• Title/Summary/Keyword: cold-stress protein

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Molecular Cloning and Expression of the Metallothionein Gene under Environmental Stresses in Sweet Potato (고구마 metallothionein 유전자의 클로닝 및 환경 스트레스 하에서 발현 분석)

  • Kim, Young-Hwa;Yu, Eun Jeong;Huh, Gyung-Hye
    • Journal of Life Science
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    • v.27 no.12
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    • pp.1415-1420
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    • 2017
  • The metallothionein (MT) gene (IbMT3) was selected from an EST library of suspension-cultured sweet potato cells. The MT gene, which is one of abundant ESTs in the library, is involved in stress regulation of cells and tissues. A full-length IbMT3 cDNA was obtained and analysis of its nucleotide sequence revealed that IbMT3 encoded a type 3 MT protein, based on its structural characteristics. The function of type 3 MT in plants is not yet known. Northern blot analysis showed stronger expression of IbMT3 in suspension-cultured cells than in sweet potato plant leaves. Since cell culture is known to impose a state of oxidative stress on cells, sweet potato plants were subjected to oxidative stress to investigate the transcriptional regulation of IbMT3. When the herbicide methyl viologen (MV) was administered for 6, 12, and 24 hr, IbMT3 transcription rapidly increased at 6 hr and then decreased. A cold treatment at $15^{\circ}C$ for 24 and 48 hr resulted in a gradual increase in IbMT3 expression. These findings indicate that IbMT3 expression is regulated in response to environmental and oxidative stress. IbMT3 isoform is expected to have antioxidant effects in sweet potato plants and may play an important role in cellular adaptation to oxidative stress.

Gastroprotective effects of the nonsaponin fraction of Korean Red Ginseng through cyclooxygenase-1 upregulation

  • Lee, Jeong-Oog;Kim, Ji Hye;Kim, Sunggyu;Kim, Mi-Yeon;Hong, Yo Han;Kim, Han Gyung;Cho, Jae Youl
    • Journal of Ginseng Research
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    • v.44 no.4
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    • pp.655-663
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    • 2020
  • Background: Korean Red Ginseng is known to exhibit immune-enhancing and anti-inflammatory properties. The immune-enhancing effects of the nonsaponin fraction (NSF) of Korean Red Ginseng have been studied in many reports. However, the gastroprotective effect of this fraction is not fully understood. In this study, we demonstrate the activities of NSF for gastrointestinal protection and its related critical factor. Methods: The in vitro and in vivo regulatory functions of NSF on cyclooxygenase-1 (COX-1) messenger RNA and protein levels were examined by reverse transcription polymerase chain reaction and immunoblotting analyses. Gastroprotective effects of NSF were investigated by histological score, gastric juice pH, and myeloperoxidase activity on indomethacin-induced, cold stress-induced, and acetylsalicylic acid-induced gastritis and dextran sulfate sodium-induced colitis in in vivo mouse models. Results: NSF did not show cytotoxicity, and it increased COX-1 messenger RNA expression and protein levels in RAW264.7 cells. This upregulation was also observed in colitis and gastritis in vivo models. In addition, NSF treatment in mice ameliorated the symptoms of gastrointestinal inflammation, including histological score, colon length, gastric juice pH, gastric wall thickness, and myeloperoxidase activity. Conclusion: These results suggest that NSF has gastroprotective effects on gastritis and colitis in in vivo mouse models through COX-1 upregulation.

LebZIP2 induced by salt and drought stress and transient overexpression by Agrobacterium

  • Seong, Eun-Soo;Kwon, Soon -ung;Ghimire, Bimal Kumar;Yu, Chang-Yeon;Cho, Dong-Ha;Lim, Jung-Dae;Kim, Kyoung-Su;Heo, Kweon;Lim, Eun-Sang;Chung, Ill-Min;Kim, Myong-Jo;Lee, Youn-Su
    • BMB Reports
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    • v.41 no.10
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    • pp.693-698
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    • 2008
  • The full-length cDNA of LebZIP2 (Lycopersicon esculentum bZIP2) encodes a protein of 164 amino acids and contains a N-terminal basic-region leucine zipper domain. Analysis of the deduced tomato LebZIP2 amino acid sequence revealed that it shares 85% sequence identity with both tobacco bZIP and pepper CcbZIP. LebZIP2 mRNA is expressed at a high level exclusively in flowers. Presently, LebZIP2 was strongly increased also following NaCl and mannitol treatments. No significant LebZIP2 expression was evident following cold treatment. Transient LebZIP2 overexpression resulted in increased NbNOA1 and NbNR transcript levels in Nicotiana benthamiana leaves. Our results indicate that LebZIP2 might play roles as an abiotic stress-signaling pathway and as a transcriptional regulator of the NbNOA1 or NbNR genes.

Isolation and characterization of Brcpi1 gene encoding phytocystatin from chinese cabbage (Brassica rapa L.) seedlings (배추 유래 phytocystatin 유전자, Brcpi1의 분리 및 발현특성 분석)

  • Jung, Yu-Jin;Cho, Yong-Gu;Kang, Kwon-Kyoo
    • Journal of Plant Biotechnology
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    • v.36 no.4
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    • pp.407-414
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    • 2009
  • A cDNA clone encoding phytocystatin was isolated from Brassica rapa seedlings, through rapid amplification of cDNA ends (RACE). This gene (name as Brcpi1; GenBank accession no.: EF079953) had a total length of 881 bp with an open reading frame of 609 bp, and encoded predicted polypeptide of 203 amino acid (aa) residues including a putative N-terminal signal peptide. Other relevant regions found its sequence included the G and PW conserved aa motifs, and the consensus LARFAV sequence for phytocystatins and the reactive site QVVAG. The BrCPI1 protein shared 95, 94, 81, 80 and 78% identity with other CPI proterins isolated from Brassica oleracea (BoCPI-1), Arabidopsis thaliana (AtCY SB), Glycine max (GmCPI), Oryza sativa (OsCYS-2) and Zea may (ZmCPI) at amino acid level, respectively. Southern blot analysis showed that Brcpi1 was a low copy gene. Expression pattern analysis revealed that Brcpi1 was a tissue-specific expressing gene during reproductive growth and strongly expressed at mature seedling stages. Furthermore, overexpression of Brcpi1 in transgenic Arabidopsis was enhanced tolerance to salt and cold stresses. Meanwhile the juvenile seedling of Brcpi1 transgenic plants was not affected by various concentrations ABA in MS medium. Taken together, the results showed that Brcpi1 functioned as a cysteine protease inhibitor and it exhibited a protective agent against diverse types of abiotic stress, which induced this gene in a tissue- and stress-specific manner.

Complete genome sequence of Variovorax sp. PMC12, a plant growth-promoting bacterium conferring multiple stress resistance in plants (다양한 스트레스에 대한 식물의 내성을 유도하는 식물생육촉진 세균Variovorax sp. PMC12 균주의 유전체 염기서열)

  • Lee, Shin Ae;Kim, Hyeon Su;Kim, Yiseul;Sang, Mee Kyung;Song, Jaekyeong;Weon, Hang-Yeon
    • 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.

Molecular Cloning and Function Analysis of an Anthocyanidin Synthase Gene from Ginkgo biloba, and Its Expression in Abiotic Stress Responses

  • Xu, Feng;Cheng, Hua;Cai, Rong;Li, Lin Ling;Chang, Jie;Zhu, Jun;Zhang, Feng Xia;Chen, Liu Ji;Wang, Yan;Cheng, Shu Han;Cheng, Shui Yuan
    • Molecules and Cells
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    • v.26 no.6
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    • pp.536-547
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    • 2008
  • Anthocyanidin synthase (ANS, leucoanthocyanidin oxygenase), a 2-oxoglutarate iron-dependent oxygenase, catalyzed the penultimate step in the biosynthesis of the anthocyanin class of flavonoids, from the colorless leucoanthocyanidins to the colored anthocyanidins. The full-length cDNA and genomic DNA sequences of ANS gene (designated as GbANS) were isolated from Ginkgo biloba for the first time. The full-length cDNA of GbANS contained a 1062-bp open reading frame (ORF) encoding a 354-amino-acid protein. The genomic DNA analysis showed that GbANS gene had three exons and two introns. The deduced GbANS protein showed high identities to other plant ANSs. The conserved amino acids (H-X-D) ligating ferrous iron and residues (R-X-S) participating in 2-oxoglutarate binding were found in GbANS at the similar positions like other ANSs. Southern blot analysis indicated that GbANS belonged to a multi-gene family. The expression analysis by real-time PCR showed that GbANS expressed in a tissue-specific manner in G. biloba. GbANS was also found to be up-regulated by all of the six tested abiotic stresses, UV-B, abscisic acid, sucrose, salicylic acid, cold and ethylene, consistent with the promoter region analysis of GbANS. The recombinant protein was successfully expressed in E. coli strain with pET-28a vector. The in vitro enzyme activity assay by HPLC indicated that recombinant GbANS protein could catalyze the formation the cyanidin from leucocyanidin and conversion of dihydroquercetin to quercetin, suggesting GbANS is a bifunctional enzyme within the anthocyanidin and flavonol biosynthetic pathway.

Accelerated Growth of Corynebacterium glutamicum by Up-Regulating Stress-Responsive Genes Based on Transcriptome Analysis of a Fast-Doubling Evolved Strain

  • Park, Jihoon;Lee, SuRin;Lee, Min Ju;Park, Kyunghoon;Lee, Seungki;Kim, Jihyun F.;Kim, Pil
    • Journal of Microbiology and Biotechnology
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    • v.30 no.9
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    • pp.1420-1429
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    • 2020
  • Corynebacterium glutamicum, an important industrial strain, has a relatively slower reproduction rate. To acquire a growth-boosted C. glutamicum, a descendant strain was isolated from a continuous culture after 600 generations. The isolated descendant C. glutamicum, JH41 strain, was able to double 58% faster (td=1.15 h) than the parental type strain (PT, td=1.82 h). To understand the factors boosting reproduction, the transcriptomes of JH41 and PT strains were compared. The mRNAs involved in respiration and TCA cycle were upregulated. The intracellular ATP of the JH41 strain was 50% greater than the PT strain. The upregulation of NCgl1610 operon (a putative dyp-type heme peroxidase, a putative copper chaperone, and a putative copper importer) that presumed to role in the assembly and redox control of cytochrome c oxidase was found in the JH41 transcriptome. Plasmid-driven expression of the operon enabled the PT strain to double 19% faster (td=1.82 h) than its control (td=2.17 h) with 14% greater activity of cytochrome c oxidase and 27% greater intracellular ATP under the oxidative stress conditions. Upregulations of genes those might enhance translation fitness were also found in the JH41 transcriptome. Plasmid-driven expressions of NCgl0171 (encoding a cold-shock protein) and NCgl2435 (encoding a putative peptidyl-tRNA hydrolase) enabled the PT to double 22% and 32% faster than its control, respectively (empty vector: td=1.93 h, CspA: td=1.58 h, and Pth: td=1.44 h). Based on the results, the factors boosting growth rate in C. gluctamicum were further discussed in the viewpoints of cellular energy state, oxidative stress management, and translation.

Changes in the Myocardial Antioxidant Enzyme System by Post-Ischemic Reperfusion During Corontory Artery Bypass Operations (관상동맥우회술시 심근허혈후 재관류에 의한 활성산소 방어효소계의 변화)

  • 김응중;김기봉
    • Journal of Chest Surgery
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    • v.29 no.8
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    • pp.850-860
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    • 1996
  • Oxygen free radicals and their metabolites have been implicated as possible causes of reperrusion injury In animal models. Their role in the clinical setting is still controversial. The aim of this study was to evaluate the degree of tissue damage, oxidative stress. and changes in the antioxidant enzyme system in patients undergoing cor nary artery bypass graft operations(CABG) with myocardial protection by cold blood cardioplegia. In patients undergoing CABG(n:10). the levels of lactate dehydrogenate(LDH), creatine phosphokinase MB fraction(CK-MB), and malondialdehyde(M DA) were measured In the coronary sinus effluent before aortic cross clamping and 20 minutes after reperfusion. At the same time, the myocardial tissue activities of superoxide dismutase(SOD). catalase(CAT), glutathione peroxiddse(GSHPX), glutathione reductase (GSSGRd), and glucose 6-phosphate dehydrogenate(GfPDH ) were determined in the right atrial auricle excised before aortic cross clamping and in the left atrial auricle excised 20 minutes after reperfuslon. The levels of increased significantly after reperrusion(p< U.05). There were no significant changes in CAT and CfPDH levels. Western blot analysis was performed to study the induction of antioxidant enzyme and demonstrated increased amount of Cu,Zn-SOD.

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Analysis of receptor like kinase (RLK) gene to stress in rice (Oryza sativa L.) using real-time PCR (Real-time PCR을 이용한 스트레스에 따른 벼의 Receptor like kinase (RLK) 유전자의 발현 변화 분석)

  • Kang, Min-Hee;Kim, Il-Wook;Han, Sang-Hoon;Yun, Choong-Hyo;Yoon, Byoung-Su
    • Journal of Plant Biotechnology
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    • v.35 no.4
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    • pp.281-290
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    • 2008
  • In plant, Receptor-like kinases (RLKs) are protein family, though its function is not yet understood, consisted of a predicted signal sequence, single transmembrane region, and cytoplasmic kinase domain. RLKs are involved in hormonal response pathways, cell differentiation, plant growth and development, self-incompatibility, and symbiont and pathogen recognition. In this study, expression levels of RLG1, RLG5, RLG6, RLG#6, RLG8, RLG10, RLG17, RLG18 and RLG20 were analyzed by Real-time PCR, when rice (Oryzae sativa) was treated abiotic stress. The expression levels of all RLGs were compared each other by analyzed value of threshold cycles ($C_T$). Consequently, RLGs were suppressed by NaCl as salinity stress, and expression of each RLK genes were showed difference treated salicylic acid and wound, respectively. However, All RLGs were induced under low temperature condition. Therefore, our results indicate protection-function of RLK genes to be an early response of rice against cold weather.

A Very Early-Maturing, Cold Tolerant and High Quality japonica Rice Variety 'Hanseol' (극조숙 고품질 내냉성 벼 신품종 '한설')

  • Lee, Jeong-Heui;Shin, Young-Seop;Jeong, O-Young;Kim, Myeong-Ki;Kim, Yeon-Gyu;Kim, Hong-Yeol;Lee, Jeom-Ho;Lee, Jeong-Il;Cho, Young-Chan;Jeon, Yong-Hee;Choi, Yong-Hwan;Yang, Chang-Ihn;Hong, Ha-Cheol;Won, Yong-Jae;Shin, Jin-Chul;Kim, Hyung-Yoon;Seo, Dae-Ha;Hwang, Hung-Goo;Yea, Jong-Doo
    • Korean Journal of Breeding Science
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    • v.42 no.6
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    • pp.632-637
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    • 2010
  • 'Hanseol' is a new very early-maturing, cold tolerant and high quality japonica rice variety developed from a cross of 'Jinbu24' and 'Jinbu25' by the rice breeding team of National Institute of Crop Science (NICS), Rural Development Administration (RDA) in 2009. The heading date of this variety is July 25, which is four days earlier than check variety, 'Jinbubyeo'. 'Hanseol' has 65 cm of culm length, 99 spikelets per panicle, 82.9% of ripened grain rate, and 21.5 g of 1,000 grain-weight of brown rice. This variety shows susceptibility to bacterial leaf blight and virus diseases, and insect pests. It is tolerant to cold stress in terms of less heading delay and high fertility in cold water irrigated cultivation. This variety shows delayed leaf senescence and considerable tolerance to viviparous germination at ripening stage. The milled rice of this variety exhibits translucent, clear non-glutinous endosperm and medium-short grain. 'Hanseol' showed low gelatinization temperature and 6.1% protein content, 19.1% amylose content and good palatability of cooked rice. The milled rice yield of this variety is about 5.43 MT/ha at ordinary culture in local adaptability test for three years. 'Hanseol' would be highly adaptable to mid-north and mid-mountainous areas, and mid-northern alpine area in Korea.