• Journal of Microbiology and Biotechnology
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• 제12권6호
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• pp.989-992
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• 2002

Sweet persimmon (Diospyros kaki Thunb.) is widely cultivated in the southern part of Korea and its cultivation is increasing. However, anthracnose disease caused by Colletotricuhum species is one of the major hinderances to the cultivation and production of sweet persimmon. Therefore, in the current study, PCR was used to specifically detect Colletotrichum spp., based on the sequences of the ITS II regions in the rDNA. Using the sequence data, CO-1 was designated to detect Colletotrichum together the with ITS 4 primer. The result showed that a single segment of ca. 500 bp was observed only in Colletotrichum, but not in any other fungal and bacterial isolates. The annealing temperatures and template DNA quantites were also investigated to identify optimal conditions for detection. Using these species-specific primers, a unique band was obtained at annealing temperatures ranging from $55^{\circ}C\;and\;61^{\circ}C$ and template DNA levels from 10 pg- $10{\mu}g$.

• The Plant Pathology Journal
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• 제34권6호
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• pp.575-579
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• 2018

Apple stem grooving virus (ASGV) is considered to cause the most economically important viral disease in pears in Korea. The current PCR-based methods used to diagnose ASGV are time-consuming in terms of target detection. In this study, a novel assay for specific ASGV detection that is based on reverse transcription-recombinase polymerase amplification is described. This assay has been shown to be reproducible and able to detect as little as $4.7ng/{\mu}l$ of purified RNA obtained from an ASGV-infected plant. The major advantage of this assay is that the reaction for the target virus is completed in 1 min, and amplification only requires an incubation temperature of $42^{\circ}C$. This assay is a promising alternative method for pear breeding programs or virus-free certification laboratories.

• The Plant Pathology Journal
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• 제31권1호
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• pp.90-96
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• 2015

Garlic generally becomes coinfected with several types of viruses belonging to the Potyvirus, Carlavirus, and Allexivirus genera. These viruses produce characteristically similar symptoms, they cannot be easily identified by electron microscopy (EM) or immunological detection methods, and they are currently widespread around the world, thereby affecting crop yields and crop quality adversely. For the early and reliable detection of garlic viruses, virus-specific sets of primers, including species-specific and genus-specific primers were designed. To effectively detect the twelve different types of garlic viruses, primer mixtures were tested and divided into two independent sets for multiplex polymerase chain reaction (PCR). The multiplex PCR assays were able to detect specific targets up to the similar dilution series with monoplex reverse transcription (RT)-PCR. Seventy-two field samples collected by the Gyeongbuk Agricultural Technology Administration were analyzed by multiplex RT-PCR. All seventy two samples were infected with at least one virus, and the coinfection rate was 78%. We conclude that the simultaneous detection system developed in this study can effectively detect and differentiate mixed viral infections in garlic.

• 식물병연구
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• 제7권3호
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• pp.123-133
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• 2001

Many pathovars of the species Pseudomonas syringae are known to produce different phytotoxins as secondary metabolites. Although phytotoxins generally enhance the virulence of P. syringae, they are not required for pathogenesis. Among the phytotoxins produced by P. syringae, lipodepsipeptides, coronatine, phaseolotoxin, and tabtoxin are the most well-known toxins which have been intensively studied for their structure, mode of action, biosynthesis, and regulation. A polymerase chain reaction (PCR) technique that amplifies a segment of the phytotoxin gene cluster using a primer set has been developed in recent years. This method offers the advantages of speed and sensitivity compared to the approaches based on physiological and biochemical methods. PCR detection of genes involved in the production of toxins could be exploited for early diagnosis of plant diseases caused by P. syringae pathovars.

• The Plant Pathology Journal
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• 제32권5호
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• pp.396-406
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• 2016

Continuous supplementation of mineral nutrients and salicylic acid (SA) as foliar application could improve efficacy in controlling basal stem rot (BSR) disease in oil palm seedling. It is revealed from the results that the highest disease severity index (58.3%) was recorded in T8 treatments at 9 months after inoculation. The best disease control was achieved by T7 treatments (calcium/copper/SA [Ca/Cu/SA]) (5.0%) followed by T1 (5.5%), T5 (5.8%), T3 (8.3%), T6 (8.3%), T4 (13.3%), and T2 (15.8%) treatments. Continuous supplementation of Ca/Cu/SA was found to be the most effective in controlling the disease and the high performance liquid chromatography results showed the detection of ergosterol at very low concentration in the treated samples. Moreover, the transmission electron microscopy analysis results clearly indicated that T7 treatment was also enhancing lignification, which was responsible for the thickness of the secondary cell walls and middle lamella compared to untreated samples. It was therefore, concluded that continuous supplementation of minerals nutrients and SA could effectively suppress disease severity by reducing ergosterol activity and also improve the process of lignification in the treated plants. Furthermore, this treatment also managed to delay the onset of BSR symptoms and promote the growth of the seedlings and eventually suppress the BSR disease.

• The Plant Pathology Journal
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• 제32권5호
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• pp.414-422
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• 2016

Eucalyptus dieback disease, caused by Cylindrocladium scoparium, has occurred in last few years in large Eucalyptus planting areas in China and other countries. Rapid, simple, and reliable diagnostic techniques are desired for the early detection of Eucalyptus dieback of C. scoparium prior to formulation of efficient control plan. For this purpose, three PCR-based methods of nested PCR, multiplex PCR, loop-mediated isothermal amplification (LAMP) were developed for detection of C. scoparium based on factor 1-alpha (tef1) and beta-tubulin gene in this study. All of the three methods showed highly specific to C. scoparium. The sensitivities of the nested PCR and LAMP were much higher than the multiplex PCR. The sensitivity of multiplex PCR was also higher than regular PCR. C. scoparium could be detected within 60 min from infected Eucalyptus plants by LAMP, while at least 2 h was needed by the rest two methods. Using different Eucalyptus tissues as samples for C. scoparium detection, all of the three PCR-based methods showed much better detection results than regular PCR. Base on the results from this study, we concluded that any of the three PCR-based methods could be used as diagnostic technology for the development of efficient strategies of Eucalyptus dieback disease control. Particularly, LAMP was the most practical method in field application because of its one-step and rapid reaction, simple operation, single-tube utilization, and simple visualization of amplification products.

• 식물병연구
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• 제27권2호
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• pp.79-83
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• 2021

The aim of the present study was to develop a sensitive and specific detection method for the rapid detection of apple scar skin viroid (ASSVd) in apple leaves. The resulting reverse transcription recombinase polymerase amplification (RT-RPA) assay can be completed in 10 min at 42℃, is 10 times more sensitive than conventional reverse transcription polymerase chain reaction, and can specifically amplify ASSVd without any cross-reactivity with other common apple viruses, including apple stem grooving virus, apple stem pitting virus, and apple chlorotic leaf spot virus. The reliability of the RT-RPA assay was assessed, and the findings suggested that it can be successfully utilized to detect ASSVd in field-collected samples. The RT-RPA assay developed in the present study provides a potentially valuable means for improving the detection of ASSVd in viroid-free certification programs, especially in resource-limited conditions.

• 식물병연구
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• 제23권1호
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• pp.65-68
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• 2017

2단계의 nested PCR 방법을 이용하여 보리 및 벼 재배 토양에서 Barley yellow mosaic virus (BaYMV)를 검출하였다. BaYMV 분절 RNA1 외피단백질 영역의 특이 프라이머로 1차 PCR을 하고 내부서열로부터 작성된 프라이머로 2차 PCR을 실시하여 확보된 372 bp의 PCR 산물이 BaYMV 외피단백질 영역과 98%-100% 염기서열이 일치하여 BaYMV를 검출할 수 있음을 확인하였다. 이 결과는 토양으로부터 BaYMV 검출에 관한 최초의 보고이며 토양전염성 바이러스의 정확한 진단과 예찰에 적용될 수 있을 것으로 생각한다.

• 품질경영학회지
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• 제48권4호
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• pp.581-596
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• 2020

Purpose: purpose of this study was to propose the web application platform which can be to detect and discriminate various diseases and pest of tomato plant based on the large amount of disease image data observed in the facility or the open field. Methods: The deep learning algorithms uesed at the web applivation platform are consisted as the combining form of Faster R-CNN with the pre-trained convolution neural network (CNN) models such as SSD_mobilenet v1, Inception v2, Resnet50 and Resnet101 models. To evaluate the superiority of the newly proposed web application platform, we collected 850 images of four diseases such as Bacterial cankers, Late blight, Leaf miners, and Powdery mildew that occur the most frequent in tomato plants. Of these, 750 were used to learn the algorithm, and the remaining 100 images were used to evaluate the algorithm. Results: From the experiments, the deep learning algorithm combining Faster R-CNN with SSD_mobilnet v1, Inception v2, Resnet50, and Restnet101 showed detection accuracy of 31.0%, 87.7%, 84.4%, and 90.8% respectively. Finally, we constructed a web application platform that can detect and discriminate various tomato deseases using best deep learning algorithm. If farmers uploaded image captured by their digital cameras such as smart phone camera or DSLR (Digital Single Lens Reflex) camera, then they can receive an information for detection, identification and disease control about captured tomato disease through the proposed web application platform. Conclusion: Incheon Port needs to act actively paying.

• The Plant Pathology Journal
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• 제34권2호
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• pp.104-112
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• 2018

Accurate and rapid detection of bacterial plant pathogen is the first step toward disease management and prevention of pathogen spread. Bacterial plant pathogens Clavibacter michiganensis subsp. nebraskensis (Cmn), Pantoea stewartii subsp. stewartii (Pss), and Rathayibacter tritici (Rt) cause Goss's bacterial wilt and blight of maize, Stewart's wilt of maize and spike blight of wheat and barley, respectively. The bacterial diseases are not globally distributed and not present in Korea. This study adopted comparative genomics approach and aimed to develop specific primer pairs to detect these three bacterial pathogens. Genome comparison among target pathogens and their closely related bacterial species generated 15-20 candidate primer pairs per bacterial pathogen. The primer pairs were assessed by a conventional PCR for specificity against 33 species of Clavibacter, Pantoea, Rathayibacter, Pectobacterium, Curtobacterium. The investigation for specificity and sensitivity of the primer pairs allowed final selection of one or two primer pairs per bacterial pathogens. In our assay condition, a detection limit of Pss and Cmn was $2pg/{\mu}l$ of genomic DNA per PCR reaction, while the detection limit for Rt primers was higher. The selected primers could also detect bacterial cells up to $8.8{\times}10^3cfu$ to $7.84{\times}10^4cfu$ per gram of grain seeds artificially infected with corresponding bacterial pathogens. The primer pairs and PCR assay developed in this study provide an accurate and rapid detection method for three bacterial pathogens of grains, which can be used to investigate bacteria contamination in grain seeds and to ultimately prevent pathogen dissemination over countries.