• Journal of Plant Biotechnology
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• v.35 no.3
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• pp.185-193
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• 2008

We evaluated the response to salt stress of two different ginseng lines, STG3134 and STG3159, which are sensitive and tolerant, respectively, to salt treatment. Plants were exposed to a 5 dS/m salt solution, and chlorophyll fluorescence was measured. STG3134 ginseng was more sensitive than STG3159 to salt stress. To characterize the cellular response to salt stress in the two different lines, changes in protein expression were investigated using a proteomic approach. Total protein was extracted from detached salt-treated leaves of STG3134 and STG3159 ginseng, and then separated by two-dimensional polyacrylamide gel electrophoresis(2-DE). Approximately 468 protein spots were detected by 2-DE and Coommassie brilliant blue staining. Twenty-two proteins were found to be reproducibly up- or down-regulated in response to salt stress. Among these proteins, twelve were identified using MALDI-TOF MS and ESI-Q-TOF and classified into several functional groups: photosynthesis-related proteins(oxygen-evolving enhancer proteins 1 and 2, rubisco and rubisco activase), detoxification proteins(polyphenol oxidase) and defense proteins($\beta$-1,3-glucanase, ribonuclease-like storage protein, and isoflavone reductase-like protein). The protein levels of ribonuclease-like storage protein, which was highly induced in STG3159 ginseng as compared to STG3134, correlated tightly with mRNA transcript levels, as assessed by reverse-transcription(RT)-PCR. Our results indicate that salinity induces changes in the expression levels of specific proteins in the leaves of ginseng plants. These changes may, in turn, playa role in plant adaptation to saline conditions.

• Journal of Life Science
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• v.31 no.2
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• pp.115-125
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• 2021

Calcium-dependent protein kinases (CDPKs) are known to be involved in regulating plant responses to abiotic stresses such as salinity, cold temperature and dehydration,. Although CDPKs constitute a large multigene family consisting of 31 genes in rice, only a few rice CDPKs' functions have been identified. Therefore, in order to elucidate the functions of OsCPK11 in rice, this study was intended to focus on the expression pattern analysis of OsCPK11 in wild type and ND0001 oscpk11 mutant plants under these abiotic stresses. For the salt, cold and drought stress treatment, seedlings were exposed to 200 mM NaCl, 4℃ and 20% PEG 6,000, respectively. RT-PCR and quantitative real-time PCR were performed to determine the expression patterns of OsCPK11 in wild type and ND0001 mutant plants. RT-PCR results showed that OsCPK11 transcripts in the wild type and heterozygous mutant were detected, but not in the homozygous mutant. Real-time PCR results showed that relative expression of OsCPK11 of wild type plants was increased and reached to the highest level at 24 hr, at 6 hr and at 24 hr under salt, cold and drought stress conditions, respectively. Relative expression of OsCPK11 of ND0001 homozygous plant was significantly reduced compared to that of wild type. These results suggested that oscpk11 homozygous mutant knocks out OsCPK11 and OsCPK11 might be involved in salt, cold and drought stress signaling by regulating its gene expression.

• Journal of Microbiology and Biotechnology
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• v.10 no.3
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• pp.415-418
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• 2000

Productivity of a rice plant is greatly influenced by salt stress. One of the ways to achieve tolerance to salinity is to transfer genes encoding protective enzymes from other organisms, such as microorganisms. The bacterial gene, mtlD, which encodes mannitol-1-phosphate dehydrogenase (Mtl-DH), was introduced to the cytosol of a rice plant by an imbibition technique to overproduce mannitol. The germination and survival rate of the imbibed rice seeds were markedly increased by transferring the mtlD gene when it was delivered in either a pBIN19 or pBmin binary vector. When a polymerase chain reaction was performed with the genomic DNAs of the imbibed rice leaves as a template and with mtlD-specific primers, several lines were shown to contain an exogenous mtlD DNA. However, a reverse transcription (RT)-PCR analysis revealed that not all of them showed an expression of this foreign gene. This paper demonstrates that the growth and germination of rice plants transiently transformed with the bacterial gene, mtlD, are enhanced and these enhancements may have resulted from the experssion of the mtlD gene. The imbibition method empolyed in this study fulfills the requirements for testing the function of such a putative gene in vivo prior to the production of a stable transgenic plant.

• BMB Reports
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• v.40 no.3
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• pp.325-332
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• 2007

The mitogen-activated protein kinase (MAPK) cascade is one of the major and evolutionally conserved signaling pathways and plays pivotal role in the regulation of stress and developmental signals in plants. Here, a novel gene, termed Gossypium hirsutum MAPK (GhMAPK), was isolated from cotton. The full-length cDNA of GhMAPK encodes for a 372 amino acid protein that contains all 11 of the MAPK conserved subdomains and the phosphorylationactivation motif, TEY. Amino acid sequence alignment revealed that GhMAPK shared high identity with group-C MAPK in plants and showed 83~89% similarities with MAPKs from Arabidopsis, apricot, pea, petunia, and tobacco. Southern blot analysis indicated that the GhMAPK belonged to a multygene family in cotton. Two introns were found within the region of genomic sequence. Northern blot analysis revealed that the transcripts of GhMAPK accumulated markedly when the cotton seedlings were subjected to various abiotic stimuli such as wounding, cold (4$^{\circ}C$), or salinity stress; Furthermore, GhMAPK was upregulated by the exogenous signaling molecules, such as salicylic acid (SA) and hydrogen peroxide ($H_2O_2C$), as well as pathogen attacks. These results indicate that the GhMAPK, which has a high degree of identity with group-C plant MAPKs, may also play an important role in response to environmental stresses.

• Journal of Life Science
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• v.19 no.10
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• pp.1456-1462
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• 2009

D-chiro-Inositol, which is the isomer of myo-inositol, is a well known drug for the treatment of type II diabetes. The methylated form of D-chiro-inositol, D-pinitol and D-chiro-inositol are synthesized when the plants are exposed to the abiotic stresses such as drought, salinity and low temperature as osmoprotectants. In soybean, myo-inositol is converted to ononitol by O-methyltransferase, and ononitol is converted to D-pinitol by ononitol epimerase and finally converted to D-chiro-inositol by demethylase. However there have been some reports that in buckwheat, myo-inositol can be converted to D-chiro-inositol directly. This study was conducted to determine the changes of soluble cyclitols in buckwheat seedlings after exposure to salt and drought stresses by GC-FID. The results indicated that myo-inositol may be the precursor of D-chiro-inositol biosynthesis.

• Journal of Crop Science and Biotechnology
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• v.10 no.4
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• pp.257-264
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• 2007

Rice is one of the world's staple crops and is a major source of carbohydrate. Rice is exported from several countries, providing a major source of income. There are many documents reporting that rice is a salt-sensitive crop in its developmental stages. The objective of this investigation is to evaluate the effective salt-tolerance defense mechanisms in aromatic rice varieties. Pathumthani 1(PT1), Jasmine(KDML105), and Homjan(HJ) aromatic rice varieties were chosen as plant materials. Rice seedlings photoautotrophically grown in-vitro were treated with 0, 85, 171, 256, 342, and 427 mM NaCl in the media. Data, including sodium ion$(Na^+)$ and potassium ion$(K^+)$ accumulation, osmolarity, chlorophyll pigment concentration, and the fresh and dry weights of seedlings were collected after salt-treatment for 5 days. $Na^+$ in salt-stressed seedlings gradually accumulated, while $K^+$ decreased, especially in the 342-427 mM NaCl salt treatments. The $Na^+$ accumulation in both salt-stressed root and leaf tissues was positively related to osmolarity, leading to chlorophyll degradation. In the case of the different rice varieties, the results showed that the HJ variety was identified as being salt-tolerant, maintaining root and shoot osmolarities as well as pigment stabilization when exposed to salt stress or $Na^+$ enrichment in the cells. On the other hand, PT1 and KDML105 varieties were classified as salt-sensitive, determined by chlorophyll degradation using Hierarchical cluster analysis. In conclusion, the HJ-salt tolerant variety should be further utilized as a parental line or genetic resource in breeding programs because of the osmoregulation defensive response to salt-stress.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.52 no.4
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• pp.458-468
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• 2007

To tolerate environmentally adverse conditions such as cold, drought, and salinity, plants often synthesize and accumulate proline in cells as compatible osmolytes. ${\Delta}^1$-Pyrroline-5-carboxylate synthetase(P5CS) catalyzes the rate-limiting step of proline biosynthesis from glutamate. Two complete genes, MtP5CS1 and MtP5CS2, were isolated from the model legume Medicago truncatula by cDNA cloning and bacterial artificial chromosome library screening. Nucleotide sequence analysis showed that both genes consisted of 20 exons and 19 introns. Alignment of the predicted amino acid sequences revealed high similarities with P5CS proteins from other plant species. The two MtP5CS genes were expressed in response to high salt and low temperature treatments. Semi-quantitative reverse transcription-polymerase chain reaction showed that MtP5CS1 was expressed earlier than MtP5CS2, indicating differential regulation of the two genes. To evaluate the reverse genetic effects of nucleotide changes on MtP5CS function, a Targeting Induced Local Lesions in Genomes approach was taken. Three mutants each were isolated for MtP5CS1 and MtP5CS2, of which a P5CS2 nonsense mutant carrying a codon change from arginine to stop was expected to bring translation to premature termination. These provide a valuable genetic resource with which to determine the function of the P5CS genes in environmental stress responses of legume crops.

• KSBB Journal
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• v.9 no.2
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• pp.140-146
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• 1994

Aureobasidium pullulans produced high concentration of polyols extracellularly in the media of sucrose, glucose and mannose as sole carbon source. Mannitol was the main polyol produced during the late exponential and stationary phases of growth together with small quantities of glycerol. Sucrose and glucose were rather rapidly metabolized to mannitol among carbon sources examined where the initial glucose concentration showed no difference in the amount of mannitol. In contrast 20%(w/v) of sucrose was the most appropriate concentration tested. However, the yield of mannitol based on substrate used($Y_{p/s}$) was independent on the initial concentration, and the mean value of mannitol yield in 10% glucose and sucrose media was 0.144 and 0.188, respectively. Mannitol production was reduced in response to an elevated water stress imposed by salts within the range from 0.25 to IM of NaCl or KCl as stress solutes. However, glycerol contents and its ratio to mannitol were increased at the conditions of high salinity. Based on the results, extracellular mannitol produced by A. pullulans probably resulted partly from osmoregulation(in case of glycerol) and mainly from, as known to occur in most of fungi, enzymatic reduction of the corresponding hexoses through phosphate pathway.

• Korean Journal of Microbiology
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• v.54 no.4
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• pp.471-473
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• 2018

Variovorax sp. PMC12 is a rhizobacterium isolated from tomato rhizosphere and enhanced the plant resistance to abiotic and biotic stresses. Here we present the complete genome sequence of strain PMC12. The genome is comprised of two circular chromosomes harboring 5,873,297 bp and 1,141,940 bp, respectively. A total of 6,436 protein-coding genes, 9 rRNAs, 64 tRNAs, 3 ncRNAs, and 80 pseudogenes were identified. We found genes involved in 1-aminocyclopropane-1-carboxylate (ACC) deaminase, antioxidant activity, phosphate solubilization, and biosynthesis of proline and siderophore. Those genes may be related to capability of improving plant resistance to various stresses including salinity, cold temperature, and phytopathogen.

• Journal of Microbiology and Biotechnology
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• v.33 no.3
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• pp.378-386
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• 2023

The CYP707A family genes encoding ABA 8'-hydroxylase catabolize abscisic acid (ABA), a plant stress hormone that plays an important role in stress condition, such as drought, heat, cold and salinity. Phaseic acid (PA) is a catabolic product of ABA. Recent studies have shown that PA is important for the physiological functions in plants. It is also a neuroprotective molecule that protects against ischemic brain injury in mice. To obtain enzymes for the PA production, four CaCYP707A genes (CaCYP707A1, CaCYP707A2, CaCYP707A3 and CaCYP707A4) were isolated from hot pepper. They were heterologously expressed in Escherichia coli. Among them, CaCYP707A2 showed significantly higher expression levels in both the membrane fraction and the soluble fraction. Preferred redox partners were investigated to improve the efficiency of CaCYP707A2's catalytic reaction, and NADPH-cytochrome P450 reductase (CPR) from hot pepper (CaCPR) was preferred over other redox partners (i.e., rat CPR and ferredoxin reductase/ferredoxin). The production of 8'-hydroxy ABA and PA by ABA hydroxylation activity was confirmed in CaCYP707A2 from both membrane and soluble fractions. Therefore, CaCYP707A2 is the first identified plant CYP protein that is expressed a soluble form in cytosolic fraction having stable activity. Taken together, we propose a new CYP707A protein with industrial applications for PA production without additional modifications in E. coli heterologous expression.