Stable transformation of the moss Physcomitrella patens

1 May 1991

journal article
Published by Springer Nature in Molecular Genetics and Genomics

Vol. 226 (3) , 418-424
https://doi.org/10.1007/bf00260654

Abstract

We report the stable transformation of Physcomitrella patens to either G418 or hygromycin B resistance following polyethylene glycol-mediated direct DNA uptake by protoplasts. The method described in this paper was used successfully in independent experiments carried out in our two laboratories. Transformation was assessed by the following criteria: selection of antibiotic-resistant plants, mitotic and meiotic stability of phenotypes after removal of selective pressure and stable transmission of the character to the offspring; Southern hybridisation analysis of genomic DNA to show integration of the plasmid DNA; segregation of the resistance gene following crosses with antibiotic-sensitive strains; and finally Southern hybridisation analysis of both resistant and sensitive progeny. In addition to stable transformants, a heterogeneous class of unstable transformants was obtained.

Keywords

This publication has 32 references indexed in Scilit:

Hybrid genes in the analysis of transformation conditions. 3. Temporal/spatial fate of NPTII gene integration, its inheritance and factors affecting these processes in Nicotiana plumbaginifolia
Plant Molecular Biology, 1990
Cloning and nucleotide sequence analysis of genes coding for the major chlorophyll-binding protein of the moss Physcomitrella patens and the halotolerant alga Dunaliella salina
Gene, 1989
Enhancement of transformation rates in higher plants by low-dose irradiation: Are DNA repair systems involved in the incorporation of exogenous DNA into the plant genome?
Plant Molecular Biology, 1989
Arabidopsis, a useful weed
Cell, 1989
Cell-Cell Interactions in Chlamydomonas
Annual Review of Plant Physiology, 1985
Plasmid-encoded hygromycin B resistance: the sequence of hygromycin B phosphotransferase gene and its expression in Escherichia coli and Saccharomyces cerevisiae
Gene, 1983
Agarose plating and a bead type culture technique enable and stimulate development of protoplast-derived colonies in a number of plant species
Plant Cell Reports, 1983
A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity
Analytical Biochemistry, 1983
Regulation of DNA ligase activity by poly(ADP-ribose)
Nature, 1982
GENETIC AND PHYSIOLOGICAL STUDIES OF THE EFFECT OF LIGHT ON THE DEVELOPMENT OF THE MOSS, PHYSCOMITRELLA PATENS
Photochemistry and Photobiology, 1978