• Title/Summary/Keyword: oxidative stress-inducible SWPA2 promoter

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Transgenic poplar expressing AtNDPK2 exhibits enhanced biomass in the LMO field

  • An, Chul-Han;Kim, Yun-Hee;Park, Sung-Chul;Jeong, Jae-Cheol;Lee, Haeng-Soon;Choi, Yong-Im;Noh, Eun-Woon;Yun, Dae-Jin;Kim, Se-Bin;Kwak, Sang-Soo
    • Journal of Plant Biotechnology
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    • v.38 no.3
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    • pp.228-233
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    • 2011
  • Nucleoside diphosphate kinase 2 (NDPK2) is known to regulate the expression of antioxidant genes and auxin-responsive genes in plants. Previously, it was noted that the overexpression of Arabidopsis NDPK2 (AtNDPK2) under the control of an oxidative stress-inducible SWPA2 promoter in transgenic poplar (Populus alba ${\times}$ P. tremular var. glandulosa) plants (referred to as SN plants) enhanced tolerance to oxidative stress and improved growth (Plant Biotechnol J 9: 34-347, 2011). In this study, growth of transgenic poplar was assessed under living modified organism (LMO) field conditions in terms of biomass in the next year. The growth of transgenic poplar plants increased in comparison with non-transgenic plants. The SN3 and SN4 transgenic lines had 1.6 and 1.2 times higher dry weight in stems than non-transgenic plants at 6 months after planting, respectively. Transgenic poplar also exhibited increased transcript levels of auxin-response genes such as IAA1, IAA2, IAA5 and IAA6. These results suggest that enhanced AtNDPK2 expression increases plant biomass in transgenic poplar through the regulation of auxin-response genes.

Transgenic Siberian Ginseng Cultured Cells That Produce High Levels of Human Lactoferrin (인체 락토페린 생산 형질전환 가시오갈피 배양세포)

  • Jo Seung-Hyun;Kwon Suk-Yoon;Kim Jae-Whune;Lee Ki-Teak;Kwak Sang-Soo;Lee Haeng-Soon
    • Journal of Plant Biotechnology
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    • v.32 no.3
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    • pp.209-215
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    • 2005
  • Lactoferrin is an iron-binding glycoprotein with many biological roles, including the protection against microbial and virus infection, stimulation of the immune system. We developed the transgenic Siberian ginseng (Acanthopanax senticosus) cell cultures producing the human lactoferrin (hLf) protein following Agrobacterium tumefaciens-mediated transformation. A construct containing a targeting signal peptide from tobacco endoplasmic reticulum fused to hLf cDNA under the control of an oxidative stress-inducible SWPA2 promoter was engineered. Transgenic Siberian ginseng cultured cells to produce a recombinant hLf protein were successfully generated and confirmed by PCR and Southern blot analysis. ELISA and western blot analysis showed that full length-hLf protein was synthesized in the transgenic cells. The production of hLf increased proportionally to cell growth and reached a maximal (up to 3% of total soluble proteins) at the stationary phase. These results suggest that the transgenic Siberian ginseng cultured cells in this study will be biotechnologically useful for the commercial production of medicinal plant cell cultures to produce hLf protein.

Enhanced Tolerance to Oxidative Stress of Transgenic Potato (cv. Superior) Plants Expressing Both SOD and APX in Chloroplasts (SOD와 APX를 동시에 엽록체에 발현시킨 형질전환 감자 (cv. Superior)의 산화스트레스 내성 증가)

  • Tang, Li;Kwon, Suk-Yoon;Kim, Myoung-Duck;Kim, Jin-Seog;Kwak, Sang-Soo;Lee, Haeng-Soon
    • Journal of Plant Biotechnology
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    • v.34 no.4
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    • pp.299-305
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    • 2007
  • Oxidative stress is a major damaging factor for plants exposed to environmental stresses. Previously, we have generated transgenic potato (cv. Superior) plants expressing both CuZnSOD and APX genes in chloroplast under the control of an oxidative stress-inducible SWPA2 promoter (referred to as SSA plants) and selected the transgenic potato plant lines with tolerance against methyl viologen (MV)-mediated oxidative stress. When leaf discs of SSA plants were subjected to $3{\mu}M$ methyl viologen (MV), they showed approximately 40% less damage than non-transgenic (NT) plants. SSA plantlets were treated with $0.3{\mu}M$ MV stress, SSA plants also exhibited reduced damage in root growth. When 350 MV was sprayed onto the whole plants, SSA plants showed a significant reduction in visible damage, which was approximately 75% less damage than leaves of NT plants. These plants will be used for further analysis of tolerance to environmental stresses, such as high temperature and salt stress. These results suggest that transgenic potato (cv. Superior) plants would be a useful plant crop for commercial cultivation under unfavorable growth conditions.

Enhanced Salt Stress Tolerance in Transgenic Potato Plants Expressing IbMYB1, a Sweet Potato Transcription Factor

  • Cheng, Yu-Jie;Kim, Myoung-Duck;Deng, Xi-Ping;Kwak, Sang-Soo;Chen, Wei
    • Journal of Microbiology and Biotechnology
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    • v.23 no.12
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    • pp.1737-1746
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    • 2013
  • IbMYB1, a transcription factor (TF) for R2R3-type MYB TFs, is a key regulator of anthocyanin biosynthesis during storage of sweet potatoes. Anthocyanins provide important antioxidants of nutritional value to humans, and also protect plants from oxidative stress. This study aimed to increase transgenic potatoes' (Solanum tuberosum cv. LongShu No.3) tolerance to environmental stress and enhance their nutritional value. Transgenic potato plants expressing IbMYB1 genes under the control of an oxidative stress-inducible peroxidase (SWPA2) promoter (referred to as SM plants) were successfully generated through Agrobacterium-mediated transformation. Two representative transgenic SM5 and SM12 lines were evaluated for enhanced tolerance to salinity, UV-B rays, and drought conditions. Following treatment of 100 mM NaCl, seedlings of SM5 and SM12 lines showed less root damage and more shoot growth than control lines expressing only an empty vector. Transgenic potato plants in pots treated with 400 mM NaCl showed high amounts of secondary metabolites, including phenols, anthocyanins, and flavonoids, compared with control plants. After treatment of 400 mM NaCl, transgenic potato plants also showed high DDPH radical scavenging activity and high PS II photochemical efficiency compared with the control line. Furthermore, following treatment of NaCl, UV-B, and drought stress, the expression levels of IbMYB1 and several structural genes in the flavonoid biosynthesis such as CHS, DFR, and ANS in transgenic plants were found to be correlated with plant phenotype. The results suggest that enhanced IbMYB1 expression affects secondary metabolism, which leads to improved tolerance ability in transgenic potatoes.

Development of transgenic cassava plants expressing IbOr gene by somatic embryogenesis (체세포배발생에 의한 IbOr 유전자 형질전환 카사바 개발)

  • Kim, Sun Ha;Kim, Myoung Duck;Park, Sung-Chul;Jeong, Jae Cheol;Lee, Haeng-Soon;Kwak, Sang-Soo
    • Journal of Plant Biotechnology
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    • v.42 no.2
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    • pp.88-92
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    • 2015
  • Cassava (Manihot esculenta Crantz) is a useful root crop for food, animal feed and various industrial materials including biofuel. Despite of its importance as an industrial crop, the genetic engineering approaches to manipulate transgenic plant development in cassava are limited. In this study, to develop new cultivar with high level of carotenoids and enhanced tolerance to environmental stresses, sweetpotato IbOr gene involved in accumulation of carotenoids was introduced into an Indonesian IDB high-yielding cassava cultivar under the control of oxidative stress-inducible SWPA2 promoter through Agrobacterium-mediated transformation of friable embryogenic calli. The 19 transgenic lines were successfully generated on the basis of gDNA-PCR and IbOr transcript levels for further characterization in terms of carotenoid contents and environmental stresses. Therefore, IbOr transgenic cassava plants may be developed for enhanced biomass production with high levels of carotenoids on marginal lands.

Selection of transgenic sweetpotato plants expressing 2-Cys peroxiredoxin with enhanced tolerance to oxidative stress (Peroxiredoxin 유전자 발현 산화스트레스 내성 형질전환 고구마의 선발)

  • Kim, Myoung-Duck;Yang, Kyoung-Sil;Kwon, Suk-Yoon;Lee, Sang-Yeol;Kwak, Sang-Soo;Lee, Haeng-Soon
    • Journal of Plant Biotechnology
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    • v.36 no.1
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    • pp.75-80
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    • 2009
  • In order to develop transgenic sweetpotato plants [Ipomoea batatas (L.) Lam. cv. Yulmi] with enhanced tolerance to oxidative stress, we constructed transformation vectors expressing 2-Cys peroxiredoxin (Prx) gene under the control of the stress-inducible SWPA2 or enhanced 35S promoter (named as SP or EP). Transgenic sweetpotato plants were attempted to generate from embryogenic calli using an Agrobacterium-mediated transformation system. Embryogenic calli gave rise to somatic embryos and then converted into plantlets on MS medium containing 100 mg/L kanamycin. Transgenic plants were regenerated in the same medium. Southern blot analysis confirmed that the Prx gene was inserted into the genome of the plants. To further study we selected the transgenic plant lines with enhanced tolerance against methyl viologen (MV). When sweetpotato leaf discs were subjected to methyl MV at $20{\mu}M$, transgenic plants showed about 40% higher tolerance than non-transgenic or empty vector-transformed plants.

High-yield Production of Functional Human Lactoferrin in Transgenic Cell Cultures of Siberian Ginseng(Acanthopanax senticosus)

  • Jo, Seung-Hyun;Kwon, Suk-Yoon;Park, Doo-Sang;Yang, Kyoung-Sil;Kim, Jae-Whune;Lee, Ki-Teak;Kwak, Sang-Soo;Lee, Haeng-Soon
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.5
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    • pp.442-448
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    • 2006
  • Human lactoferrin (hLf) is an iron-binding glycoprotein that has been considered to play many biological roles in the human, including the stimulation of the immune system, antimicrobial and anti-inflammatory effects, and regulation of iron absorption. We generated transgenic Siberian ginseng (Acanthopanax senticosus) cell cultures producing a functional hLf protein using the signal peptide sequence from the endoplasmic reticulum and driven by an oxidative stress-inducible SWPA2 promoter which is highly expressed in plant cell cultures. The production of hLf increased proportionally to cell growth and showed a maximal level (up to 3.6% of total soluble protein) at the stationary phase in suspension cultures. Full-length hLf protein was identified by immunoblot analysis in transgenic cell cultures of Siberian ginseng. Recombinant hLf (rhLf) was purified from suspension cells of Siberian ginseng by ammonium sulfate precipitation, cation-exchange and gel filtration chromatography. N-terminal sequences of rhLf were identical to native hLf (nhLf). The overall monosaccharide composition of rhLf showed the presence of plant specific xylose while sialic acid is absent. Antibacterial activity of purified rhLf was higher than that of nhLf. Taken together, we anticipate that medicinal Siberian ginseng cultured cells, as demonstrated by this study, will be a biotechnologically useful source for commercial production of functional hLf not requiring further purification.

Characterization of Transgenic Tall Fescue Plants Expressing Two Antioxidant Genes in Response to Environmental Stresses (두 가지 항산화유전자를 동시에 발현시킨 형질전환 톨 페스큐 식물체의 환경스트레스에 대한 내성 특성 해명)

  • Lee, Sang-Hoon;Lee, Ki-Won;Kim, Ki-Yong;Choi, Gi-Jun;Seo, Sung;Kwak, Sang-Soo;Kwon, Suk-Yoon;Yun, Dae-Jin;Lee, Byung-Hyun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.27 no.2
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    • pp.109-116
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    • 2007
  • Environmental stress is the major limiting factor in plant productivity. As an effort to solve the global food and environmental problems using the plant biotechnology, we have developed transgenic tall fescue (Festuca arundinacea Schreb.) plants via Agrobacterium-mediated gene transfer method. To develop transgenic tall fescue plants with enhanced tolerance to the environmental stresses, both CuZn superoxide dismutase (CuZnSOD) and ascorbate peroxidase (APX) genes were incorporated in a pIG121 binary vector and the both of the genes were controlled separately by an oxidative stress-inducible sweet potato peroxidase 2 (SWPA2) premoter expressed in chloroplasts. Leaf discs of transgenic plants showed 10-30% less damage compared to the wild-type when they exposed to a wide range of environmental stresses including methyl viologen (MV), $H_2O_2$ and heavy metals. In addition, when $200{\mu}M$ MV was sprayed onto the whole plants, transgenic plants showed a significant reduction of visible damage compared to wild-type plants that were almost damaged. These results suggest that over expression of CuZnSOD and APX genes in transgenic plants might be a useful strategy to protect the crops against a wide range of environmental stresses.

Chloroplast-targeted Expression of PsAPX1 Enhances Tolerance to Various Environmental Stresses in Transgenic Rice (엽록체에 과발현된 PsAPX1 유전자 형질전환 벼의 다양한 환경스트레스 내성 증진)

  • Park, Hyang-Mi;Kim, Yul-Ho;Choi, Man-Soo;Lee, Jae-Eun;Choi, In-Bea;Choi, Im-Soo;Shin, Dong-Bum;Lee, Jang-Yong;Kwak, Sang-Soo;Kwon, Suk-Yoon
    • Korean Journal of Breeding Science
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    • v.41 no.3
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    • pp.261-270
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    • 2009
  • Oxidative stress is a major damaging factor for plants exposed to environmental stresses. In order to develop transgenic rice plants with enhanced tolerance to various environmental stresses, PsAPX1, the gene of ascorbate peroxidase isolated from Pisum sativum was expressed in chloroplast under the control of an oxidative stress inducible sweet potato peroxidase2 (SWPA2) promoter (referred to as PsAPX1 plants). PsAPX1 transgenic plants showed enhanced tolerance to various environmental stresses, such as 170 mM NaCl, UV-B, ozone, 20% PEG, and drought in compared with non-transgenic (NT) plants. These results suggest that chloroplast-targeted over-expression of PsAPX1 gene could be very useful strategy for developing transgenic rice plants with increased tolerance to environmental stresses.