• Journal of Ginseng Research
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• 제47권6호
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• pp.714-725
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• 2023

Background: Our previous investigation indicated that the preparation of Panax ginseng Meyer (P. ginseng) inhibited melanogenesis. It comprised salicylic acid (SA), protocatechuic acid (PA), p-coumaric acid (p-CA), vanillic acid (VA), and caffeic acid (CA). In this investigation, the regulatory effects of P. ginseng phenolic acid monomers on melanin production were assessed. Methods: In vitro and in vivo impact of phenolic acid monomers were assessed. Results: SA, PA, p-CA and VA inhibited tyrosinase (TYR) to reduce melanin production, whereas CA had the opposite effects. SA, PA, p-CA and VA significantly downregulated the melanocortin 1 receptor (MC1R), cycle AMP (cAMP), protein kinase A (PKA), cycle AMP-response element-binding protein (CREB), microphthalmia-associated transcription factor (MITF) pathway, reducing mRNA and protein levels of TYR, tyrosinase-related protein 1 (TYRP1), and TYRP2. Moreover, CA treatment enhanced the cAMP, PKA, and CREB pathways to promote MITF mRNA level and phosphorylation. It also alleviated MITF protein level in α-MSH-stimulated B16F10 cells, comparable to untreated B16F10, increasing the expression of phosphorylation glycogen synthase kinase 3β (p-GSK3β), β-catenin, p-ERK/ERK, and p-p38/p38. Furthermore, the GSK3β inhibitor promoted p-GSK3β and p-MITF expression, as observed in CA-treated cells. Moreover, p38 and ERK inhibitors inhibited CA-stimulated p-p38/p38, p-ERK/ERK, and p-MITF increase, which had negative binding energies with MC1R, as depicted by molecular docking. Conclusion: P. ginseng roots' phenolic acid monomers can safely inhibit melanin production by bidirectionally regulating melanin synthase transcription. Furthermore, they reduced MITF expression via MC1R/cAMP/PKA signaling pathway and enhanced MITF post-translational modification via Wnt/mitogen-activated protein kinase signaling pathway.

• BMB Reports
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• 제40권3호
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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.

• Molecules and Cells
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• 제22권1호
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• pp.58-64
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• 2006

WRKY family proteins are a class of plant-specific transcription factors involved in stress response signaling pathways. In this study a gene encoding a putative WRKY protein was isolated from a pepper EST database (http://genepool.kribb.re.kr). The cDNA, named Capsicum annuum WRKY2 (CaWRKY2), encodes a putative polypeptide of 548 amino acids, containing two WRKY domains with zinc finger motifs and two potential nuclear localization signals. Northern blot analyses showed that CaWRKY2 mRNA was preferentially induced during incompatible interactions of pepper plants with PMMoV, Pseudomonas syringae pv. syringae 61, and Xanthomonas axonopodis pv. vesicatoria race 3. Furthermore, CaWRKY2 transcripts were strongly induced by wounding and ethephon treatment, whereas only moderate expression was detected following treatment with salicylic acid and jasmonic acid. CaWRKY2 was translocated to the nucleus when a CaWRKY2-smGFP fusion construct was expressed in onion epidermal cells. CaWRKY2 also had transcriptional activation activity in yeast. Taken together our data suggest that CaWRKY2 is a pathogen-inducible transcription factor that may have a role in early defense responses to biotic and abiotic stresses.

• Plant Biotechnology Reports
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• 제4권2호
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• pp.165-172
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• 2010

Genes that are expressed early in specific response to high salinity conditions were isolated from rapeseed plant (Brassica napus L.) using an mRNA differential display method. Five PCR fragments (DD1.5) were isolated that were induced by, but showed different response kinetics to, 200 mM NaCl. Nucleotide sequence analysis and homology search revealed that the deduced amino sequences of three of the five cDNA fragments showed considerable similarity to those of ${\beta}$-mannosidase (DD1), tomato Pti-6 proteins (DD5), and the tobacco harpin-induced protein hin1 (DD4), respectively. In contrast, the remaining clones, DD3 and DD2, did not correspond to any substantial existing annotation. Using the DD3 fragment as a probe, we isolated a full-length cDNA clone from the cDNA library, which we termed BnSWD1 (Brassica napus salt responsive WD40 1). The predicted amino-acid sequence of BnSWD1 contains eight WD40 repeats and is conserved in all eukaryotes. Notably, the BnSWD1 gene is expressed at high levels under salt-stress conditions. Furthermore, we found that BnSWD1 was upregulated after treatment with abscisic acid, salicylic acid, and methyl jasmonate. Our study suggests that BnSWD1, which is a novel WD40 repeat-containing protein, has a function in salt-stress responses in plants, possibly via abscisic acid-dependent and/or -independent signaling pathways.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2022년도 추계학술대회
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• pp.232-232
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• 2022

We reported in our recent studies that the combined treatment of Paenibacillus yonginensis DCY84^T (DCY84^T) and Silicon (Si) promotes initial plant growth and increases resistance to biotic and abiotic stress. To understand the molecular background of these phenotypes, Liquid Chromatography Mass Spectrometry (LC-MS) analysis was performed, and it was confirmed that unsaturated fatty acid metabolites such as oleic acid and linoleic acid decreased in response to the combined treatment of DCY84^T and Si. The stearoyl-acyl carrier protein desaturase (SACPD) introduces the cis double bond into the acyl-ACPs at C9, resulting in the production of unsaturated fatty acid. We identified OsSSI2 encoding SACPD in rice and found that the expression of OsSSI2 was reduced under DCY84^T and Si treatment. Furthermore, qRT-PCR analysis revealed that the expression of OsWRKY45, which is downstream of OsSSI2, was upregulated in response to DCY84^T and Si treatment. These results enable the speculation that activation of the salicylic acid (SA)-responsive gene, OsWRKY45, may contribute to enhancing biological stress resistance. Based on this, we propose a probable model for the rice defense pathway following DCY84^T and Si treatment. This model retains a WRKY45-dependent but NH1(NPR1)-independent SA signaling pathway.

• Journal of Plant Biotechnology
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• 제44권1호
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• pp.61-68
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• 2017

GASA는 GA에 의해 조절되는 식물 유전자로서, 여러 식물에 보존되어 있고 다양한 조직에서 식물의 생장과 발달 및 스트레스 반응에 관여하는 것으로 알려져 있다. 본 연구에서는 GASA 유전자를 현사시나무(Populus alba ${\times}$ P. glandulosa)에서 분리하여 이를 PagGASA라 명명하고, 유전자의 구조와 발현특성을 조사하였다. PagGASA 유전자는 95개의 아미노산으로 구성된 단백질을 암호화하며, 아미노 말단에 시그널 펩티드 영역과 카르복시 말단에 12개 시스테인 잔기가 보존되어 있다. PagGASA는 현사시나무의 염색체에 1 ~ 2 copy 존재하며, 꽃과 뿌리에서 높게 발현하였다. 또한 PagGASA는 GA 뿐 아니라 ABA와 JA, SA와 같은 스트레스 관련 식물 호르몬의 처리에 의해서 발현이 증가하는 것으로 나타났다. 현사시나무에 형질전환하여 PagGASA를 과발현시킨 결과 건조 내성에 효과가 있음을 확인하였다. 따라서 PagGASA는 스트레스 관련 식물 호르몬 신호전달과 연결되어 건조 스트레스 방어기작에서 중요한 역할을 할 것으로 생각된다.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.164-164
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• 2017

High throughput transcriptome investigations of immunity in plants highlight the complexity of gene networks leading to incompatible interaction. To identify genes crucial to resistance against Xanthomonas oryzae pv oryzae, functional genetic analysis of selected differentially expressed genes from our microarray data set was carried out. A total of 13 overexpression vector constructs were made using 35S CaMV promoter which drive constitutive expression in rice. Most of the genes are developmentally expressed especially during maximum tillering stage and are commonly highly expressed in the leaves. When screened against Xoo strain K2, the transgenic plants displayed shorter lesion length compared with wild type Dongjin which indicates partial resistance. The levels of ROS continuously magnified after inoculation which indicates robust cellular sensing necessary to initiate cell death. Elevated transcripts levels of several defense-related genes at the downstream of defense signal network also corroborate the phenotype reaction of the transgenic plants. Moreover, expression assays revealed regulation of these genes by cross-communicating signal-transductions pathways mediated by salicylic and jasmonic acid. These collective findings revealed the key immune signaling conduits critical to mount full defense against Xoo.

• Animal cells and systems
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• 제9권4호
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• pp.239-244
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• 2005

A cDNA clone for a wound- and pathogen-induced gene in Chinese cabbage (Brassica rapa subsp. pekinensis) was isolated and characterized. The cabbage gene, designated BrPR4, encodes a pathogenesis-related protein 4 (PR4) of 140 amino acids. The BrPR4 protein shows high similarity with wound-inducible antifungal proteins of tobacco, potato, barley, and wheat. The BrPR4 gene is locally induced by a nonhost pathogen, Pseudomonas syringae pv. tomato, that elicits a hypersensitive response in Chinese cabbage. Treatment of the cabbage leaves with benzothiadiazole (BTH), methyl jasmonate or ethephon showed that the BrPR4 gene expression is strongly induced by ethylene, but not by methyl jasmonate or BTH. The BrPR4 gene is also activated by wounding. Interestingly, however, the wound-inducible BrPR4 gene expression is repressed by salicylic acid or BTH, suggesting that there is cross-talk between salicylate-dependent and -independent signaling pathways.

• Journal of Plant Biotechnology
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• 제31권1호
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• pp.83-88
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• 2004

스트레스 관련 화합물 및 병원균을 고구마 (Ipomoea batatas)잎에 처리하여 고구마 배양세포에서 분리한 산성 퍼옥시다제 swpa2 유전자의 발현을 RT-PCR로 분석하였다. Swpa2유전자는 건강한 식물체 조직에서는 발현되지 않았으나 각종 스트레스에 의해 발현이 유도되었다. Swpa2는 과산화수소 (440mM)처리에 의하여 강하게 발현이 유도되었고 NaCl (100mM), ABA (0.1mM), methyl jasmonate (MeJA, 0.1mM) 처리에 의하여도 발현이 유도되었다. 그러나 salicylic acid (SA)처리에 의해서는 발현이 유도되지 않아, 이 유전자는 SA와 MeJA의 신호전달에는 서로 다르게 관여함이 시사되었다. 또한 swpa2유전자는 무름병을 유발하는 박테리아(Pectobacterium chrysanthemii)의 감염에 의하여 강하게 발현되므로, 이 POD는 병저항성에 관여함을 알 수 있었다. 따라서 swpa2유전자는 각종 비생물학적 스트레스 뿐 아니라 생물학적 스트레스에 수반되는 산화 스트레스를 극복하는데 관여함이 강하게 제시되었다.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.69.1-69
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• 2003

RSH (relA/spoT homolog) has been known to determine the level of guanosine tetraphosphate (ppGpp) and guanosine pentaphosphate (pppGpp), which are the effector nucleotide of the prokaryotic stringent response and also play a role in antibiotic production and differentiation in Streptomyces species but not a little in eukaryotic organism, especially in plant. Salicylic acid (SA), a critical signal molecule of establishing systemic acquired resistance (SAR), could induce SAR in Pepper (Capcicum annuum) against Phytophthora capsici. And the extent of SAR induction was in proportion to the dosage of SA (or BTH). Suppression subtractive hybridization (SSH), a PCR-based method for cDNA subtraction, was carried out between SA-treated and non-SA-treated pepper leaves to isolate genes which may be responsible for defense signaling against pathogens. Early upregulated gene was selected from reverse northern and kinetics of SSH-genes transcripts in SA-treated pepper leaves upon SA treatment. Full-length cDNA of the gene (PepRSH； Pepper RelA / SpoT homolog) had an open reading frame (ORF) of 2166 bp encoding a protein of 722 amino acids and a significant homology with (p)ppGpp phosphohydrolase or synthetase. Genomic DNA gel blot analysis showed that pepper genome has at least single copy of PepRSH. PepRSH transcripts was very low in untreated pepper leaves but strongly induced by SA and methyljasmonic acid (MeJA), indicating that PepRSH may share common SA and MeJA-mediated signal transduction pathway Functional analysis in E. coli showed PepRSH confers phenotypes associated with (p)ppGpp synthesis through a complementation using active site mutagenesis.