ISSN 1021-4437, Russian Journal of Plant Physiology, 2008, Vol. 55, No. 2, pp. 241–245. © Pleiades Publishing, Ltd., 2008.
In vitro ovary and ovule cultures have been used for
the production of haploids in a wide range of taxa [1–
4]. Gynogenesis may be the only efﬁcient means of pro-
ducing haploids in
, in which other means of haploid
production are not possible. Haploids derived via gyno-
genesis have a considerable potential in plant breeding
because of time saved by the reduction of classical
selection cycle period and the genetic value of homozy-
gous lines. Also the presence of a single set of chromo-
somes allows the detection of mutations controlled by
recessive genes and recovery of unique recombinants.
(L. f.) Cass.) (Aster-
aceae family) is an important oil seed crop widely
grown in India and Ethiopia; it is also cultivated in
Bangladesh, Pakistan, Kenya, Uganda, and Malawi.
Niger seeds contain 30–50% edible, semi-drying oil
with pleasant nutty taste . Improvement of niger was
hindered by self-incompatibility mechanism, and it
causes serious difﬁculty for inbred line development
and maintenance . Production of haploids and subse-
quently homozygous lines has a great potential in niger.
In the present study, we have attempted unpollinated
ovule culture of niger and succeeded in direct haploid
ADE—adenine; BA—benzyladenine; B5—Gam-
borg nutrient medium; IBA—indole-3-butyric acid; 2-IP—
2-isopentylpurine; KN—kinetin; MS—Murashige and Skoog
nutrient medium; NN—Nitsch and Nitsch nutrient medium;
N6—Chu nutrient medium; TDZ—thidiazuron.
This text was submitted by the authors in English.
plant regeneration from cultured ovules. We evaluated
the growth regulators, culture media, and suitability of
developmental stage (age) of the ovule on direct plant
regeneration from cultured ovules. This is the ﬁrst
report about successful production of niger gynogenic
MATERIALS AND METHODS
Plant material and sterilization procedure.
(L. f.) Cass. cvs. JNC-6
and Ootacamund were obtained from Project Coordina-
tor, All India Coordinated Research Project on Sesame
and Niger (Jawaharlal Nehru Agricultural University,
Jabalpur, India) and used as experimental donor mate-
rials in the present investigation. Plants were grown in
the experimental plots using standard agronomic prac-
tices. Capitulla (ﬂower buds) were collected before
opening of the ﬂorets (two/one days before anthesis or
on the day of anthesis) and were washed in 1% (v/v)
Laboline (detergent) and 0.5% Carbendazim (fungi-
cide) for 10 min and surface-sterilized with 5% sodium
hypochorite solution for 20 min. The ﬁnal step of ster-
ilization was carried out in a horizontal laminar ﬂow
using 0.1% mercuric chloride solution for 5 min.
Finally, the ﬂower buds were rinsed several times in
sterile distilled water. Ovules were removed from
ﬂower buds and inoculated onto culture medium.
Culture medium and culture conditions.
medium employed for ovule culture was Murashige
and Skoog (MS)  medium containing 30 g/l
Haploid Plant Regeneration from Unpollinated Ovule Cultures
of Niger (
(L. f.) Cass.)
J. G. Bhat and H. N. Murthy
Department of Botany, Karnatak University, Dharwad, 580 003 India;
Received February 9, 2007
—Haploid plants were regenerated in vitro from unpollinated ovules of niger (
f.) (Cass.) on Murashige and Skoog nutrient medium (MS) supplemented with 10
M naphthaleneacetic acid
M NAA + 1.5
M kinetin and 30 g/l sucrose. Gamborg (B5) medium was the best for plant regeneration
(in comparison with MS, Nitsch and Nitsch (NN), and Chu (N6) media) from cultured ovules, and 6.66 and
7.33 ovules of JNC-6 and Ootacamund cultivars were involved in direct plant regeneration on this medium.
Matured ovules (ovules collected one day before anthesis or on the day of anthesis) only responded to cultural
regimes and involved in direct plantlet development. Cytological preparation of root tips and chloroplast counts
in the guard cells of leaf stomata of regenerated plants conﬁrmed their haploid nature.
Key words: Guizotia abyssinica - gynogenesis - haploids - ovule culture - parthenogenesis - plant regeneration