Hybrid genes in the analysis of transformation conditions : I. Setting up a simple method for direct gene transfer in plant protoplasts.

Plant molecular biology ·Vol. 8 ·No. 5 ·1987-09-00 ·Pages 363-73

Negrutiu I, Shillito R, Potrykus I, Biasini G, Sala F

Abstract

Direct gene transfer into plant protoplasts has been recently developed, and conditions for high frequency transformation of SR1 tobacco protoplasts established. In this paper we analyse numerous transformation parameters in a comparative study on SR1 Nicotiana tabacum and N. plumbaginifolia, and report on a simple chemical technique for very efficient protoplast transformation. It is based on the synergistic interaction of MgCl2 and PEG. The technique yielded up to 1400 transformants per 3×10(5) treated N. tabacum protoplasts (up to 4.8% of the survivors, late selected clones). Using N. plumbaginifolia, the frequencies were 10-fold lower, indicating that the 'competence' for transformation has a species-specific component.

Authors & Affiliations

5 authors, click to expand affiliations / ORCID

Negrutiu I

Friedrich Miescher-Institut, P.O. Box 2543, CH-4002, Basel, Switzerland.

Shillito R

Potrykus I

Biasini G

Sala F

References (12)

12 references, click to expand

Transformation of plant protoplasts with DNA: cotransformation of non-selected calf thymus carrier DNA and meiotic segregation of transforming DNA sequences.
Plant Mol Biol. 1985 Jul;5(4):235-46 PMID: 24306764
Agarose plating and a bead type culture technique enable and stimulate development of protoplast-derived colonies in a number of plant species.
Plant Cell Rep. 1983 Oct;2(5):244-7 PMID: 24258119
Fusion of plant protoplasts: a study using auxotrophic mutants of Nicotiana plumbaginifolia, Viviani.
Theor Appl Genet. 1986 Mar;72(2):279-86 PMID: 24247847
Nutritional requirements of protoplast-derived, haploid tobacco cells grown at low cell densities in liquid medium.
Planta. 1980 Jun;149(1):7-18 PMID: 24306186
Stable transformation of maize after gene transfer by electroporation.
Nature. 1986 Feb 27-Mar 5;319(6056):791-3 PMID: 3005872
Fate of selectable marker DNA integrated into the genome of Nicotiana tabacum.
DNA. 1986 Apr;5(2):101-13 PMID: 3519133
A general method for polyethylene-glycol-induced genetic transformation of bacteria and yeast.
Gene. 1983 Nov;25(2-3):333-41 PMID: 6363214
Expression of genes transferred into monocot and dicot plant cells by electroporation.
Proc Natl Acad Sci U S A. 1985 Sep;82(17):5824-8 PMID: 3862099
Molecular and general genetics of a hybrid foreign gene introduced into tobacco by direct gene transfer.
Mol Gen Genet. 1985;199(2):169-77 PMID: 3860712
Streptomycin resistant and sensitive somatic hybrids of Nicotiana tabacum + Nicotiana knightiana: correlation of resistance to N. tabacum plastids.
Theor Appl Genet. 1981 Mar;59(3):191-5 PMID: 24276446
Direct gene transfer to plants.
EMBO J. 1984 Dec 1;3(12):2717-22 PMID: 16453573
Regeneration of fully nitrate reductase - deficient mutants from protoplast culture of Nicotiana plumbaginifolia (Viviani).
Theor Appl Genet. 1983 Sep;66(3-4):341-7 PMID: 24263936

Article Info

Journal

Plant molecular biology

Abbr.

Plant Mol Biol

ISSN

0167-4412

DOI

10.1007/BF00015814

Published

1987-09-00

Pages

363-73

Language

English

Region

Netherlands

NLM ID

9106343

External Links

PubMed source DOI: 10.1007/BF00015814 View journal details

Analysis Services

Hybrid genes in the analysis of transformation conditions : I. Setting up a simple method for direct gene transfer in plant protoplasts.

Contact

WeChat Official Account