• Research in Plant Disease
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• v.11 no.1
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• pp.48-55
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• 2005

Cylindrocarpon destructans is a soil-borne plant pathogenic fungus causing root rot on ginseng and trees. Rapid and exact detection of this pathogen was practiced on ginseng seedlings by nested PCR using speciesspecific primer set. The second round of PCR amplification by Dest 1 and Dest 4 primer set formed 400 bp of species-specific fragment of C. destructans from the product of first round of amplification by ITS 1 and ITS 4 primer set. In the PCR sensitivity test based on DNA density, nested PCR detected to the limit of one fg and it meant the nested PCR could detect up to a few spores of C. destructans. Also, nested PCR made it possible to detect the pathogen from ginseng seedlings infected by replantation on artificial infested soil. Our nested PCR results using species-specific primer set could be utilized for diagnosis of root rot disease in ginseng cultivation.

• Journal of Electrochemical Science and Technology
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• v.13 no.4
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• pp.431-437
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• 2022

As one of the most hazardous and deadliest pathogens, Listeria monocytogenes (LM) posed various serious diseases to the human being, thus designing effective strategy for its detection is of great significance. In this work, by preparing Ti₃C₂T_x MXenes nanoribbon (Ti₃C₂T_xR) as carrier and selecting thionine (Th) acted simultaneously as signal probe and functional monomer, a LM pathogen-imprinted polymers (PIP) integrated probe electrochemical sensor was design to monitor LM for the first time, that was carried out through the electropolymerization of Th on the Ti₃C₂T_xR/GCE surface in the existence of LM. Upon eluting the templates from the LM imprinted cavities, the fabricated PIP/Ti₃C₂T_xR/GCE sensor can rebound LM cells effectively. By recording the peak current of Th as the response signal, it can be weakened when LM cell was re-bound to the LM imprinted cavity on PIP/Ti₃C₂T_xR/GCE, and the absolute values of peak current change increase with the increasement of LM concentrations. After optimizing three key parameters, a considerable low analytical limit (2 CFU mL^-1) and wide linearity (10-10⁸ CFU mL^-1) for LM were achieved. In addition, the experiments demonstrated that the PIP/Ti₃C₂T_xR sensor offers satisfactory selectivity, reproducibility and stability.

• Mycobiology
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• v.51 no.4
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• pp.195-209
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• 2023

The seed borne disease such as bakanae is difficult to control. Crop yield loss caused by bakanae depending on the regions and varieties grown, ranging from 3.0% to 95.4%. Bakanae is an important disease of rice worldwide and the pathogen was identified as Fusarium fujikuroi Nirenberg (teleomorph: Gibberella fujikuroi Sawada). Currently, four Fusaria (F. fujikuroi, F. proliferatum, F. verticillioides and F. andiyazi) belonging to F. fujikuroi species complex are generally known as the pathogens of bakanae. The infection occurs through both seed and soil-borne transmission. When infection occurs during the heading stage, rice seeds become contaminated. Molecular detection of pathogens of bakanae is important because identification based on morphological and biological characters could lead to incorrect species designation and time-consuming. Seed disinfection has been studied for a long time in Korea for the management of the bakanae disease of rice. As seed disinfectants have been studied to control bakanae, resistance studies to chemicals have been also conducted. Presently biological control and resistant varieties are not widely used. The detection of this pathogen is critical for seed certification and for preventing field infections. In South Korea, bakanae is designated as a regulated pathogen. To provide highly qualified rice seeds to farms, Korea Seed & Variety Service (KSVS) has been producing and distributing certified rice seeds for producing healthy rice in fields. Therefore, the objective of the study is to summarize the recent progress in molecular identification, fungicide resistance, and the management strategy of bakanae.

• Journal of Microbiology and Biotechnology
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• v.25 no.9
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• pp.1401-1409
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• 2015

The aim of this study was to develop a SYBR Green-based real-time PCR assay for the rapid, specific, and sensitive detection of Acidovorax avenae subsp. citrulli, which causes bacterial fruit blotch (BFB), a serious disease of cucurbit plants. The molecular and serological methods currently available for the detection of this pathogen are insufficiently sensitive and specific. Thus, a novel SYBR Green-based real-time PCR assay targeting the YD-repeat protein gene of A. avenae subsp. citrulli was developed. The specificity of the primer set was evaluated using DNA purified from 6 isolates of A. avenae subsp. citrulli, 7 other Acidovorax species, and 22 of non-targeted strains, including pathogens and non-pathogens. The AC158F/R primer set amplified a single band of the expected size from genomic DNA obtained from the A. avenae subsp. citrulli strains but not from the genomic DNA of other Acidovorax species, including that of other bacterial genera. Using this assay, it was possible to detect at least one genomeequivalents of the cloned amplified target DNA using 5 × 10⁰ fg/µl of purified genomic DNA per reaction or using a calibrated cell suspension, with 6.5 colony-forming units per reaction being employed. In addition, this assay is a highly sensitive and reliable method for identifying and quantifying the target pathogen in infected samples that does not require DNA extraction. Therefore, we suggest that this approach is suitable for the rapid and efficient diagnosis of A. avenae subsp. citrulli contaminations of seed lots and plants.

• Research in Plant Disease
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• v.24 no.4
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• pp.342-347
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• 2018

Bacterial canker caused by Clavibacter michiganensis is considered to be one of the most serious diseases, leading to economic damage to tomato worldwide. Diagnosis of the bacterial canker on tomato is known to be difficult because the causal pathogen is slow-growing on artificial media as well as causes latent infection in tomato. In this study, as a less time-consuming method, a specific primer set was newly designed for rapid detection of C. michiganensis. The method presented here is so simple, easy, and fast that it can be useful and practical in direct detection of the bacterial canker pathogen from tomato plants.

• Journal of Korean Society on Water Environment
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• v.39 no.1
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• pp.102-110
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• 2023

Wastewater Based Epidemiology (WBE) provides useful information not only on the use of illegal drugs in the community, but also on the presence of hygiene and health products and infectious pathogens in sewage facilities. As a consequence of the SARS-CoV-19 virus epidemic in 2019, monitoring the status of the infection is of utmost importance. SARS-CoV-19 was also detected in sewage, and the number and trend of infections in the community suggest that the application of the WBE system would be useful and appropriate. This study introduces a pre-treatment concentration method including viruses in sewage samples. A total of seven methods which were subdivided into methods for adsorption-extraction, ultra-filtration, PEG precipitation, and ultra-centrifugation, and the results for analyzing the recovery rates were included. Meanwhile, it is necessary to pay attention to rapid detection technologies which analyze infectious pathogens at the site of sewage facilities. These can include ELISA, FTIR, SERS, and biosensor based on the detection principle, and the characteristics, advantages, and disadvantages of each were summarized herein. If rapid detection technologies and accurate quantitative analyses are further developed, the use of sewage mechanics in response to pandemic viruses is expected to expand further.

• Journal of Microbiology
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• v.44 no.1
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• pp.92-97
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• 2006

We developed an mPCR assay for the simultaneous detection, in one tube, of Escherichia coli O157:H7, Salmonella spp., Staphylococcus aureus and Listeria monocytogenes using species-specific primers. The mPCR employed the E. coli O157:H7 specific primer Stx2A, Salmonella spp. specific primer Its, S. aureus specific primer Cap8A-B and L. monocytogenes specific primer Hly. Amplification with these primers produced products of 553, 312, 405 and 210 bp, respectively. All PCR products were easily detected by agarose gel electrophoresis, and the sequences of the specific amplicons assessed. Potential pathogenic bacteria, in laboratory-prepared and four commercially available kimchi products, were using this mPCR assay, and the amplicons cloned and sequenced. The results correlated exactly with sequences derived for amplicons obtained during preliminry tests with known organisms. The sensitivity of the assay was determined for the purified pathogen DNAs from four strains. The mPCR detected pathogen DNA at concentrations ranging from approximately 0.45 to $0.05\;pM/{\mu}l$. Thus, this mPCR assay may allow for the rapid, reliable and cost-effective identification of four potentially pathogens present in the mixed bacterial communities of commercially available kimchi.

• ALGAE
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• v.35 no.2
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• pp.133-144
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• 2020

Pythium porphyrae is responsible for devastating outbreaks in seaweed farms of Pyropia, the most valuable cultivated seaweed worldwide. While the genus Pythium contains many well studied pathogens, the genome of P. porphyrae has yet to be sequenced. Here we report the first available gene repertoire of P. porphyrae and a preliminary analysis of pathogenicity-related genes. Using ab initio detection strategies, similarity based and manual annotation, we found that the P. porphyrae gene repertoire is similar to classical phytopathogenic Pythium species. This includes the absence of expanded RxLR effector family and the detection of classical pathogenicity-related genes like crinklers, glycoside hydrolases, cellulose-binding elicitor lectin-like proteins and elicitins. We additionally compared this dataset to the proteomes of 8 selected Pythium species. While 34% of the predicted proteome appeared specific to P. porphyrae, we could not attribute specific enzymes to the degradation of red algal biomass. Conversely, we detected several cellulases and a cutinase conserved with plant-pathogenic Pythium species. Together with the recent report of P. porphyrae triggering disease symptoms on several plant species in lab-controlled conditions, our findings add weight to the hypothesis that P. porphyrae is a reformed plant pathogen.

• The Korean Journal of Pesticide Science
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• v.4 no.2
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• pp.1-10
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• 2000

Fungicide resistance study in Korea is still in its infancy, and most of those resistance studies are largely limited to newness of the detected resistant strains. In future, detection of fungicide-resistant strains has to be based on sensitivity distribution of pathogen populations to certain fungicides, and standard levels of certain fungicides for resistance should be determined under the basis of this data. Most of the early research on fungicide resistance in Korea has overlooked this point, and resulted in inconsistency and confusion for monitoring sensitivity shift of pathogen population among individual researchers. Fungicide resistance detected in vitro tests has to be documented in field trials by examining control efficacy against resistant and wild-type pathogen populations. Resistance detection in wife has to be correlated with lower activity in practice. Using this process, fungicide resistance will have a practical meaning. Fitness evaluation of resistant strains for survival is, in particular, of importance to determine the future stability of the resistance in the pathogen population. In fields, sensitivity change of pathogen populations should be carefully monitored with and without fungicide selection pressures to establish long-term management strategies against fungicide resistance. It is becoming an urgent task to provide information through research for designing and implementing successful counter-measures against fungicide resistance problems in Korea.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.135.1-135
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• 2003

A technique for detection of Xanthomonas axonopodis pv. citri, a causal bacterium of canker on Unshiu orange fruits, was developed using bacteriophage. Procedure for the detection was designed on the basis of the previous reports that one group(CPI) of X. axonopodis pv. citri bacteriophage and corresponding two Iysotypes distributed in Korea. First, fruit surface was washed with sterile distilled water and pellet was obtained from centrifugation. The pellet was resuspended in Wakimoto's potato semi-synthetic broth medium and divided equally into two parts. One part was heated in boiling water to kill bacterial cells. Bacteriophages(CP$_1$) were respectively added into two parts and 0.1 ml from each part was mixed with soft agar medium. After incubation for 18 hrs at 25$^{\circ}C$, the causal bacterium of canker was determined based on plaques formed on the medium. This procedure can be effectively used for detection of living bacterial pathogen on fruit surfaces of Unshiu orange.