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Genetic transformation of cauliflower ( Brassica oleracea var. botrytis ) by direct DNA uptake into mesophyll protoplasts
Author(s) -
Radchuk Volodymyr V.,
Ryschka Ulrich,
Schumann Günter,
Klocke Evelyn
Publication year - 2002
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1034/j.1399-3054.2002.1140313.x
Subject(s) - brassica oleracea , protoplast , botrytis , transformation (genetics) , biology , cauliflower mosaic virus , plasmid , gene , dna , botany , phosphotransferase , horticulture , biochemistry , genetically modified crops , botrytis cinerea , transgene
Mesophyll protoplasts of Brassica oleracea var. botrytis were successfully transformed using polyethylene glycol (PEG). The success of plant transformation depended on both gene transfer and plant regeneration. Parameters, such as PEG and vector concentrations and heat shock conditions were tested in experiments on transient expression of the β ‐glucuronidase (EC 3.2.1.31) gene and the most suitable conditions for DNA uptake were determined. Two antibiotic resistance marker genes for neomycin phosphotransferase (EC 2.7.1.95) and hygromycin phosphotransferase (EC 2.7.1.104), and three vector plasmids with different lengths were used to obtain stable transformants.