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

Plant cell reports ·Vol. 14 ·No. 4 ·1995-01-00 ·Pages 221-6

Mathur J, Koncz C, Szabados L

Abstract

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 Ca(2+)-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.

Authors & Affiliations

3 authors, click to expand affiliations / ORCID

Mathur J

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

Koncz C

Szabados L

References (13)

13 references, click to expand

Activation of a plant gene by T-DNA tagging: auxin-independent growth in vitro.
Science. 1992 Nov 20;258(5086):1350-3 PMID: 1455228
Agrobacterium tumefaciens-mediated transformation of Arabidopsis thaliana root explants by using kanamycin selection.
Proc Natl Acad Sci U S A. 1988 Aug;85(15):5536-40 PMID: 16593964
High-frequency T-DNA-mediated gene tagging in plants.
Proc Natl Acad Sci U S A. 1989 Nov;86(21):8467-71 PMID: 2554318
Transient expression in Arabidopsis thaliana protoplasts derived from rapidly established cell suspension cultures.
Plant Cell Rep. 1993 Mar;12(5):241-4 PMID: 24197149
GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants.
EMBO J. 1987 Dec 20;6(13):3901-7 PMID: 3327686
Direct gene transfer to protoplasts of Arabidopsis thaliana.
Plant Cell Rep. 1991 Feb;9(10):571-4 PMID: 24220714
Systemic Endopolyploidy in Arabidopsis thaliana.
Plant Physiol. 1991 Jul;96(3):985-9 PMID: 16668285
Facile transformation of Arabidopsis.
Plant Cell Rep. 1991 Aug;10(5):235-9 PMID: 24221586
A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
Anal Biochem. 1976 May 7;72:248-54 PMID: 942051
Plant regeneration fromArabidopsis thaliana protoplasts.
Plant Cell Rep. 1990 Jan;8(9):534-7 PMID: 24226280
T-DNA insertional mutagenesis in Arabidopsis.
Plant Mol Biol. 1992 Dec;20(5):963-76 PMID: 1463832
Arabidopsis thaliana: protocol for plant regeneration from protoplasts.
Plant Cell Rep. 1991 Feb;9(10):575-8 PMID: 24220715
Efficient transformation of Arabidopsis thaliana using direct gene transfer to protoplasts.
Mol Gen Genet. 1989 May;217(1):6-12 PMID: 2549369

Article Info

Journal

Plant cell reports

Abbr.

Plant Cell Rep

ISSN

0721-7714

DOI

10.1007/BF00233637

Published

1995-01-00

Pages

221-6

Language

English

Region

Germany

NLM ID

9880970

External Links

PubMed source DOI: 10.1007/BF00233637 View journal details

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

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