Abstract
Experimental conditions influencing DNA uptake efficiency by maize protoplasts in polyethyleneglycol (PEG) mediated transfection experiments have been studied systematically. The data provide evidence that the extracellular DNA is precipitated efficiently by combined action of PEG together with divalent cations and DNA is taken up by the plant protoplasts in the precipitated form. The particle size is strongly effected by the pH of the PEG solution. At optimal pH 6– 6.5 a very fine and homogenous precipitate forms in presence of Ca2+ and Mg2+ ions and is efficiently incorporated by maize and rice protoplasts.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- CAT:
-
Chloramphenicolacetyl transferase
- DAPI:
-
4′,6-Diamidino-2-Phenylindole
- HEPES:
-
N-2-Hydroxyethylpiperazine-N′-2- ethanesulfonic acid
- MES:
-
2(N-Morpholino)ethanesul fonic acid
- PEG:
-
Polyethyleneglycol
- SW:
-
seawater
References
Bradford MM (1976) Anal. Biochem. 72:248–254
Deshayes A, Herrera Estrella I and Caboche M (1985) EMBO J. 4:2731–2738
Fromm M, Taylor LP and Walbot V (1985) Proc. Natl. Acad. Sci. USA 82:5824–5828
Gorman CM, Moffat LF and Howard BH (1982) Mol. Cell. Biol. 2:1044–1051
Graham FL and Van Der Eb (1973) Virology 52:456–467
Junker B, Zimny J, Lührs R and Lörz H (1987) Plant Cell Reports 6:329–332
Klein TM, Gradziel T, Fromm ME and Sanford JC (1988) Bio/Technology 6:559–563
Krens FA, Molendijk L, Wullems GJ and Schilperoort RA (1982) Nature 296:72–74
Loyter A, Scangos GA and Ruddle F (1982) Proc. Natl. Acad. Sci. USA 79:422–426
Lörz H, Baker B and Schell J (1985) Mol. Gen. Genet. 199:178–182
Lörz H, Hartke S and Stolarz A (1988) in: Plant Breeding and Genetic Engineering (Zakri AH ed) Sabrao/Malaysia 295–313
Meyer P, Walgenbach E, Bussmann K, Hombrecher G and Saedler H (1985) Mol. Gen. Genet. 201:513–518
Möller A, Nordheim A, Kozlowski SA, Patel DJ and Rich A (1984) Biochem. 23:54–62
Murashige T, Skoog F (1962) Physiol. Plant. 15:473–497
Nagy JI, Maliga P (1976) Z. Pflanzenphysiol. 78:453–455
Negrutiu I, Shillito R, Potrykus I, Biasini G, Sala F (1987) Plant Molecular Biology 8:363–373
Pröls M, Töpfer M, Schell J and Steinbiß H-H (1988) Plant Cell Reports 7:221–224
Saenger W, Hunter WN and Kennard O (1986) Nature 324:385–388
Shillito RD, Saul MW, Pazkowski J, Müller M and Potrykus I (1985) Bio/Technology 3:1099–1103
Author information
Authors and Affiliations
Additional information
Communicated by H. Lörz
Rights and permissions
About this article
Cite this article
Maas, C., Werr, W. Mechanism and optimized conditions for PEG mediated DNA transfection into plant protoplasts. Plant Cell Reports 8, 148–151 (1989). https://doi.org/10.1007/BF00716828
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00716828