ISSN 1021-4437, Russian Journal of Plant Physiology, 2007, Vol. 54, No. 5, pp. 653–658. © Pleiades Publishing, Ltd., 2007.
Published in Russian in Fiziologiya Rastenii, 2007, Vol. 54, No. 5, pp. 738–743.
contains many species of agro-
nomic interest, and
transformation was actively studied in them. Since the
ﬁrst study presented , many more reports soon fol-
lowed [2–6]. The most interesting features for transfer
into cabbage species are resistance to herbicides [7, 8],
pathogens , and insects .
In the cabbage group, a strong inﬂuence of plant
genotype on the success of transformation was demon-
strated [11, 12], indicating difﬁculties of obtaining a
common transformation procedure. In our previous
study, we have developed the protocol for simple, fast,
and highly efﬁcient
tion of resistance to Basta
herbicide in Savoy cabagge
) . Here we
present results obtained by application of the same pro-
tocol for two domestic cabbage genotypes, discussing
the effects of genotype and bacterial strain on transfor-
MATERIALS AND METHODS
Plant material, inoculation, and plant regeneration.
Experiments were performed using
two well known cabbage inbred lines selected at the
Centre for Vegetable Crops, Smederevska Palanka
(Serbia). Seeds were surface-sterilized 20 min in 15%
commercial bleach (4% NaOCl), thoroughly rinsed in
autoclaved water, and germinated on basal MS 
supplemented with 2% sucrose and 0.64% agar (BM).
Segments of 10-day-old seedlings, precultured for 48 h
on BM supplemented with 1.0 mg/l BA and 0.5 mg/l
IBA (SR—Shoot Regeneration medium), were
immersed for 10–15 min in the
sion, washed for 20 min in 500 mg/l Tolycar (cefo-
taxime, Jugoremedija, Zrenjanin, Serbia), blotted with
ﬁlter paper, and placed back on the preculturing
medium . After 2 days, explants were transferred to
the SR medium containing 500 mg/l Tolycar and sub-
cultured at 6-week intervals on media with decreasing
concentration of Tolycar (100 50 0 mg/l). In
the ﬁrst subculture, SR medium was also supplemented
with 2.0 mg/l
. Regenerated shoots were multi-
plied by several subcultures on BM with 0.5 mg/l BA
and 0.1 mg/l IBA and then rooted on BM with 4.0 mg/l
IBA and sucrose increased to 4%.
Effects of Plant Genotype and Bacterial Strain
T. Sretenovi -Raji
, S. Ninkovi
, B. Uzela
, B. Vinterhalter
, and D. Vinterhalter
Genome Diversity Research Group, Institute of Plant Genetics and Crop Plant Research (IPK),
Correns str. 3, D-06466 Gatersleben, Germany
Department of Plant Physiology, Institute for Biological Research “S. Stankovi ”,
Bulevar Despota Stefana 142, 11000 Belgrade, Serbia;
fax: (+381) 11 2761 433, e-mail: email@example.com
Received January 10, 2007
—Two inbred lines of
were transformed with two
strains harboring resistance to herbicide Basta: AGL1/pDM805 and LBA4404/pGKB5 (LB5-1).
Inoculated cotyledons and hypocotyls provided equally good explants and manifested a high percentage of
shoot regeneration on MS medium supplemented with 1 mg/l benzyladenine and 0.5 mg/l indole-3-butyric acid.
genotype was superior to
in shoot regeneration (48.1 vs. 26.9%), multiplication, and acclimation
in the greenhouse (76 vs. 40%).
AGL1/pDM805 provided more regenerated shoots per explant,
especially in the case of cotyledon explants, and the higher transformation rate (up to 35% vs. up to 12%) as
compared to LB5-1. Putative transformants survived spraying with 10–30 mg/l phosphinothricin. Transforma-
tion was conﬁrmed by GUS assay and PCR analysis in T0 and T1 generations.
Key words: Brassica oleracea var. capitata - Agrobacterium tumefaciens - bar gene - cabbage transformation -
: BA—6-benzyladenine; BM—basal medium;
-glucuronidase; IBA—indole-3-butyric acid; L-PPT—
phosphinothricin; MS—Murashige and Skoog nutrient medium.
The text was submitted by the authors in English.