• Journal of Veterinary Science
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• 제21권2호
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• pp.22.1-22.9
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• 2020

Rabid raccoon dogs (Nyctereutes procyonoides koreensis) have been responsible for animal rabies in South Korea since the 1990s. A recombinant rabies vaccine strain, designated as ERAGS, was constructed for use as a bait vaccine. Therefore, new means of differentiating ERAGS from other rabies virus (RABV) strains will be required in biological manufacturing and diagnostic service centers. In this study, we designed two specific primer sets for differentiation between ERAGS and other RABVs based on mutation in the RABV glycoprotein gene. Polymerase chain reaction analysis of the glycoprotein gene revealed two DNA bands of 383 bp and 583 bp in the ERAGS strain but a single DNA band of 383 bp in the field strains. The detection limits of multiplex reverse transcription polymerase chain reaction (RT-PCR) were 80 and 8 FAID₅₀/reaction for the ERAGS and Evelyn-Rokitnicki-Abelseth strains, respectively. No cross-reactions were detected in the non-RABV reference viruses, including canine distemper virus, parvovirus, canine adenovirus type 1 and 2, and parainfluenza virus. The results of multiplex RT-PCR were 100% consistent with those of the fluorescent antibody test. Therefore, one-step multiplex RT-PCR is likely useful for differentiation between RABVs with and those without mutation at position 333 of the RABV glycoprotein gene.

• The Plant Pathology Journal
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• 제34권6호
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• pp.490-498
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• 2018

Panicle blight and seed rot disease caused mainly by Burkholderia glumae and Burkholderia gladioli is threatening rice cultivation worldwide. The bacteria have been reported as seed-borne pathogens from rice. Accurate detection of both pathogens on the seeds is very important for limiting the disease dissemination. Novel primer pairs targeting specific molecular markers were developed for the robust detection of B. glumae and B. gladioli. The designed primers were specific in detecting the target species with no apparent cross-reactions with other related Burkholderia species at the expected product size. Both primer pairs displayed a high degree of sensitivity for detection of B. glumae and B. gladioli separately in monoplex PCR or simultaneously in duplex PCR from both extracted gDNA and directly preheated bacterial cell suspensions. Limit of detection was as low as 0.1 ng of gDNA of both species and $3.86{\times}10^2cells$ for B. glumae and $5.85{\times}10^2cells$ for B. gladioli. On inoculated rice seeds, the designed primers could separately or simultaneously detect B. glumae and B. gladioli with a detection limit as low as $1.86{\times}10^3cells$ per rice seed for B. glumae and $1.04{\times}10^4cells$ per rice seed of B. gladioli. The novel primers maybe valuable as a more sensitive, specific, and robust tool for the efficient simultaneous detection of B. glumae and B. gladioli on rice seeds, which is important in combating rice panicle blight and seed rot by early detection and confirmation of the dissemination of pathogen-free rice seeds.

• Journal of Veterinary Science
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• 제21권4호
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• pp.63.1-63.9
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• 2020

Background: Canine adenovirus type 2 (CAV-2) induces infectious laryngotracheitis in members of the family Canidae, including dogs. To date, no ELISA kits specific for CAV-2 antibody have been commercialized for dogs in Korea. Objectives: We aimed to develop new indirect enzyme-linked immunosorbent assay (I-ELISA) to perform rapid, accurate serological surveys of CAV-2 in dog serum samples. Methods: In total, 165 serum samples were collected from dogs residing in Chungbuk and Gyeongbuk provinces between 2016 and 2018. The Korean CAV-2, named the APQA1701-40P strain, was propagated in Madin-Darby canine kidney cells and purified in an anion-exchange chromatography column for use as an antigen for I-ELISA. The virus-neutralizing antibody titers of CAV-2 in the dog sera were measured by virus neutralization (VN) test. Results: We compared the results obtained between the VN and new I-ELISA tests. The sensitivity, specificity, and accuracy of new I-ELISA were 98.6%, 86.4% and 97.0% compared with VN test, respectively. New I-ELISA was significantly correlated with VN (r = 0.91). Conclusions: These results indicate that new I-ELISA is useful for sero-surveillance of CAV-2 in dog serum.

• The Plant Pathology Journal
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• 제38권3호
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• pp.182-193
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• 2022

Turfgrass, the most widely grown ornamental crop, is severely affected by fungal pathogens including Sclerotinia homoeocarpa, Rhizoctonia solani, and Magnaporthe poae. At present, turfgrass fungal disease management predominantly relies on synthetic fungicide treatments. However, the extensive application of fungicides to the soil increases residual detection frequency, raising concerns for the environment and human health. The bacterial volatile compound, 2,3-butanediol (BDO), was found to induce plant resistance. In this study, we evaluated the disease control efficacy of a combination of stereoisomers of 2,3-BDO and commercial fungicides against turfgrass fungal diseases in both growth room and fields. In the growth room experiment, the combination of 0.9% 2R,3R-BDO (levo) soluble liquid (SL) formulation and 9% 2R,3S-BDO (meso) SL with half concentration of fungicides significantly increased the disease control efficacy against dollar spot and summer patch disease when compared to the half concentration of fungicide alone. In field experiments, the disease control efficiency of levo 0.9% and meso 9% SL, in combination with a fungicide, was confirmed against dollar spot and large patch disease. Additionally, the induction of defense-related genes involved in the salicylic acid and jasmonic acid/ethylene signaling pathways and reactive oxygen species detoxification-related genes under Clarireedia sp. infection was confirmed with levo 0.9% and meso 9% SL treatment in creeping bentgrass. Our findings suggest that 2,3-BDO isomer formulations can be combined with chemical fungicides as a new integrated tool to control Clarireedia sp. infection in turfgrass, thereby reducing the use of chemical fungicides.

• The Plant Pathology Journal
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• 제31권1호
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• pp.25-32
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• 2015

Pseudomonas coronafaciens causes halo blight on oats and is a plant quarantine bacterium in many countries, including the Republic of Korea. Using of the certificated seed is important for control of the disease. Since effective detection method of P. coronafaciens is not available yet, PCR and TaqMan PCR assays for specific detection of P. coronafaciens were developed in this study. PCR primers were designed from the draft genome sequence of P. coronafaciens LMG 5060 which was obtained by the next-generation sequencing in this study. The PCR primer set Pc-12-F/Pc-12-R specifically amplified 498 bp from the 13 strains of P. coronafaciens isolated in the seven different countries (Canada, Japan, United Kingdom, Zimbabwe, Kenya, Germany, and New Zealand) and the nested primer set Pc-12-ne-F/Pc-12-ne-R specifically amplified 298 bp from those strains. The target-size PCR product was not amplified from the non-target bacteria with the PCR and nested primer sets. TaqMan PCR with Pc-12-ne-F/Pc-12-ne-R and a TaqMan probe, Pc-taqman, which were designed inside of the nested PCR amplicon, generated Ct values which in a dose-dependent manner to the amount of the target DNA and the Ct values of all the P. coronafaciens strains were above the threshold Ct value for positive detection. The TaqMan PCR generated positive Ct values from the seed extracts of the artificially inoculated oat seeds above 10 cfu/ml inoculation level. PCR and TaqMan PCR assays developed in this study will be useful tools to detect and identify the plant quarantine pathogen, P. coronafaciens.

• Journal of Biosystems Engineering
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• 제33권6호
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• pp.446-452
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• 2008

Machine vision system was used for analyzing leaf color disorders of tomato plants in a greenhouse. From the day when a few leave of tomato plants had started to wither, a series of images were captured by 4 times during 14 days. Among several color image spaces, Saturation frame in HSI color space was adequate to eliminate a background and Hue frame was good to detect infected disease area and tomato fruits. The processed image ($G{\sqcup}b^*$ image) by OR operation between G frame in RGB color space and $b^*$ frame in $La^*b^*$ color space was useful for image segmentation of a plant canopy area. This study calculated a ratio of the infected area to the plant canopy and manually analyzed leaf color disorders through an image segmentation for Hue frame of a tomato plant image. For automatically analyzing plant leave disease, this study selected twenty-seven color patches on the calibration bars as the corresponding to leaf color disorders. These selected color patches could represent 97% of the infected area analyzed by the manual method. Using only ten color patches among twenty-seven ones could represent over 85% of the infected area. This paper showed a proposed machine vision system may be effective for evaluating various leaf color disorders of plants growing in a greenhouse.

• 식물병연구
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• 제18권2호
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• pp.129-132
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• 2012

Citrus bacterial canker is an economically important disease affecting citrus production in many citrusgrowing areas and several pathotypes have been recognized within the Xanthomonas pathogens causing canker. In view of the containment of the disease, accurate identification of the causal bacterium is important. In this study, triplex PCR method was developed by using the previously reported primers. Two groups of primer combination, such as, one group including primers 2/3, J-pth1/J-pth2 and XACF/XACR, and another group 2/3, J-pth1/J-pth2 and Xac01/Xac02, were suitable for the detection and differentiation of X. a. pv. citri $A^w$, X. a. pv. aurantifolii B and C, and X. a. pv. citrumelo E strains. Moreover, the primer combination of Xac01 and J-pth2 promised us to use as a specific primer set to detect X. a. pv. citrumelo E strain. The PCR methods developed in this study could be used for the rapid differentiation of Xanthomonas pathotypes of citrus.

• 한국동물위생학회지
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• 제37권4호
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• pp.263-271
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• 2014

In this study, we developed a cost and time saving one-step multiplex RT-PCR for the simultaneous detection and differentiation of swine influenza viruses (SIV) and 2009 pandemic influenza H1N1 virus (pH1N1). The one-step multiplex RT-PCR using four sets of primer was confirmed to be capable of detection of all SIV subtypes and differential diagnosis of major SIV subtype H1, H3 and pH1N1 on individual or mixed viral culture samples. The sensitivity of the multiplex RT-PCR was determined to be at least $2^{-6}$ $HA/25{\mu}L$ of the presented SIVs, providing sufficient efficacy for a routine SIV monitoring in diagnostic laboratories. In addition, compared with the conventional RT-PCR methods that cannot avoid the carryover DNA contamination, the developed RT-PCR applied with the uracil DNA glycosylase (UNG) system was proven to prevent a false positive reaction by carryover contamination of the pre-amplified DNA. In conclusion, the one-step RT-PCR with UNG system could be applicable to detect and differentiate of SIV from the viral cultures without worry of carryover DNA contamination in clinical laboratories.

• International Journal of Computer Science & Network Security
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• 제23권9호
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• pp.77-90
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• 2023

Today, crops face many characteristics/diseases. Insect damage is one of the main characteristics/diseases. Insecticides are not always effective because they can be toxic to some birds. It will also disrupt the natural food chain for animals. A common practice of plant scientists is to visually assess plant damage (leaves, stems) due to disease based on the percentage of disease. Plants suffer from various diseases at any stage of their development. For farmers and agricultural professionals, disease management is a critical issue that requires immediate attention. It requires urgent diagnosis and preventive measures to maintain quality and minimize losses. Many researchers have provided plant disease detection techniques to support rapid disease diagnosis. In this review paper, we mainly focus on artificial intelligence (AI) technology, image processing technology (IP), deep learning technology (DL), vector machine (SVM) technology, the network Convergent neuronal (CNN) content Detailed description of the identification of different types of diseases in tomato and potato plants based on image retrieval technology (CBIR). It also includes the various types of diseases that typically exist in tomato and potato. Content-based Image Retrieval (CBIR) technologies should be used as a supplementary tool to enhance search accuracy by encouraging you to access collections of extra knowledge so that it can be useful. CBIR systems mainly use colour, form, and texture as core features, such that they work on the first level of the lowest level. This is the most sophisticated methods used to diagnose diseases of tomato plants.

• 식물병연구
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• 제17권3호
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• pp.376-380
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• 2011

P. syringae pv. tomato는 토마토에서 bacterial speck병을 일으키는 종자전염 세균으로, 감수성 품종에서 주로 발병하여 경제적으로 큰 손실을 입힌다. 따라서 P. syringae pv. tomato는 한국을 비롯한 많은 나라에서 식물 검역대상 세균으로 지정하여 관리되고 있다. 본 연구에서 우리는 토마토 종자로부터 PCR을 이용하여 Pst를 검출할 수 있는 방법을 개발하였다. P. syringae pv. tomato의 hrpZ 유전자에서 특이적인 프라이머를 개발하였다. 개발된 프라이머는 P. syringae pv. tomato에서만 501 bp 크기의 특이적 DNA를 증폭하였으며, P. syringae pv. glycinea, P. syringae pv. maculicola, P. syringae pv. atropurpurea, P. syringae pv. morsprunorum와 같은 다른 토마토 세균병원균과 P. syringae pathovar 균주들에서는 증폭되지 않았다. Nested PCR 프라이머를 이용한 PCR에서도 오직 P. syringae pv. tomato에서만 119 bp 크기의 특이적 DNA가 증폭되었고, 토마토 종자에서 P. syringae pv. tomato을 정확하고 민감하게 검출하였다. 본 연구는 현재까지 사용되고 있는 Pst의 검출방법의 민감도를 비교한 최초의 보고로 본 연구에서 개발된 PCR방법들은 토마토 종자에서 Pst을 검출하는 매우 유용한 방법으로 생각된다.