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Immunocytolocalization of tryptophan decarboxylase in Catharanthus roseus hairy roots

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Abstract

We investigated the intracellular distribution of tryptophan decarboxylase (TDC) (EC 4.1.1.28) in Catharanthus roseus hairy roots using immunofluorescence and immunogold techniques. TDC was detected by immunofluorescence localization in the cytosol and in the apoplastic region of the meristematic cells of the roots, with a slight enrichment in the epidermal cells of the root cap and in the meristematic region. In the enlargement zone, TDC was localized only in the first three layers of the cortex. In the maturation zone, the enzyme was not present. Immunogold studies confirmed that the enzyme was localized in the cytosol of the meristematic region, and intense gold labeling was found in the apoplastic zone. A protein fraction isolated from the apoplastic zone and assayed for TDC activity showed high activity.

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Abbreviations

BCA:

bicinchoninic acid

BSA:

bovine serum albumin

PBS:

phosphate buffered saline

PBS-BSA:

phosphate buffered saline-1% bovine serum albumin

TDC:

tryptophan decarboxylase

SSS:

strictosidine synthase

G10H:

geraniol-10-hydroxylase

NMT:

S-adenosyl-L-methionine: 16-methoxy-2, 3-dihydro-3-hydroxytabersonine-N-methyltransferase

DAT:

acetyl coenzyme A: deacetylvindoliine-O-acetyltransferase

IAA:

indole acetic acid

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Authors and Affiliations

  1. Unidad de Bioquímica y Biología Molecular de Plantas, Centro de investigacíon Científica de Yucatán, Calle 43, númerol 30, CP 97200, Mérida, Yucatán, México

    Víctor M. Loyola-Vargas

Authors
  1. Oscar A. Moreno-Valenzuela
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  2. Yereni Minero-García
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  3. Ligia Brito-Argáez
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  4. Eloísa Carbajal-Mora
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  5. Olga Echeverría
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  6. Gerardo Vázquez-Nin
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  7. Víctor M. Loyola-Vargas
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Correspondence to Víctor M. Loyola-Vargas.

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Moreno-Valenzuela, O.A., Minero-García, Y., Brito-Argáez, L. et al. Immunocytolocalization of tryptophan decarboxylase in Catharanthus roseus hairy roots. Mol Biotechnol 23, 11–18 (2003). https://doi.org/10.1385/MB:23:1:11

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