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Transcriptional regulation and mutational analysis of a dctA encoding organic acid transporter protein from Pseudomonas chlororaphis O6.

  • Nam, Hyo-Song;Cho, Baik-Ho;Kim, Young-Cheol
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.100.1-100
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    • 2003
  • A dctA gene encoding a protein with identity to a C4-dicarboxylate/H+ was cloned from a beneficial biocontrol bacterium, P. chororaphis O6. Expression of the dctA was induced in minimal medium by several organic acids and was repressed by glucose. Highest expression was observed in early-log cells grown on fumarate and succinate with decline as cells approached late-log phase. The dctA transcript accumulated weakly when cells were grown on malate but strong expression was observed with benzoate. Expression of the dctA transcript was repressed in early-log cells upon addition of glucose to fumarate, but was detected as the cell culture aged. A dctA-deficient mutant of O6, constructed by marker exchange mutagenesis, did not grow on minimal medium containing succinate, benzoate, or fumarate, and growth on malate was delayed. The dctA mutant and wild type grew equally on glucose. The dctA mutant on cucumber roots in sterilized potting soil was colonized at levels comparable to those of the wild type, but induction level of disease resistance by the mutant against target leaf spot disease was decreased. These results may indicate that the dctA is essential for utilization of certain organic acids and its expression is controlled by the availability of sugars. In addition, the dctA is not essenitial for cucumber root colonization, but important for induction of disease resistance.

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A Study on the Test Bed Evaluation for the Ecological Restoration of Unused Road - Focused on the Experimental Construction Site in Young Dong Province of GyungBu Expressway(Seven years after construction.) - (폐도로 생태복원을 위한 시험시공지 평가 연구 - 경부선 영동군 황간지역 시험시공지에 대한 시공 후 7년 시점의 추적조사 -)

  • Jeon, Gi-Seong
    • Journal of the Korean Society of Environmental Restoration Technology
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    • v.19 no.5
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    • pp.47-57
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    • 2016
  • In the recent years, the incidence of abandoned concrete roads have been increasing rapidly due to road improvements and facilities route construction, these abandoned expressway and unused roads are scattered throughout the country but the management system is not clear, they does not be an effectively managed because there is no maintenance cost. In response to these social concern and expectation, the Korea Expressway Corporation that is management authority of the expressway is developing policy initiatives and various projects to restore the closed road to ecological for eco-friendly projects of the existing expressway. And as part of these projects, Hwanggan IC unmanaged abandoned concrete roads restoration project was done and it was conducting monitoring for ecological restoration that is not one-off but sustainable. After test construction over seven years at the time, test construction of four ways enforced but the boundaries have become blurred over time. And Pinus koraiensis, Callicarpa dichotoma and Sorbaria stellipilla var. typica planted in the site are dominating, else Amorpha fruticosa and Indigofera pseudotinctoria sowed are dominating. Invasive species is that Robinia pseudoacacia was growing in the colony. Over monitoring result time, the vegetation was taking root and the ecosystem was being restore. But the disturbance of vegetation happens due to planting of tree species that doesn't fit in the restoration area and advent of ecosystem Disturbing Species. The study will be providing a basic data that identify change of plant environment by monitoring of the site and soil environment during June to November 2016 and secure an objective evaluation data by analyzing the effects of ecological restoration for revegetation test bed evaluation for ecological restoration of expressway in ecological restoration construction in later.

Spore Associated Bacteria (SAB) of Arbuscular Mycorrhizal Fungi (AMF) and Plant Growth Promoting Rhizobacteria (PGPR) Increase Nutrient Uptake and Plant Growth Under Stress Conditions

  • Gopal, Selvakumar;Chandrasekaran, Murugesan;Shagol, Charlotte;Kim, Ki-Yoon;Sa, Tong-Min
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.4
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    • pp.582-592
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    • 2012
  • Microorganisms present in the rhizosphere soil plays a vital role in improving the plant growth and soil fertility. Many kinds of fertilizers including chemical and organic has been approached to improve the productivity. Though some of them showed significant improvement in yield, they failed to maintain the soil properties. Rather they negatively affected soil eventually, the land became unsuitable for agricultural. To overcome these problems, microorganisms have been used as effective alternative. For past few decades, plant growth promoting rhizobacteria (PGPR) and arbuscular mycorrhizal fungi (AMF) have been used as effective inoculants to enhance the plant growth and productivity. PGPR improves the plant growth and helps the plant to withstand biotic and abiotic stresses. AM fungi are known to colonize roots of plants and they increase the plant nutrient uptake. Spore associated bacteria (SAB) are attached to spore wall or hyphae and known to increase the AMF germination and root colonization but their mechanism of interaction is poorly known. Better understanding the interactions among AMF, SAB and PGPR are necessary to enhance the quality of inoculants as a biofertilizers. In this paper, current knowledge about the interactions between fungi and bacteria are reviewed and discussed about AMF spore associated bacteria.

Diversity of Fungal Endophytes in Various Tissues of Panax ginseng Meyer Cultivated in Korea

  • Park, Young-Hwan;Lee, Soon-Gu;Ahn, Doek-Jong;Kwon, Tae-Ryong;Park, Sang-Un;Lim, Hyoun-Sub;Bae, Han-Hong
    • Journal of Ginseng Research
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    • v.36 no.2
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    • pp.211-217
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    • 2012
  • Endophytic fungi were isolated from various tissues (root, stem, petiole, leaf, and flower stalk) of 3- and 4-year-old ginseng plants (Panax ginseng Meyer) cultivated in Korea. The isolated endophytic fungi were identified based on the sequence analysis of the internal transcribed spacer (ITS), 1-5.8-ITS 2. A morphological characterization was also conducted using microscopic observations. According to the identification, 127 fungal isolates were assigned to 27 taxa. The genera of Phoma, Alternaria and Colletotrichum were the most frequent isolates, followed by Fusarium, Entrophospora and Xylaria. Although 19 of the 27 taxa were identified at the species level, the remainder were classified at the genus level (6 isolates), phylum level (Ascomycota, 1 isolate), and unknown fungal species (1 isolate). Endophytic fungi of 13 and 19 species were isolated from 3- and 4-year-old ginseng plants, respectively, and Phoma radicina and Fusarium solani were the most frequently isolated species colonizing the tissues of the 3- and 4-year-old ginseng plants, respectively. The colonization frequency (CF%) was dependant on the age and tissue examined: the CFs of the roots and stems in the 3-year-old ginseng were higher than the CF of tissues in the 4-year-old plants. In contrast, higher CFs were observed in the leaves and petioles of 4-year-old plants, and endophytic fungi in the flower stalks were only detected in the 4-year-old plants. In conclusion, we detected diverse endophytic fungi in ginseng plants, which were distributed differently depending on the age and tissue examined.

Characteristic of Orchid Mycorrhizal Fungi from Roots of Cypripedium japonicum and C. macranthum (광릉요강꽃과 복주머니란의 뿌리에 감염된 난균근균의 특성)

  • Sim, Mi-Yeong;Youm, Jae-Young;Chung, Jae-Min;Lee, Byung-Chun;Koo, Chang-Duck;Eom, Ahn-Heum
    • The Korean Journal of Mycology
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    • v.38 no.1
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    • pp.1-4
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    • 2010
  • Orchid mycorrhizal fungi(OMF) were examined in roots of the two threatened orchid species, Cypripedium japonicum and C. macranthum. The morphological characteristics of mycorrhizal colonization in the roots of two orchid species were observed. OMF colonized in the roots of two species were identified using molecular analysis. DNA from the root was extracted and amplified internal transcribed spacer(ITS) region using basidiomycete ITS primers, ITS1-OF and ITS4-OF. Four species belonging to Tulasnellaceae in roots of C. japonicum and two species of Tulasnellaceae and one basidiomycetous species was found in roots of C. macranthum.

The effect of Glomus intraradices on the physiological properties of Panax ginseng and on rhizospheric microbial diversity

  • Tian, Lei;Shi, Shaohua;Ma, Lina;Zhou, Xue;Luo, Shasha;Zhang, Jianfeng;Lu, Baohui;Tian, Chunjie
    • Journal of Ginseng Research
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    • v.43 no.1
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    • pp.77-85
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    • 2019
  • Background: Glomus intraradices is a species of arbuscular mycorrhizal fungi that, as an obligate endomycorrhiza, can form mutually beneficial associations with plants. Panax ginseng is a popular traditional Chinese medicine; however, problems associated with ginseng planting, such as pesticide residues, reduce the ginseng quality. Methods: In this experiment, we studied the effect of inoculating G. intraradices on several physiological properties and microbial communities of ginseng. UV-Visible Spectrum method was used to detect physical properties. Denaturing gradient gel electrophoresis method was used to analyze microbial communities. Results: The results indicated that inoculation with G. intraradices can improve the colonization rate of lateral ginseng roots, increase the levels of monomeric and total ginsenosides, and improve root activity as well as polyphenol oxidase and catalase activities. We also studied the bacterial and fungal communities in ginseng rhizospheric soil. In our study, G. intraradices inoculation improved the abundance and Shannon diversity of bacteria, whereas fungi showed a reciprocal effect. Furthermore, we found that G. intraradices inoculation might increase some beneficial bacterial species and decreased pathogenic fungi in rhizospheric soil of ginseng. Conclusion: Our results showed that G. intraradices can benefit ginseng planting which may have some instructive and practical significance for planting ginseng in farmland.

Diversity of Root-Associated Paenibacillus spp. in Winter Crops from the Southern Part of Korea

  • CHEONG HOON;PARK SOO-YOUNG;RYU CHOONG-MIN;KIM JIHYUN F.;PARK SEUNG-HWAN;PARK CHANG SEUK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1286-1298
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    • 2005
  • The genus Paenibacillus is a new group of bacilli separated from the genus Bacillus, and most of species have been isolated from soil. In the present study, we collected 450 spore-forming bacilli from the roots of winter crops, such as barley, wheat, onion, green onion, and Chinese cabbage, which were cultivated in the southern part of Korea. Among these 450 isolates, 104 Paenibacillus-like isolates were selected, based on their colony shape, odor, color, and endospore morphology, and 41 isolates were then finally identified as Paenibacillus spp. by 16S rDNA sequencing. Among the 41 Paenibacillus isolates, 23 were classified as P. polymyxa, a type species of the genus Paenibacillus, based on comparison of the 16S rDNA sequences with those of 32 type strains of the genus Paenibacillus from the GenBank database. Thirty-five isolates among the 41 Paenibacillus isolates exhibited antagonistic activity towards plant fungal and bacterial pathogens, whereas 24 isolates had a significant growth-enhancing effect on cucumber seedlings, when applied to the seeds. An assessment of the root-colonization capacity under gnotobiotic conditions revealed that all 41 isolates were able to colonize cucumber roots without any significant difference. Twenty-one of the Paenibacillus isolates were shown to contain the nifH gene, which is an indicator of $N_{2}$ fixation. However, the other 20 isolates, including the reference strain E681, did not incorporate the nifH gene. To investigate the diversity of the isolates, a BOX-PCR was performed, and the resulting electrophoresis patterns allowed the 41 Paenibacillus isolates to be divided into three groups (Groups A, B, and C). One group included Paenibacillus strains isolated mainly from barley or wheat, whereas the other two groups contained strains isolated from diverse plant samples. Accordingly, the present results showed that the Paenibacillus isolates collected from the rhizosphere of winter crops were diverse in their biological and genetic characteristics, and they are good candidates for further application studies.

Diversity of Endophytic Fungi from Different Verticillium-Wilt-Resistant Gossypium hirsutum and Evaluation of Antifungal Activity Against Verticillium dahliae In Vitro

  • Li, Zhi-Fang;Wang, Ling-Fei;Feng, Zi-Li;Zhao, Li-Hong;Shi, Yong-Qiang;Zhu, He-Qin
    • Journal of Microbiology and Biotechnology
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    • v.24 no.9
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    • pp.1149-1161
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    • 2014
  • Cotton plants were sampled and ranked according to their resistance to Verticillium wilt. In total, 642 endophytic fungi isolates representing 27 genera were recovered from Gossypium hirsutum root, stem, and leaf tissues, but were not uniformly distributed. More endophytic fungi appeared in the leaf (391) compared with the root (140) and stem (111) sections. However, no significant difference in the abundance of isolated endophytes was found among resistant cotton varieties. Alternaria exhibited the highest colonization frequency (7.9%), followed by Acremonium (6.6%) and Penicillium (4.8%). Unlike tolerant varieties, resistant and susceptible ones had similar endophytic fungal population compositions. In three Verticillium-wilt-resistant cotton varieties, fungal endophytes from the genus Alternaria were most frequently isolated, followed by Gibberella and Penicillium. The maximum concentration of dominant endophytic fungi was observed in leaf tissues (0.1797). The evenness of stem tissue endophytic communities (0.702) was comparatively more uniform than the other two tissues. Eighty endophytic fungi selected from 27 genera were evaluated for their inhibition activity against highly virulent Verticillium dahliae isolate Vd080 in vitro. Thirty-nine isolates exhibited fungistasis against the pathogen at varying degrees. Seven species, having high growth inhibition rates (${\geq}75%$), exhibited strong antifungal activity against V. dahliae. The antifungal activity of both volatile and nonvolatile metabolites was also investigated. The nonvolatile substances produced by CEF-818 (Penicillium simplicissimum), CEF-325 (Fusarium solani), CEF-714 (Leptosphaeria sp.), and CEF-642 (Talaromyces flavus) completely inhibited V. dahliae growth. These findings deepen our understanding of cotton-endophyte interactions and provide a platform for screening G. hirsutum endophytes with biocontrol potential.

Identification of Arbuscular Mycorrhizal Fungi Colonizing Panax ginseng Using 18S rDNA Sequence (18S rDNA를 이용한 인삼(Panax ginseng)의 내생균근 균의 동정)

  • Eo, Ju-Kyeong;Kim, Dong-Hun;Jeong, Hyeon-Suk;Eom, Ahn-Heum
    • Applied Biological Chemistry
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    • v.47 no.2
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    • pp.182-186
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    • 2004
  • Morphological observation of roots and molecular technique were used to investigate the symbiotic relationships between arbuscular mycorrhizal (AM) fungi and ginseng roots. Korean ginseng, Panax ginseng, was collected from 8 sites in Korea. Colonization pattern of AM fungi in ginseng roots was determined as an Arum type under light microscopes. Nested PCR using AM fungal specific primers was employed to amplify a partial region on 18s rDNA of AM fungi from the root extracted mixed DNA. The amplified DNA was cloned and analyzed by random fragment length polymorphism (RFLP) with restriction enzymes, AluI, HinfI and AsuC21. One from each RFLP pattern was selected for sequencing. A total 16 clones were sequenced and identified as 2 species of AM fungi; Paraglomus brasilianum and Glomus spurcum. Paramglomus brasilianum was found from most of the ginseng roots, in this syudy suggesting that this species of AM fungi could have specific relationship with the ginseng root. Possible roles of AM fungal species in ginseng roots are discussed.

Quantitative Changes of PR Proteins and Antioxidative Enzymes in Response to Glomus intraradices and Phytophthora capsici in Pepper (Capsicum annuum L.) Plants

  • Zheng, Hu-Zhe;Kim, Yong-Woong;Lee, Hyun-Jin;Park, Ro-Dong;Jung, Woo-Jin;Kim, Young-Cheol;Lee, Sang-Hyun;Kim, Tae-Hwan;Kim, Kil-Yong
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.553-562
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    • 2004
  • To investigate protective activity in pepper plants, which were pre-inoculated with arbuscular mycorrhizal (AM) fungi Glomus intra radices (Gi), against pathogenic strain Phytophthora capsici (Pc), pathogenesis-related (PR) proteins and antioxidant enzymes were examined. The growth of root and shoot was the highest in peppers inoculated with G. intraradices, compared with non-inoculated control plants and those challenged by the pathogen with and without mycorrhizae after nine days of infection. Mycorrhizal colonization rate was reduced by about 10% in pathogen-challenged plants, but disease pressure was reduced. The activities of PR proteins, $\beta$-1- 3-glucanase and chitinase, were increased in Pc-treated plants compared to Gi+Pc-treated plants in leaves, but those in roots were suppressed. Superoxide dismutase activity and $H_2O_2${/TEX> content in Gi+Pc and Pc-treated plants were gradually increased in leaves. However, those in roots continuously increased up to 5 days, and then decreased dramatically. Peroxidase activity in leaves and roots increased after P. capsici infection both in plants inoculated with or without G. intraradices. These results suggest that AM fungi, G. intra radices, potentially act as one of the protective agents against plant pathogens. Changes of PR proteins and antioxidative enzymes in mycorrhizae-inoculated pepper appear to be regulated differently in leaves and roots by pathogen infection.