• Title/Summary/Keyword: Tobacco Plant Peroxidase

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Expression of Catalase (CAT) and Ascorbate Peroxidase (APX) in MuSI Transgenic Tobacco under Cadmium Stress

  • Kim, Kye-Hoon;Kim, Young-Nam;Lim, Ga-Hee;Lee, Mi-Na;Jung, Yoon-Hwa
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.1
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    • pp.53-57
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    • 2011
  • The MuSI is known as a multiple stress resistant gene with several lines. A previous study using RT-PCR showed that the expression of MuSI gene in tobacco plant induced its tolerance to Cd stress. This study was conducted to examine the enhanced Cd tolerance of the MuSI transgenic tobacco plant through germination test and to understand the role of the involved antioxidant enzymes for the exhibited tolerance. Germination rate of MuSI transgenic tobacco was more than 10% higher than that of wild-type tobacco, and seedlings of MuSI transgenic tobacco grew up to 1.6 times larger and greener than seedlings of wild-type tobacco at 200 and 300 ${\mu}M$ Cd. From the third to the fifth day, CAT activities at 100 and 200 ${\mu}M$ Cd and APX activities at 100, 200 and 300 ${\mu}M$ Cd of MuSI transgenic tobacco were up to two times higher than those of wild-type tobacco. MuSI gene is shown to enhance the activities of antioxidant enzymes resulting in higher tolerance to oxidative stress compared with the control plant.

Transformation of A Plant by Ascorbate Peroxidase Gene using Agrobacterium tumefaciens (Ascorbate Peroxidase 유전자의 도입에 의한 식물의 형질전환)

  • 이인애;이효신;배은경;김기용;이병현;손대영;조진기
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.22 no.2
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    • pp.101-106
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    • 2002
  • This study was conducted to obtain the transformed tobacco (Nicotiana tubacum) plants with cytosolic ascorbate peroxidase gene(ApxSC7) using Agrobacterium tumefaciens LBA4404. A cDNA encoding the cytosolic ascorbate peroxidase of strawberry, ApxSC7, was introduced into tobacco plants via Agrobacterium-mediated gene transfer system. The expression vector, pIG-AP8, harboring ApxSC7 gene was used for production of transgenic tobacco plants. A large number of transgenic plants were regenerated on a medium containing hygromycin. Integration of ApxSC7 gene was confirmed by PCR and Southern blot analyses with genomic DNA. Northern blot analyses revealed that the pIGap8 gene was constitutively expressed.

Production of Antibacterial Substance, and Changes in Peroxidase nd Polyphenoloxidase Activities in Tobacco Plants Inoculated with Avirulent Isolate of Pseudomonas solanacearum (비병원성 Pseudomonas solanacearum 접종에 의한 담배내 항균물질생성과 Peroxidase 및 Polyphenoloxidase의 변화)

  • Yi Young Keun;Min Tae Gi;Park Won Mok
    • Korean Journal Plant Pathology
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    • v.3 no.3
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    • pp.203-209
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    • 1987
  • The substances obtained from the leaf, stem and root of tobacco plants inoculated with avirulent and virulent isolates of Pseudomonas solanacearum were at R_f\;0.6$ and R_f\;0.9$ on TLC plate, respectively. Both substances showed antibacterial activities not only on P. solanacearum but also on Erwinia carotovora subsp. carotovora and Escherichia coli in vitro. However, the antibacterial substances were not detectable from the filtrate of the autoclaves tobacco sap medium, in which the avirulent or virulent bacterium was cultured for 3 days. Peroxidase and poly phenoloxidase activities and their isozyme patterns did not differ significantly between plants treated with the virulent and avirulent isolates, or between the susceptible cultivar BY 4 and the resistant cultivar NC 82. However, activities of the two enzymes were increased in leaves of the susceptible cultivar BY 4 treated with either the virulent or the avirulent isolate.

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A Novel Oxidative Stress-inducible Peroxidase Promoter and Its Applications to Production of Pharmaceutical Proteins in Transgenic Cell Cultures

  • Lee, Ok-Sun;Park, Sun-Mi;Kwon, Suk-Yoon;Lee, Haeng-Soon;Kim, Kee-Yeun;Kim, Jae-Whune;Kwak, Sang-Soo
    • Journal of Plant Biotechnology
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    • v.4 no.4
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    • pp.143-150
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    • 2002
  • A strong oxidative stress-inducible peroxidase promoter (referred to as SWPA2 promoter) was cloned from tell cultures of sweetpotato (Ipomoea batatas) and characterized in transgenic tobacco cultured cells in terms of biotechnological applications. Employing a transient expression assay in tobacco protoplasts, with five different 5'-deletion mutants of the SWPA2 promoter fused to the $\beta$-glucuronidase (GUS) reporter gene, the 1314 bp deletion mutant showed approximately 30 times higher GUS expression than the CaMV 35S promoter. The expression of GUS activity in suspension cultures of transgenic cells derived from transgenic tobacco leaves containing the -1314 bp SWPA2 promoter-GUS fusion was strongly expressed following 15 days of subculture compared to other deletion mutants, suggesting that the 1314 bp SWPA2 promoter will be biotechnologically useful for the development of transgenic cell lines engineered to produce key pharmaceutical proteins. In this respect, we developed transgenic cell lines such as tobacco (Nicotiana tabacum L. BY-2), ginseng (Panax ginseng) and Siberian ginseng (Acanthopanax senticosus) using a SWPA2 promoter to produce a human lactoferrin (hLf) and characterized the hLf production in cultured cells. The hLf production monitored by ELISA analysis in transgenic BY-2 cells was directly increased proportional to cell growth and reached a maximal level (up to 4.3% of total soluble protein) at the stationary phase in suspension cultures. The SWPA2 promoter should result in higher productivity and increased applications of plant cultured cells for the production of high-value recombinant proteins.

Effect of Plant Growth Regulators on Minimizing Ozone Injury in Tobacco(Nicotiana tabacum L.) (식물생장조절제(植物生長調節劑) 처리(處理)가 담배의 오존 피해경감(被害輕減)에 미치는 영향(影響))

  • Park, K.S.;Cho, J.H.;Sohn, J.K.;Lee, S.C.
    • Korean Journal of Weed Science
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    • v.18 no.1
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    • pp.54-62
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    • 1998
  • This experiment was conducted to find out the effects of ABA and IAA on activities of antioxidant enzymes, antioxidant content, and growth of tobacco under exposure to ozone. The exposure to ozone in tobacco plant significantly decreased plant height, but it did not show any difference in vegetative characteristics except plant height of IAA $10^{-3}$M treated. Total chlorophyll content of NC 82 was dramatically decreased with increase in days after ozone treatment. However, reduction of chlorophyll was minimized when plant growth regulators were treated before ozone exposure. Three days treament of ozone in tobacco increased ascorbic acid of oxidised form, while slightly decreased `in reduced ascorbic acid by IAA treatment. But seven days of ozone treatment showed increase in ascorbic acid and decrease in dehydroascorbic acid. Ozone treatment did not show any difference in glutathione content and glutathione reductase activity when plant growth regulators were treated. Activities of superoxide dismutase(SOD), ascorbate peroxidase(AP) and guaiacol peroxidase(GP) were increased by the exposure to ozone for three days. However, there were no difference in activities of SOD, AP and GP due to exposure to ozone for seven days. These reactions may be interpreted as protective responses to prevent or alleviate the damage of tobacco plant by ozone exposure.

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Responses of Transgenic Tobacco Plants Overexpressing Superoxide Dismutase and Ascorbate Peroxidase in Chloroplasts to Water Stress (Superoxide Dismutase와 Ascorbate Peroxidase를 엽록체에 과발현하는 형질전환 담배의 수분스트레스에 대한 반응)

  • 최선미;권석윤;곽상수;박용목
    • Journal of Environmental Science International
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    • v.10 no.1
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    • pp.79-84
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    • 2001
  • To assess resistance of transgenic tobacco plants which overexpress superoxide dismutase (SOD) and ascorbate peroxidase (APX) in chloroplasts to water stress, changes in leaf water potential, turgor potential, stomatal conductance and transpiration rate were measured. Leaf water potential in all plants remained high up to day 4 after withholding water but thereafter decreased markedly. In spite of a remarkable decrease in leaf water potential, some of transgenic plants maintained higher turgor potential compared with control plant on day 12. In particular, the transgenic plant expressing MnSOD showed an outstanding maintenance in turgor pressure by osmotic adjustment throughout the experiment, resulting in high stomatal conductance and transpiration rate. However, among transgenic plants, osmotic potential was reduced more effectively in multiple transformants such as the double transformant expressing both MnSOD and APX, and the triple transformant expressing CuznSOD, MnSOD and APX than single transformants. Consequently, further research is needed to get general agreement on the tolerance of transgenic plants to water stress at different growth stages for each transgenic plant.

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Responses of Transgenic Tobacco Plants Expressing Sweet Potato Peroxidases to Gamma Radiation (감마선에 대한 고구마 Peroxidase 형질전환 담배식물체의 반응)

  • 윤병욱;이행순;권석윤;김재성;곽상수
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.4
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    • pp.265-269
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    • 1999
  • Transgenic tobacco plants expressing either a sweet potato anionic peroxidase (POD) (swpal) or neutral POD (swpnl) were irradiated by gamma radiation, and the gamma radiation-induced biochemical changes in antioxidant enzymes and plant growth inhibition were investigated at 30 days after treatment. Gamma radiation significantly inhibited the growth of all plants regardless of transgenic or nontransformed plants, showing a dose-dependent inhibition. In high dosage of 50 and 70 Gy, plant heights were severely retarded and new leaves does not emerged. No significant changes in antioxidant enzymes such as POD, superoxide dismutase and catalase were observed in all plants regardless of irradiation dosages ranging from 10 to 50 Gy. These results suggest that sweet potato PODs may be not involved in the protection against the oxidative stress induced by gamma radiation.

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Studies on Peroxidase Isozymes of Tobacco and Korean Radish: implication of Specific Isoperoxidase in Shoot Formation (담배와 무 Peroxidase Isozyme들의 세포 생화학적 연구: 조직 특이성 발현과의 연관성)

  • 김승수
    • Proceedings of the Botanical Society of Korea Conference
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    • 1987.07a
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    • pp.357-389
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    • 1987
  • In an attempt to clarify the physiological functions of individual isoperoxidases, we have studied enzymatic and immunological properties as well as cellular distribution of isoperoxidases from tobacco callus and Korean radish. The gene expression patterns of isoperoxidases in shoot and non-shoot-forming tobbaco callus were also examined by rabbit reticulocyte lysatein vitro translation system. These results indicate that fraction of translatable poly(A)-isoperoxidase mRNA was increased considerably in shoots. At the present time, at least 6-7 isoperoxidases could be detected from the translation mixture of total cellular RNA, among which only one cell wall localized anodic isoperoxidase (named A3) mRNA was bimorphic mRNA. These data suggest the possible regulation of peroxidase activity during shoot formation by altering the polyadenylation state of mRNA. In case of Korean radish seedlings, poly(A)- peroxidase mRNA were also increased depending upon aging.

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Antioxidative Responses of Transgenic Tobacco Plants Expressing both Superoxide Dismutase and Ascorbate Peroxidase in Chloroplasts to Several Herbicides (Superoxide Dismutase와 Ascorbate Peroxidase가 엽록체내로 동시에 과대발현된 형질전환 담배의 제초제들에 대한 항산화 반응)

  • Kim Jin-Seog;Lee Byung-Hoi;Kwon Suk-Yoon;Kim Yun-Hee;Kim So-Hee;Cho Kwang-Yun
    • Journal of Plant Biotechnology
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    • v.32 no.2
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    • pp.97-103
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    • 2005
  • Antioxidative responses of transgenic tobacco plants expressing both superoxide dismutase (SOD) and ascorbate peroxidase (APX) in chloroplasts was investigated with several herbicides. In greenhouse test, tolerance of SOD/APX-overexpressed tobacco (CA) to photosystem (PS) I inhibitor paraquat was increased by about 40%. However, any response differences between CA and wild type (WT) tobacco was not observed in a treatment with PS II inhibitors (bromoxynil, diuron and bromacil), chlorophyll biosynthesis inhibitor(oxyfluorfen), carotenoid biosynthesis inhibitor (fluridone) and 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase inhibitor (glyphosate). This tendency was also similar in the growth chamber test of low light intensity, using paraquat and diuron. That is, increased antioxidant activity of CA was shown only in paraquat treatment. When paraquat was foliar-treated to 6 to 9-leaf stage plant, the third to fourth placed leaf from shoot tip showed relatively higher antioxidant activity. Ascorbate supplemented to paraquat solution alleviated the phytotoxicity with a similar range in both CA and WT. In conclusion, CA specifically responded to oxidative stress induced by paraquat among tested herbicides in a whole plant assay.

A comparison of individual and combined $_L$-phenylalanine ammonia lyase and cationic peroxidase transgenes for engineering resistance in tobacco to necrotrophic pathogens

  • Way, Heather M.;Birch, Robert G.;Manners, John M.
    • Plant Biotechnology Reports
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    • v.5 no.4
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    • pp.301-308
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    • 2011
  • This study tested the relative and combined efficacy of ShPx2 and ShPAL transgenes by comparing Nicotiana tabacum hybrids with enhanced levels of $_L$-phenylalanine ammonia lyase (PAL) activity and cationic peroxidase (Prx) activity with transgenic parental lines that overexpress either transgene. The PAL/Prx hybrids expressed both transgenes driven by the 35S CaMV promoter, and leaf PAL and Prx enzyme activities were similar to those of the relevant transgenic parent and seven- to tenfold higher than nontransgenic controls. Lignin levels in the PAL/Prx hybrids were higher than the PAL parent and nontransgenic controls, but not significantly higher than the Prx parent. All transgenic plants showed increased resistance to the necrotrophs Phytophthora parasitica pv. nicotianae and Cercospora nicotianae compared to nontransgenic controls, with a preponderance of smaller lesion categories produced in Prx-expressing lines. However, the PAL/Prx hybrids showed no significant increase in resistance to either pathogen relative to the Prx parental line. These data indicate that, in tobacco, the PAL and Prx transgenes do not act additively in disease resistance. Stacking with Prx did not prevent a visible growth inhibition from PAL overexpression. Practical use of ShPAL will likely require more sophisticated developmental control, and we conclude that ShPx2 is a preferred candidate for development as a resistance transgene.