• Proceedings of the Zoological Society Korea Conference
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• 2000.10a
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• pp.129.2-130
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• 2000

No Abstract, See Full Text

• Korean Journal of Microbiology
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• v.37 no.4
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• pp.265-272
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• 2001

The diversity of acid-tolerant epiphytic bacterial communities on deciduous oak tree (Quercus dentate Thunb.) leaves was examined both in the natural forest area with a clean air and in the industrial estate to assess effects of acidic depositions to the phyllosphere using 16S rDNA sequence data. A total of 444 acid-tolerant epiphytic bacterial clones were obtained, resulting in 17 phylotypes by performing a analysis of restriction fragment length polymorphism (RFLP) for PCR-amplified 16S rDNA products. A very low diversity of dominating acid tolerant bacterial communities in both areas was found, just 2 subphyla groups, $\gamma$-Proteobacteria and low-G+C gram-positive bacteria. As tree leaves grow older, diversities of acid-tolerant bacteria on them significantly increased. The community structure of acid-tolerant epiphytic bacteria consisted of Pseudomonas and Enterobacteriaceae groups in the $\gamma$-Proteobacteria subphylum, and Streptococcaceae and Staphylococcus groups in the low-G+C gram-positive bacteria subphylum. The direct influence of acidic depositions on bacterial phylogenetic composition could not be detected especially when higher taxonomic levels such as subphylum, but at narrower or finer levels it could be observed by a detection of Xanthomonadales group belonged to the $\gamma$-Proteobacteria only in the industrial area and of Acetobacteraceae group belonged to the $\alpha$-Proteobacteria. There remains that these specific acid-tolerant epiphytic bacterial groups could be used as indicators for assessing effects of acidic depositions on the phyllosphere.

• Korean Journal of Microbiology
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• v.37 no.1
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• pp.61-65
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• 2001

Epifluorescence microscopy was used to observe epiphytic bacteria directly on plant leaf surfaces as well as indirectly in the leaf liberating solution by staining with fluorochromes of 4',6-diamidino-2-phenylindole (DAPI) and acridine orange(AO). Epiphytic bacteria could not be well observed on the leaf surface by staining with AO due to an intrusive orange or red background fluorescence. However, DAPI gave us clear epifluorescent images of the bacteria on the leaf. On the contrary, epiphytic bacteria in the liberating leaf solution were well observed on filters stained by both types of fluorochrome, although DAPI showed better fluorescent images than AO and not necessarily required a washing step of the filters stained. The optimum conditions of the DAPI stains were 5 $\mu$g/ml for 5 min both for leaves and for filters of the liberating solution. It was confirmed that a critical step in the epifluorescence microscopy of leaf surfaces was to minimize release of water from the leaf. For this, the stained leaf samples were put on a filter paper, kept in a dry oven at $70^{\circ}C$ for 2 min instead of air-drying, and then immediately observed by epifluorescence microscopy. The established technique was applied to enumerate epiphytic bacteria on oak tree leaf surfaces.

• Proceedings of the Microbiological Society of Korea Conference
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• 2002.10a
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• pp.100-105
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• 2002

The diversity of epiphytic bacterial communities on deciduous oak tree (Quercus dentate Thunb.) leaves was examined both in the natural forest area with a clean air and in the industrial estate to assess effects of acidic deposition to the phyllosphere using 16S rDNA sequence data. In addition, acid-tolerant epiphytic bacterial communities were compared. A total of 78 epiphytic and 444 acid-tolerant clones were obtained from clone libraries, resulting in 20 and 17 phylotypes by analysis of restriction fragment length polymorphism (RFLP) for PCR-amplified 16S rDNA products. A low bacterial diversity in both areas was found. As tree leaves grow older, bacterial diversities were slightly increased in the level of subphylum. The community structure of epiphytic bacteria in both areas in April consisted of only two subphyla, $\beta-and\;\gamma-Proteobacteria$. In August two additional subphyla in both areas were found, but the composition was a little different, Acidobacteria and Cytophaga-Flexibacter-Bacteroids (CFB) group in the industrial estate and a -Proteobacteria and CFB group in the natural area, respectively. Acidobacteria could be an indicator of epiphytic bacteria for acidic deposition on plant leaves, whereas a -Proteobacteria be one of epiphytic bacteria that naturally survive on leaves that are not affected by acidic deposition. The acid-tolerant bacterial communities in April were composed of two subphyla, $\gamma-Proteobacteria$ and Low G+C gram-positive bacteria in both areas, and in August a-Proteobacteria was added to the community just in the natural forest area. The direct influence of acidic deposition on the acid-tolerant bacterial phylogenetic composition could not be detected in higher taxonomic levels such as subphylum, but at narrower or finer levels it could be observed by a detection of Xanthomonadales group of $\gamma-Proteobacteria$ just in the industrial estate.

• Korean Journal of Microbiology
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• v.33 no.4
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• pp.262-266
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• 1997

Total, direct viable count, and acid-tolerant epiphytic bacterial population sizes were quantified on leaves of chestnut tree (Castanea crenata S. et Z.) near Taejon Industrial Estate affected by acid precipitation and deposition as well as in the clean natural forest area, Mt. Kyejok, in Taejon city from August 1996 to August 1997. Geometric mean numbers of total, direct viable count, and acid-tolerant epiphytic bacteria were $9.9{\times}10^5cell/cm^2$, $1.6{\times}10^6cell/cm^2$, and $7.1{\times}10^3cfu/cm^2$ respectively, being 1.5, 2, and 2.6 times those in the clean area. Acid-tolerant epiphytic bacterial numbers at pH 5.6 by MPN method were $3.3{\times}10^4$ in the industrial area, about the same as the number, $3.4{\times}10^4MPN/cm^2$, of the clean area. Acid-tolerant bacterial number at pH 4.0 was $1.9{\times}10^{-1}MPN/cm^2$ in the industrial area, whereas none was detected in the clean area. Acid-tolerant bacteria at pH 3.0 were not detected at all in the industrial area as well as in the clean area. Epiphytic bacterial population sizes were generally the greatest in May when leaves are emerged and grew hut the lowest in November when defoliation occurs. These results showed that air pollutant deposition on leaves did not cause a decrease of epiphytic bacteria at least and acid deposition on leaves did cause an increase of acid-tolerant bacteria.

• Journal of Microbiology and Biotechnology
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• v.16 no.10
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• pp.1622-1628
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• 2006

Bacteria from the Methylobacterium genus, called pink-pigmented facultative methylotrophic bacteria (PPFMs), are common inhabitants of plants, potentially dominating the phyllosphere population, and are also encountered in the rhizosphere, seeds, and other parts of plants, being versatile in nature. The consistent success of the Methylobacterium plant association relies on methylotrophy, the ability to utilize the one-carbon compound methanol emitted by plants. However, the efficiency of Methylobacterium in plant growth promotion could be better exploited and thus has attracted increasing interest in recent years. Accordingly, the present study investigated the inoculation effects of Methylobacterium sp. strains CBMB20 and CBMB 110 on seed imbibition to tomato and red pepper on the growth and accumulation of phytohormone levels under gnotobiotic conditions. Seeds treated with the Methylobacterium strains showed a significant increase in root length when compared with either the uninoculated control or Methylobacterium extorquens $miaA^-$ knockout mutanttreated seeds. Extracts of the plant samples were used for indole-3-acetic acid (IAA), trans-zeatin riboside (t-ZR), and dihydrozeatin riboside (DHZR) assays by immunoanalysis. The treatment with Methylobacterium sp. CBMB20 or CBMB 110 produced significant increases in the accumulation of IAA and the cytokinins t-ZR and DHZR in the red pepper extracts, whereas no IAA was detected in the tomato extracts, although the cytokinin concentrations were significantly increased. Therefore, this study proved that the versatility of Methylobacterium as a plant-growth promoting bacteria could be better exploited.

• Journal of Microbiology and Biotechnology
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• v.17 no.2
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• pp.226-233
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• 2007

Members of Methylobacterium, referred as pink-pigmented facultative methylotrophic bacteria, are frequently associated with terrestrial and aquatic plants, tending to form aggregates on the phyllosphere. We report here that the production of autoinducer molecules involved in the cell-to-cell signaling process, which is known as quorum sensing, is common among Methylobacterium species. Several strains of Methylobacterium were tested for their ability to produce N-acyl-homoserine lactone (AHL) signal molecules using different indicators. Most strains of Methylobacterium tested could elicit a positive response in Agrobacterium tumefaciens harboring lacZ fused to a gene that is regulated by autoinduction. The synthesis of these compounds was cell-density dependent, and the maximal activity was reached during the late exponential to stationary phases. The bacterial extracts were separated by thin-layer chromatography and bioassayed with A. tumefaciens NTI (traR, tra::lacZ749). They revealed the production of various patterns of the signal molecules, which are strain dependent. At least two signal molecules could be detected in most of the strains tested, and comparison of their relative mobilities suggested that they are homologs of N-octanoyl-$_{DL}$-homoserine lactone ($C_8-HSL$) and N-decanoyl-$_{DL}$-homoserine lactone ($C_{10}-HSL$).

• Microbiology and Biotechnology Letters
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• v.47 no.1
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• pp.1-10
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• 2019

Saline soils comprise more than half a billion hectares worldwide. Thus, they warrant attention for their efficient, economical, and environmentally acceptable management. Halophytes are being progressively utilized for human benefits. The halophyte microbiome contributes significantly to plant performance and can provide information regarding complex ecological processes involved in the osmoregulation of halophytes. Microbial communities associated with the rhizosphere, phyllosphere, and endosphere of halophytes play an important role in plant health and productivity. Members of the plant microbiome belonging to domains Archaea, Bacteria, and kingdom Fungi are involved in the osmoregulation of halophytes. Halophilic microorganisms principally use compatible solutes, such as glycine, betaine, proline, trehalose, ectoine, and glutamic acid, to survive under salinity stress conditions. Plant growth-promoting rhizobacteria (PGPR) enhance plant growth and help to elucidate tolerance to salinity. Detailed studies of the metabolic pathways of plants have shown that plant growth-promoting rhizobacteria contribute to plant tolerance by affecting the signaling network of plants. Phytohormones (indole-3-acetic acid and cytokinin), 1-aminocyclopropane-1-carboxylic acid deaminase biosynthesis, exopolysaccharides, halocins, and volatile organic compounds function as signaling molecules for plants to elicit salinity stress. This review focuses on the functions of plant microbiome and on understanding how the microorganisms affect halophyte health and growth.

• Research in Plant Disease
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• v.22 no.2
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• pp.81-93
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• 2016

Out of plant-associated bacteria, certain plant growth-promoting bacteria (PGPB) have been reported to increase plant growth and productivity and to elicit induced resistance against plant pathogens. In this study, our objective was to broaden the range of applications of leaf-colonizing PGPB for foliar parts of road tress and pepper. Total 1,056 isolates of endospore-forming bacteria from tree phylloplanes were collected and evaluated for the enzymatic activities including protease, lipase, and chitinase and antifungal capacities against two fungal pathogens, Colletotrichum graminicola and Botrytis cinerea. Fourteen isolates classified as members of the bacilli group displayed the capacity to colonize pepper leaves after spraying inoculation. Three strains, 5B6, 8D4, and 8G12, and the mixtures were employed to evaluate growth promotion, yield increase and defence responses under field condition. Additionally, foliar application of bacterial preparation was applied to the road tress in Yuseong, Daejeon, South Korea, resulted in increase of chlorophyll contents and leaf thickness, compared with non-treated control. The foliar application of microbial preparation reduced brown shot-hole disease of Prunus serrulata L. and advanced leaf abscission in Ginkgo biloba L. Collectively, our results suggest that leaf-colonizing bacteria provide potential microbial agents to increase the performance of woody plants such as tree and pepper through spray application.

• Journal of Microbiology and Biotechnology
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• v.28 no.2
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• pp.227-235
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• 2018

Foodborne illness represents a major threat to public health and is frequently attributed to pathogenic microorganisms on fresh produce. Recurrent outbreaks often come from vegetables that are grown close to or within the ground. Therefore, the first step to understanding the public health risk of microorganisms on fresh vegetables is to identify and describe microbial communities. We investigated the phyllospheres on Chinese cabbage (Brassica rapa subsp. pekinensis, N = 54). 16S rRNA gene amplicon sequencing targeting the V5-V6 region of 16S rRNA genes was conducted by employing the Illumina MiSeq system. Sequence quality was assessed, and phylogenetic assessments were performed using the RDP classifier implemented in QIIME with a bootstrap cutoff of 80%. Principal coordinate analysis was performed using a weighted Fast UniFrac matrix. The average number of sequence reads generated per sample was 34,584. At the phylum level, bacterial communities were composed primarily of Proteobacteria and Bacteroidetes. The most abundant genera on Chinese cabbages were Chryseobacterium, Aurantimonadaceae_g, Sphingomonas, and Pseudomonas. Diverse potential pathogens, such as Pantoea, Erwinia, Klebsiella, Yersinia, Bacillus, Staphylococcus, Salmonella, and Clostridium were also detected from the samples. Although further epidemiological studies will be required to determine whether the detected potential pathogens are associated with foodborne illness, our results imply that a metagenomic approach can be used to detect pathogenic bacteria on fresh vegetables.