• 제목/요약/키워드: Isolated phloem cells

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Streptanthus tortus의 培養細胞로부터 사부 세포의 분리와 분리된 篩部 및 柔組織 細胞에서 설탕의 능동수송 (Isolation of Phloem Cells and Active Transport of Sucrose by Isolated Phloem and Parenchyma Cells of Streptanthus tortus Suspension Cultures)

  • 조봉희
    • 식물조직배양학회지
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    • 제25권1호
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    • pp.7-11
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    • 1998
  • Streptanthus 조직 배양 세포를 사용하여 사부를 순수분리 시키고, 분리된 사부에서 사부 하적에 대한 기작을 규명하기 위해 다음 연구를 하였다. 유조직 세포는 0.2% macerase 와 0.03% cellulase의 가수분해 효소로 처리하여 원형질체를 얻었고, 분화된 세포에서는 0.03% cellulase + 0.02% pectinase + 0.2% macerase + 0.025% rohamet PC로 가수분해시켜서 순수한 사부 세포와 사부 원형질체와 반세포의 원형질체를 분리하였다. 분리된 유조직 세포와 반세포의 원형질체에서는 단당류인 포도당을 수송시키나, 설탕은 수송시키지 못했다. 반면 분리된 사부 세포는 설탕을 능동수송 시키나 포도당은 수송시키지 못했다. 이는 설탕의 사부 하적은 반세포 없이도 가능하며, 반세포는 설탕 운반체가 없어서 설탕을 직접 수송할 수 있는 능력이 없다는 것을 보여주는 것이다. 그리고 사부 세포에서 설탕의 수송은 에너지대사에 의존하는 능동수송으로 나타났다.

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A pathogen-induced osmotin-like protein gene, CAOSMl, from pepper: Differential expression and in situ localization in pepper tissues during pathogen infection and abiotic stresses

  • Hong, J.K.;Jung, H.W.;Lee, B.K.;Lee, S.C.;Hwang, B.K.
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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    • pp.78.1-78
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    • 2003
  • An osmotin-like protein (CAOSMl) gene was isolated from pepper leaves infected with the avirulent strain Bv5-4a of Xmthomonas campestris pv. vesicatoria. The cDNA encodes a polypeptide of 250 amino acids with a molecular mass of 27, 361 Da. Its amino acid sequence is highly homologous to various osmotin-like proteins from other plant species. The CAOSMl gene expression was organ- and tissue-specifically regulated In pepper plants. The CAOSMl mRNA was intensely localized in the endodermis area of root tissue and in the phloem cells of vascular bundles of red fruit tissue, but not in leaf, stem, and green fruit tissues of healthy pepper plants. Infection by X. c. pv vesintoria, Colletotrichum coccodes, or Phytopkhora capsici iinduced CAOSMl transcription in the leaf or stem tissues. Expression of the CAOSMl gene was somewhat higher in the incompatible than the compatible interactions of pathogens with pepper. The CAOSMl mRNA was prevalently localized in the phloem cells of the vascular bundle of leaf tissues infected by C. coccodes. The CAOSMl gene was activated in leaf tissues by treatment with ethylene, methyl jasmonate, high salinity, cold acclimation and mechanical wounding, but not by abscisic acid (ABA) and drought. These results indicate that the pepper CAOSMl protein functions in response to Pathogens and some abiotic stresses in pepper plants

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Generation and Expression in Plants of a Single-Chain Variable Fragment Antibody Against the Immunodominant Membrane Protein of Candidatus Phytoplasma Aurantifolia

  • Shahryari, F.;Safarnejad, M.R.;Shams-Bakhsh, M.;Schillberg, S.;Nolke, G.
    • Journal of Microbiology and Biotechnology
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    • 제23권8호
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    • pp.1047-1054
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    • 2013
  • Witches' broom of lime is a disease caused by Candidatus Phytoplasma aurantifolia, which represents the most significant global threat to the production of lime trees (Citrus aurantifolia). Conventional disease management strategies have shown little success, and new approaches based on genetic engineering need to be considered. The expression of recombinant antibodies and fragments thereof in plant cells is a powerful approach that can be used to suppress plant pathogens. We have developed a single-chain variable fragment antibody (scFvIMP6) against the immunodominant membrane protein (IMP) of witches' broom phytoplasma and expressed it in different plant cell compartments. We isolated scFvIMP6 from a naïve scFv phage display library and expressed it in bacteria to demonstrate its binding activity against both recombinant IMP and intact phytoplasma cells. The expression of scFvIMP6 in plants was evaluated by transferring the scFvIMP6 cDNA to plant expression vectors featuring constitutive or phloem specific promoters in cassettes with or without secretion signals, therefore causing the protein to accumulate either in the cytosol or apoplast. All constructs were transiently expressed in Nicotiana benthamiana by agroinfiltration, and antibodies of the anticipated size were detected by immunoblotting. Plant-derived scFvIMP6 was purified by affinity chromatography, and specific binding to recombinant IMP was demonstrated by enzyme-linked immunosorbent assay. Our results indicate that scFvIMP6 binds with high activity and can be used for the detection of Ca. Phytoplasma aurantifolia and is also a suitable candidate for stable expression in lime trees to suppress witches' broom of lime.