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Direct gene transfer to plants
Author(s) -
Paszkowski Jerzy,
Shillito Raymond D.,
Saul Michael,
Mandák Václáv,
Hohn Thomas,
Hohn Barbara,
Potrykus Ingo
Publication year - 1984
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1002/j.1460-2075.1984.tb02201.x
Subject(s) - biology , selectable marker , gene , genetics , cauliflower mosaic virus , nicotiana tabacum , protoplast , plasmid , transformation (genetics) , chimeric gene , microspore , microbiology and biotechnology , gene expression , genetically modified crops , botany , transgene , stamen , pollen
Evidence for direct, gene‐mediated stable genetic transformation of plant cells of Nicotiana tabacum is presented. A selectable hybrid gene comprising the protein coding region of the Tn5 aminoglycoside phosphotransferase type II gene under control of cauliflower mosaic virus gene VI expression signals was introduced into plant protoplasts as part of an Escherichia coli plasmid. The gene was stably integrated into plant genomic DNA and constitutively expressed in selected, drug resistant, protoplast‐derived cell clones. The mode of integration of the foreign gene into the plant genome resembled that observed for DNA transfection of mammalian cells. Plants regenerated from transformed cell lines were phenotypically normal and fertile, and they maintained and expressed the foreign gene throughout the development of vegetative and generative organs. Microspores, grown in anther culture, developed into resistant and sensitive haploid plantlets. Genetic crossing analysis of one of the transformed plants revealed the presence of one dominant trait for kanamycin resistance segregating in a Mendelian fashion in the F 1 generation.