Summary: suspension culture in Arabidopsis
sshiu at students.wisc.edu
Tue Dec 5 23:50:29 EST 1995
The following messages are the response I get for a question regarding the use
of suspension culture for enzyme assay in Arabidopsis. There are certain
references being cited more than once. In order to retain the original
message, I didn't do too much editing. Thanks for those who provide
information about my question.
1. Dr Chris Cobbett/chris_cobbett at muwayf.unimelb.edu.au/11.22.95
*See May and Leaver 1993 Plant Physiol 103 : 621-627
2. Dr Jed H. Deolling/doelling at biodec.wustl.edu/11.21.95
*Plant Cell Reports (1993) 12:241-244. Transient expression in
Arabidopsis thaliana protoplasts derived from rapidly established
suspension cultures. Doelling and Pikaard. The suspension cultures
grow very well but it is difficult to get good protoplasts for
transient expression studies.
*Also see Plant Science Letters (1975) 5: 293-304. Arabidopsis
thaliana as a model system in somatic cell genetics I. Cell and
Tissue culture. Negrutiu, Beeftink and Jacobs.
3. Dr Craig Pikkard/pikaard at biodec.wustl.edu/11/24/95
*We make suspension cultures as needed, and also regularly isolate
protoplasts from mesophyll cells for transient expression assays.
See the following papers:
(1) Doelling, Jed H. and Craig S. Pikaard (1993). Transient expression
in Arabidopsis thaliana protoplasts derived from rapidly established
cell suspension cultures. Plant Cell Reports 12: 241-244.
(2) Doelling, Jed H., Reginald Gaudino, and Craig S. Pikaard (1993).
Functional analysis of Arabidopsis thaliana rRNA gene and spacer
promoters by transient expression. Proc. Natl. Acad. Sci. USA,
(3) Doelling, Jed H. and Craig S. Pikaard (1995). The minimal ribosomal
RNA gene promoter of Arabidopsis thaliana includes a critical element
at the transcription initiation site. The Plant Journal (November
4. Dr I. A. Graham/gbca59 at pop-server.cent.gla.ac.uk/11.22.95
*Check out the following ref. May and Leaver: 1993. Plant Physiol,
103: 621-627. The cell culture described in this paper grows like
5. Dr.Rafael Pont-Lezica/lezica at cict.fr/11.19.95
*Take a look to Axelos et al, 1992, "A protocol for transient gene
expression in Arabidopsis thaliana protoplasts isolated from cell
suspension cultures.Plant Physiol Biochem 30, 123-128."
6. Dr Jamie McLaughlin/ZO1JCM at strines.shef.ac.uk/11.22.95
*I find a common way to get suspension cultures form arabidopsis is to use
shoots and induce the formation of callus by transfer on to a callus
induction media (CIM). After callus formation transfer to suspension media
(SM). From May, M.J. and Leaver, C.J. (1993). Oxidative stimulation of
glutathione synthesis in Arabidopsis thaliana suspension cultures. Plant
Physiol. 103, 621-627.
Gamborg's B5, 2% (w/v) Glucose, MES 0.5 g/l, 2,4-D 0.5 mg/ml Kinetin 0.05
mg/ml, 0.8% Agar
Murashige and Skoog, 3% sucrose, 0.5 mg/ml NAA, 0.05mg/ml kinetin
7. Dr Carl Douglas/cdouglas at pop.unixg.ubc.ca/11.23.95
*Try the Hahlbrock lab at the Max-Planck Institute in Cologne, Germany
(for example, contact Dr. Imre Somssich or Dr. Bernt Weisshaar) -
they have a suspension.
*Dr. Keith Davis at Ohio State University may also have one.
8. Dr Andreas Mordhorst/Andreas.Mordhorst at MAC.MB.WAU.NL/11.28.1995
*For suspension cultures (or better liquid in vitro cultures) of
Arabidopsis look into the following puplications:
(1) Ford: Plant Cell reports (1990) 8: 534-537,
(2) Mathur et al.: Plant Cell reports (1995) 14: 221-226,
(3) Wenck and Marton: BioTechniques (1995) 18: 640-643.
*We also have an (unpublished) protocol starting from seedling tissue
in MS-medium with 2,4-D. If you are interested please let me know.
*I also found in the meanwhile a number of other publications
concerning suspension cultures in Arabidopsis or where the authors
used in liquid cultured cells to eg. isolate protoplasts or to study
gene expression. Note, that in some cases the cultures seem not to
consist of fine dispered cell colonies of even single cells, like for
instance in carrot cell suspensions.
(1) Xuan and Menczel (1980): Z. Pflanzenphysiol. 96: 77-80.
(2) Braam (1992): PNAS USA: 89 3213-3216.
(3) Axelos et al. (1992) Plant Physiol. Biochem. 30: 123-128.
(4) de Virville et al. (1994) Plant Physiol. Biochem. 32: 159-166.
(5) Kaldenhoff et al. (1994) Planta 195: 182-187.
(6) Soni et al. (1995) Plant Cell 7: 85-103.
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