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A simple method for isolation, liquid culture, transformation and regeneration of Arabidopsis thaliana protoplasts

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Summary

An efficient technique was developed for the isolation, culture, transformation and regeneration of protoplasts derived from auxin conditioned Arabidopsis root cultures. On an average 30% of root protoplasts underwent cell division in liquid culture and formed somatic embryolike structures which regenerated to plants without embedding in Ca2+-alginate. The protoplast protocol was applicable to different landraces of Arabidopsis thaliana (L.) Heynh., such as RLD, Columbia or C24. PEG-mediated DNA uptake into protoplasts using different uidA reporter gene constructs yielded transient gene expression in over 25% of treated cells indicating that root-derived protoplasts are suitable recipients for transformation.

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Abbreviations

BA:

6-benzylaminopurine

2,4D:

2,4dichlorophenoxyacetic acid

IAA:

indole-3-acetic acid

ISA:

indole-3buryric acid

IPAR:

6-(γ,γ-dimethylallylamino)purine riboside

NAA:

αnaphthaleneacetic acid

uidA:

ß-glucuronidase gene

GUS:

ß-glucuronidase enzyme

CaMV:

Cauliflower Mosaic Virus

nos:

nopaline synthase

MES:

2[N-morpholino]ethane-sulfonicacid

PEG:

polyethylene glycol

X-gluc:

5bromo-4-chloro-3-indolyl glucuronide

MUG:

4-methyl umbelliferyl glucuronide

MU:

4-methylumbelliferone

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Authors and Affiliations

  1. Institute of Plant Biology, Biological Research Center of Hungarian Academy of Sciences, H-6726 Szeged, P.O. Box 521, 62, Temesvári krt., Hungary

    Jaideep Mathur, Csaba Koncz & László Szabados

  2. Max-Planck Institut für Züchtungsforschung, D-50829, Köln, Carl-von-Linné-Weg 10, Germany

    Jaideep Mathur & Csaba Koncz

  3. Department of Botany, University of Gorakhpur, 273 009, Gorakhpur, India

    Jaideep Mathur

Authors
  1. Jaideep Mathur
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  2. Csaba Koncz
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  3. László Szabados
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Communicated by K. Hahlbrock

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Mathur, J., Koncz, C. & Szabados, L. A simple method for isolation, liquid culture, transformation and regeneration of Arabidopsis thaliana protoplasts. Plant Cell Reports 14, 221–226 (1995). https://doi.org/10.1007/BF00233637

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  • DOI: https://doi.org/10.1007/BF00233637

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