The feeding study indicated that both EAA-adjusted diets resulted in a lower odor emission especially in p-cresol and skatole. Both protease and urease activity in feces were also closely related to odor emissions from nitrogen metabolism compounds. Conclusion: Dialysis digestion in the gastric-intestinal phase followed by fresh fecal inoculation fermentation is suitable for in vitro diet evaluation. The enzyme activity in the fermentation and the fecal samples might provide a simple and effective estimation tool for nitrogen-related odor emission prediction in both in vitro and in vivo experiments.

• Horticultural Science & Technology
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• v.33 no.6
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• pp.883-890
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• 2015

The soilborne fungus Fusarium oxysporum f. sp. lilii (Fol) is a serious threat to all lily cultivars, especially infecting bulbs and flowers. It has become increasingly important to develop varieties resistant against the bulb rot disease. Genetic diversity of cultivars and reliable screening methods are required for this purpose. Here, an efficient in vitro screening system for evaluating resistance to Fol in 38 in vitro-grown lily plants was investigated. Various factors including culture conditions of Fol, inoculum density, appropriate plant materials, inoculation method and duration, and incubation period of plant materials after inoculation were combined to optimize the screening method. As a result, we suggest optimal conditions for an in vitro screening system for the selection of Fol-resistant lily cultivars as follows. Fol was grown on potato dextrose agar (PDA) medium for 6 days at $25^{\circ}C$ in darkness and used as working inoculation. Spore suspensions were prepared (inoculum density: $1.0{\times}10^4$ $spores{\cdot}mL^{-1}$), and then leaf segments $1.5{\times}2.0cm^2$ were inoculated by dipping for 22 hours at $25^{\circ}C$ in dark. Later, leaves were cultured on 0.6% agar plates at $25^{\circ}C$ and 50% humidity with a photoperiod of 16 hours light/8 hours dark (fluence rate of $40{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$) to examine the progress of bulb rot. After 7 days, disease levels were classified into indices 1 (no symptom) to 6 (serious bulb rot). Soil inoculation of Fol carried out with resistant or susceptible lily cultivars that had been selected through in vitro screening confirmed the reproducibility of results. Therefore, the in vitro screening method established in this study is efficient and reliable for selection of lily cultivars resistant against bulb rot disease.

• Korean Journal Plant Pathology
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• v.14 no.1
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• pp.46-51
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• 1998

A method for inoculum production of Botryosphaerisa dothidea was developed using cucumber disks and cereal media. Disks of cucumber fruits, and cereal media of barley, wheat, and rice seeds were inoculated with mycelial plugs of B. dothidea and incubated at 27$^{\circ}C$. Pycnidia were produced on the surface of cucumber disks and seeds after 5 days of inoculation. When the inoculated barley seeds were immersed in sterilized distilled water for 5 minutes, abundant conidia of B. dothidea were exuded from mature pycnidia. Conidia were held together by mucilage as they were released from an ostiole. Compared with the conventional method for inoculum preparation using agar media, such as potato-dextrose agar and oatmeal agar, this method could minimize the tedious work required for inoculum preparation within a shorter period of time.

• Research in Plant Disease
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• v.22 no.3
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• pp.152-157
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• 2016

Root-dipping inoculation method has been used to investigate resistance of radish plants to Fusarium wilt. However, the method requires a lot of labor and time because of complicate procedure. This study was conducted to establish a simple and effective mass-screening method for resistant radish to Fusarium wilt. Radish seedlings of susceptible and resistant cultivars were used to investigate wounding method by scalpel, inoculum concentration, and pathogen-inoculated growth stage of seedlings. We established an efficient mass-screening method based on our results as following: Roots of 14-day-old seedlings of radish are cut with a scalpel at a $90^{\circ}$ angle to a 2 cm-depth at a 1 cm-distance from main stem and then inoculated by pouring with a 10 ml-aliquot of a fungal spore suspension ($1.0{\times}10^7conidia/ml$) on soil. The inoculated plants are cultivated in a growth room at $25^{\circ}C$ for about 4 weeks with 12-hour light a day. The proposed screening method enables to effectively select resistant from mass radish plants cultivars to Fusarium wilt.

• Korean Journal of Microbiology
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• v.12 no.3
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• pp.131-137
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• 1974

It is a well-known fact that an isolation of non-sporeforming anaerobes, considered normal flora in man ordinarily but causes serious infections sometimes, is a dificult procedure because of their great oxygen sensitivity. Among the many techniques employed in clinical laboratories, despite of its high expenses, the GasPak method has been most widely used because of its relative simplicity. On the other hand, the steel wool method has gained a good reputation recently. This technique makes it possible to treat individual plate so that any single specimen can be promptly cultured anaerobically. The procedure is quite simple and the expenses are negligible. In the present study it is to compare these two methods as to their efficiency of anaerobic cultivation using 13 VPI strains of non-sporeforming amaerobic bacteria. Among the 13 species the following 11, Bacteroides fragilis ss. fragilis, B. fragilis ss. thetaiotaomicron, propionibacterium acnes, Eubacterium limosum, E. lentum, peptococcus asaccharolyticus, Pc. prevotii, Pc. magnus, peptostreptococcus anaerobius, Ps. intermedius nad Veillonella parvula, grew well with the steel wool method whose colony numbers reaching 57 to 119% of those with GasPak method. The remaining two species, Fusobacterium nucleatum and F.necrophorum, did not grow well with the steel wool method showing the colony numbers were only 0.4% of those with GasPak method in the case of Fusobacterium nucleatum. In the case of Fusobacterium necrophorum, very few colonies developed even with a heavy inoculation. As to the size of colonies, there were no significant difference between these two methods.

• Journal of Microbiology and Biotechnology
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• v.21 no.10
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• pp.1069-1072
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• 2011

A real-time PCR assay method was established to monitor Leuconostoc spp. populations via specific amplification of the dextransucrase gene. Quantification of L. mesenteroides B-512F using both genomic DNA and cell suspensions yielded a log-linear correlation spanning approximately 5 log units. By using this method, monitoring changes of Leuconostoc spp. during sauerkraut fermentation was successfully accomplished with accuracy after inoculation of starter and sugars (sucrose and maltose).

• Korean Journal of Veterinary Research
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• v.30 no.3
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• pp.309-315
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• 1990

The present experiment was done to identify newcastle disease virus(NDV) antigens in frozen sections of various oragns from experimentally NDV-infected with indirect immunoperoxidase method. Section were incubated with rabbit anti-NDV polyclonal as first antibody, followed by incubation with goat anti-rabbit or protein A peroxidase conjugate. Positive reactions were often detected in the epithelium of trachea and in the lymphocyte of spleen at 24 hours after virus inoculation. the viral antigen was localized mainly in the cytoplasm of infected cells. The method approved to be highly specific for the identification of NDV and allowed a precise localization of the viral antigens in infected cells.

• Korean Journal of Food Science and Technology
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• v.52 no.3
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• pp.296-303
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• 2020

Hop-resistant lactic acid bacteria (LAB) are well-known, major beer-spoilage bacteria. The hop-gradient agar containing ethanol (c-HGA+E) method has been used to examine hop-resistance of beer-spoilage LAB. However, the selection of beer-spoilage bacteria by the c-HGA+E method is either too selective or too inclusive. Furthermore, it is accompanied by a complicated experimental procedure, high-cost and time. To overcome these disadvantages, the modified hop-gradient agar with ethanol (m-HGA+E) method was developed. The most remarkable modifications were the shape of the petri dish and the inoculation method for bacteria. The efficiency and validation of the m-HGA+E approach were proven by the formation of colonies at different hop concentrations in the bottom layer, co-culture with the bacteriocin producer and by PCR detection of hop-resistant genes. This study demonstrated that m-HGA+E is a rapid, economical, and easy method to monitor potential hop-resistant beer-spoilage LAB during the beer brewing process.

• Research in Plant Disease
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• v.23 no.4
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• pp.334-347
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• 2017

Bacterial wilt caused by Ralstonia solanacearum is an important disease in cultivation of chili pepper, causing plant death and significant yield losses. Cultivation of disease-resistant varieties is the most suitable measure to control bacterial wilt of chili pepper. To establish an efficient screening method for resistant chili pepper to R. solanacearum, six resistant or susceptible cultivars to the R. solanacearum were selected and the development of bacterial wilt on the cultivars according to several conditions was investigated. Drenching bacterial suspension into the cut roots using a scalpel was more simple and effective to distinguish resistant and susceptible cultivars than inoculation methods of root-dipping or soil-drenching without wounding. A resistant pepper, 'MC4' to R. solanacearum showed high resistance under the developed conditions which were 21- to 28-day-old pepper inoculated with $1{\times}10^8cfu/ml$ of bacterial suspension. On the other hands, the susceptible cultivars represented high disease severity under the conditions. These results indicated that we developed an efficient method to evaluate resistance of chili pepper cultivars against bacterial wilt. In addition, we successfully evaluated resistance degree of 140 commercial chili pepper cultivars to R. solanacearum using the developed method.