ISSN 1021-4437, Russian Journal of Plant Physiology, 2006, Vol. 53, No. 3, pp. 413–417. © MAIK “Nauka /Interperiodica” (Russia), 2006.
Published in Russian in Fiziologiya Rastenii, 2006, Vol. 53, No. 3, pp. 462–467.
L.) is one of the most
important ﬁber crops, which is widely cultivated
throughout the world. Therefore, it is attractive for
genetic engineering. Cotton transformation has been
reported through injection of DNA directly into
embryos in immature cotton bolls. The putative trans-
formed plants were identiﬁed by genotypes and pheno-
types, including boll size, ﬁber length, disease and
insect resistance [1, 2].
Recent advances in
formation have made it possible to introduce foreign
genes into various cotton species [2–7]. The transfor-
mation of cotton was ﬁrstly achieved by Umbeck et al.
, and transgenic Coker 312 cotton seedlings were
regenerated from hypocotyls and cotyledons that were
Successfully developing transgenic cot-
ton plants were obtained only from a few varieties with
a few transformation systems [2, 3, 6].
This study was conducted to establish an efﬁcient
method for high-frequency shoot production via
organogenesis and transformation for cotton
cv. Cukurova 1518.
MATERIALS AND METHODS
Plant material and media.
The seeds of cotton (
L., cv. Cukurova 1518) were delinted
with sulfuric acid for 1 h in a rotary shaker. Seeds were
washed with water and dried, then dipped in 70% etha-
nol for 1 min and 15% (v/v) Clorox (Commercial
bleach containing 5.25% sodium hypochlorite) with a
drop of Tween-20 for 15 min. The seeds were then
rinsed four times in sterile distilled water, dried with
sterile ﬁlter paper, and transferred to half-strength MS
medium supplemented with 30% glucose, Gamborg
B5 vitamins, 500 mg/l Mes, 0.2% phytogel, and phyto-
hormones for 5, 8, and 10 days at 25
in the dark.
The pH was adjusted to 5.8. Zeatin (0.5, 1, 2, 3, and
5 mg/l) was ﬁlter-sterilized and added to the medium
Effect of explants on shoot regeneration.
were prepared from cotyledons, hypocotyls, and
excised shoot meristems or their halves from in vitro
grown 5-, 8-, and 10-day-old seedlings. The cotyledons
were cut transversely and longitudinally into two
halves, hypocotyls were of 0.5 or 2 cm in length, the
shoot meristems were 1–2cm-long and some of them
and Regeneration of Cotton Plants
S. Unlu Yuceer and N. K. Koc
Department of Plant Protection, Faculty of Agriculture, University of Cukurova, Balcali, Adana, 01330 Turkey;
fax: +90 322 338 60 47; e-mail: firstname.lastname@example.org
Received June 29, 2005
L.) was transformed by the EHA101 strain of
harboring a binary vector pGA482GG plasmid carrying the marker genes for neomycin phosphotrans-
ferase II (
) determining resistance to kanamycin and
). The cotyledons, hypocotyls,
shoot meristem tissue, and its segments taken from in vitro growing seedlings were used as explants. Explants
were cultured in a Murashige and Skoog (MS) medium containing various hormone combinations to induce
shoot regeneration. The highest frequency of shoot formation was obtained from the shoot meristem. After
selection in the MS medium containing kanamycin (50 mg/l), these tissues were tested by histochemical GUS
assay. Shoots regenerated from excised shoot meristems or their halves were cultured for 4–6 weeks to obtain
rooted plants, which then produced fully-developed plants and seeds in pots. Genomic integration of the kanamy-
cin-resistance gene was detected by the PCR analysis. Seed germination percentage was 95% after the F1 seeds
of transgenic cotton plants were cultured on half-strength MS medium supplemented with 50 mg/l kanamycin.
Thus, a protocol for effective
-mediated genetic transformation of cotton was optimized.
Key words: Agrobacterium tumefaciens - Gossypium hirsutum - transgenic plants
: BA—benzyladenine; GUS—
IBA—indole-3-butyric acid; MS—Murashige and Skoog nutrient
medium; PCR—polymerase chain reaction; X-gluc—5-bromo-4-
The text was submitted by the authors in English.