• The Plant Pathology Journal
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• v.33 no.2
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• pp.184-192
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• 2017

Tylenchulus semipenetrans is an important and widespread plant-parasitic nematode of citrus worldwide and can cause citrus slow decline disease leading to significant reduction in tree growth and yield. Rapid and accurate detection of T. semipenetrans in soil is important for the disease forecasting and management. In this study, a loop-mediated isothermal amplification (LAMP) assay was developed to detect T. semipenetrans using DNA extracted from soil. A set of five primers was designed from the internal transcribed spacer region (ITS1) of rDNA, and was highly specific to T. semipenetrans. The LAMP reaction was performed at $63^{\circ}C$ for 60 min. The LAMP product was visualized directly in one reaction tube by adding SYBR Green I. The detection limit of the LAMP assay was $10^{-2}J2/0.5g$ of soil, which was 10 times more sensitive than conventional PCR ($10^{-1}J2/0.5g$ of soil). Examination of 24 field soil samples revealed that the LAMP assay was applicable to a range of soils infested naturally with T. semipenetrans, and the total assay time was less than 2.5 h. These results indicated that the developed LAMP assay is a simple, rapid, sensitive, specific and accurate technique for detection of T. semipenetrans in field soil, and contributes to the effective management of citrus slow decline disease.

• Journal of Ginseng Research
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• v.43 no.1
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• pp.1-9
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• 2019

Background: Korean ginseng is an important cash crop in Asian countries. However, plant yield is reduced by pathogens. Among the Ilyonectria radicicola-species complex, I. mors-panacis is responsible for root-rot and replant failure of ginseng in Asia. The development of new methods to reveal the existence of the pathogen before cultivation is started is essential. Therefore, a quantitative real-time polymerase chain reaction method was developed to detect and quantify the pathogen in ginseng soils. Methods: In this study, a species-specific histone H3 primer set was developed for the quantification of I. mors-panacis. The primer set was used on DNA from other microbes to evaluate its sensitivity and selectivity for I. mors-panacis DNA. Sterilized soil samples artificially infected with the pathogen at different concentrations were used to evaluate the ability of the primer set to detect the pathogen population in the soil DNA. Finally, the pathogen was quantified in many natural soil samples. Results: The designed primer set was found to be sensitive and selective for I. mors-panacis DNA. In artificially infected sterilized soil samples, using quantitative real-time polymerase chain reaction the estimated amount of template was positively correlated with the pathogen concentration in soil samples ($R^2=0.95$), disease severity index ($R^2=0.99$), and colony-forming units ($R^2=0.87$). In natural soils, the pathogen was recorded in most fields producing bad yields at a range of $5.82{\pm}2.35pg/g$ to $892.34{\pm}103.70pg/g$ of soil. Conclusion: According to these results, the proposed primer set is applicable for estimating soil quality before ginseng cultivation. This will contribute to disease management and crop protection in the future.

• Korean Journal of Microbiology
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• v.43 no.3
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• pp.201-209
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• 2007

The CFDA (6-carboxyfluorescein diacetate) direct viable count method and plate count (PC) method using conventional nutrient broth (NB) medium and $10^{-2}$ diluted NB (DNB) medium were applied to samples collected from Mt. Yongdoo In Andong, in an effect to determine the number of living bacteria pine mushroom forest soil. The number of living bacteria determined via plate count in NB medium comprised $5{\sim}8%$ of the CFDA direct viable count, and the bacteria in the DNB medium comprised $40{\sim}47%$. This result indicated that viable but nonculturable (VBNC) bacteria existed in the pine mushroom forest soil at a high percentage. The phylogenetic characteristics of the VBNC bacterial populations in the samples of pine mushroom (Tricholoma matsutake) forest soil were analyzed via the direct extraction of DNA and 16S rDNA-ARDRA. The 115 clones from pine mushroom forest soil were clustered into 31 different RFLP phylotypes by ARDRA. Based on the 16S rDNA sequences, the 31 ARDRA clusters were classified into 6 phylogenetic groups: ${\alpha}-,\;{\beta}-,\;{\gamma}-Proteobacteria$, Acidobacteria, Actinobacteria and Firmicutes. Among these bacterial populations, approximately 85% were classified as members of phylum Acidobacteria. The Acidobacteria phylum was shown to exist abundantly in the pine mushroom forest soil.

• Journal of Species Research
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• v.12 no.4
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• pp.307-312
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• 2023

Oxytricha multilineata, Mixophrya pantanalensis pantanalensis, and Caudiurostyla sinensis were isolated from soil samples collected from Cheongju-si and Yeoju-si, confirmed as new to South Korea. Oxytricha multilineata was distinguished from other congeners by seven dorsal kineties and dorsal bristles about 15 ㎛ long. Mixophrya pantanalensis pantanalensis was characterized by five to seven lithosomes and six dorsal kineties. Caudiurostyla sinensis was characterized by colorless cortical granules present, 10-14 midventral pairs, 7-9 left and 6-9 right marginal rows and four or five dorsal kineties. We determined the ribosomal DNA sequences (including 18S rDNA, ITS1, 5.8S rDNA, ITS2, and partial 28S rDNA) from above three species. And the genetic distances were compared with their congeners.

• Journal of Korean Society of Forest Science
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• v.96 no.4
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• pp.425-431
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• 2007

To investigate genetic diversity and PCR detection of Rhizina undulata, PCR detection and sequence analysis of rDNA ITS region of R. undulata in soil were analyzed and developed. The length of partial 18S rDNA from four R. undulata isolates were 1,375 nt. The sequence similarity of R. undulata isolates was 100%. The rDNA ITS regions of R. undulata isolates were 585 nt long. Nucleotide sequencing of the ITS regions showed that PDK-1, PTT-1 and PDJ-9 isolates had 100% sequence identity. But, PDS-5 isolate differed from the three isolates by two nucleotide substitution. R. undulata-specific primers designed by the sequence of ITS region were used in PCR detection of R. undulata. PCR products about 525 bp size, which is specific to R. undulata, were amplified from total DNAs of R. undulata isolates. To assay the sensitivity of PCR detection by R. undulata ITS-specific primer, purely cultured mycelial suspension of R. undulata was serially diluted and mixed with 100g of sterile sandy loam soil, respectively. And then, PCR products of total DNAs extracted from each mycelium-soil mixtures were analysed. The PCR protocol could detected up to 1ng mycelium of R. undulata within 100g of soil.

• Korean Journal of Heritage: History & Science
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• v.47 no.4
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• pp.92-105
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• 2014

Microbial characteristics of bacterial population were investigated in human remains and soil inside the bones in excavated grave no.4 and no.5 at Keundokgol site, Osu-ri, Buyeo. Phylogenetic characteristics of bacterial populations were analyzed by direct extracting of ancient DNA. In this study, based on the 16S rDNA sequences, in case of grave no.4, 319s from human remain were classified into 11 phyla, and 462s from soil were classified into 16 phyla. In case of grave no.5, 271s from human remain were classified into 10 phyla, and 497s from soil were classified into 11 phyla. Especially, Actinobacteria phylogenetic group are dominant group of bacterial populations in grave no.4 and no.5. Also, most of these were analyzed uncultured group. Thus, the discovery of a diversely microbial community and uncultured group was thought to be due to the specificity of the sample. Conclusively the general excavated human bones were contaminated with soil bacteria species their near around. This results contribute to preservation and management of ancient human bone from archaeological sites.

• Journal of the Korea Organic Resources Recycling Association
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• v.13 no.2
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• pp.85-90
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• 2005

Changes of biochemical and genetic diversity of microbial communities in the soil damaged by a forest fire were analyzed. Soil samples were collected from Gangnung area where a forest fire was broken out in 2000. Two soil samples were from the burnt area, one from the naturally restoring soil (NS) and the other from the artificially restoring soil (AS). A normal, unaffected soil sample (US) was also included as a control. For the biochemical diversity, each sample was directly applied to the BIOLOG system, and the cluster analysis through a statistic process (SPSS) were performed. Genetic diversity was analyzed through DGGE using 16S-rDNA amplified from soil DNA. Among the samples tested, top soils of US and NS, and sub soil of NS revealed more than 70% of the similarity value in biochemical diversity. In case of genetic diversity, however, the similarity value was found to be in the range of 53% to 68% in all samples. This result indicates that the biochemical diversity is not always correlated with the genetic diversity in the analysis of microbial communities.

• Korean Journal of Heritage: History & Science
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• v.41 no.1
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• pp.99-108
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• 2008

The paleogenetic analysis has become an increasingly important subject of archaeological, anthropological, biological as well as public interest. Recently, scientific research for human skeletal remains was more activated because of increasing awareness of the valuable archaeological information by the ancient DNA analysis. State of preservation of organic remains vary in different soil and burying environmental condition. Almost all available tissue disappear to analysis ancient DNA of bone in acidic soil caused by climate and geological features in Korea. Many preserved human remains excavated in the 'Heogwakmyo'(limelayered tomb of Chosun Dynasty Period) is able to explain through the relationship between burial conditions and bone survival form the burial method and ceremony. Ancient DNA analysis of excavated human bone form ancient tomb requires to remove contaminants such as microorganism's DNA and soil components that affect authentic results. Particularly, contamination control of contemporary human DNA is major serious problem and should verified by criteria of authenticity. In order to understand migration and culture of ancient population, when possible, ancient DNA studies needs to go abreast both radiocarbon and stable isotope studies because the dietary inferences will suggest ancient subsistence and settlement patterns. Also when the paleogenetic research supported with the arts and humanities research such as physical anthropology and archaeology, more valuable ancient genetic information is providing a unique results about evolutionary and population genetics studies to reconstruct the past.

• Proceedings of the Korean Society of Soil Zoology Conference
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• 1998.10a
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• pp.24-25
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• 1998

• The Plant Pathology Journal
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• v.38 no.3
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• pp.229-238
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• 2022

Fusarium wilt caused by Fusarium oxysporum f. sp. niveum (Fon) is the most serious soil-borne disease in the world and has become the main limiting factor of watermelon production. Reliable and quick detection and quantification of Fon are essential in the early stages of infection for control of watermelon Fusarium wilt. Traditional detection and identification tests are laborious and cannot efficiently quantify Fon isolates. In this work, a real-time polymerase chain reaction (PCR) assay has been described to accurately identify and quantify Fon in watermelon plants and soil. The FONRT-18 specific primer set which was designed based on identified specific sequence amplified a specific 172 bp band from Fon and no amplification from the other formae speciales of Fusarium oxysporum tested. The detection limits with primers were 1.26 pg/µl genomic DNA of Fon, 0.2 pg/ng total plant DNA in inoculated plant, and 50 conidia/g soil. The PCR assay could also evaluate the relationships between the disease index and Fon DNA quantity in watermelon plants and soil. The assay was further used to estimate the Fon content in soil after disinfection with CaCN₂. The real-time PCR method is rapid, accurate and reliable for monitoring and quantification analysis of Fon in watermelon plants and soil. It can be applied to the study of disease diagnosis, plant-pathogen interactions, and effective management.