Yong Ju Jin's research while affiliated with Institute of Agricultural Sciences and other places

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Publications (9)


Fig. 1. Sampling of adjacent pepper plant rhizosphere soils. A map of Seven pepper farms (4, 5, 6, 10, 11, 13, and 15) in two regions (Imsil and Sunchang) where sampling was performed. Images of neighboring healthy plants (H) and diseased plants (D).
Fig. 3. Bacterial community structure in pepper rhizosphere. (A) A percentage of bacterial community abundance at class level in both soil groups with (D) and without (H) disease outbreaks of pepper rhizosphere. Relative abundance is calculated by averaging the abundance of duplicate samples from each soil group in the pepper rhizosphere. (B) Differences in abundance of major microbiome in pepper rhizosphere microbiome. Data is displayed as a relative abundance (%). Each point corresponds to a single sample. (C) Changes in the alpha diversity of the pepper rhizosphere microbiome.
Changes in the Composition and Microbial Community of the Pepper Rhizosphere in Field with Bacterial Wilt Disease
  • Article
  • Full-text available

December 2022

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73 Reads

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1 Citation

The Plant Pathology Journal

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Ju Hee An

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Bacterial wilt caused by Ralstonia solanacearum is considered one of the most harmful diseases of pepper plants. Recently, research on plant disease control through the rhizosphere microbiome has been actively conducted. In this study, the relationship with disease occurrence between the neighboring plant confirmed by analyzing the physicochemical properties of the rhizosphere soil and changes in the microbial community. The results confirmed that the microbial community changes significantly depending on the organic matters, P<sub>2</sub>O<sub>5</sub>, and clay in the soil. Despite significant differences in microbial communities according to soil composition, Actinobacteriota at the phylum level was higher in healthy plant rhizosphere (mean of relative abundance, D: 8.05 ± 1.13; H: 10.06 ± 1.59). These results suggest that Actinobacteriota may be associated with bacterial wilt disease. In this study, we present basic information for constructing of healthy soil in the future by presenting the major microbial groups that can suppress bacterial wilt.

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Fig. 1. Polymerase chain reaction amplification of the HAD family hydrolase gene with the primers. Lane M is the size marker (1 kb DNA plus ladder; Inclon Co., Ltd.) while lanes 1 and 6 were Pectobacterium odoriferum strains, and lanes 7 to 29 included strains from other Pectobacterium, Dickeya, Pantoea, Erwinia, and Musicola species as listed in Table 1.
Fig. 2. Species-specificity assay of the PeOd360F/R primer using real-time polymerase chain reaction. The bacterial strains in lanes 1-29 are described in Table 1. Lane 30, distilled water (n = 3). (A) Amplification curves. (B) Melting curve. (C) Melting peak. The results of the melting peak analysis indicated that the amplified products have a melting temperature (Tm) of approximately 88°C. RFU, relative fluorescence units.
Fig. 3. Standard curve analysis of real-time polymerase chain reaction performed using a dilution series. A standard curve was generated from the threshold cycles (Ct) also known as crossing points of the Pectobacterium oderiferum standard dilutions. (A) Genomic DNA. (B) Cloned DNA. (C) Bacterial cell suspension. The R 2 value of each curve was >0.99.
Primer sequences, their targets, and the annealing temperatures used in Pectobacterium odoriferum PCR screens
A New Approach Using the SYBR Green-Based Real-Time PCR Method for Detection of Soft Rot Pectobacterium odoriferum Associated with Kimchi Cabbage

December 2022

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80 Reads

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6 Citations

The Plant Pathology Journal

i>Pectobacterium odoriferum is the primary causative agent in Kimchi cabbage soft-rot diseases. The pathogenic bacteria Pectobacterium genera are responsible for significant yield losses in crops. However, P. odoriferum shares a vast range of hosts with P. carotovorum , P. versatile , and P. brasiliense , and has similar biochemical, phenotypic, and genetic characteristics to these species. Therefore, it is essential to develop a P. odoriferum- specific diagnostic method for soft-rot disease because of the complicated diagnostic process and management as described above. Therefore, in this study, to select P. odoriferum -specific genes, species-specific genes were selected using the data of the P. odoriferum JK2.1 whole genome and similar bacterial species registered with NCBI. Thereafter, the specificity of the selected gene was tested through blast analysis. We identified novel species-specific genes to detect and quantify targeted P. odoriferum and designed specific primer sets targeting HAD family hydrolases. It was confirmed that the selected primer set formed a specific amplicon of 360 bp only in the DNA of P. odoriferum using 29 Pectobacterium species and related species. Furthermore, the population density of P. odoriferum can be estimated without genomic DNA extraction through SYBR Green-based real-time quantitative PCR using a primer set in plants. As a result, the newly developed diagnostic method enables rapid and accurate diagnosis and continuous monitoring of soft-rot disease in Kimchi cabbage without additional procedures from the plant tissue.


Novel Detection and Quantification Approach of Erwinia amylovora In Vitro and In Planta Using SYBR Green-Based Real-Time PCR Assay

July 2022

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31 Reads

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3 Citations

Plant Disease

Fire blight, caused by the bacterial pathogen Erwinia amylovora, is a highly destructive disease of apples and pears, Since the apple tree entity infected by E. amylovora eventually systemically dies, the E. amylovora is a considerably important pathogen in the orchard that requires long-term management. And, it is crucial to prevent the spread of the pathogen by expeditious diagnosis. In this study, via comparative approaches to the genome sequences of the strains of various Erwinia species, we designed specific primers targeting a hypothetical gene that is single-copy and located in the chromosomal DNA of E. amylovora. This primer set specifically amplified the DNA of E. amylovora but no other bacteria, including Erwinia pyrifoliae, Pectobacterium spp., Pantoea spp., and Dickeya chrysanthemi. Furthermore, the SYBR Green-based real-time PCR using the primer set allowed estimating the population of E. amylovora accurately. Developing a rapid and accurate diagnostic method using the novel primer set enables effective defense against pathogen spread through continuous monitoring and quick response.


Fig. 1. Comparative analysis of Erwinia amylovora strains of specific binding sitez in hypothetical protein (WP_162010770.1) gene. (A) The BLASTN search of hypothetical protein gene from E. amylovora CFBP1430. (B) Comparison and alignment of primer binding sites among E. amylovora strains.
Fig. 3. Binding site of the RS24580-205F/R primer to the Erwinia pyrifoliae genomic DNA template.
Fig. 4. Sensitivity of the RS24580-205 primer set using serially diluted bacterial genomic DNA and suspension cells. Genomic DNA (gDNA) of Erwinia amylovora TS3128 (A) and E. pyrifoliae CP12327 (B). M, 1 kb size marker; lane 1, 5 ng gDNA; lane 2, 500 pg gDNA; lane 3, 50 pg gDNA; lane 4, 5 pg gDNA; lane 5, 500 fg gDNA; lane 6, 50 fg gDNA; lane 7, 5 fg gDNA; -, negative control. Bacterial cells of E. amylovora TS3128 (C) and E. pyrifoliae CP12327 (D). M, 1 kb size marker; lane 1, 10 9 cfu/ml; lane 2, 10 8 cfu/ml; lane 3, 10 7 cfu/ml; lane 4, 10 6 cfu/ml; lane 5, 10 5 cfu/ml; lane 6, 10 4 cfu/ml; lane 7, 10 3 cfu/ml; lane 8, 10 2 cfu/ml; -, negative control.
Bacterial strains used in this study
Discrimination and Detection of Erwinia amylovora and Erwinia pyrifoliae with a Single Primer Set

June 2022

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62 Reads

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3 Citations

The Plant Pathology Journal

Erwinia amylovora and Erwinia pyrifoliae cause fire blight and black-shoot blight, respectively, in apples and pears. E. pyrifoliae is less pathogenic and has a narrower host range than that of E. amylovora. Fire blight and black-shoot blight exhibit similar symptoms, making it difficult to distinguish one bacterial disease from the other. Molecular tools that differentiate fire blight from black-shoot blight could guide in the implementation of appropriate management strategies to control both diseases. In this study, a primer set was developed to detect and distinguish E. amylovora from E. pyrifoliae by conventional polymerase chain reaction (PCR). The primers produced amplicons of different sizes that were specific to each bacterial species. PCR products from E. amylovora and E. pyrifoliae cells at concentrations of 10<sup>4</sup> cfu/ml and 10<sup>7</sup> cfu/ml, respectively, were amplified, which demonstrated sufficient primer detection sensitivity. This primer set provides a simple molecular tool to distinguish between two types of bacterial diseases with similar symptoms.


Species-Specific Detection and Quantification of Erwinia pyrifoliae in Plants by a Direct SYBR Green Quantitative Real-Time PCR Assay

May 2022

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18 Reads

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3 Citations

PhytoFrontiers™

The present study describes a SYBR Green real-time quantitative (q) PCR assay to detect Erwinia pyrifoliae in plants. E. pyrifoliae, first described in South Korea, is a phytopathogenic bacterial species in the genus Erwinia. In particular, specific detection, quantitation, and identification of E. pyrifoliae is still challenging, as symptoms resulting from its colonization of Asian pear blossoms are very similar to those caused by E. amylovora. E. pyrifoliae has biochemical, phenotypic, and genetic properties similar to those of E. amylovora. Moreover, other Erwinia species, including E. tasmaniensis and E. billingiae, are also detected by currently available molecular methods and with traditional methods as well. Therefore, in this study, previously published genome sequences of the genera Erwinia and Pantoea were compared to exploit species-specific genes for use as improved qPCR targets to detect E. pyrifoliae. In silico analyses of the selected gene and designed primer sequences, in conjunction with bio-SYBR Green real-time qPCR, confirmed the robustness of this newly developed assay. Consequently, the bio-SYBR Green real-time qPCR-based protocols developed here can be used for rapid and specific detection of E. pyrifoliae. They will potentially simplify and facilitate diagnosis and monitoring of this pathogen and guide plant disease management. [Formula: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .


Figure 2. Specificity, melting peak and standard curve analysis of the SYBR Green qPCR assay with the Bam249F/R and BS310F/R primer sets. (A) B. velezensis. (a) Fluorescence intensity as a function of template concentration. For each assay, a series of 10-fold dilutions of cloned DNA (ranging from 1.42 × 10 3 to 1.42 × 10 9 copies/µl) was used as the template (1-7, sample dilutions). (b) Standard curve derived from the amplification plot. (c) Melting curve analysis (1-7, sample dilutions). (d) Melting peak analysis (1-7, sample dilutions). The amplified product derivatives of the relative fluorescence units [−d(RFU)/dT] were plotted as a function of temperature (amplified product, 86.0 °C). The large peak indicates the amplified product, while the small peak indicates the no-template control. (B) B. subtilis subsp. subtilis. (a) Fluorescence intensity as a function of template concentration. For each assay, a series of 10-fold dilutions of cloned DNA (ranging from 1.39 × 10 3 to 1.39 × 10 9 copies/µl) was used as the template (1-7, sample dilutions). (b) Standard curve derived from the amplification plot. (c) Melting curve analysis (1-7, sample dilutions). (d) Melting peak analysis (1-7, sample dilutions). The amplified product derivatives of the relative fluorescence units [−d(RFU)/dT] were plotted as a function of temperature (amplified product, 83.5 °C). The large peak indicates the amplified product, while the small peak indicates the no-template control. 
Table 2 , and lane 33 is the negative control (distilled water). (B) B. subtilis subsp. subtilis. Lane M is the size marker (1 kb DNA plus ladder; Gibco BRL). Lanes 1-6 are B. subtilis strains. Lanes 7-32 are strains of other Bacillus species, as listed in Table 3 and lane 33 is the negative control (distilled water).
Figure 3. Changes in the real-time PCR Ct values during the quantification of Bacillus velezensis (A) and B. subtilis subsp. subtilis (B) using total DNA isolated from salted Chinese cabbage kimchi fermented at 4 °C (a), 15 °C and 25 °C (b). 
Specific PCR amplification of Bacillus velezensis (A) and B. subtilis subsp. subtilis (B) with the Bam249F/R and BS310F/R primer sets. (A) B. velezensis. Lane M is the size marker (1 kb DNA plus ladder; Gibco BRL). Lanes 1–16 are B. velezensis strains. Lanes 17–32 are strains of other Bacillus species, as listed in Table 2, and lane 33 is the negative control (distilled water). (B) B. subtilis subsp. subtilis. Lane M is the size marker (1 kb DNA plus ladder; Gibco BRL). Lanes 1–6 are B. subtilis strains. Lanes 7–32 are strains of other Bacillus species, as listed in Table 3 and lane 33 is the negative control (distilled water).
Understanding the ontogeny and succession of Bacillus velezensis and B. subtilis subsp. subtilis by focusing on kimchi fermentation

May 2018

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1,157 Reads

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40 Citations

Scientific Reports

Bacillus subtilis and B. velezensis are frequently isolated from various niches, including fermented foods, water, and soil. Within the Bacillus subtilis group, B. velezensis and B. subtilis subsp. subtilis have received significant attention as biological resources for biotechnology-associated industries. Nevertheless, radical solutions are urgently needed to identify microbes during their ecological succession to accurately confirm their action at the species or subspecies level in diverse environments, such as fermented materials. Thus, in this study, previously published genome data of the B. subtilis group were compared to exploit species- or subspecies-specific genes for use as improved qPCR targets to detect B. velezensis and B. subtilis subsp. subtilis in kimchi samples. In silico analyses of the selected genes and designed primer sequences, in conjunction with SYBR Green real-time PCR, confirmed the robustness of this newly developed assay. Consequently, this study will allow for new insights into the ontogeny and succession of B. velezensis and B. subtilis subsp. subtilis in various niches. Interestingly, in white kimchi without red pepper powder, neither B. subtilis subsp. subtilis nor B. velezensis was detected.



Figure 2. Specificity, melting peak and standard curve of the T1PL186F/R and LPA187F/R primer set by SYBR Green qPCR. (A) Lactobacillus plantarum subsp. plantarum. (a) Fluorescence intensity as a function of the template concentration. For each assay, a series of 10-fold dilutions of cloned DNA (ranging from 1.43 × 10 3 to 1.43 × 10 9 copies/µl) were used as template (1-7, sample dilutions). (b) Standard curve derived from the amplification plot. (c) Melting curve analysis (1-7, sample dilutions). (d) Melting peak analysis (1-7, sample dilutions). The derivatives of the relative fluorescence units of the amplified products [-d(RFU)/dT] were plotted as a function of temperature (amplified product, 86.5 °C). The high peak indicates the amplified product, and the low peak is the no-template control. (B) Lactobacillus plantarum subsp. argentoratensis. (a) Fluorescence intensity as a function of the template concentration. For each assay, a series of 10-fold dilutions of cloned DNA (ranging from 1.42 × 10 3 to 1.42 × 10 9 copies/µl) were used as template (1-7, sample dilutions). (b) Standard curve derived from the amplification plot. (c) Melting curve analysis (1-7, sample dilutions). (d) Melting peak analysis (1-7, sample dilutions). The derivatives of the relative fluorescence units of the amplified products [−d(RFU)/dT] were plotted as a function of temperature (amplified product, 81.5 °C). The intense peak indicates the amplified product. 
Figure 3. Changes in the real-time PCR Ct values during the quantification of Lactobacillus plantarum subsp. plantarum (A) and Lactobacillus plantarum subsp. argentoratensis (B) from total DNA isolated from salted Chinese cabbage kimchi fermented at 4 °C (a), 15 °C and 25 °C (b). 
Specific PCR amplification of Lactobacillus plantarum subsp. plantarum and L. plantarum subsp. argentoratensis with the T1PL186F/R and LPA187F/R primer set. Lane M shows the size marker (1 kb Plus DNA ladder; Gibco BRL); lanes 1 to 8 contain L. plantarum subsp. plantarum samples; lane 9 contains a L. plantarum subsp. argentoratensis sample; lanes 10 to 36 contain samples of strains from other Lactobacillus species along with samples of strains from species of Leuconostoc, Pediococcus, Streptococcus and Weissella, as listed in Table 2.
New insight and metrics to understand the ontogeny and succession of Lactobacillus plantarum subsp. plantarum and Lactobacillus plantarum subsp. argentoratensis

April 2018

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197 Reads

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13 Citations

Scientific Reports

Lactobacillus plantarum is one of the most extensively studied Lactobacillus species because of its presence in a variety of environmental niches, versatility, and metabolic capabilities, resulting in the use of this organism in many industrial applications. However, although extensive effort has been invested in screening this species from a variety of habitats, a reliable and accurate method for studying the succession and ontogeny of this organism in complex ecosystems is still required to confirm the activity of L. plantarum at the subspecies level. Therefore, in this study, novel subspecies-specific genes for the quantitative detection of two L. plantarum subspecies were identified by comparative genomic analysis. The specificity of primer sets for selected genes specific to each targeted microbe was confirmed in kimchi samples. Interestingly, in all the kimchi samples at 4 °C, the presence of L. plantarum subsp. argentoratensis was not observed. Hence, we found that low temperatures markedly affected the ontogeny of L. plantarum subsp. argentoratensis during kimchi fermentation. Subsequently, this touchstone method will offer new insight and metrics to understand the ontogeny and succession of L. plantarum subsp. plantarum and L. plantarum subsp. argentoratensis in various niches.


Citations (6)


... Studies have shown that actinomycetes have the ability to biocontrol various plant pathogens through various mechanisms (Hong-Thao et al. 2016). Actinomycetes are relatively abundant in the rhizosphere of healthy peppers, where they are known to inhibit the growth of several plant pathogens in the rhizosphere and produce extracellular enzymes that promote crop production and immunity by breaking down the complex mixture in the soil (Kong et al. 2022). In our study, an aeration rate of 15 mg/L showed a higher correlation with Actinobacteria. ...

Reference:

Relationship between pepper (Capsicum annuum L.) root morphology, inter-root soil bacterial community structure and diversity under water–air intercropping conditions
Changes in the Composition and Microbial Community of the Pepper Rhizosphere in Field with Bacterial Wilt Disease

The Plant Pathology Journal

... In the last few decades, the taxonomy of Pectobacterium has undergone major modifications and reclassifications. A total of 22 species have been validly reported: P. actinidiae , P. aquaticum , P. aroidearum (Xu et al. 2021), P. atrosepticum (Ismiyatuningsih et al. 2016;Toth et al. 2022), P. betavasculorum (Rastgou et al. 2022), P. brasiliense (Oulghazi et al. 2021), P. cacticida (Xu et al. 2021), P. carotovorum , P. colocasium (Zhou et al. 2022), P. fontis (Oulghazi et al. 2019), P. jejuense (Hong et al. 2023), P. odoriferum (Jin et al. 2022), P. parmentieri (Khayi et al. 2016), P. parvum (Pasanen et al. 2020), P. peruviense (Waleron et al. 2018), P. polaris (Dees et al. 2017), P. polonicum (Waleron et al. 2019a), P. punjabense (Sarfraz et al. 2018), P. quasiaquaticum (Moussa et al. 2021), P. versatile (Kravchenko et al. 2021), P. wasabiae (Khayi et al. 2016), and P. zantedeschiae (Waleron et al. 2019b). Among these species, Pectobacterium brasiliense is considered the most virulent and highly aggressive soft-rot pathogen. ...

A New Approach Using the SYBR Green-Based Real-Time PCR Method for Detection of Soft Rot Pectobacterium odoriferum Associated with Kimchi Cabbage

The Plant Pathology Journal

... These molecular detection protocols were confirmed using isolated DNA or cell cultures of E. amylovora. PCR can be inhibited by plant-derived phenolic compounds; nevertheless, plant extracts obtained by grinding the tissue or tissue suspensions of host plants with distinct symptoms can be used as templates for real-time PCR (Jin et al., 2022). In general, real-time PCR provides advantages over traditional PCR; these advantages include accurate detection, immediate information availability, and reduced time. ...

Novel Detection and Quantification Approach of Erwinia amylovora In Vitro and In Planta Using SYBR Green-Based Real-Time PCR Assay
  • Citing Article
  • July 2022

Plant Disease

... Several qPCR-based methods and specific primers have been developed for the quantification of E. pyrifoliae (Jin et al., 2022;Lehman et al., 2008;Wensing et al., 2012). The sensitivity of our ddPCR method was similar to or better than that of these previously reported techniques. ...

Species-Specific Detection and Quantification of Erwinia pyrifoliae in Plants by a Direct SYBR Green Quantitative Real-Time PCR Assay

PhytoFrontiers™

... B. velezensis DSM 33864 was initially isolated from feces of a healthy human donor and showed an increased ability to reduce the growth of C. difficile in vitro compared to multiple related members of the Bacillus genus 16 . B. velezensis is abundant in the traditional Korean food kimchi and has recently gained qualified presumption of safety (QPS) status for food and feed production by the European Food Safety Authority (EFSA) 17,18 . Bacillus spores can also be advantageous as they are generally resistant to stomach-and bile acids 19 . ...

Understanding the ontogeny and succession of Bacillus velezensis and B. subtilis subsp. subtilis by focusing on kimchi fermentation

Scientific Reports

... The specific primers used for L. plantarum were T1PL186F 5 ′ -ACC CCC GTT CCG TCA GA − 3 ′ , and T1PL186R 5 ′ -ATC ACC GCT TCC CCG CTC ATT − 3 ′ . The amplicon lengths were 186 base pairs, and they were annealed at 65 • C. The target gene was LPXTGa motif cell wall anchor domain protein, and the Genbank accession number is EFK28973.1 (Jin, Park, Cho, Lee, & Park, 2018). For B. subtilis, the pair of primers is BS310F 5 ′ -GGC CTA TTG AAC ACC CTG ATT TA − 3 ′ , and BS310R 5 ′ -CGG ATG CGG CCT TCT TTT TC -3 ′ . ...

New insight and metrics to understand the ontogeny and succession of Lactobacillus plantarum subsp. plantarum and Lactobacillus plantarum subsp. argentoratensis

Scientific Reports