• Title, Summary, Keyword: 애기장대

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Development of a Kit for Diagnosing AtCYP78A7 Protein in Abiotic-tolerant Transgenic Rice Overexpressing AtCYP78A7 (AtCYP78A7 과발현 환경스트레스 내성 형질전환 벼의 단백질 진단 키트 개발)

  • Nam, Kyong-Hee;Park, Jung-Ho;Pack, In-Soon;Kim, Ho Bang;Kim, Chang-Gi
    • Journal of Life Science
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    • v.28 no.7
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    • pp.835-840
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    • 2018
  • Quantitative determination of the protein expression levels is one of the most important parts in assessment of the safety of foods derived from genetically modified (GM) crops. Overexpression of AtCYP78A7, a gene encoding cytochrome P450 protein, has been reported to improve tolerance to abiotic stress, such as drought and salt stress, in transgenic rice (Oryza sativa L.). In the present study, an enzyme-linked immunosorbent assay (ELISA) kit for diagnosing AtCYP78A7 protein including AtCYP78A7-specific monoclonal antibody was developed. GST-AtCYP78A7 recombinant protein was induced and purified by affinity column. Four monoclonal antibodies (mAb 6A7, mAb 4C2, mAb 11H6, and mAb 7E8) against recombinant protein were also produced and biotinylated with avidin-HRP. After pairing test using GST-AtCYP78A7 protein and lysate of rice samples, mAb 4C2 and mAb 7E8 were selected as a capture antibody and a detecting antibody, respectively, for ELISA kit. Product test using rice samples indicated that percentages of detected protein in total protein were greater than 0.1% in AtCYP78A7-overexpressing transgenic rice (Line 10B-5 and 18A-4), whereas those in negative control non-transgenic rice (Ilpum and Hwayoung) were less than 0.1%. The ELISA kit developed in this study can be useful for the rapid detection and safety assessment of transgenic rice overexpressing AtCYP78A7.

Herbicidal Properties of 5,8-dihydroxy-1,4-naphthoquinone and Their Possible Mode of Action (천연물 유래 5,8-dihydroxy-1,4-naphthoquinone의 살초특성과 작용기구)

  • Choi, Jung-Sup;Kim, Ji-Yeon;Seo, Bo-Ram;Ko, Young-Kwan;Cha, Mi-Ran;Kim, Young-Sup;Ryu, Shi-Yong;Hwang, In-Taek
    • Korean Journal of Weed Science
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    • v.31 no.3
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    • pp.250-259
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    • 2011
  • This study was conducted to assess the possibility of 5,8-dihydroxy-1,4-naphthoquinone (DHNQ) as a environmental friendly herbicide candidate. Foliar application of DHNQ showed excellent herbicidal effect to the 3 grasses and 5 broad-leaved weeds. Among them, Digitaria sanguinalis and Solanum nigrum were completely controlled by $250{\mu}g\;mL^{-1}$ of DHNQ with main symptoms of desiccation or burndown within 24 hours. Aeschynomene indica was also sensitive to DHNQ treatment. All of the eight weed species were controlled by 90~100% at a concentration of $1000{\mu}g\;mL^{-1}$. However, soil application of DHNQ to Digitaria sanguinalis did not show any herbicidal symptoms. DHNQ strongly inhibited KAPAS activities in vitro and the $IC_{50}$ was $4.4{\mu}M$. Cellular leakage from cucumber leaf squares treated with DHNQ increased depending on the concentrations increased from 6.25 to $100{\mu}M$ after 24 hours incubation with or without light. However, chlorophyll loss in cucumber leaf squares was negligible. Biotin supplements significantly rescued the inhibition of germination rate of Arabidopsis thaliana seeds previously inhibited by the DHNQ. According to above results, DHNQ is a good natural herbicide candidate having a new target KAPAS, which is involved in biotin biosynthesis pathway, with environmental friendly.

The vacuolar processing enzyme (VPE) mutation suppresses an HR-like cell death induced by the double knockout mutant of vacuolar Ca2+-ATPases in Arabidopsis (애기장대에서 두 액포막 칼슘펌프 돌연변이에 의하여 유도되는 세포사멸 표현형의 액포수식효소(VPE) 돌연변이에 의한 억제)

  • Park, Hyeong-Cheol;Lee, Sang-Min;Kim, Ho-Soo;Chung, Woo-Sik
    • Journal of Plant Biotechnology
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    • v.38 no.2
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    • pp.169-175
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    • 2011
  • Calcium ($Ca^{2+}$) signals have been implicated in regulating plant development and responses to the environmental stresses including a programmed cell death pathway. In animals and plants, cytosolic $Ca^{2+}$ signals have been involved in the activation of programmed cell death (PCD). Recently, we reported that disruption of Arabidopsis vacuolar $\b{A}$utoinhibited $\underline{C}a^{2+}$-$\b{A}$TPases (ACAs), ACA4 and ACA11, resulted in the activation of a salicylic acid-dependent programmed cell death pathway. Although extensive studies have revealed various components of a PCD in plants, executors to directly induce PCD are well unknown. Here, we provide that the vacuolar processing enzymes (VPEs) are involved in a PCD induced by the double knockout mutant of vacuolar $Ca^{2+}$-ATPases in Arabidopsis. The gene expression of VPE was rapidly up-regulated and the enzyme activity of VPE was increased in the double mutant plants. We also generated aca4/aca11/avpe, aca4/aca11/${\gamma}$vpe and aca4/aca11/avpe/${\gamma}$vpe mutant plants. Although cell death phenotype of the double mutant plants was not completely disappeared in the triple and quadruple mutant plants, the triple and quadruple mutant plants showed to significantly delay cell death phenotype of the double mutant plants. These results suggest that the VPE is involved in the HR-like cell death in the double mutant of vacuolar $Ca^{2+}$-ATPases in Arabidopsis.

Characterization of SID2 that is required for the production of salicylic acid by using β-GLUCURONIDASE and LUCIFERASE reporter system in Arabidoposis (리포트 시스템을 이용한 살리실산 생합성 유전자 SID2의 발현 해석)

  • Hong, Mi-Ju;Cheong, Mi-Sun;Lee, Ji-Young;Kim, Hun;Jeong, Jae-Cheol;Shen, Mingzhe;Ali, Zahir;Park, Bo-Kyung;Choi, Won-Kyun;Yun, Dae-Jin
    • Journal of Plant Biotechnology
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    • v.35 no.3
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    • pp.169-176
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    • 2008
  • Salicylic acid(SA) is a phytohormone that is related to plant defense mechanism. The SA accumulation is triggered by abiotic and biotic stresses. SA acts as a signal molecular compound mediating systemic acquired resistance and hypersensitive response in plant. Although the role of SA has been studied extensively, an understanding of the SA regulatory mechanism is still lacking in plants. In order to comprehend SA regulatory mechanism, we have been transformed with a SID2 promoter:GUS::LUC fusion construct into siz1-2 mutant and wild plant(Col-0). SIZ1 encodes SUMO E3 ligase and negatively regulates SA accumulation in plants. SID2(SALICYLIC ACID INDUCTION DEFICIENT2) is a crucial enzyme of SA biosynthesis. The Arabidopsis SID2 gene encodes isochorismate synthase(ICS) that controls SA level by conversion of chorismate to isochorismate. We compared the regulation of SID2 in wild-type and siz1-2 transgenic plants that express SID2 promoter:GUS::LUC constructs respectively. The expressions of $\beta$-GLUCURONIDASE and LUCIFERASE were higher in siz 1-2 transgenic plant without any stress treatment. SID2 promoter:GUS::LUC/siz1-2 transgenic plant will be used as a starting material for isolation of siz1-2 suppressor mutants and genes involved in SA-mediated stress signaling pathway.

Organization and function of shoot apical meristem affecting growth and development in plants (식물의 생장과 발달에 영향을 미치는 슈트 정단분열조직의 체제와 기능)

  • Lee, Kyu Bae
    • Journal of Plant Biotechnology
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    • v.41 no.4
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    • pp.180-193
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    • 2014
  • In plants, a shoot apex has a small region known as the shoot apical meristem (SAM) having a group of dividing (initiating) cells. The SAM gives rise to all the groundabove structures of plants throughout their lifetime, and thus it plays important role in growth and development of plants. This review describes theories to explain the SAM organization and function developed over the last 250 years. Since in 1759 German botanist C. F. Wolff has described firstly the SAM, in 1858 Swiss botanist C. N${\ddot{a}}$geli proposed the apical cell theory from the observation of a large single apical cell in the SAM of seedless vascular plants: however, this view was recognized to be unsuitable to seed plants. In 1868, German botanist J. Hanstein suggested the histogen theory: this concept subdividing the SAM into dermatogen, periblem, and plerome was unable to generally apply to seed plants. In 1924, German botanist A. Schmidt proposed the tunica-corpus theory from the examination of angiosperm SAM in which two parts show different planes of cell division: this theory was proved to be not suitable to gymnosperm SAM, not have stable surface tunica layer. In 1938, American botanist A. Foster described zones in gymnosperm SAM based on the cytohistologic differentiation and thus called it a cytohistological zonation theory. With works by E. Gifford, in 1954, this zonation pattern was demonstrated to be also applicable to angiosperm SAM. As another theory, in 1952 French botanist R. Buvat proposed the m${\acute{e}}$rist${\grave{e}}$me d'attente (waiting meristem) theory: however, this concept was confuted because of its negation of function during vegetative growth phase to central initial cells. Rescent studies with Arabidopsis thaliana have found that formation and maintenance of the SAM are under the control of selected genes: SHOOTMERISTEMLESS (STM) gene forms the SAM, and WUSCHEL (WUS) and CLAVATA (CLV) genes function in maintaining the SAM; signaling between WUS and CLV genes act through a negative feedback loop.

Screening of salt-tolerance plants using transgenic Arabidopsis that express a salt cress cDNA library (Salt cress 유전자의 형질전환을 통한 내염성 식물체 선별)

  • Baek, Dongwon;Choi, Wonkyun;Kang, Songhwa;Shin, Gilok;Park, Su Jung;Kim, Chanmin;Park, Hyeong Cheol;Yun, Dae-Jin
    • Journal of Plant Biotechnology
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    • v.41 no.2
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    • pp.81-88
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    • 2014
  • Salt cress (Thellungiella halophila or Thellungiella parvula), species closely related to Arabidopsis thaliana, represents an extremophile adapted to harsh saline environments. To isolate salt-tolerance genes from this species, we constructed a cDNA library from roots and leaves of salt cress plants treated with 200 mM NaCl. This cDNA library was subsequently shuttled into the destination binary vector [driven by the cauliflower mosaic virus (CaMV) 35S promoter] designed for plant transformation and expression via recombination- assisted cloning. In total, 305,400 pools of transgenic BASTA-resistant lines were generated in Arabidopsis using either T. halophila or T. parvula cDNA libraries. These were used for functional screening of genes involved in salt tolerance. Among these pools, 168,500 pools were used for primary screening to date from which 7,157 lines showed apparent salt tolerant-phenotypes in the initial screen. A secondary screen has now identified 165 salt tolerant transgenic lines using 1,551 (10.6%) lines that emerged in the first screen. The prevalent phenotype in these lines includes accelerated seed germination often accompanied by faster root growth compared to WT Arabidopsis under salt stress condition. In addition, other lines showed non-typical development of stems and flowers compared to WT Arabidopsis. Based on the close relationship of the tolerant species to the target species we suggest this approach as an appropriate method for the large-scale identification of salt tolerance genes from salt cress.

Flower and Microspore Development in 'Campbell Early' (Vitis labruscana) and 'Tamnara' (V. spp.) Grapes ('캠벨얼리'와 '탐나라' 포도의 꽃과 소포자 발달)

  • Yim, Bomi;Mun, Jeong-Hwan;Jeong, Young-Min;Hur, Youn Young;Yu, Hee-Ju
    • Horticultural Science & Technology
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    • v.33 no.3
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    • pp.420-428
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    • 2015
  • The majority of cultivated varieties of grape have perfect flowers that are clustered in an individual inflorescence. Grape flower has a single pistil, five stamens, a protective flower cap (calyptra), and a calyx. After fertilization, an individual flower develops into a single berry. Although there are a number of reported studies focusing on berry formation, berry enlargement, and sugar accumulation in grape, the morphological studies of flower, including gametophyte morphogenesis and structural change in floral organs, have not yet been studied in detail. In this study, we investigated the flower structure and development characteristics of grape using microscopy and defined the floral development stages 9 to 13 based on microspore or male gametophyte development stage from tetrad to mature pollen. We used seeded diploid table grapes 'Campbell Early' (Vitis labruscana) and 'Tamnara' (V. spp.) as plant materials. At floral development stage 9, pollen mother cells develop to tetrads. During floral development stages 10 to 11, unicellular microspore develop to mid bicellular pollen. At the end of floral stage 12, male gametophyte develops to mature tricelluar pollen. In floral stage 13, the flower cap falls off and flower bud opens. During floral development stages 9 to 12, there were no major changes in calyx length, whereas the length of the flower cap continuously increased. The flower cap-to-calyx length ratio was 2.0, 3.0, 4.5, and 6.5 at floral stages 9, 10, 11, and 12, respectively. The flower cap-to-calyx length ratio was consistent in the two grape cultivars, suggesting that the ratio is a morphological character representing floral development stage. This study provides a reference for determining floral development stage of the two grape cultivars. It will be useful for the determination of optimum time for microspore culture needed to generate doubled haploid lines and appropriate gibberellic acid treatment needed to induce parthenocarpic fruit development in 'Tamnara' grape.

Effect of LED mixed light conditions on the glucosinolate pathway in brassica rapa (배추 유묘의 글루코시놀레이트 합성 기작에 미치는 LED 혼합광의 효과)

  • Moon, Junghyun;Jeong, Mi Jeong;Lee, Soo In;Lee, Jun Gu;Hwang, Hyunseung;Yu, Jaewoong;Kim, Yong-Rok;Park, Se Won;Kim, Jin A
    • Journal of Plant Biotechnology
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    • v.42 no.3
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    • pp.245-256
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    • 2015
  • In the agricultural industries, LEDs are used as supplementary, as well as main lighting sources in closed cultivation systems. In cultivation using artificial light sources, various light qualities have been tried to supplement fluorescent lamps to promote plant growth and metabolism. Microarray analysis of Brassica rapa seedlings under blue and fluorescent mixed with blue light conditions identified changes in three genes of the glucosinolate pathway. This attracted attention as functional materials highly expressed 3.6-4.6 fold under latter condition. We selected four more genes of the glucosinolate pathway from the Brassica database and tested their expression changes under fluorescent light mixed with red, green, and blue, respectively. Some genes increased expression under red and blue mixed conditions. The Bra026058, Bra015379, and Bra021429; the orthologous genes of CYP79F1, ST5a, and FMOGS-OX1 in Arabidopsis, are highly expressed in Brassica rapa under fluorescent mixed with blue light conditions. Further, Bra029355, Bra034180, Bra024634, and Bra022448; the orthologous genes of MAM1, AOP3, UGT74B1, and BCAT4 in Arabidopsis, are highly expressed in Brassica rapa under fluorescent mixed with red light conditions. The various light conditions had unique effects on the varieties of Brassica, resulting in differences in glucosinolate synthesis. However, in some varieties, glucosinolate synthesis increased under mixed blue light conditions. These results will help to construct artificial light facilities, which increase functional crops production.

Expression Analysis of Oryza sativa Ascorbate Peroxidase 1 (OsAPx1) in Response to Different Phytohormones and Pathogens (벼 ascobate peroxidase 단백질의 병원균 및 식물호르몬에 대한 발현 분석)

  • Wang, Yiming;Wu, Jingni;Choi, Young Whan;Jun, Tae Hwan;Kwon, Soon Wook;Choi, In Soo;Kim, Yong Chul;Gupta, Ravi;Kim, Sun Tae
    • Journal of Life Science
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    • v.25 no.10
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    • pp.1091-1097
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    • 2015
  • We have isolated and characterized an ascorbate peroxidase (APx) gene, OsAPx1 from rice. Northern and Western blot analyses indicated that at young seedling stage, OsAPx1 mRNA was expressed highly in root, shoot apical meristem (SAM) and leaf sheath than leaf. In mature plant, OsAPx1 gene expressed highly in root, stem and flower but weakly in leaf. OsAPx1 gene and protein expression level was induced in leaves inoculated with Magnaporthe oryzae (M. oryzae) and Xanthomonas oryzae pv. oryzae (Xoo). Phytohormones treatment showed that OsAPx1 was up-regulated by jasmonic acid (JA), but was down regulated by ABA and SA co-treatments with JA, resulting that they have antagonistic effect on pathogen responsive OsAPx1 expression. Phylogenetic analysis illustrated that Arabidopsis AtAPx1 has a close relationship with OsAPx1. In AtAPx1 knock out lines, the accumulation of O2- and H2O2 are all highly detected than wild type, revealing that the high concentration of exogenous H2O2 cause the intercellular superoxide anion and hydrogen peroxide accumulation in AtAPx1 knockout plant. These results suggested that OsAPx1 gene may be associated with the pathogen defense cascades as the mediator for balancing redox state by acting ROS scavenger and is associated with response to the pathogen defense via Jasmonic acid signaling pathway.