• Molecules and Cells
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• v.39 no.2
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• pp.111-118
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• 2016

MiR399f plays a crucial role in maintaining phosphate homeostasis in Arabidopsis thaliana. Under phosphate starvation conditions, AtMYB2, which plays a role in plant salt and drought stress responses, directly regulates the expression of miR399f. In this study, we found that miR399f also participates in plant responses to abscisic acid (ABA), and to abiotic stresses including salt and drought. Salt and ABA treatment induced the expression of miR399f, as confirmed by histochemical analysis of promoter-GUS fusions. Transgenic Arabidopsis plants overexpressing miR399f (miR399f-OE) exhibited enhanced tolerance to salt stress and exogenous ABA, but hypersensitivity to drought. Our in silico analysis identified ABF3 and CSP41b as putative target genes of miR399f, and expression analysis revealed that mRNA levels of ABF3 and CSP41b decreased remarkably in miR399f-OE plants under salt stress and in response to treatment with ABA. Moreover, we showed that activation of stress-responsive gene expression in response to salt stress and ABA treatment was impaired in miR399f-OE plants. Thus, these results suggested that in addition to phosphate starvation signaling, miR399f might also modulates plant responses to salt, ABA, and drought, by regulating the expression of newly discovered target genes such as ABF3 and CSP41b.

• Journal of Microbiology and Biotechnology
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• v.28 no.7
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• pp.1156-1167
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• 2018

The aim of this study was to isolate and characterize lead (Pb)-solubilizing bacteria from heavy metal-contaminated mine soils and to evaluate their inoculation effects on the growth and Pb absorption of Brassica juncea. The isolates were also evaluated for their plant growth-promoting characteristics as well as heavy metal and salt tolerance. A total of 171 Pb-tolerant isolates were identified, of which only 15 bacterial strains were able to produce clear haloes in solid medium containing PbO or $PbCO_3$, indicating Pb solubilization. All of these 15 strains were also able to dissolve the Pb minerals in a liquid medium, which was accompanied by significant decreases in pH values of the medium. Based on 16S rRNA gene sequence analysis, the Pb-solubilizing strains belonged to genera Bacillus, Paenibacillus, Brevibacterium, and Staphylococcus. A majority of the Pb-solubilizing strains were able to produce indole acetic acid and siderophores to different extents. Two of the Pb-solubilizing isolates were able to solubilize inorganic phosphate as well. Some of the strains displayed tolerance to different heavy metals and to salt stress and were able to grow in a wide pH range. Inoculation with two selected Pb-solubilizing and plant growth-promoting strains, (i.e., Brevibacterium frigoritolerans YSP40 and Bacillus paralicheniformis YSP151) and their consortium enhanced the growth and Pb uptake of B. juncea plants grown in a metal-contaminated soil. The bacterial strains isolated in this study are promising candidates to develop novel microbe-assisted phytoremediation strategies for metal-contaminated soils.

• Microbiology and Biotechnology Letters
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• v.45 no.1
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• pp.12-18
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• 2017

Microbial strains exhibiting proteolytic activity were isolated from kimchi, one of traditional fermented foods in Korea. Eight strains formed clear zones around their colonies when grown on TSA plates supplemented with skim milk. MBE/L865 exhibited 2.6-fold higher protease activity than that of control strain (Bacillus subtilis KCTC13112). MBE/L865 was identified as B. subtilis and deposited in the Korean Collection for Type Cultures under the accession number of KCCM43059. The optimum growth conditions for B. subtilis KCCM43059 were determined to be $37^{\circ}C$ and pH 8. The strain showed maximum protease activity ($429.37{\pm}18.65U/mg$ protein) at $60^{\circ}C$ and pH 6. Further, B. subtilis KCCM43059 had a higher salt (NaCl) tolerance than that of the control strain.

• BMB Reports
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• v.42 no.8
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• pp.486-492
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• 2009

OsDREB1D, a special DREB (dehydration responsive element binding protein) homologous gene, whose transcripts cannot be detected in rice (Oryza sativa L), either with or without stress treatments, was amplified from the rice genome DNA. The yeast one-hybrid assay revealed that OsDREB1D was able to form a complex with the dehydration responsive element/C-repeat motif. It can also bind with a sequence of LTRE (low temperature responsive element). To analyze the function of OsDREB1D, the gene was transformed and over-expressed in Arabidopsis thaliana cv. Columbia. Results indicated that the over-expression of OsDREB1D conferred cold and high-salt tolerance in transgenic plants, and that transgenic plants were also insensitive to ABA (abscisic acid). From these data, we deduced that this OsDREB1D gene functions similarly as other DREB transcription factors. The expression of OsDREB1D in rice may be controlled by a special mechanism for the redundancy of function.

• Plant Resources
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• v.8 no.3
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• pp.202-208
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• 2005

Osmotic stress is one of major limiting factors in crop production. In particular, seasonal drought often causes the secondary disease in the field, resulting in severe reduction in both quality and productivity. Recent efforts have revealed that many genes encoding protein kinases play important roles in osmotic stress signal transduction pathways. Previously, the AtSIK (Arabidopsis thaliana Stress Inducible Kinase) mutants have shown to enhance tolerance to abiotic stresses, accompanying with higher expression of abiotic stress-related genes than did the wild-type plants. In this study, we have transformed potato (cv. Taedong Valley) with the AtSIK expression cassette. Both PCR and RT-PCR using AtSIK-specific primers showed stable integration and expression of the AtSIK gene in individual transgenic lines, respectively. Foliar application of herbicide ($Basta^{(R)}$) at commercial application rate (0.3% (v/v)) revealed another evidence of stable gene introduction of T-DNA which includes the bar gene for herbicide resistance. Overexpression of the AtSIK gene under dual CaMV35S promoter increased sensitivity to salt stress (300 mM NaCl), which was demonstrated by the reduction rate of chlorophyll contents in leaves of transgenic potato lines. These results suggest that possible increase of osmotic tolerance in potato plants may be achieved by antisense expression of AtSIK gene.

• Korean Journal of Environmental Agriculture
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• v.34 no.3
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• pp.244-251
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• 2015

BACKGROUND: Recent studies have described the importance of bacteria that can degrade polycyclic aromatic hydrocarbons (PAHs). Here we screened bacterial isolates from commercial gasoline for PAH degraders and characterized their ability to degrade PAHs, lipids and proteins as well as their enantioselective epoxide hydrolase activity, salt tolerance, and seawater survival. METHODS AND RESULTS: One hundred two bacteria isolates from commercial gasoline were screened for PAH degraders by adding selected PAHs on to the surface of agar plates by the sublimation method. A clear zone was found only around the colonies of PAH degraders, which accounted for 13 isolates. These were identified as belonging to Bacillus sp., Brevibacterium sp., Micrococcus sp., Corynebacterium sp., Arthrobacter sp., and Gordonia sp. based on 16S rRNA sequences. Six isolates belonging to Corynebacterium sp., 3 of Micrococcus sp., Arthrobacter sp. S49, and Gordonia sp. H37 were lipid degraders. Arthrobacter sp. S49 was the only isolate showing high proteolytic activity. Among the PAH-degrading bacteria, Arthrobacter sp. S49, Brevibacterium sp. S47, Corynebacterium sp. SK20, and Gordonia sp. H37 showed enantioselective epoxide hydrolase activity with biocatalytic resolution of racemic styrene oxide. Among these, highest enantioselective hydrolysis activity was seen in Gordonia sp. H37. An intrinsic resistance to kanamycin was observed in most of the isolates and Corynebacterium sp. SK20 showed resistance to additional antibiotics such as tetracycline, ampicillin, and penicillin. CONCLUSION: Of the 13 PAH-degraders isolated from commercial gasoline, Arthrobacter sp. S49 showed the highest lipid and protein degrading activity along with high active epoxide hydrolase activity, which was the highest in Gordonia sp. H37. Our results suggest that bacteria from commercial gasoline may have the potential to degrade PAHs, lipids, and proteins, and may possess enantioselective epoxide hydrolase activity, high salt tolerance, and growth potential in seawater.

• Journal of Microbiology and Biotechnology
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• v.29 no.3
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• pp.347-356
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• 2019

Bacillus sp. BS2 showing strong fibrinolytic activity was isolated from sea squirt (munggae) jeotgal, a traditional Korean fermented seafood. BS2 was identified as B. velezensis by molecular biological methods. B. velezensis BS2 grows well at 15% NaCl and at $10^{\circ}C$. When B. velezensis BS2 was cultivated in TSB broth for 96 h at $37^{\circ}C$, the culture showed the highest fibrinolytic activity ($131.15mU/{\mu}l$) at 96 h. Three bands of 27, 35 and 60 kDa were observed from culture supernatant by SDS-PAGE, and fibrin zymography showed that the major fibrinolytic protein was the 27 kDa band. The gene (aprEBS2) encoding the major fibrinolytic protein was cloned, and overexpressed in heterologous hosts, B. subtilis WB600 and E. coli BL21 (DE3). B. subtilis transformant showed 1.5-fold higher fibrinolytic activity than B. velezensis BS2. Overproduced AprEBS2 in E. coli was purified by affinity chromatography. The optimum pH and temperature were pH 8.0 and $37^{\circ}C$, respectively. $K_m$ and $V_{max}$ were 0.15 mM and $39.68{\mu}M/l/min$, respectively, when N-succinyl-Ala-Ala-Pro-Phe-pNA was used as the substrate. AprEBS2 has strong ${\alpha}$-fibrinogenase and moderate ${\beta}$-fibrinogenase activity. Considering its high fibrinolytic activity, significant salt tolerance, and ability to grow at $10^{\circ}C$, B. velezensis BS2 can be used as a starter for jeotgal.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2002.04a
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• pp.41-47
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• 2002

Adverse environmental conditions such as drought, high salt and cold/freezing are major factors that reduces crop productivity worldwide. According to a survey, $50-80\%$ of the maximum potential yield is lost by these 'environmental or abiotic stresses', which is approximately ten times higher than the loss by biotic stresses. Thus, Improving stress-tolerance of crop plants is an important way to improve agricultural productivity. In order to develop such stress-tolerant crop plants, we set out to identify key stress signaling components that can be used to develop commercially viable crop varieties with enhanced stress tolerance. Our primary focus so far has been on the identification of transcription factors that regulate stress responsive gene expression, especially those involved in ABA-mediated stress response. Be sessile, plants have the unique capability to adapt themselves to the abiotic stresses. This adaptive capability is largely dependent on the plant hormone abscisic acid (ABA), whose level increases under various stress conditions, triggering adaptive response. Central to the response is ABA-regulated gene expression, which ultimately leads to physiological changes at the whole plant level. Thus, once identified, it would be possible to enhance stress tolerance of crop plants by manipulating the expression of the factors that mediate ABA-dependent stress response. Here, we present our work on the isolation and functional characterization of the transcription factors.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2002.04b
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• pp.41-47
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• 2002

Adverse environmental conditions such as drought, high salt and cold/freezing are major factors that reduces crop productivity worldwide. According to a survey, 50-80% of the maximum potential yield is lost by these 'environmental or abiotic stresses', which is approximately ten times higher than the loss by biotic stresses. Thus, improving stress-tolerance of crop plants is an important way to improve agricultural productivity. In order to develop such stress-tolerant crop plants, we set out to identify key stress signaling components that can be used to develop commercially viable crop varieties with enhanced stress tolerance. Our primary focus so far has been on the identification of transcription factors that regulate stress responsive gene expression, especially those involved in ABA-mediated stress response. Be sessile, plants have the unique capability to adapt themselves to the abiotic stresses. This adaptive capability is largely dependent on the plant hormone abscisic acid (ABA), whose level increases under various stress conditions, triggering adaptive response. Central to the response is ABA-regulated gene expression, which ultimately leads to physiological changes at the whole plant level. Thus, once identified, it would be possible to enhance stress tolerance of crop plants by manipulating the expression of the factors that mediate ABA-dependent stress response. Here, we present our work on the isolation and functional characterization of the transcription factors.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.6
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• pp.818-823
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• 2012

Tall fescue (Festuca arundinacea Schreb.) is an important cool season forage plant that is not well suited to extreme heat, salts, or heavy metals. To develop transgenic tall fescue plants with enhanced tolerance to abiotic stress, we introduced an alfalfa Hsp23 gene expression vector construct through Agrobacterium-mediated transformation. Integration and expression of the transgene were confirmed by polymerase chain reaction, northern blot, and western blot analyses. Under normal growth conditions, there was no significant difference in the growth of the transgenic plants and the non-transgenic controls. However, when exposed to various stresses such as salt or arsenic, transgenic plants showed a significantly lower accumulation of hydrogen peroxide and thiobarbituric acid reactive substances than control plants. The reduced accumulation of thiobarbituric acid reactive substances indicates that the transgenic plants possessed a more efficient reactive oxygen species-scavenging system. We speculate that the high levels of MsHsp23 proteins in the transgenic plants protect leaves from oxidative damage through chaperon and antioxidant activities. These results suggest that MsHsp23 confers abiotic stress tolerance in transgenic tall fescue and may be useful in developing stress tolerance in other crops.